Application of

Biotechnology

for
Functional Foods
Pew Initiative on Food
and Biotechnology
© 2007 Pew Initiative on Food and Biotechnology. All rights reserved.
No portion of this paper may be reproduced by any means, electronic or mechanical,
without permission in writing from the publisher. This report was supported by a
grant from The Pew Charitable Trusts to the University of Richmond. The opinions
expressed in this report are those of the authors and do not necessarily refect the
views of The Pew Charitable Trusts or the University of Richmond.
Application of Biotechnology for Functional Foods
3

Contents
Preface...........................................................................................................................................................................5
Part.1:.Applications.of.Biotechnology.for.Functional.Foods...................................................................7
Part.2:.Legal.and.Regulatory.Considerations.Under.Federal.Law...................................................... 37
Summary.................................................................................................................................................................... 63
Selected.References............................................................................................................................................... 65
Application of Biotechnology for Functional Foods
Application of Biotechnology for Functional Foods
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Preface
S
ince the earliest days of agricultural biotechnology development, scientists
have envisioned harnessing the power of genetic engineering to enhance
nutritional and other properties of foods for consumer beneft. The frst
generation of agricultural biotechnology products to be commercialized,
however, were more geared towards so-called input traits, genetic
modifcations that make insect, virus and weed control easier or more effcient.
These frst products have been rapidly adopted by U.S. farmers, and now account
for the majority of soybeans, cotton and corn grown in the United States.
Agricultural biotechnology innovations aimed directly towards consumers,
sometimes collectively referred to as output traits, have been a longer time in
development. As the technology advances, and we learn more about the genes and
biochemical pathways that control those attributes that could offer more direct
consumer benefts, the long-awaited promise of genetically engineered food with
more direct consumer benefts moves closer to reality.
One category of potential products aimed at consumers is those products with
added health benefts, also known as “functional foods.” The term functional food
means different things to different people, but generally refers to foods that provide
health benefts beyond basic nutrition.
This report looks at the potential to develop functional foods through the
application of modern biotechnology. The frst section describes some recent
scientifc advances that could lead to functional foods on grocery store shelves,
and the second section analyzes the legal authorities that could govern the use of
biotechnology-derived functional foods.
The range of work being done on functional foods described in this report—from
oils that product no trans fats or contain heart healthy omega-3 fatty acids, to
cassava with increased protein content to help fght malnutrition in developing
nations, to foods with enhanced levels of antioxidants—is impressive. This report
is not intended to be an exhaustive catalog, however, but is rather a snapshot in
time to give readers a sense of the kinds of products that may one day be available.
Application of Biotechnology for Functional Foods
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It should also be noted that much of the work described here is still in preliminary
stages, and may never make its way into consumer products for technical, economic
or other reasons.
The analysis of relevant statutory authorities suggests that there is ample legal
authority to cover the kinds of functional foods currently being explored in
laboratories, but that different authorities may come into play for different kinds
of foods and that the application of different authorities can have signifcant
consequences for product developers, food manufacturers and consumers.
Different authorities impose different safety and labeling standards, have different
requirements for regulatory review and clearance or approval, and could result in
different levels of transparency to the public. The use of modern biotechnology to
produce functional foods will not likely fundamentally challenge existing regulatory
structures, but may challenge the boundaries of some regulatory classifcations.
The Pew Initiative on Food and Biotechnology’s frst report, Harvest on the
Horizon (2001), provided a broad overview of what could be the “next generation”
of genetically engineered agricultural products. It is ftting that this, the last of
the Initiative’s reports, turns again to look at a category of new products on the
horizon.
We would like to acknowledge the contributions of Joyce A. Nettleton, who
created the scientifc review used in the development of this paper; and of Edward
L. Korwek, for the review of regulatory authorities that could govern future
functional foods.
Michael Fernandez
Executive Director
April 2007
Application of Biotechnology for Functional Foods
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PART 1
Applications of Modern


Biotechnology to Functional Food
Applications of Biotechnology
for Functional Foods

I. Background
A. Functional Foods
A relatively recent concept in the U.S. to describe the broad healthfulness of foods is the
term “functional foods.” These foods are defned as foods that provide health benefts
beyond basic nutrition (International Food Information Council 2004). The Food and
Nutrition Board of the National Academy of Sciences described a functional food as,
“any modifed food or food ingredient that may provide a health beneft beyond that of the
traditional nutrients it contains” (Food and Nutrition Board 1994). The original concept of
functional foods originated in Japan from its development of a special seal to denote Foods
for Specifed Health Use (FOSHU). More than 270 foods have FOSHU status in Japan.
Foods qualify as “functional foods” because they contain non-essential substances with
potential health benefts. Examples of the diverse foods and their bioactive substances that
are considered “functional foods” are: psyllium seeds (soluble fber), soy foods (isofavones),
cranberry juice (proanthocyanidins), purple grape juice (resveratrol), tomatoes (lycopene),
and green tea (catechins). The broad classifcation of functional foods carries some irony,
as John Milner, Chief of the Nutrition Science Research Group at the National Cancer
Institute noted, “It is unlikely that a non-functional food exists.”
Bioactive components of functional foods may be increased or added to traditional
foods through genetic engineering techniques. An example would be the high lycopene
tomato, a genetically modifed tomato with delayed ripening characteristics that is high in
lycopene, which has potent antioxidant capabilities. This report focuses on biotechnology
applications in functional and improved foods, using the National Academy of Sciences
defnition as a guideline.
B. Applications of Biotechnology in Food Crops
In 1990, the U.S.Food and Drug Administration (FDA) approved the frst genetically
engineered food ingredient for human consumption, the enzyme chymosin, used in cheese-
making. It is estimated that today 70% or more of cheese made in the U.S. uses genetically
engineered chymosin. The frst genetically engineered food, the FlavrSavr™ tomato, was
approved for human consumption in the U.S. in 1994.
Application of Biotechnology for Functional Foods
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C. Transgenic Acreage Expands Steadily
Seven million farmers in 18 countries now grow genetically engineered crops. Leading
countries are the U.S., Argentina, Canada, Brazil, China, and South Africa. Cultivation
of genetically engineered crops globally has expanded more than 10% per year for the
past seven years, according to the International Service for the Acquisition of Agri-biotech
Applications (ISAAA, James 2004). Such an expansion rate amounts to a 40-fold increase
in the global area of transgenic crops from 1996 to 2003. Thus, in spite of continuing
controversy, the technology continues to be adopted by farmers worldwide. ISAAA
highlighted its key fndings this way:
In 2003, GM crops were grown in 18 countries with a combined population
of 3.4 billion, living on six continents in the North and the South: Asia,
Africa and Latin America, and North America, Europe and Oceania….
the absolute growth in GM crop area between 2002 and 2003 was almost
the same in developing countries (4.4 million hectares) and industrial
countries (4.6 million hectares) … the three most populous countries in
Asia—China, India, and Indonesia, the three major economies of Latin
America—Argentina, Brazil and Mexico, and the largest economy in
Africa, South Africa, are all offcially growing genetically engineered crops.
The leading genetically engineered crops globally and in the U.S. are soy, maize (corn),
cotton, and canola. In the U.S., transgenic virus-resistant papaya and squash are also
cultivated.
D. Agronomic Traits Prevail
Research in plant biotechnology has focused primarily on agronomic traits—characteristics
that improve resistance to pests, reduce the need for pesticides, and increase the ability of
the plant to survive adverse growing conditions such as drought, soil salinity, and cold.
Biotechnology traits developed and commercialized to date have largely focused on pest
control (primarily Bt crops) or herbicide resistance. Many plant pests have proven either
diffcult or uneconomical to control with chemical treatment, traditional breeding, or other
agricultural technologies and in these instances in particular, biotechnology has proven to
be an effective agronomic tool. Herbicide resistance allows farmers to control weeds with
chemicals that would otherwise damage the crop itself.
Varieties combining two different traits, such as herbicide tolerance and insect resistance,
have been introduced in cotton and corn. The addition of new traits, such as resistance
to rootworm in maize, and the combinations of traits with similar functions, such as two
genes for resistance to lepidopteran pests in maize, are expected to increase. In its 2003
report, ISAAA suggested that fve new Bt and novel gene products for insect resistance
in maize could be introduced.
While the improvement of agronomic characteristics in major crops has been highly
successful, few products genetically engineered to meet the specifc needs of either
food processors or consumers have yet been commercialized. Recently, however, a
renewed emphasis on developing agricultural biotechnology applications more relevant
to consumers has accompanied continuing efforts to develop crops with improved
agronomic traits. Although genetically engineered crops with enhanced health, nutrition,
functional, and consumer benefts have lagged behind agronomic applications, research
on many such products is in the advanced stages of development. These applications
Application of Biotechnology for Functional Foods
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Fats and Fatty Acids – Like Oil for Water
Fats are slippery substances that usually do not dissolve in water. We see them in foods in marbled
meat, salad and cooking oils, and spreads such as margarine and butter. Substantial amounts also
hide in foods such as cheese, mayonnaise, peanut butter, doughnuts, and chips.
What distinguishes fats from one another is their fatty acids. Each fat contains three fatty acids, which
may be a combination of three different types. People have been warned for years to limit their intake
of saturated fat, the kind rich in saturated fatty acids. These warnings relate to the ability of most
saturated fatty acids to raise blood cholesterol levels, thereby increasing the risk of heart disease.
Butter, cheese and other dairy foods, and meats are rich in saturated fatty acids.
So-called “good fats” are rich in unsaturated fatty acids. These fats or oils are usually liquid at room
temperature. Unsaturation refers to the presence of “double bonds” in the fatty acid. The more double
bonds there are, the more unsaturated the fatty acid is. Fatty acids with just one double bond are
called “monounsaturated” and the amount in a food appears on the nutrition label. Olive oil and high
oleic sunflower oil contain mainly monounsaturated fatty acids.
Other vegetable and fish oils are abundant in polyunsaturated fatty acids with two to six double
bonds. The amount of polyunsaturated fat is also listed on the nutrition label. Heart healthy foods are
those having a majority of mono- and polyunsaturated fatty acids.
could improve human and livestock nutrition and health, the nutritional quality of food
animals for human consumption, and create ingredients with superior properties for food
manufacturing and processing.
II. Food applIcatIons For Human HealtH
A. Quantity and Quality of Food Oils
Food oils have both nutritional and functional qualities. From a nutritional perspective,
fats and oils contribute more energy (calories) than any other nutrient category, about nine
calories per gram. This compares with about four calories per gram from carbohydrates
and protein. At the same time, specifc fatty acids that comprise most of what we call
“fat” can affect a person’s risk of developing certain chronic diseases such as heart disease.
Research over the past several decades has shown that some categories of fatty acids, such
as saturated fatty acids, increase the risk of heart disease and other chronic diseases when
consumed in excess. Fatty acids also infuence how foods behave during manufacturing and
processing. For example, saturated fatty acids add stability, texture, and favor to foods, so
they are not simple to replace.
To reduce the saturated fatty acid content of foods, plant breeders and food manufacturers
increased their use of vegetable oils rich in polyunsaturated fatty acids and developed food
oils low in saturated fatty acids. One example is canola oil with 6% to 7% total saturated
Application of Biotechnology for Functional Foods
10

fatty acids. To improve the stability of vegetable oils rich in polyunsaturated fatty acids,
food manufacturers developed partially hydrogenated oils. The process of hydrogenation
reduced the polyunsaturated fatty acid content and increased oil stability, but created trans
fatty acids, which were subsequently associated with adverse health effects. As a result,
hydrogenated fats, the main source of dietary trans fatty acids, are now being eliminated
from foods. Food manufacturers are developing other ways to reduce undesirable saturated
fat content while maintaining stability such as using short chain saturated fatty acids and
monounsaturated fatty acids.
To date, one functional food oil created with the tools of biotechnology has been
commercialized. Calgene’s high lauric acid canola, Laurical™, containing 38% lauric
acid, is used in confectionary products, chocolate, and non-food items such as shampoo.
Conventional canola oil does not contain lauric acid. Laurical™ is a substitute for coconut
and palm oils. FDA approved its use in foods in 1995 (FDA 1995). The following section
describes research to date focused on developing crop varieties with other unique
oil profles.
B. Strategic Aims of Altered Fatty Acid Profile
Improving the healthfulness and functionality of food oils can be accomplished in several
ways. Where traditional plant breeding reaches its limits, biotechnology may be used to:
n Reduce saturated fatty acid content for “heart-healthy” oils
n Increase saturated fatty acids for greater stability in processing and frying
n Increase oleic acid in food oils for food manufacturing
n Reduce alpha-linolenic acid for improved stability in food processing
n Introduce various omega-3 polyunsaturated fatty acids including long-chain forms
n Enhance the availability of novel fatty acids, e.g., gamma-linoleic acid
C. Achievements in Altered Fatty Acid Profile
Reduced saturated fatty acid content: Genetically modifed soybeans have been developed
that contain about 11% saturates compared with 14% in conventional soybeans (Table
1). In May 2003, scientists reported the development of transgenic mustard greens
(Brassica juncea) containing 1% to 2% saturated fatty acids, a level signifcantly less
than in the control plants (Yao et al. 2003). The transgenic plants also contained slightly
higher amounts of oleic acid, a monounsaturated fatty acid, and higher levels of the
polyunsaturates, linoleic and alpha-linolenic acids than the control plants. These results
illustrate that alterations in one type of fatty acid may affect the levels of others, suggesting
that combined strategies or genetic transformations may be necessary to achieve specifc
fatty acid profles.
Palm oil low in saturated fatty acids is currently in development. This tropical oil contains
about half saturated fatty acids (49.3%), primarily palmitic acid (16:0, 43.5%). However,
with the recent success of biotechnology techniques in palm, transgenic palm oil enriched
in oleic and stearic acids is under development (Parveez et al. 2000). Because of the long life
cycle of palm and the time required to regenerate the plants in tissue culture, genetically
engineered palm is not anticipated for another two decades (Parveez et al. 2000).
Application of Biotechnology for Functional Foods
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Increased saturated fatty acid content: Because saturated fatty acids confer certain
functional properties to food fats and oils and are more stable to heat and processing than
unsaturated fatty acids, their use in cooking and baking is essential. To avoid the use of
animal fats and hydrogenated vegetable oils with trans fatty acids, genetic engineering
techniques have been used in canola and soy to develop oils with more short chain saturated
fatty acids—12 to 18 carbons long—mainly lauric (12:0), myristic (14:0), palmitic (16:0),
and stearic (18:0) acids. For example, Calgene’s high lauric acid canola, Laurical™,
containing 37% lauric acid, was developed using the enzyme acyl-ACP thioesterase isolated
from the California Bay Laurel (Umbellularia californica). Conventional canola oil contains
no lauric acid, and only about 6% short chain saturated fatty acids. This was the frst
transgenic oilseed crop produced commercially. High laurate canola is used in confectionary
products, chocolate, and non-food items such as shampoo as a substitute for coconut and
palm oils. FDA approved its use in 1995.
Enrichment of canola with even shorter chain saturated fatty acids, those with eight and
ten carbons, has also been accomplished (Dehesh et al. 1996). Using a palmitoyl-acyl carrier
protein thioesterase gene from a Mexican shrub, Cupea hookeriana, Dehesh and colleagues
developed lines of canola with as much as 75% caprylic (8:0) and capric acids (10:0). These
fatty acids are absent in conventional canola oil. When consumed, these water-soluble fatty
acids are mainly oxidized for energy.
Soybeans have been genetically modifed to produce oil enriched in stearic acid (18:0),
a saturated fatty acid that scientists believe does not raise serum cholesterol levels. The
stearic acid-rich oil shown in Table 1 had 28% stearic and 20% oleic acids, with lower
linoleic acid (18:2) than the conventional oil. Gene transfer technology also boosted the
stearic acid content of canola (Hawkins and Kridl 1998). Researchers at Calgene, Inc.,
Davis, CA, cloned three thioesterase genes from mangosteen, a tropical tree that stores up
to 56% of its seed oil as stearate. One of these genes led to the accumulation of up to 22%
stearate in transgenic canola seed oil, an increase of more than 1,100% over conventional
varieties (Hawkins and Kridl 1998).
Increased oleic acid content: The most recent approach to developing more healthful
food oils is increased oleic acid content. High oleic acid oils are lower in saturated and
polyunsaturated fatty acids compared with conventional oil. Oleic acid, the predominant
monounsaturated fatty acid in seed oils, is abundant in olive (72%), avocado (65%), and
canola (56%) oils, but not in others. Like saturated fatty acids, high oleic acid oils are
useful in food processing and manufacturing for maintaining functionality and stability
during baking and frying. Unlike saturated fatty acids, however, they do not raise blood
cholesterol concentrations and are therefore considered more healthful.
Biotechnology offers a means to increase the oleic acid content of vegetable oils, usually at
the expense of polyunsaturated fatty acids, and sometimes, saturated fatty acids, depending
on the particular transformations used. The concomitant reduction in polyunsaturated
fatty acids has the added advantage of increasing the stability of the oil and ultimately the
processed food. While traditional plant breeding allowed a modest increase in oleic acid,
biotechnology has been necessary to achieve the high levels desired. For example, canola
oil moderately high in oleic acid was developed using traditional plant breeding techniques.
With the application of biotechnology, oleic acid content increased to 75% (Corbett 2002).
Others have developed canola oil with more than 80% oleic acid (Wong et al. 1991, Scarth
and McVetty 1999).
Application of Biotechnology for Functional Foods
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More recently, Buhr and colleagues at the University of Nebraska used genetic engineering
to increase oleic acid levels in soybeans by inhibiting the ability of the plant to convert oleic
acid to polyunsaturated fatty acid (Buhr et al. 2002). When the conversion enzyme was
inhibited, the level of oleic acid increased from 18% in the wild-type seed to 57% in the
transgenic seed. When two gene transformations were applied, oleic acid content increased
to 85%, with saturated fatty acids reduced to 6%.
Using a different approach, scientists at DuPont used the technique of cosuppression to
reduce the production of polyunsaturated fatty acids in soybeans. Cosuppression occurs
when the presence of a gene silences or turns off the expression of a related gene. Like
Buhr and colleagues, these scientists were able to turn off the production of the enzyme
that converts oleic acid to polyunsaturated fatty acids. The result was greatly increased
production of oleic acid and reduced production of polyunsaturated fatty acids. Examples
of genetically modifed high oleic acid oils compared with their conventional counterparts
are shown in Table 1.
Gene silencing has also been used to produce high oleic and high stearic acid cottonseed
oils (Liu et al. 2002). Cottonseed oil is high in palmitic acid, very high in linoleic acid, and
free of alpha-linolenic acid. Conventional cottonseed oil has about 13% oleic acid. When
gene silencing was used to transform cotton, the resulting oil had 78% oleic and only 4%
linoleic acids, respectively, with palmitic acid reduced from 26% to 15%. Cotton was also
genetically modifed to produce high stearic oil having 40% stearic and 39% linoleic acids,
with 15% palmitic acid. A combination was also developed to have 40% stearic, 37% oleic
and only 6% linoleic and 14% palmitic acid. These examples illustrate the power and
specifcity of this technology to develop tailored seed oils.
Reduced alpha-linolenic acid: Several genetic transformations designed to increase oleic or
stearic acid content do so at the expense of the polyunsaturated fatty acids alpha-linolenic
and linoleic acids. These fatty acids have desirable nutritional characteristics, but their
presence reduces the stability of oils for baking, processing, and frying and increases their
susceptibility to oxidation or rancidity. Oils with appreciable amounts of alpha-linolenic
acid such as canola and soybean, with about 10% and 8% alpha-linolenic acid, respectively,
have been genetically modifed to reduce this fatty acid. Such oils would be desirable for the
commercial uses mentioned. Pioneer Hi-Bred, a DuPont company, developed low alpha-
linolenic acid soybean seeds through conventional breeding techniques with less than 3%
alpha-linolenic acid in its oil. Marketed under the brand TREUS™ the company claims
that the oil eliminates the need for hydrogenation in food processing. A similar product
from Monsanto, Vistive™, offers a similar level of reduction in alpha-linolenic acid.
Omega-3 fatty acids: There is extensive interest in increasing Americans’ consumption
of omega-3 fatty acids, because they are associated with many health benefts, but are
consumed only in small amounts. In 2002, the National Academy of Sciences’ Institute of
Medicine recognized that omega-3 fatty acids are essential in the diet and established an
estimated adequate intake for them (Institute of Medicine 2002). The main food sources
of the long-chain omega-3 fatty acids are fsh, especially fatty species such as salmon,
rainbow trout, mackerel, herring, and sardines. Some plants—mainly canola, soybean, and
fax oils—provide the 18-carbon omega-3 fatty acid, alpha-linolenic acid. However, higher
plants lack the enzymes to make 20- and 22-carbon polyunsaturated fatty acids needed by
mammals. Humans can convert alpha-linolenic acid to the more biologically active long-
chain forms, but they do so very ineffciently. Thus, plant foods with alpha-linolenic acid
may be insuffcient to supply the need for long-chain omega-3 fatty acids, especially during
pregnancy and lactation (Pawlosky et al. 2001).
Application of Biotechnology for Functional Foods
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Western diets contain predominately omega-6 polyunsaturated fatty acids found in
soybean, corn, sunfower, canola, and cottonseed oils. It is now recognized that diets high
in omega-6 fatty acids and low in omega-3 fatty acids may exacerbate several chronic
diseases (Simopoulos et al. 2000). Because of the many health benefts associated with the
regular consumption of omega-3 fatty acids, several health organizations, including the
American Heart Association and the 2005 Dietary Guidelines for Americans, have called
for increased consumption of these substances. One limitation to boosting consumption is
that they occur naturally mainly in fatty fsh and some seeds. Ironically, reducing the level
of alpha-linolenic acid in soy and canola oils used in food processing, may actually reduce
consumption of this fatty acid, although product developers are working to combine high
omega-3 and low alpha-linolenic traits in one product.
Although aquaculture has increased the availability of some fsh and shellfsh species,
increasing worldwide demand has put severe pressure on wild aquatic resources and limited
seafood availability. Thus, it would be desirable to increase the availability of these fatty
acids or their precursors in a variety of other foods, especially plants. Such foods would
also be useful for animal and fsh feed.
taBle 1. selected fatty acid content of vegetable oils with
modified fatty acid profiles compared with the commodity oil.

oIl
oleic
(18:1)
linoleic
(18:2)
alpha-linolenic
(18:3)
total
saturates
canola
Conventional 60 20 10 7
High oleic 75 14 3 <7
High oleic 84 5 3 5
Low linolenic 65 22 4 7
Low linolenic P6 78 11–13 2–3 N/A
High myristate/palmitate 34 15 4 43
High laurate (37%) 34 12 7 45
sunFlower
Conventional 20 65 <1 10
High oleic 82 10 <1 8
Mid oleic 56 33 <1 9
soyBean
Conventional soy 23 51 7 14
Low linolenic 23 60 2 15
High palmitic (17%) 17 55 8 20
High stearic (28%) 20 35 7 35
High oleic soybean 83 2 3 12
otHers
Conventional safflower 14 75 0 6
High oleic safflower 75 14 0 6
Conventional corn 24 58 <1 13
High oleic corn 70 - - -
Olive 75 8 <1 14
Avocado 65 15 1 14
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One strategy to increase the availability of long-chain omega-3 fatty acids is to develop
oilseed crops such as canola and soybean that contain stearidonic acid (18:4n-3). This
omega-3 fatty acid occurs naturally in only a few plants such as black currant seed oil and
echium. Stearidonic acid is the frst product formed when alpha-linolenic acid is converted
to eicosapentaenoic acid (EPA), a desirable long-chain omega-3 fatty acid. Usually, this frst
step limits the amount of EPA produced, but increasing the level of stearidonic acid helps
overcome this limitation. Then the body’s enzymes convert stearidonic acid to 20-carbon
polyunsaturated fatty acids.
Dr. Virginia Ursin and colleagues at Calgene studied the metabolism of stearidonic acid in
people (James et al. 2003). Her studies showed that when either stearidonic acid or EPA was
consumed the amount of EPA in red blood cells increased signifcantly. This fnding meant
that the stearidonic acid was converted to EPA and appeared in red cells just as readily as
the preformed EPA. In contrast, when the study volunteers consumed alpha-linolenic acid,
there was no change in their red cell EPA content. None of the fatty acids consumed had
any effect on cell DHA levels, another long-chain omega-3 fatty acid associated with health
benefts. Although the study used supplements, not stearidonic acid from transgenic plants,
the fndings suggest that plants with stearidonic acid would have potential to provide EPA.
Toward this end, scientists at Calgene, have successfully transformed canola so that it
makes stearidonic acid. This genetic engineering feat required two genes from the fungus
Mortierella alpina and one from canola for the three enzymes needed to produce suffcient
stearidonic acid (Ursin 2003). The engineered plants accumulated up to 23% stearidonic
acid in the seed oil with a reduction in oleic acid content from about 60% to about 22%.
By breeding the transgenic lines with various lines of canola the investigators were able to
develop a line of canola containing more than 55% of alpha-linolenic acid and stearidonic
acid. Total omega-6 fatty acids remained about 22%, a level similar to conventional canola.
Calgene scientists have also developed soybean that contains stearidonic acid (Ursin,
personal communication 2004).
The implications of Calgene’s work with stearidonic acid are substantial. This is the frst
demonstration of the incorporation into edible plants of a biologically potent source of
long-chain omega-3 fatty acids. This work marks an important advance in the development
of plant-based sources of long-chain omega-3 fatty acids that could be consumed directly
or incorporated into food products. However, because stearidonic acid contains four double
bonds, it is vulnerable to oxidation and would require antioxidant protection. One can
imagine that transgenic canola and soybean could be developed using additional traits to
boost antioxidant protection, possibly from vitamin E.
In May 2004, a landmark paper announced the production of long-chain polyunsaturated
fatty acids—both omega-6 and omega-3 types—in Arabidopsis thaliana, a type of cress
widely used as a model plant in biotechnology research. Dr. Baoxiu Qi and colleagues
at the University of Bristol, United Kingdom, transferred to Arabidopsis thaliana three
genes encoding for different enzymes in the metabolic pathway from linoleic and alpha-
linolenic acids to arachidonic and eicosapentaenoic acids, respectively (Qi et al. 2004). The
additional genes were necessary to provide the enzymes to make these long-chain fatty
acids. Yields of EPA (13%) and arachidonic acid (29%) in leaves were signifcantly higher
than in conventional cress, which usually does not produce these fatty acids, and accounted
for 43% of the total 20-carbon polyunsaturated fatty acids. In addition to the production
of EPA and arachidonic acid, the concentration of alpha-linolenic acid was reduced from
48% to 14%. This achievement was also remarkable because it used a pathway seldom
found in plants.
Application of Biotechnology for Functional Foods
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This work is important in several regards. One is that it demonstrates the feasibility of
developing plants capable of synthesizing long-chain polyunsaturated fatty acids. Another
is the relatively high effciency of conversion of the precursor fatty acids to the long-chain
forms. A third advantage is the improved balance of omega-6 and omega-3 fatty acids,
with signifcant reduction in the amounts of the 18-carbon precursors linoleic and alpha-
linolenic acid compared with conventional plants. Yet another is the demonstration that
plants can be engineered not only with respect to the outcome of fnal products, but also the
pathways for achieving the desired ends. A likely next step will be to apply this technology
to seed oil crops such as canola and soybean to see if the long-chain polyunsaturated fatty
acids will accumulate in the seed.

Although production of EPA in plants represents an enormous scientifc achievement, the
question of making Docahexenoic Acid (DHA), a 22-carbon polyunsaturated omega-3 fatty
acid important in retina and brain function and other body systems remained unsolved.
In mammals, the conversion of EPA to DHA is ineffcient and requires several steps. It
is possible, in theory, to perform this conversion in a direct manner, but the enzymes to
do so are not present in mammals. Several research groups have examined many algae
and identifed the specifc enzymes for this conversion (Sayanova and Napier 2004, Meyer
et al. 2004). Once the genes for these enzymes were identifed and cloned they could be
incorporated into model organisms to see whether DHA would be produced. In late 2004,
Amine Abbadi at the University of Hamburg, Germany, working with colleagues in the U.K.
and the U.S., reported the successful transformation of yeast that yielded small amounts of
DHA (Abbadi et al. 2004). This accomplishment required four gene transformations. The
team then went on to develop transgenic fax, a plant with abundant alpha-linolenic acid
for conversion to long-chain fatty acids.
Several steps remain before long-chain polyunsaturated fatty acids will be available in
commercial crops. However, the demonstration that plants can be modifed to make these
important nutrients means that many of the scientifc hurdles have been conquered. This
work gives a large boost to the potential for plants to be an important dietary source of
these fatty acids.
Gamma-linolenic acid: This fatty acid is the frst step in the conversion of linoleic acid to
arachidonic acid in the omega-6 fatty acid pathway. When consumed in evening primrose
or borage oils, it is poorly converted to arachidonic acid. For that and other reasons, it may
have potential beneft in cardiovascular disease (Fan and Chapkin 1998). Gamma-linolenic
acid has been associated with improved skin conditions in human subjects, improved liver
function in patients with liver cancer, and with anti-cancer effects in cell culture studies. It
was also shown to enhance the effectiveness of tamoxifen, an anti-estrogenic medication
used to prevent the recurrence of breast cancer. It is believed to suppress the production of
estrogen receptors in cells.
Gamma-linolenic acid is naturally present in appreciable amounts in few plants, notably
borage (Borago offcinalis), evening primrose (Oenothera biennis), black currant oil (Ribes
nigrum) and echium (Echium plantagineum). The ability to increase the production of
gamma-linolenic acid in tobacco plants by transferring the gene for the delta-6 desaturase
enzyme from various sources was frst shown in 1996 by Reddy and Thomas at Texas A&M
University, and by others in 1997 (Reddy and Thomas 1996, Sayanova et al. 1997). A recent
study reported that gamma-linolenic acid content in transgenic canola ranged from 22%
to 45% (Wainright et al. 2003). Evening primrose has also been genetically modifed for
enhanced gamma-linolenic acid content (Wainright et al. 2003). Arcadia Biosciences, Davis,
CA, has also reported transgenic saffower plants with 65% gamma-linolenic acid in the oil.
Application of Biotechnology for Functional Foods


Ursin’s study of transgenic canola enriched in stearidonic acid discussed above also
reported that a cross between the transgenic line of canola producing stearidonic acid and
a canola line high in gamma-linolenic acid yielded a canola containing about 11% gamma-
linolenic acid and about 14% stearidonic acid (Ursin 2003). This example illustrates the
variety of fatty acids that can be developed in seed oils using a combination of genetic
engineering and traditional plant breeding techniques.
III. QuantIty and QualIty oF plant proteIn
Efforts to improve the protein content and quality of staple foods have been underway for
decades. The main focus is crops grown in developing countries, where nutrient shortfalls
are widespread and dietary diversity limited. Foods such as potato and cassava, staple foods
in several parts of South America and Africa, have less than one percent protein.
Efforts to improve protein quality strive to increase the amount of limiting essential amino
acids provided by the protein in the food. The amino acids most often present in inadequate
amounts are lysine, tryptophan, and methionine. Improvements in protein quality beneft
both human and animal nutrition and increase the feed effciency of crops fed to food
animals. For example, corn is widely fed to cattle but it is limiting in lysine and methionine.
Corn with higher levels of these amino acids would signifcantly improve feed effciency and
lower input costs to farmers. Improved corn varieties consumed by humans would also have
nutritional benefts.
There are various ways of improving protein quantity and quality. One is to increase the
total amount of protein produced by selecting germplasm with an altered balance of seed
proteins. This may be done by traditional cross breeding or genetic engineering. Another
approach is to introduce genes from other sources for proteins that have a favorable balance
of essential amino acids. An example is the introduction into potato of a gene for seed
albumin protein from amaranth. A third approach seeks to increase the production of
specifc amino acids such as lysine. This approach was used in the development of Quality
Protein Maize, discussed below.
William Folk and his team at the University of Missouri, Columbia, MO, pioneered another
approach to improve seed protein quality. Their strategy was to substitute more desirable
and scarce amino acids for more abundant ones in certain seed proteins (Chen et al. 1998,
Wu et al. 2003). They applied this concept to rice by increasing the production of lysine, an
essential amino acid, at the expense of the non-essential amino acids, glutamine, asparagine
and glutamic acid.
Cassava: A staple food for some 500 million people in tropical and sub-tropical parts of
the world, cassava (Manihot esculenta Crantz), also known as yucca or manioc, thrives in
marginal lands having little rain and nutrient-poor soils. It is widely consumed in Africa,
and parts of Asia and South America. Cassava root has less than 1% protein and poor
nutritional value. However, the leaves are also consumed and these are a good source of
beta-carotene, the precursor of vitamin A.
In 2003, Zhang and colleagues reported using a synthetic gene to increase the protein
content in cassava (Zhang et al. 2003). The gene is for a storage protein rich in nutritionally
essential amino acids. When the gene was expressed in cassava, transformed plants
expressed the gene in roots and leaves, both of which are consumed in human diets. The
experiment demonstrated the feasibility of increasing the quantity and quality of protein
in cassava.
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Cassava also contains cyanogenic glucosides that can produce chronic toxicity if not
eliminated or reduced by grating, sun-drying, or fermenting. Efforts to develop cassava
varieties low in these toxicants is a high research priority.

Corn: Corn (Zea mays) is the predominant staple food in much of Latin America and
Africa. Although some varieties may contain appreciable quantities of protein, its quality
is poor because of low lysine and tryptophan content. In 1964, it was discovered that
corn bearing a gene known as opaque-2 contained increased concentrations of lysine
and tryptophan and had signifcantly improved nutritional quality (Food and Agriculture
Organization 1992). However, opaque-2 corn proved to have low yields, increased
susceptibility to diseases and pests, and inferior functional characteristics.
At the International Maize and Wheat Improvement Center (CIMMYT) in Mexico, work
with the opaque-2 gene continued using both traditional breeding and molecular methods.
After at least 12 years’ work, CIMMYT researchers succeeded in developing hardy corn
varieties that contained twice the lysine and tryptophan content as traditional varieties,
but were disease-resistant and high-yielding. Scientists Surinder K. Vasal and Evangelina
Villegas of CIMMYT were awarded the World Food Prize in 2000 for their work developing
‘Quality Protein Maize’. Quality Protein Maize varieties have been adapted to and released
in over 40 countries in Latin America, Africa, and Asia.
Recent researchers at CIMMYT reported the development of transgenic corn with multiple
copies of the gene from amaranth (Amaranthus hypochondriacus) that encodes for the seed
storage protein amarantin (Rascon-Cruz et al. 2004). Total protein in the transgenic corn
was increased by 32% and some essential amino acids were elevated 8% to 44%.
In 2004, a team of researchers at the University of California, Riverside, reported that
transgenic corn with increased production of the plant regulating hormone, cytokinin, had
nearly twice the content of protein and oil as conventional corn (Young et al. 2004). This
development resulted from an unusual change in the way the plant developed. Normally,
corn ears develop fowers in pairs, one of which usually dies. Under the infuence of the
additional cytokinin, both fowers developed but yielded only a single kernel. These kernels
contained more protein and oil than conventional corn.
Pursuing a different strategy to improve protein quality, researchers at Monsanto, St. Louis,
MO, used genetic engineering techniques to reduce the amount of zein storage proteins.
These storage proteins constitute over half the protein in corn and are defcient in lysine
and tryptophan. Increased production of other proteins in the corn led to higher levels of
lysine, tryptophan, and methionine (Huang et al. 2004). The agronomic and nutritional
properties of these lines are currently being evaluated.
Researchers at the Max Planck Institute, Germany, have focused on methionine, another
limiting amino acid. They elucidated several key steps in methionine metabolism in
plants. This work, currently in the preliminary stage, could pave the way for using
genetic engineering techniques to improve the methionine content of plants (Hesse
and Hoefgen 2003).
Potato: Potato (Solanum tuberosum) is a dietary staple throughout parts of Asia, Africa,
and South America. Typically, potatoes contain about 2% protein and 0.1% fat. It was
reported in 2000 that, as in cassava, transfer of the gene for seed albumin protein from
Amaranthus hypochondriacus to potato resulted in a “striking” increase in protein content
of the transgenic potatoes (Chakraborty et al. 2000). In 2004, researchers at the National
Application of Biotechnology for Functional Foods
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Centre for Plant Genome Research, India, reported the development of a nutritionally
improved potato line with 25% higher yields of tubers and 35%–45% greater protein
content (ISAAA 2004). Dubbed the “protato,” the protein-rich potato had signifcant
increases in lysine and methionine, which enhance the quality of the additional protein
(Council for Biotechnology Information 2004). In February 2004, this potato was reported
“approaching release” to farming communities.
It should be noted that while potatoes are known for their high starch content, it has been
possible to genetically engineer potatoes that contain fat (triglycerides). In July 2004, Klaus
and colleagues at the Max Planck Institute of Molecular Plant Physiology demonstrated
increased fatty acid synthesis in potatoes (Klaus et al. 2004).
Rice: Almost half the world’s population eats rice (Oryza sativa L.), at least once a day
(IRRI undated). Rice is the staple food among the world’s poor, especially in Asia and parts
of Africa and South America. It is the primary source of energy and nutrition for millions.
Thus, improving the nutritional quality of rice could potentially improve the nutritional
status of nearly half the world’s population, particularly its children. Commodity rice
contains about 7% protein, but some varieties, notably black rice, contain as much as 8.5%
(Food and Agriculture Organization 2004). The most limiting amino acid in rice is lysine.
Efforts to increase the nutritional value of rice target protein content and quality along
with key nutrients often defcient in rice-eating populations, such as vitamin A and iron.
The International Rice Research Institute (IRRI), Philippines, is a primary center for rice
research and development of improved varieties.
In 1999, Dr. Momma and colleagues at Kyoto University, Japan, reported a genetically
engineered rice having about 20% greater protein content compared with control rice
(Momma et al. 1999). Transgenic plants containing a soybean gene for the protein glycinin
contained 8.0% protein and an improved essential amino acid profle compared with 6.5%
protein in the control rice.
As mentioned briefy above, Dr. William Folk and his team genetically modifed rice to
increase its content of the amino acid lysine (Wu et al. 2003). They did so by modifying
the process of protein synthesis, rather than by gene transfer or the expression of new
proteins. They achieved an overall 6% increase in lysine content in the grain (Chen et al.
1998). Although lysine content remained below optimum levels, the scientists suggested that
additional transformations and modifcations could further boost lysine levels.
Perhaps the most famous genetic transformations in rice are those in “Golden Rice”
involving the vitamin A precursor, beta-carotene, and iron. The lead scientist in the golden
rice project, Dr. Ingo Potrykus, now retired from the Swiss Federal Institute of Technology,
was also involved in applying biotechnology for the improvement of rice protein. Although
details are sparse, Potrykus described the work of Dr. Jesse Jaynes, who synthesized a
synthetic gene coding for an ideal high-quality storage protein with a balanced mixture
of amino acids. The gene, named Asp-1, was transferred to rice with the appropriate
genetic instructions for its production in the endosperm or starchy part of the rice grain.
The transgenic rice plants accumulated the Asp-1 protein in their endosperm in a range
of concentrations and provided essential amino acids but data are not yet available on the
concentrations achieved or their nutritional relevance. Precedent for the expression of a
synthetic gene in rice grown in cell culture suggests that Jaynes’ approach is viable (Huang
et al. 2002).
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IV. modIFIed carBoHydrate
A. Starch
Starch from cereals, grains, and tubers contribute a substantial share of dietary calories
and in many poor countries, provide the majority of food energy. Starch is also important
for feed and industrial purposes. Its use as paste goes back at least 4000 years BCE to the
Egyptians who cemented strips of papyrus stems together with starch paste for writing
paper.
Besides providing energy, starch confers functional characteristics to foods: texture,
viscosity, solubility, gelatinization, gel stability, clarity, etc. These characteristics depend
on the proportion of amylose and amylopectin, the main components of starch. Amylose
and amylopectin differ from each other in chain length, branching, and degree of
polymerization. Amylose is linear and amylopectin is highly branched. How a particular
starch will be used in foods, determines what ratio of amylose to amylopectin is most
suitable. High amylose starches include high amylose corn (70%), corn (28%), wheat (26%)
and sago (26%). In contrast, waxy rice and waxy sorghum contain no amylose. Members
of the potato family—potato, sweet potato, cassava—have 17% to 20% amylose.
Many lines of corn have been developed with different characteristics derived from modifed
starch ratios and increased amylose content. Transgenic high amylose potatoes developed
by inhibiting two branching enzymes were reported to yield more tubers and have lower
starch content, smaller granules, and increased reducing sugars (Hofvander et al. 2004).
Biotechnology has also been directed to increasing starch content (Geigenberger et al. 2001,
Regierer et al. 2002). Potatoes were genetically altered to increase the activity of adenylate
kinase, an enzyme involved in the plant’s energy metabolism and starch production.
The resulting transgenic potatoes had substantially increased adenylates and a 60%
increase in starch compared with wild-type plants (Regierer et al. 2002). Unexpectedly, the
concentrations of several amino acids were increased 2- to 4-fold, and tuber yield increased.
Considerable publicity was given to potatoes engineered by Monsanto in the early 1990s
to have increased starch content. These were touted as more desirable for French fries
because they would absorb less fat during frying. They are an example of the type of starch
modifcation that may have secondary health benefts as a consequence of how they are used.
B. Fructan
Fructans are polymers (repeating units) of the sugar fructose. They serve in food
products as a low-calorie sweetener, source of dietary fber, and bulking agent. They
may also stimulate the growth of desirable colonic bacteria, such as bifda. Fructans have
environmentally friendly non-food applications in the manufacture of biodegradable
plastics, cosmetics, and detergents. Fructans are naturally occurring in Jerusalem artichokes
(sunchokes) and chicory, but agronomic shortcomings in growing these crops have limited
their use.
Inulin, a fructan found in Jerusalem artichokes, was successfully synthesized in potatoes
following the transfer of two genes from globe artichokes (Cynara scolymus) (Hellwege et
al. 2000). The full spectrum of inulin molecules present in artichokes was expressed in the
transgenic potatoes. Inulin comprised 5% of the dry weight of the transgenic tubers and
did not infuence sucrose concentration. However, starch content was reduced.
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In a program called the Agriculture and Fisheries Programme, or FAIR, the European
Commission funded multidisciplinary research programs in agriculture and fsheries,
including a project on fructans for food and non-food uses. Research to date includes the
isolation of several genes for fructosyl transferase enzymes involved in the production of
fructans. The feasibility of using these enzymes has been demonstrated in model plants and
target crops such as sugar beet (Anonymous 2000). In addition, it was reported in 2004 that
genes encoding for fructosyl transferases in onion were isolated and transferred to sugar
beet, a plant that does not normally synthesize fructans (Weyens et al. 2004). Following
the transfer of the genes, onion-type fructans were produced from sucrose without loss in
storage carbohydrate.
V. Increased VItamIn content In plants
A. Beta-carotene and Other Carotenoids
Beta-carotene belongs to the family of carotenoids and is abundant in plants of orange
color. It is the precursor of vitamin A and can be converted to the active vitamin during
digestion. Other carotenoids do not have potential vitamin A activity. Humans cannot
synthesize carotenoids and therefore depend on foods to supply them. Many staple foods,
particularly rice, contain no beta-carotene or its precursor carotenoids. Diets lacking other
food sources of vitamin A or beta-carotene are associated with vitamin A defciency which
can result in blindness, severe infections, and sometimes death. According to the World
Health Organization, vitamin A defciency is the leading cause of preventable blindness
worldwide. The defciency affects some 134 million people, particularly children, in 118
countries. Overcoming this nutrient defciency is an urgent global health challenge.
The development of “Golden Rice,” so named because of its yellow color conferred by the
presence of beta-carotene, was a landmark achievement in the application of biotechnology
to nutrition and public health. Peter Burkhardt, working with Ingo Potrykus and colleagues
in Switzerland, was the frst to show that transgenic rice, carrying a gene from daffodil,
could express phytoene, a key intermediate in the synthesis of beta-carotene (Burkhardt et
al. 1997). Subsequently, the Potrykus group reported the application of three transgenes in
the development of rice expressing the entire pathway for the production of beta-carotene
(Ye et al. 2000, Beyer et al. 2002). Additional work with Golden Rice included the insertion
of a gene to increase the iron content (Potrykus 2003). IRRI is currently cross-breeding the
nutrient-enhanced transgenic rice with local rice varieties from Asia and Africa, and feld-
testing the new lines for nutritional value and agronomic performance. Varieties of Golden
Rice are not expected to be ready for farmers for several more years.
The development of transgenic plants able to produce a variety of carotenoids is an active
area of research. It is clear that production of phytoene, the frst product in the pathway for
carotenoid synthesis, is the rate-limiting step in generating carotenoids (Cunningham 2002).
Using gene transfer technology to increase the expression of phytoene synthase, the enzyme
that makes phytoene, increases the synthesis of carotenoids substantially. For example,
Shewmaker and colleagues (1999) from Monsanto reported an increase up to 50-fold in
carotenoids, mainly alpha and beta-carotene, in canola (Brassica napa). However, vitamin
E levels decreased signifcantly, oleic acid content increased, and linoleic and alpha-linolenic
acids were reduced compared with non-transgenic seeds. These other changes would have
to be modifed or evaluated to determine whether they might have meaningful nutrition
implications.
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In a separate study on canola, the Monsanto group transferred to canola three genes from
bacteria that affect the phytoene synthesis pathway. When they included a triple construct—
genes for three different enzymes, phytoene synthase, phytoene desaturase and lycopene
cyclase—the resulting transgenic canola seeds maintained the same amount of total
carotenoids, but increased the ratio of beta to alpha-carotene from 2:1 to 3:1 (Ravanello
et al. 2003).
Stalberg and colleagues in Sweden also studied the effects of phytoene synthase on
carotenoid synthesis in transgenic Arabidopsis thaliana. They examined three keto-
carotenoids; transformed seeds had a 4.6-fold increase in total pigment and a 13-fold
increase in these three carotenoids (Stalberg et al. 2003). They also reported a 43-fold
average increase in beta-carotene (Lindgren et al. 2003). Lutein, another nutritionally
important carotenoid, was signifcantly increased, but zeaxanthin was only increased by a
factor of 1.1. They also observed substantial levels of lycopene and alpha-carotene in the
seeds, whereas only trace amounts were found in the control plants. However, germination
was delayed in proportion to the increased levels of carotenoids.
Others have examined the effect of transgenes affecting phytoene metabolism on carotenoid
synthesis. Dr. Peter Bramley’s group at the University of London, United Kingdom,
transformed tomatoes using a bacterial gene encoding for an enzyme that converts
phytoene to lycopene, the precursor of beta-carotene. Tomatoes carrying the bacterial
gene had about a 3-fold increase in beta-carotene content, but total carotenoids were not
increased (Romer et al. 2000). The altered carotene content did not affect plant growth and
development.
Lutein and zeaxanthin are nutritionally important carotenoids for protection of the retina
and reduced risk of age-related macular degeneration (Krinsky et al. 2003, Gale et al. 2003).
Lutein is found in dark green leafy vegetables such as spinach and collards, and zeaxanthin
occurs in yellow foods such as mangoes, corn, and peaches. The latter is not particularly
abundant in Western diets. Romer and colleagues at Universitat Konstanz, Germany, were
able to use biotechnology to block the conversion of zeaxanthin to another carotenoid
and thereby increase its content in potatoes (Romer et al. 2002). With this approach they
obtained increased levels of zeaxanthin in potatoes ranging from 4- to 130-fold. Total
carotenoids were increased by 5.7-fold, but in some, lutein content was decreased. Alpha-
tocopherol (vitamin E) was increased 2- to 3-fold in the transgenic potatoes. Fine-tuning
these alterations has the potential to signifcantly enhance the nutritional value of potatoes.
In another study, Bramley’s group transferred the gene that increases carotenoid synthesis
from a bacterium to tomatoes and measured total and specifc carotenoids in the transgenic
fruits (Fraser et al. 2002). Total carotenoids were 2- to 4-fold higher in transgenic fruits than
in nontransformed plants, with increases in phytoene, lycopene, beta-carotene, and lutein
of ranging from 1.8- to 2.4-fold.
Tomatoes with delayed ripening were produced as a result of inserting a gene encoding for
S-adenosylmethionine decarboxylase, an enzyme involved in the ripening process (Mehta
et al. 2002). An additional consequence of this transgenic modifcation was a several-
fold increase in lycopene content. Lycopene is normally converted to beta-carotene, but
tomatoes with increased lycopene content may have enhanced nutritional value. Lycopene
consumption has been linked to reduced risk and spread of prostate cancer, though
defnitive data are lacking (Etminan 2004, Kristal 2004).
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The carotenoid astaxanthin is synthesized by algae and plants and is responsible for the
pink color in shrimp and salmon. Humans absorb astaxanthin poorly, but absorption is
increased in the presence of fat (Mercke Odeberg et al. 2003). Astaxanthin is of interest
because of its strong antioxidant properties in vitro. It is less certain whether it is an
antioxidant in human health. Astaxanthin is used commercially in feed for cultured
salmon and trout.
Production of astaxanthin in fowers and fruits has also been accomplished with the
techniques of biotechnology. Using a gene from the alga Haematococcus pluvialis,
researchers at The Hebrew University of Jerusalem, Israel, transferred the gene into tobacco
(Nicotiana tabacum). Transgenic tobacco plants produced astaxanthin and changed color
(Mann et al. 2000). This ability to manipulate pigmentation in fruits and fowers may have
commercial potential and possible implications for increasing the availability of carotenoids
for human health.
B. Vitamin E
There is strong interest in vitamin E because of studies linking it to decreased occurrence
of several degenerative diseases and cancers, although effcacy remains unproven and data
are inconsistent (71, 72). Some recent trials with vitamin E supplementation reported no
protection against cardiovascular disease or cancer and some chance of increased risk
of heart failure (Eidelman et al. 2004, Lonn et al. 2005, Miller et al. 2005). Also, because
vitamin E is an anti-oxidant, it is useful in foods and oils to provide oxidative stability.
Vitamin E is found mainly in vegetable oils, wheat germ, and a few other foods not widely
consumed. The most potent form of the vitamin is alpha-tocopherol, but the less potent
gamma, beta, and delta forms are more widespread in plants. Efforts to increase the content
of vitamin E in food plants, particularly cereals and grains, which may have low amounts,
have sought to increase the amount of precursor substances by overexpressing the genes
for various enzymes involved in the biosynthetic pathway. Vitamin E biosynthesis involves
complicated pathways so that multiple genetic manipulations are required.
In September 2003, Dr. Edgar Cahoon of the U.S. Department of Agriculture (USDA) and
co-researchers at the Donald Danforth Plant Science Center, St. Louis, MO, announced
the development of transgenic corn with increased levels of vitamin E. Insertion of a gene
from barley into corn increased the conversion of vitamin E precursors to vitamin E itself
(Cahoon et al. 2003). The content of vitamin E and tocotrienol, a closely related substance,
was increased up to 6-fold. However, much of the antioxidant produced was tocotrienols
rather than vitamin E (Aijawi and Shintani 2004). Tocotrienols, although potent
antioxidants in vitro, are poorly absorbed in humans; however, they may have cholesterol
lowering properties (Theriault et al. 1999). Besides enhancing the potential therapeutic and
nutritional value of corn, this alteration may increase its oxidative stability after harvest.
In soybeans, Van Eenennaam and colleagues developed transgenic plants that were able to
increase the conversion of the weaker forms of tocopherol typically present in soybeans
to the more potent alpha-tocopherol (vitamin E) form. The result was a 5-fold increase
in vitamin E activity (Van Eenennaam et al. 2003). This work paves the way for the
development of vitamin E-rich oils and plants with potential health benefts.
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C. Vitamin C
Vitamin C, or ascorbic acid, is abundant in citrus and other fruits such as strawberry and
kiwi, but is very low in cereals and grains. Moreover, it cannot be synthesized by humans
nor stored to any appreciable extent. Thus, we depend on regular dietary consumption
to meet our vitamin C needs. In areas of the world where foods containing vitamin C
are not widely consumed, strategies to increase the vitamin C in cereals and grains hold
considerable potential to improve health.

In 2003, Gallie and colleagues at the University of California, Riverside, announced the
successful transformation of corn and tobacco that resulted in a 20% increase in vitamin
C (Chen et al. 2003). They accomplished this increase by transferring a gene from wheat
for an enzyme that recycles vitamin C and prevents its breakdown. Increased expression of
this enzyme in transgenic corn and transgenic tobacco resulted in a 2- to 4-fold increase in
vitamin C content in the kernel. Application of this approach to other food plants remains
to be developed and evaluated.
The ability to increase the vitamin C content of strawberries, a good source of the vitamin,
was reported in 2003 by a research team at the University of Málaga, Spain (Agius et al.
2003). Using a gene for the enzyme D-galacturonate reductase transferred to strawberry
(Fragaria spp) this group showed that vitamin C content in the modifed strawberry fruit
increased with the expression of the transgene. This study demonstrated the feasibility of
using this enzyme to raise the level of vitamin C in plants.
D. Folic Acid
Inadequate intake of the vitamin folic acid, one of a family of folates, is associated with
megaloblastic anemia, birth defects, impaired cognitive development, and some chronic
diseases. Folates are available in small amounts in a variety of fruits and vegetables, but
intakes tend to be low. For this reason, it would be desirable to increase the concentration of
folates in dietary staples and foods widely consumed. In the U. S. several foods are fortifed
with folic acid. In 2004, Hossain and colleagues at Tufts University, Medford, MA, and the
Donald Danforth Plant Science Center reported a 2- to 4-fold increase in folates and pterins,
the precursors of folates, in transgenic Arabidopsis thaliana modifed by the incorporation
of the transgene for the frst step in the synthesis of folic acid (Hossain et al. 2004). Other
investigators have developed transgenic tomatoes, also enriched in the same gene, that had
twice the amount of folate as control fruit (Diaz de la Garza et al. 2004). This group was
able to boost folate content 10-fold by including a second gene transformation to increase
the content of another substance, para-aminobenzoate, needed for folate synthesis. These
studies provide good evidence of the potential to increase the availability of this vitamin in
widely consumed foods.
E. Antioxidants
Vitamins C and E function as antioxidants in the body. However, many other substances
widely distributed in foods in small amounts also provide protection against potentially
damaging breakdown products arising from oxidation during normal metabolism.
Oxidation breakdown products, such as reactive oxygen species, oxidized lipids, and free
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radicals have been associated with chronic diseases, so there has been considerable interest
in the availability of antioxidants. Caution should be sounded, however, because in small
amounts many of these substances are protective; in high doses, they can act as prooxidants
and may be harmful. Several examples of the applications of biotechnology for enhanced
antioxidant capacity in foods are described below.
Phenolic compounds are the most widespread antioxidants in foods. They include such
substances as favanols, tocopherols, quercetin, resveratrol, and many others. They have
become familiar to consumers because they are touted in foods as diverse as berries,
wine, tea, olive oil, and many others. Potatoes are a source of antioxidant favanoids and
vitamin C. To enhance the antioxidant content of potatoes, Lukaszewicz and colleagues
conducted a series of transformations using one or multiple genes encoding enzymes in
the biofavanoid synthesis pathway (Lukaszewicz et al. 2004). Transgenic plants exhibited
signifcantly increased levels of phenolic acids and anthocyanins, plus improved antioxidant
capacity. However, starch and glucose levels were decreased. These fndings point to
complex relationships between antioxidant content and other compounds, but indicate
that antioxidant levels in potatoes can be altered using biotechnology.
Another phenolic antioxidant, chlorogenic acid, accumulates in some crops and is
found in apples, green coffee beans, tomatoes, and tea. It is synthesized by the enzyme
hydroxycinnamoyl transferase in solanaceous plants (e.g., potato, tomato, eggplant).
In 2004, it was reported that transgenic tomatoes carrying the gene for this enzyme
accumulated higher levels of chlorogenic acid with no side effects on levels of other
phenolics (Niggeweg et al. 2004). The transgenic tomatoes also showed improved
antioxidant capacity, suggesting that such enhanced tomatoes might provide additional
antioxidants.
Yet another transformation in tomatoes was recently reported to result in the synthesis
of resveratrol, an antioxidant not normally found in tomatoes. Resveratrol is usually
associated with grapes and wines where it is abundant. In this study, tomatoes
incorporating the gene for stilbene synthase, an enzyme in the pathway for resveratrol
synthesis, had a resveratrol content of 53 mg/g fresh tomato upon ripening (Giovinazzo
et al. 2005). The contents of two other antioxidants, vitamin C and glutathione, were also
increased.
VI. trace mIneral content and BIoaVaIlaBIlIty
Improving human nutrition by increasing the availability of trace minerals in crops is
potentially highly effcient and effective. This strategy may reach more people in developing
countries than fortifcation of foods, because many subsistence farmers grow their own
food and are outside the market system. If they have access to and consume improved
crop varieties, they will not only improve their nutrient intake, they may improve their
crop yields and consequently their economic wellbeing. This is because trace minerals
are essential to the plant’s ability to resist disease and other environmental stresses (Bouis
2002). Further, plants with improved ability to take up minerals from the soil will not
deplete nutrient-poor soils. Such plants are able to unbind minerals in the soil and make
them available to the plant, thus making use of an abundant resource in the soil that is
otherwise unavailable. Mineral-effcient plants are also more drought resistant and require
fewer chemical inputs (Bouis 2002).
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A. Iron
Iron defciency anemia is one of the most widespread nutritional defciencies in the world.
The United Nations estimates that over three billion people in developing countries are
iron-defcient (Administrative Committee 2000). The problem for women and children
is more severe because of their greater need for iron. For this reason, the enrichment of
staple foods, especially those consumed in poor countries, is one of the top priorities in
international agricultural and nutrition research. In rice-eating populations, iron defciency
anemia is caused by insuffcient dietary iron, absorption inhibitors such as phytate, and
lack of enhancing factors for iron absorption such as ascorbic acid.

Although much is known about the uptake of iron and zinc in roots and transport of
minerals to and from vegetative parts of the plant, some plants accumulate very little trace
minerals in the grain (Holm et al. 2002). For example, in wheat only 20%, and in rice just
5% of the iron in leaves is transported to the grain. In cereals, much is stored in the husk
and subsequently lost during milling and polishing. Thus, strategies to increase the iron
content of cereals and grains face the challenge of targeting iron storage in a form and
location in the plant where it will be bioavailable when consumed.
A signifcant breakthrough in improving the iron content of cereals was achieved by Ingo
Potrykus and colleagues. One of the genes transferred to Golden Rice came from the
common bean, Phaseolus vulgaris. This gene encoded for the iron storage protein, ferritin,
and when expressed in the transgenic rice increased the iron content twofold (46). The
bioavailability of iron in transgenic rice varieties containing ferritin was shown to be as
good as ferrous sulphate, commonly used in iron supplements, as refected in biochemical
indices of iron status in iron-defcient laboratory rats (Murray-Kolb et al. 2002).
A different source of ferritin genes, soybean, was used in the transformation of rice to
increase iron content (Goto et al. 1999). Researchers at the Central Research Institute
Electric Power Industry, Japan, transferred the gene for ferritin from soybean into rice and
confrmed the stable incorporation of the ferritin subunit in the rice seed. Iron content in
the transgenic rice seeds was up to threefold greater than in non-transgenic control plants.
Others have used recombinant soybean ferritin under a different promotor to increase the
iron content in wheat and rice. In this case, iron was signifcantly increased in vegetative
tissues but not in seeds (Drakakaki et al. 2000). Thus, the experimental conditions, type
of promotor used, mineral transport and storage in the plant, and other conditions have
substantial effects on the outcome of genetic engineering experiments to increase mineral
content.
Iron transport and uptake in plants is carefully regulated. This is because iron has low
solubility and is toxic in excess. Recent studies have examined the function of iron
transporter proteins in transgenic plants. These proteins have been shown to increase iron
uptake into roots when iron is defcient (Eide et al. 1996). The iron transporter protein,
IRT1, frst isolated from Arabidopsis thaliana, also transports other metals such as zinc,
manganese, lead, and cadmium; the latter two can be toxic. Researchers in the laboratory
of Dr. Mary Lou Guerinot at Dartmouth College, Hanover, NH, have shown that slight
changes in the amino acid composition of the transporter protein affects the selectivity of
metals transported into the plant (Rogers et al. 2000). This fnding introduces the possibility
of engineering plants with the ability to take up desirable minerals while excluding toxic
and undesirable ones.
Application of Biotechnology for Functional Foods


A second iron transporter protein, IRT2, has also been identifed in the roots of
Arabidopsis. When the gene for IRT2 was incorporated into iron- and zinc-defcient plants,
iron uptake was increased (Vert et al. 2001). Unlike the IRT1 transporter, however, IRT2 did
not transport manganese and cadmium when it was expressed in yeast. This observation
suggests ways in which selective genetic transformations might be used to enhance the
uptake of some minerals while excluding others.
It should be noted that iron and zinc levels tend to be present together in many plants,
although the average content of each differs. For example, in screening over 1,000 varieties
of common beans, a nutritional staple in many countries, scientists at the International
Institute of Tropical Agriculture, Nigeria, found that iron content averaged about 55 mg/
g iron, but some varieties from Peru averaged more than 100 mg/g iron (Gregorio 2002).
Zinc content, averaged 35 mg/g. When varieties were selected for their iron content, higher
zinc levels were obtained as well. These observations suggest that genetic modifcation of
selected varieties to further increase iron content might boost zinc levels too.
As in beans, iron and zinc concentrations differ across varieties of rice. Aromatic rice
tends to have the highest iron levels and several varieties have been successfully crossed
with elite rice lines having excellent agronomic characteristics and grain qualities. These
micronutrient traits were shown to be stable across different growing environments and
could be crossed with high yielding varieties to improve the nutrient density. High iron
rice developed from traditional breeding is currently being tested for iron bioavailability
and effects on iron nutrition status in young women in the Philippines (World Bank 2000).
Results are not yet available.
Another approach to improving the availability of iron for infants was reported by Suzuki
and colleagues (2003) at the University of California, Davis. These investigators developed
transgenic rice in cell culture that expressed the gene for human lactoferrin, a milk protein
that binds iron. When they compared the recombinant lactoferrin with native human
lactoferrin both proteins retained functional activity after mild heat treatment, high acidity,
and in vitro digestion. Their fndings suggest that recombinant lactoferrin grown in plant
culture may be a functional alternative to human lactoferrin in infant formula and provide
another way to improve iron availability during infancy.
B. Zinc
Zinc is one of several trace minerals that can be defcient in human diets, especially
where meat is not consumed. Zinc defciency is associated with impaired growth and
reproduction, anorexia, immune disorders, and a variety of other symptoms. Zinc is also
an important constituent of more than 100 enzymes. Absorption of zinc from cereals and
grains can be impaired or blocked by the presence of some substances such as phytate.
Increasing the zinc content of cereals and grains, especially where soils are low in zinc, may
be an effective way to improve human nutrition and at the same time increase plant yields.
Ramesh and colleagues (2004) in Australia, studied the effect on seed zinc content in barley
(Hordeum vulgare cv Golden Promise) in plants transformed to increase the expression of
zinc transporter enzymes. Multiple transgenic lines exhibited higher zinc and iron contents
in their seeds compared with control plants (Ramesh et al. 2004). When grown under
zinc defcient conditions, zinc uptake in the transgenic lines was higher in the short term
compared with control plants. When zinc content was restored, uptake of zinc decreased
Application of Biotechnology for Functional Foods
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in both transgenic and control plants, suggesting that the transporter proteins may be
degraded when zinc is adequate. This study suggests that increasing the production zinc
transporter proteins may be one approach to increasing the zinc content of cereals.
C. Selenium
Selenium is an essential trace mineral incorporated into plants from soil. Consumption
of selenium has been linked to reduced risk of all cancers, but particularly those of the
lung, colo-rectum and prostate (El-Bayoumy and Sinha, 2004, Combs 2004). Selenium is
also important for specifc enzymes and proteins in the brain and is necessary for proper
immune function. However, selenium is toxic at levels only a little greater than those
required in a healthy diet, so caution is warranted with supplementation and increased
intakes. Areas where soils are defcient in selenium are well known and low to defcient
intakes have been observed among human and animal populations in these regions.
Genetic engineering technology offers considerable potential for increasing the uptake
of selenium from soils and incorporating the mineral into non-toxic compounds in the
edible parts of plants. Plants genetically modifed to absorb above average quantities of
minerals could be used to improve human or animal nutrition. For example, a study from
the University of California, Berkeley reported that genetically engineered Arabidopsis
thaliana and Indian mustard (Brassica juncea) were able to incorporate more selenium from
soil and convert it to non-toxic methylselenocysteine than wild type plants (LeDuc et al.
2004). Researchers at Purdue University, West Lafayette, IN, also showed that plants not
normally accumulating selenium, such as Arabidopsis, can be transformed to do so (Ellis et
al. 2004). These studies demonstrate the feasibility of developing crop plants with improved
ability to take up selenium from the soil and store it in a non-toxic form. Thus, selenium at
appropriate concentrations would be safe for the plant and for human consumption.
VII. pHytonutrIents and noVel suBstances
Intense research activity is being devoted to the identifcation and study of phytonutrients in
plants with an eye to their ability to protect health, improve immune function, and reduce
the risk of chronic diseases ranging from heart disease and cancer to age-related macular
degeneration. Examples of phytonutrients include: phytoestrogens, polyphenols, and
isothyocyanates. In spite of their cumbersome technical names, these various categories of
substances appear to hold signifcant potential health benefts when consumed in modest
amounts. Because they are widely distributed in fruits and vegetables, diets rich in these
foods are likely to furnish generous amounts of many of these phytonutrients.
Unfortunately, there is insuffcient scientifc data from carefully controlled studies that
adequately demonstrate safety and effcacy of substances with potential promise. For many
substances—lycopene, isofavones, resveratrol—to name a few, data appear promising, but
are not consistent or conclusive. Extensive media and manufacturer publicity about many
of these compounds generated expectations exceeding scientifc justifcation. For these
reasons, the enhancement of foods with particular phytonutrients usually lacks suffcient
scientifc grounding to justify the development of foods with enhanced levels. However, a
few examples can be cited.
Isofavones: Isofavones are a type of phytoestrogen, so named because they bind to
estrogen receptors and mimic some of the effects of the hormone estrogen. However, the
biological effects of isofavones differ markedly from estrogen and many are non-hormonal.
Application of Biotechnology for Functional Foods
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Soybeans are the richest food source of isofavones, but isofavones occur in other legumes
such as broadbeans, and in many vegetarian (“meatless”) foods made from soy products
(USDA 2002). They have been linked with easing menopausal symptoms, improving bone
health, reducing cardiovascular risk, and possibly reducing the risk of prostate cancer. The
main soy isofavones are genistein, diadzein, and glycitein. Their concentration in legumes
is greatly affected by growing conditions and climate, and these variables could potentially
override genetic modifcations.
Currently, isofavones are abundant and readily available, so genetic modifcations to
increase isofavone content might not be expected to be a high priority. However, Yu and
colleagues (2003) at the DuPont Company, Wilmington, DE, reported the application of
genetic engineering techniques to increase the isofavone content of soybeans. By activating
genes in the phenylpropanoid pathway, diadzein levels increased and genistein levels fell.
By blocking the anthocyanins branch of this synthetic pathway, the investigators obtained
higher concentrations of isofavones. Thus, it is possible to increase the level of isofavones
in soybeans.
Phytosterols and Phytostanols: Phytosterols and their saturated derivatives, phytostanols,
are plant sterols found in small quantities in vegetable oils. When consumed in suffcient
amount, they are effective in reducing blood cholesterol levels. Suffcient data of their
effcacy and safety exist that the FDA has permitting food manufacturers to claim a role
for plant sterols or stanols or their esters in reducing the risk of coronary heart disease (see
section on health claims below for a further description).
In 2003, researchers at Unilever Research, Netherlands, reported the generation of
transgenic tobacco seeds with enhanced total seed sterol level (Harker et al. 2003).
Following the insertion of the gene for a key enzyme in sterol biosynthesis, total seed sterol
levels increased by 2.4-fold. The additional sterol was present as fatty acid esters and several
intermediate sterols accumulated. This group also developed transgenic tobacco carrying
two genes involved in regulating carbon fux through the sterol biosynthesis pathway
(Holmberg et al. 2003). The two transgenes increased total seed sterol content more than
with either gene expressed singly (2.5-fold vs. 1.6-fold).
Researchers at Monsanto reported the application of genetic engineering to modify the
ratio of phytosterols to phytostanols in rapeseed (Brassic napus) and soybean (Glycine
max). Plants were transformed with a gene from yeast for the enzyme 3-hydroxysteroid
oxidase (Venkatramesh et al. 2003). Seeds from both types of plants exhibited conversion of
the major phytosterols to phytostanols and no other functionalities were affected. Several
novel phytostanols were obtained as well. Because these substances are hydrogenated they
would be expected to be more stable during food processing, yet still confer cholesterol-
lowering benefts.
Most recently, Enfssi et al. (2005) reported the development of transgenic tomatoes with
a 2.4-fold increase in phytosterols. Increases were greatest for phytoene and beta-carotene.
Such an alteration in a widely consumed food would potentially increase the consumption
of these substances in a large share of the population.
Application of Biotechnology for Functional Foods


Probiotics: The term “probiotics” refers to live microorganisms that have a health beneft
when consumed in adequate amounts. They are usually bacteria selected from species found
in the intestinal tract. Probiotic microorganisms may be concentrated and added directly
to a food or added to a milk product in small amounts and allowed to grow. The most
common foods having probiotic organisms are fermented dairy products such as yogurt
containing Lactobacillus acidophilus.
Many health benefts have been attributed to probiotics, including resistance to
infectious diseases, prevention of vaginitis, production of antimicrobial metabolites and
nutraceuticals, immunomodulation, and others (Ahmed 2003). Foods containing probiotic
bacteria are abundant in Japan and parts of Europe, but are less developed in the U.S.
Probiotics have been used as dietary supplements and oral agents for intestinal disorders.
A genetically engineered strain of L. lactis subsp. diacetyllactis is used as a buttermilk
starter culture in the U.S. (Renault 2002).
Probiotics have been used to treat infammatory bowel disease by creating more host-
friendly gut fora. Selective use of probiotic bacteria can create an environment where
stimulation of the immune system is restrained and intestinal infammation reduced
(Guarner et al. 2002). Some strains of Lactobacilli have prevented the development of
colitis in genetically susceptible mice. Other genetically engineered bacteria have been used
to secrete the anti-infammatory cytokine IL-10 (Guarner et al. 2002). Considerable research
is being devoted to the identifcation and effects of probiotic bacteria and their use as
therapeutic agents, but few products have reached application in the U.S.
Genetically engineered probiotic bacteria have been used to overcome problems associated
with more traditional technologies for developing such bacteria. In foods, genetically
engineered bacteria have been used to improve the favor and stability, or to block the
formation of unwanted favors. Genetic engineering should make it possible to strengthen
the effects of existing bacterial strains and create new ones (Steidler 2003).
VIII. antI-nutrItIonal suBstances
Many plants contain substances that inhibit nutrient uptake in various ways or are toxic
themselves. They may interfere with intestinal cell function, reduce the ability to break
down complex molecules such as proteins and starch, or may be toxic if consumed
in suffcient quantity. Some of these substances are rendered harmless with cooking
or processing, but others are resistant to digestion, heat treatment, or other forms of
processing. Examples of anti-nutritional substances are shown in Table 2.
Although anti-nutrient substances reduce nutrient availability, many beneft human
health and plants. For example, dietary fber improves colon function and reduces plasma
cholesterol levels; polyphenols and other substances have anti-carcinogenic properties.
Many of these substances are important in plant metabolism and provide resistance to
environmental stress and pests. Further, some of the adverse effects on nutrient availability
may diminish over time or with different levels of consumption, and this suggests that
humans may be better able to adapt to these substances than was once thought (Welch
2002). The question of improving micronutrient availability by reducing anti-nutritional
substances through plant breeding, biotechnology, and other means requires careful
consideration of the complexities.
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taBle 2. anti-nutrients in plant foods that reduce
nutrient bioavailability, or impair health
antI-nutrIent eFFect dIetary source
Phytic acid (phytate) Binds minerals, K, Mg, Ca, Fe, Zn Whole legume seeds, cereal
grains
Fiber, e.g. cellulose,
hemicellulose, lignin
Decreases fat and protein digestibility;
may decrease vitamin & mineral
absorption
Whole cereal grains, e.g., wheat,
corn, rice
Trypsin inhibitor Reduces the activity of the enzyme
trypsin and other closely related
enzymes that help digest protein
Legumes, e.g., soy; cereals,
potatoes
Polyphenolics, tannins, Form complexes with iron, zinc, copper
that reduces mineral absorption
Tea, coffee, beans, sorghum;
Hemaglutinins, e.g. lectins Interfere with cells lining the
gastrointestinal tract causing acute
symptoms; can bind metals and some
vitamins; can be toxic
Legumes
Cyanogens or glycoalkaloids Inhibit acetylcholinesterase activity which
impairs nerve transmission; can damage
cell membranes
Cassava, linseed, peas, beans
Glucosinolates May adversely affect thyroid activity Cabbage, broccoli
Saponins May irritate the gastrointestinal tract and
interfere with nutrient absorption
Soybeans, peanuts, sugar beets
Goitrogens Suppress thyroid function Brassica and Allium foods, e.g.,
broccoli, garlic
Heavy metals, e.g.,
cadmium, mercury, lead
May have toxic effects, e.g., high levels of
Hg impair fetal brain developmnet
Contaminated leafy vegetables
Gossypol May harm kidney function and reduce
sperm counts; can be toxic
Cottonseed
Oxalic acid Binds calcium to prevent its absorption Spinach leaves, rhubarb
Phytotoxins, e.g. solanine Can be toxic; affects gastrointestinal and
nervous systems
Green parts of potato
Mycotoxins, e.g., aflatoxin,
fumonisin
Toxins produced by certain molds;
toxic to humans and animals; can be
carcionogenic
Grain, peanuts, other crops
There are several ways of reducing anti-nutritional substances, including plant breeding,
heat, processing, fermentation and drying, and genetic engineering. Examples of
the various anti-nutritional substances that have been reduced by the application of
biotechnology in different plants are described below.
Phytate: Phytic acid is a phosphorus storage compound found in the seeds of many edible
crops, e.g., wheat, corn, barley, rice. Phytic acid forms salts (phytates) of potassium,
magnesium, calcium, iron, zinc, and other minerals that cannot be absorbed. Phytic acid-
containing foods bind minerals in the intestinal tract rendering them unavailable. When
Application of Biotechnology for Functional Foods
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these minerals are limiting in the diet, the presence of phytic acid can contribute to mineral
defciencies, particularly in the case of iron and zinc. This is a particularly important
consideration in the diets of women and children where legumes and cereals are staple
foods. In animal nutrition, especially for poultry and swine, phosphorus may have to be
supplied in a more available form to overcome the loss due to binding with phytic acid. An
additional consequence is the production of high phosphorus animal waste with adverse
environmental effects.
Lines of corn, barley, rice, and soybean with slightly different phytic acid characteristics
have been used to develop varieties with reduced seed phytic acid (Raboy 2002). Reduction
in phytate in the range of 50% to 66% has been achieved with these mutant lines. In
soybeans and corn, 80% reduction has been achieved. However, several hybrids developed
with the mutant strains exhibited lower yields. It has now been shown that low phytate
mutant corn is linked to the reduced expression of the enzyme myoinositol phosphate
synthase, the frst enzyme in the synthesis pathway for phytic acid (Shukla et al. 2004).
This fnding was confrmed recently by Italian researchers who developed a mutant corn
with 90% less phytic acid and a 10-fold increase in seed-free phosphate (Pilu et al. 2003).
Proof that zinc absorption from low phytate corn compared with wild-type varieties was
signifcantly greater was reported by Hambridge et al. (2004) who fed corn tortillas to six
healthy adults. Zinc absorption from the 80% phytate-reduced corn was three times greater
than from the wild-type corn, 4.9 mg/day compared with 1.5 mg/day.
Although genetically engineered low phytate crops have not been commercialized,
biotechnology has been used to express the enzyme phytase in plants (Chier et al.
2004). This enzyme allows animals to metabolize phytic acid and eliminates the need
to supplement feed with phosphorous. Consequently, using animal feed engineered to
produce phytase addresses many of the problems associated with high phytate levels in
animal feed. Phytase has been successfully incorporated into soybean and wheat and is
biologically active when the plants are used as animal feed (Brinch-Pedersen et al. 2000).
In a study of broiler chickens, consumption of transgenic soybeans containing phytase led
to a 50% reduction in phosphorus excretion compared with a diet supplemented with an
intermediate level of nonphytate phosphorus (Denbow et al. 1998). Feeding the transgenic
soybeans resulted in an 11% greater reduction in phosphorus excretion than feeding with
conventional soybeans to which the enzyme is added. Similarly, low-phytate corn and barley
fed to broiler chicks resulted in 33% lower phosphorus excretion compared with wild-type
grain diets and reduced the need for supplemental phosphorus (Jang et al. 2003).

Transgenic phytase-containing wheat was developed by Holm and colleagues at the Danish
Institute of Agricultural Sciences, Denmark (Brinch-Pedersen 2000, Holm et al. 2002).
Transgenic plants exhibited up to 4-fold higher phytase activity compared with wild-type
seeds. However, a drawback of such a transformation for human consumption is that the
enzyme is inactivated above 60°C and would be destroyed by cooking (Holm et al. 2002). It
is possible that more heat-stable forms of the enzyme could be used, but when more heat-
stable enzymes were incorporated into rice, which was then cooked, only 8% of the activity
remained (Holm et al. 2002).
Transgenic rice expressing phytase derived from modifed yeast genes has also been reported
(Hamada et al. 2005). By selectively modifying the genes, the investigators were able to
increase the enzyme activity above that of the original yeast gene.
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Reducing the phytate content of plants, particularly soybean, has direct implications for
human nutrition. For example, soy protein used in infant nutrition may limit mineral
absorption because of its phytate content. To investigate this question, Davidsson and
colleagues at the Swiss Federal Institute of Technology compared regular and dephytinized
soy formula in nine infants 69 to 191 days old. Regular and dephytinized formula contained
300 and 6 mg phytic acid/kg liquid, respectively. The investigators reported that zinc
absorption was signifcantly greater from dephytinized formula compared with regular
formula, 22.6% compared with 16.7% absorption (Davidsson et al. 2004). Absorption
of iron, manganese, copper and calcium did not differ between the two formulas. These
fndings suggest that use of dephytinized soy protein improves zinc absorption and can be
recommended for infant foods.
Another approach to solving the phosphorous uptake problem in animal production was
undertaken by researchers at the University of Guelph, Canada. In this case, swine were
engineered to produce the enzyme phytase (Golovan et al. 2001). These pigs expressed
the enzyme phytase in their saliva and exhibited complete phytate digestion, required no
dietary inorganic phosphorus and excreted 75% less phosphorus.
Gossypol: Cottonseed contains the polyphenolic compound gossypol, long known to
be toxic to humans and animals. Interestingly, gossypol also appears to have anti-cancer
properties toward several human prostate and breast cancer cell lines (Liu et al. 2002,
Jiang et al. 2004). Martin and colleagues at Texas A&M University, College Station, TX,
created a transgenic cotton (Gossypium vitifolium) using the antisense gene technology for
a key enzyme in the synthesis of gossypol. Transformed cotton plants had up to 70% less
gossypol in their seeds compared with non-transformed plants (Martin et al. 2003). These
fndings suggest that biotechnology can be used to reduce the gossypol levels to render the
seed oil more suitable for feed and food.
Cyanogens: Cassava (Manihot esculenta Crantz) produces various cyanogenic glycosides
such as, linamarin, lotaustralin, and acetone cyanohydrin in its roots and leaves. These
potentially toxic substances are only present in small amounts in “sweet” varieties of
cassava, but are suffciently abundant in “bitter” varieties to require removal (Padmaja
1996). Boiling and drying reduce these substances to safe levels in low cyanogen varieties,
but those with higher levels require, soaking, grating or maceration, fermenting, and sun-
drying to reduce cyanogens adequately. The toxicity of these cyanogens is exacerbated by
low protein intakes, a characteristic of countries where cassava is a staple.
In 2003, Drs. Siritunga and Sayre at the Ohio State University, Columbus, OH, reported
the development of transgenic cassava with a 99% reduction in linamarin in the roots and
between 60% and 94% reduction in leaves (Siritunga and Sayre 2003). These plants were
transformed by inhibiting the expression of two genes that catalyze the frst step in the
synthesis of linamarin. The following year, this group reported that transgenic cassava roots
expressing a different gene contained signifcantly less acetone cyanohydrin levels compared
with wild-type plants (Siritunga et al. 2004 ). These accomplishments open the door to the
development of cassava truly safe for human and animal consumption.

Steroidal glycoalkaloids: Potatoes (Solanum tuberosum spp) contain potentially toxic
steroidal glycoalkaloids, the best known of which is solanine. This substance is found in the
green tissue of potato just under the skin. While these compounds protect the plant from
pests, they reduce food quality and safety. Glycoalkaloids are synthesized from cholesterol.
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Recently, Arnqvist et. al. (2003) showed that the inhibition of plant sterol synthesis in
transgenic potatoes reduced the synthesis of glycoalkaloids by 41% in leaves and 63% in
tubers. Other investigators used antisense technology to create transgenic potatoes with
up to 40% less steroidal glycoalkaloids in the tubers (McCue et al. 2003). These studies
indicate that substantial improvements in the reduction of steroidal glycoalkaloids in
potatoes may be close at hand. It remains to be seen whether pest resistance in these
transgenic potatoes is affected by the reduction in glycoalkaloids.
Mycotoxins in corn: An important risk to the safety of grains (corn, wheat, barley),
groundnuts (peanuts), tree nuts (almonds, walnuts, pistachios) and cottonseed is the
production of toxins by fungi. Certain types of fungi—Aspergillus favus and Fusarium—
are notorious for their deadly products. The substances produced by these organisms
cause disease in plants and potentially serious illness in people and animals consuming the
infected crops. Afatoxin, a particularly dangerous cancer-causing mycotoxin produced
by the fungus Aspergillus favus, is sometimes found in peanuts and corn. Fumonosins
produced by Fusarium fungi are thought to be carcinogenic and harmful to the immune
system. Agricultural practices and grain storage conditions help to minimize growth of
these fungi, but weather also contributes to their development. Breeding crop varieties with
increased resistance to fungi may reduce the production fungal toxins. Researchers at the
U.S. Department of Agriculture developed transgenic walnut trees that displayed increased
resistance to afatoxin synthesis (USDA 2004). Partial resistance to Fusarium disease was
reported in transgenic wheat (Okubara et al. 2002) and in transgenic bacillus thuringiensis
(Bt) corn (Bakan et al. 2002). The potential for genetic engineering strategies to reduce
the production of fungal toxins in food crops has been discussed by Duvic (2001) and
Munkvold (2003).
Oxalates: Oxalic acid is present in spinach, tomato, groundnut, soybean, and chick pea.
It binds several minerals including calcium and prevents their absorption. Scientists at the
National Centre for Plant Genome Research (NCPGR), India, seek to reduce the oxalic acid
content in these foods through the transfer of the gene for oxalate decarboxylase (OXDC),
the enzyme that degrades oxalic acid. Genetically engineered tomatoes bearing the OXDC
gene have been successfully grown and are currently undergoing feld and biosafety tests
(ISAAA, 2004).
IX. allergens
Food allergy, although relatively rare, can provoke severe, sometimes fatal, responses in
susceptible people. Specifc food proteins can trigger the immune system and provoke an
allergic response. Risk of such reaction is greatest in the frst two years of life, and by age
fve about 80% of allergic infants will lose their food allergies.
Eight types of foods account for nearly 90% of all food allergies. These are: milk, eggs,
fsh, crustacea, wheat, peanuts, tree nuts, and soy. People allergic to one type of food are
frequently, but not always, allergic to others. Many proteins associated with food allergies
in these foods have been identifed, but many remain unknown.
Soybean: In people with soy allergy, as many as 28 proteins may bind with IgE, a type of
antibody involved in allergic responses, suggesting that many soy proteins have allergenic
potential. More than half of soybean allergic reactions are attributable to a single protein,
known as P34 (Cordle 2004, Wilson et al. 2005).
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Scientists at USDA and the Donald Danforth Plant Science Center have succeeded in
silencing the gene for P34 and created soybeans without this protein (Herman et al. 2003).
However, two other proteins that trigger allergic reactions may have to be removed before
soybeans could be sold as hypoallergenic. Dr. Anthony Kinney, a researcher involved in the
project, commented that removing the other proteins should not be diffcult because wild
species lacking the genes for the other proteins are already known. Careful plant breeding
with the genetically modifed soybeans may be able to produce the hypoallergenic soybeans.
Other genetic engineering strategies that alter the composition of the allergenic proteins
may render the proteins harmless to sensitive people. The ability to eliminate the most
hazardous allergens in soy would have substantial beneft for infant formula feeding and for
those who are allergic to soy.
Peanut: Allergy to peanut proteins can be fatal. The major allergens in peanut have been
identifed as Ar h1, Ar h2, and Ar h3, and their genes have been isolated. The protein Ar h2,
a Kunitz trypsin inhibitor, is believed to be the most potent of these three (Koppelman et al.
2004). Recombinant versions of these allergens produced in bacteria were heat-killed and
used in a vaccine (Li et al. 2003). The vaccine was given in three different doses to allergen-
susceptible mice. Animals were challenged after 2, 6, and 10 weeks. Treated mice produced
no anaphylactic or histamine response when challenged. Animals given the medium
and high doses remained protected for 10 weeks. This particularly encouraging research
suggests that protection against peanut allergy may be in the foreseeable future.
Rice: Rice is generally consider a hypoallergenic food, and for that reason is one of
the solid foods frst introduced to infants. Nonetheless, some people, particularly in
Japan, are allergic to rice proteins. The major allergen(s) in rice have been identifed and
hypoallergenic varieties of rice developed using antisense genetic engineering techniques
to suppress the synthesis of the predominant allergen (Nakamuro and Matsuda 1996).
Allergen content in the transgenic seeds was reduced from about 300 micrograms/seed
to about 60 to 70 micrograms/seed. However, when the hypoallergenic rice was tested in
sensitive patients, not all allergenic potential had been eliminated. Extremely sensitive
patients were also sensitive to other proteins, so that a single genetic transformation was
insuffcient to overcome their allergic responses. EuropaBio (2002) reported that several
laboratories in Japan are working to develop hypoallergenic rice, but efforts have provided
only partial success, as it is not possible to eliminate all allergens.
Shrimp: Allergic reactions to shrimp and other crustacea (lobster, crayfsh) are among the
most common food allergies. People with hypersensitivity responses to crustacea may also
be allergic to mollusks, and some arthropods such as house dust mites and cockroaches. To
date there have been no ways to overcome these allergies. The major allergen in shrimp is
the muscle protein tropomyosin, known as Pen a 1. Recent analysis of this allergen revealed
fve IgE-binding regions or epitopes (Ayuso et.al. 2002). These regions contained 15 to 38
amino acids from whose sequence the corresponding gene sequence can be determined. Dr.
Samuel Lehrer of Tulane University, New Orleans, LA, has located the gene sequence that
encodes these epitopes and has suggested that by altering these epitopes by a single amino
acid, IgE binding could be halted. This work suggests that safe recombinant allergens
could be synthesized for immunotherapy of those who are allergic to shrimp and related
substances.
Application of Biotechnology for Functional Foods
35

X. mIscellany
A. Beer and Wine
Applications of biotechnology are fnding their way into brewing and wine-making. One
application in grapevines (Vitis vinifera L.) has been increased content of the antioxidant
resveratrol. This substance is of special interest in human health for the reduction of
oxidized lipids. It is also thought to improve plant resistance to fungal disease. Gonzalez-
Candelas et al. (2000) also reported increased resveratrol in wine through the use of
transgenic yeast. In a different approach, Giorcelli et al. (2005) developed transgenic
grapevines using the gene for stilbene synthase, an enzyme that leads to the production of
resveratrol. The plants increased their production of resveratrol, but showed no improved
resistance to a leading fungal pathogen.
Transgenic yeast also has potential for modifying the favor of beer (Vanderhaegen et al.
2003) and sake (Aritomi et al. 2004). Genetic engineering also holds considerable potential
for the production and regulation of diverse favors and aromatic substances, as recently
described (Dudareva and Negre 2005).
B. Decaffeinated Coffee
Japanese scientists have succeeded in silencing the gene responsible for caffeine production
in coffee plants (Jameel 2003, Ogita et al. 2003). Caffeine content in the transgenic plants
was reduced by 70%. The researchers aim to modify Arabica coffee plants, the most
popular coffee grown. Toward this end, Ogita et al. (2004) reported additional progress on
modifying the pathways in Arabica and canephora coffee varieties which resulted in caffeine
reductions ranging from 30% to 50%. This genetic modifcation would eliminate the need
for decaffeination processing.
Application of Biotechnology for Functional Foods
Application of Biotechnology for Functional Foods
37

PART 2
Applications of Modern


Biotechnology to Functional Food
Legal and Regulatory
Considerations Under Federal Law
I. IntroductIon
“Functional food” is touted as a convenient means for consumers to promote optimal
health, including the prevention of disease. The focus of much attention in recent
years, functional food has been the subject of numerous articles, reports, and consumer
education materials, such as a “Functional Food Guide Pyramid”—a modifed version
of the USDA Food Guide Pyramid that identifes functional food from each major food
group. A considerable array of food has been described as “functional” in one or more
respects, including calcium-fortifed orange juice, whole grains, fruits and vegetables (and
components thereof, including lycopene, polyphenols, indoles, and other phytochemicals),
soybeans, omega-3 fatty acids, phytosterols, and cocoa.
Despite the widespread attention, the “functional food” concept eludes precise defnition.
As previously noted, the Food and Nutrition Board of the National Academy of Sciences
has suggested that a “functional food” is “any modifed food or food ingredient that may
provide a health beneft beyond the traditional nutrients it contains” (Food and Nutrition
Board 1994). Others argue that a functional food is any food promoted or consumed for a
specifc health effect, regardless of whether the food has been modifed in some fashion. A
frequent criticism is that all food is, in some sense, functional.
From a legal perspective, there is no separate regulatory category for functional food in the
United States. Food that is deemed functional, therefore, is subject to the same regulatory
requirements as any other food. This means that a functional food may be regulated as
“conventional food,” a “dietary supplement,” a food for “special dietary use” (including
infant formula) a “medical food,” or a “drug,” depending upon its positioning in the
marketplace, including claims made for it. Signifcantly, federal requirements for “functional
food” apply regardless of how the food is produced, such as through mechanical or genetic
methods (e.g., product formulation, modern biotechnology techniques, or other means).
Thus, rice that has been genetically enhanced to provide beta carotene is subject to basically
the same statutory and regulatory framework as rice to which beta carotene is added
through product formulation.
1
The diffculties associated with FDA regulation of functional
food historically have been related to whether such food is subject to the more onerous drug
provisions of the law.
1 The use of modern biotechnology is subject to regulation by several federal agencies, including the Animal
and Plant Health Inspection Service of the U.S. Department of Agriculture (USDA), the Environmental
Protection Agency (EPA), and the Food and Drug Administration (FDA). This report focuses on safety and
labeling requirements enforced by FDA and USDA’s Food Safety and Inspection Service (FSIS) pursuant to
certain food laws administered by those agencies. Food produced through the use of modern biotechnology
is handled somewhat differently by FDA in that it is subject to a voluntary consultation process. See 57 Fed.
Reg. 22984 (1992).
Application of Biotechnology for Functional Foods
38

This section of the report describes how functional food is regulated under certain
federal laws governing food in general.
2
To place the regulation of functional food in an
appropriate context, this report begins with an overview of the basic statutory framework
pertinent to food. It then examines the application of this basic framework to functional
food, frst in the context of food generally, and then in the context of meat and poultry
products, eggs and egg products, and animal feed, including pet food. As appropriate, the
discussion also identifes select areas of criticism and issues concerning functional food
regulation in the United States.
II. tHe FFdca Framework—oVerVIew and BrIeF HIstory
In the United States, food products other than meat and poultry and alcoholic beverages are
regulated primarily by the Food and Drug Administration’s (FDA) Center for Food Safety
and Applied Nutrition (CFSAN) pursuant to the Federal Food, Drug, and Cosmetic Act
(FFDCA).
3
In addition to providing for the regulation of “food,” the FFDCA establishes
comprehensive requirements for the marketing of drugs, medical devices, and cosmetics.
Food and other articles regulated by FDA under the FFDCA cannot be adulterated or
misbranded. Adulteration refers generally to aspects of a product that typically relate to
quality or safety and misbranding refers generally to false or misleading labeling.
Although the concept of “functional food” was not contemplated at the time Congress
enacted the FFDCA, the law has evolved—through fexible agency interpretations as well
as statutory amendments—to accommodate new products and advances in science. The
emergence of functional food as a unique and important category for marketing purposes
has led some to question whether the current legal and regulatory framework is adequate.
A. The Statutory Foundation: “Food,” “Drug,”
and Food for “Special Dietary Use”
At the time of its enactment in 1938, the FFDCA established requirements for only three
product categories of relevance to functional food regulation. In addition to covering
“food” and “drugs,” the 1938 Act introduced the concept of food for “special dietary uses.”
Although this law contained no defnition of “special dietary uses,” the legislative history
reveals that such uses were considered, at that time, to include “infant foods, invalid foods,
slenderizing foods, and other dietary [products] intended for special nutritional require-
ments” (S. Rep. No. 493, 73d Cong., 2d Sess. 12 (1934)). FDA later defned “special dietary
uses” by regulation to mean “particular (as distinguished from general) uses of food,”
including uses that may arise from disease or certain health-related conditions, age-related
nutritional needs (e.g., infancy), or a desire to supplement or fortify the diet with any vita-
min, mineral, or other dietary property. A food for special dietary use is deemed mislabeled
or misbranded under the law unless its label bears information adequate to inform purchas-
ers of its vitamin, mineral, or other dietary content.
4

2 Under applicable law, food refers to use for human beings or other animals and includes (non-human)
animal feed. “Functional food” is primarily intended for human use, as other animal feed is subject to
typically different requirements.
3 As discussed more fully below, products with meaningful amounts of meat or poultry (generally 2–3%)
are regulated by the U.S. Department of Agriculture (USDA) pursuant to the Federal Meat Inspection Act
(FMIA) or the Poultry Products Inspection Act (PPIA). Alcoholic beverages are regulated primarily by the
Bureau of Alcohol, Tobacco and Firearms of the Department of Treasury.
4 This report does not address infant formula, a “special dietary use” food for which Congress and FDA
have established unique and prescriptive requirements. “Infant formula” is defned to mean “a food
which purports to be or is represented for special dietary use solely as a food for infants by reason of its
simulation of human milk or its suitability as a complete or partial substitute for human milk.” FFDCA
§ 201(z), 21 U.S.C. § 321(z).
Application of Biotechnology for Functional Foods


B. Accommodations of Advancing Science
For nearly thirty years after enactment of the FFDCA, the basic categories of “food,”
“drug,” and food for “special dietary use” remained largely unchanged. During this time
period, FDA interpreted almost any use of food for a targeted nutritional purpose to be a
“special dietary use.” Indeed, as recently as 1971, FDA’s framework for the regulation of
food for special dietary uses (commonly referred to as “special dietary food” at that time)
included requirements addressing all uses of fortifcation (i.e., the addition of nutrients to
food or ingredients thereof), vitamin and mineral supplements, and food purported for use
in sodium-restricted diets, among other food or uses deemed to be “special.” Underlying
this framework was an assumption that food for targeted nutritional purposes (i.e.,
“special” uses) was of little or no relevance to the general population. Paradoxically, certain
products that seemed to more naturally ft the special dietary use category, such as specialty
infant formulas, were strictly regulated as drugs.
With advances in nutrition and related sciences, the special dietary use concept was
gradually reformed to accommodate evolving views concerning the relationship between
diet and health. These reforms ultimately led to new and distinct categories of “food”
and new types of claims. Three regulatory developments are of particular relevance to
the regulation of functional food, and demonstrate the progressive blurring of the legal
distinction between “food” and a “drug” since 1938: (1) the advent of “medical food”
in 1972, (2) the authorization of certain nutrition and health-related claims for food in
the Nutrition Labeling and Education Act (NLEA) of 1990, and (3) authorization of a
separate regulatory scheme for dietary supplements in the Dietary Supplement Health and
Education Act (DSHEA) of 1994.
In 1972, FDA created the “medical food” category by deciding on its own initiative to
reclassify the specialty infant formula Lofenalac®, which had been regulated as a drug,
as a distinct type of food for special dietary use. Intended for the dietary management of
phenylketonuria (PKU), an inborn error of metabolism, Lofenalac® was recognized by FDA
to meet distinctive nutritional requirements—namely, an impaired ability to metabolize
the essential amino acid phenylalanine—and therefore more appropriately regulated as a
food. In a regulation issued shortly thereafter, which exempted medical food from nutrition
labeling requirements, FDA described the new category as “foods represented for use solely
under medical supervision in the dietary management of specifc diseases and disorders”
(38 Fed. Reg. 2124, 2126 (1973)). As discussed more fully in Part III, FDA today treats
“medical food” as a wholly unique category from food for special dietary use, and has
developed very specifc criteria for the marketing of such products.
Second, in 1990, Congress passed the NLEA, which establishes a framework for the
regulation of “health claims” and “nutrient content claims,” among other requirements.
“Health claims” are statements that characterize the relationship between a food (or a
substance in the food) and a disease or health-related condition (21 C.F.R. § 101.14(a)(1)).
“Health claims” are permitted only if approved or otherwise authorized by FDA, the claim
addresses the ability of the food, as part of an appropriate diet, to reduce the risk of disease
or a health-related condition (as opposed to disease treatment, mitigation, or similar
concepts), and the food meets certain qualifying criteria established by FDA. NLEA also
authorized “nutrient content claims,” which are statements characterizing the level of a
nutrient in food, such as “sugar free” and “low sodium.” Following NLEA, FDA determined
that many nutrition-related claims were of general use to the public, and thus were no
longer indicative of “special dietary uses.”
Application of Biotechnology for Functional Foods
40

Third, in 1994, Congress enacted DSHEA, which establishes comprehensive requirements
for dietary supplements, including safety and labeling requirements. Although DSHEA
provides that dietary supplements are still “food” for many purposes, the new law creates a
unique framework for supplement regulation.
These important developments—DSHEA, NLEA, and FDA’s “medical food” policy—
helped to shape FDA’s regulation of “food” and nutritional claims, and therefore functional
food. Particularly, they created new regulatory categories and permitted many products to
bear claims that would ordinarily have triggered “drug” regulation. These new categories
and claims are described more in Section III and relate to dietary supplements, medical
food, health claims, and nutrient content claims.
C. Requests for Reform
The emergence of functional food as a unique and commercially important marketing
category has led some to question whether the current regulatory framework is adequate.
Some groups, including the Government Accountability Offce (GAO) and the Center for
Science in the Public Interest (CSPI), have criticized FDA’s oversight of ingredient safety
and labeling. GAO and CSPI have suggested that companies should be required to notify
FDA before using certain “functional” or “novel” ingredients and before making certain
claims about the effect of such ingredients on the structure or function of the body (GAO,
2000; CSPI 2002). Other groups, such as the Institute for Food Technologists (IFT) have
recommended increased fexibility and policy changes to better allow for claims based
on scientifc advances (IFT 2005). For example, IFT has proposed that FDA adopt a
notifcation procedure for health claims. As proposed by IFT, companies wishing to use a
health claim could convene an expert panel to evaluate the science supporting the claim;
if the panel found the claim to be “generally recognized as effcacious,” the claim could be
notifed to FDA and used within 90 days if the agency did not object.
These and other calls for reform led FDA, in October 2006, to announce that the agency
would hold a hearing to allow consumers, industry, and other interested parties to provide
comments on approaches to the regulation of functional food. Affrming its fexible
approach to the statute, FDA stated that “[a]lthough we are confdent that the existing
provisions of the act are adequate to ensure that conventional foods being marketed as
‘functional foods’ are safe and lawful, we believe that it would be in the best interest of
public health to begin a dialog with industry, consumers, and other stakeholders regarding
the regulation of these products” (71 Fed. Reg. 62400, 62401 (Oct. 25, 2006))
5
FDA’s views
of the current framework for regulating functional food are described in detail next.
III. classIFIcatIon oF FunctIonal Food under tHe Federal Food,
drug, and cosmetIc act
The regulatory classifcation of a product has substantial implications for its marketing
and use, as it infuences the legal requirements pertaining to safety, labeling, and whether
5 FDA did not specifcally seek comment regarding the regulation of dietary supplements. Although there
are important legal distinctions between conventional food and dietary supplements, the functional food
concept is often considered to include both categories; accordingly, both are addressed in this report.
Application of Biotechnology for Functional Foods
41

the product is subject to regulatory pre-market clearances or approval. Accordingly,
to determine how a specifc “functional food” is regulated under the FFDCA, careful
consideration of the pertinent regulatory categories is required. Following a brief review
of the pivotal defnitions of “food” and “drug” (Section A), which form the basis for much
of the regulatory landscape for functional foods, a review of each regulatory category into
which a functional food may fall is provided. These regulatory categories are drug (Section
B), conventional food (Section C), dietary supplement (Section D), food for special dietary
uses (Section E), and medical food (Section F). An overview of the basic categories and
claims available for each is presented in Table III-1.
taBle III-1 regulatory categories and general summary
category scope safety standard claims
Drug Product intended to
diagnose, prevent, treat, or
mitigate disease
New drugs must be
demonstrated to be safe and
effective
Must be approved by FDA
Conventional
Food
Product used primarily for
taste, aroma, or nutritive
value, and in a conventional
food form
Components must present
“reasonable certainty of no
harm”; must be FDA-cleared
unless determined to be
generally recognized as safe
(GRAS)
Health and nutrient content
claims must be FDA-
authorized; other claims must
not be false or misleading in
any particular
Dietary
Supplement
Product intended to
supplement the diet; must
contain one or more “dietary
ingredients” (e.g., vitamins,
herbs, substances found
in the diet), be intended
for ingestion, labeled as
a dietary supplement, not
represented for use as a
conventional food, and not
approved as a drug, among
other criteria
Must not present a
“significant or unreasonable
risk of illness or injury”; new
ingredients must be notified
to FDA
Health and nutrient content
claims must be FDA-
authorized; structure/function
claims must be notified to
FDA; other claims must not
be false or misleading in any
particular
Food for Special
Dietary Use
Food intended for supplying
particular dietary needs
(e.g., those relating to
disease, pregnancy, lactation,
underweight, or overweight)
See conventional food
standard above
Claims of special dietary
usefulness (e.g., usefulness
in reducing body weight)
Medical Food Formulated food intended for
use under the supervision
of a physician for the dietary
management of disease
with distinct nutritional
requirements
See conventional food
standard above
Claims for the dietary
management of disease
Application of Biotechnology for Functional Foods
42

A. Key Definitions: “Food” and “Drug”
Two key defnitions form the basis for much functional food regulation: “food” and “drug.”
An understanding of these overarching regulatory categories is particularly important to
appreciate the scope of permissible claims.
“Food” is defned to mean—
(1) articles used for food or drink for man or other animals,
(2) chewing gum, and
(3) articles used for components of any such article (FFDCA § 201(f), 21 U.S.C. § 321(f)).
The food defnition is based on a product’s actual (as opposed to intended) use. It has been
interpreted to mean that articles used primarily, but not exclusively, for taste, aroma, or
nutritive value are “food”:
When the statute defnes ‘food’ as ‘articles used for food,’ it means that
the statutory defnition of ‘food’ includes articles used by people in
the ordinary way most people use food—primarily for taste, aroma, or
nutritive value. To hold . . . that articles used as food are articles used solely
for taste, aroma, or nutritive value is unduly restrictive since some products
such as coffee or prune juice are undoubtedly food but may be consumed
on occasion for reasons other than taste, aroma, or nutritive value.
6
FDA actions against weight loss products marketed as starch blockers illustrate the scope
of the “food” defnition. In one such case, Nutrilab, Inc. v. Schweiker, FDA asserted a
starch blocking product to be a “drug” within the meaning of section 201(g)(1)(C). The
court agreed that the product, which contained substances that were claimed to inhibit
enzymes necessary for starch digestion, was a drug. The court reasoned that the product
was intended to affect digestion, a bodily function, and was not “food” because it was
not consumed primarily for taste, aroma, or nutritive value. In other words, the product
affected a bodily structure or function, not by way of its consumption as food, but through
a pharmaceutical mechanism of action.
7
Unlike food, the drug category is based on intended use. A product is deemed to be a drug,
in most relevant part, on the basis of its intended use to diagnose, cure, mitigate, treat, or
prevent disease, or intended use to affect a bodily structure or function. Specifcally, the
term “drug” is defned in section 201(g)(1) of the FFDCA to include—
(B) articles intended for use in the diagnosis, cure, mitigation, treatment, or
prevention of disease in man or other animals; and
(C) articles (other than food) intended to affect the structure or any
function of the body of man or other animals; and
(D) articles intended for use as a component of any article specifed in
clause (A), (B), or (C).
6 Nutrilab, Inc. v. Schweiker, 713 F.2d 335, 338 (7
th
Cir. 1983) (emphasis added).
7 Today, starch blocker products are positioned as dietary supplements, which are authorized to bear
structure/function claims without regard to their nutritive content or positioning as “food.” See infra
section III.C.
Application of Biotechnology for Functional Foods
43

Intended use is determined objectively, based on labeling claims made for a product,
instructions for use, advertising, and other materials as appropriate. Thus, by controlling
claims made for a product, a manufacturer may exercise substantial control over a product’s
classifcation. The “food” and “drug” categories are not mutually exclusive, meaning that
a product that appears to be a “food” but that is promoted for therapeutic purposes falls
within the drug defnition and can be regulated as such. Examples include phytosterol-
enhanced rapeseed oil (i.e., canola oil) promoted for cholesterol lowering properties and
yogurt promoted for probiotic properties that are suggested to prevent or treat infectious
disease.
Signifcantly, the structure/function prong of the drug defnition provides a broad exception
for food.
8
In other words, because food obviously affects bodily structures and functions
through nutrition, a food product is not subject to regulation as a “drug” merely because it
bears a structure/function claim (e.g., “Calcium builds strong bones”). Rules for structure/
function claims for conventional food and dietary supplements are explained more fully in
Sections III.C.2 and III.D.2.
B. “Drug”
1. scope
Based on the statutory defnitions described above, including the exemption for structure/
function claims that appear on food, the “disease” prong of the “drug” defnition is the
primary factor determining whether a functional food will be regulated as a “drug.” The
line between “disease claims” that render a food a “drug” and structure/function claims can
be diffcult to discern.
To help identify impermissible disease claims that cannot be used on food, FDA has defned
“disease” to mean “damage to an organ, part, structure or system of the body such that it
does not function properly (e.g., cardiovascular disease), or state of health leading to such
dysfunctioning (e.g., hypertension),” excluding diseases resulting from nutrient defciencies.
(21 C.F.R. § 101.93(g))
9
FDA has further identifed by regulation eight types of statements
that would cause a product—whether positioned in the marketplace as a conventional food
or as a supplement—to be regulated as a drug. These so-called “disease claim” categories
are featured in Table III-2.
8 The defnition specifcally provides that the term “drug” includes “articles (other than food) intended to
affect the structure or any function of the body of man or other animals.” FFDCA § 201(g)(1)(C) (emphasis
added). Thus, for example, a food could bear a structure/function claim without FDA approval, but a
cosmetic bearing a structure/function claim would be regulated as a drug because there is no “cosmetic”
exemption.
9 Although this defnition was adopted to aid FDA in the regulation of dietary supplement claims, it is
commonly used in evaluating claims for conventional food as well.
Application of Biotechnology for Functional Foods
44

taBle III-2 disease claims that may trigger “drug” status
type of claim example of Impermissible claim
Effect on a specific disease or class of diseases “Reduces the pain and stiffness of arthritis”
Effect on an abnormal condition associated with a natural
state or process, if the abnormal condition is uncommon
or can cause significant or permanent harm
“Support for Cystic Acne”
Effect on the characteristic signs or symptoms of a
specific disease or class of diseases, using scientific or
lay terminology
“Reduces joint pain”
Effect on disease through statements regarding product
name, formulation, supportive scientific references,
symbols, or use of the term disease
Name: “Hepatacure”
Formulation: Contains digoxin
Scientific references: Serial Coronary
Angiographic Evidence That Antioxidant Vitamin
Intake Reduces Progression of Coronary
Artery Atherosclerosis (if placement in labeling
suggests disease use)
Symbols: EKG
Disease: “Prevent the onset of disease”
Product belongs to a class of products that is intended to
diagnose, mitigate, treat, cure, or prevent a disease
“Antidepressant”
Product substitutes for a product that is a therapy for a
disease
“Herbal Prozac”
Product augments a particular therapy or drug action
that is intended to diagnose, mitigate, treat, cure, or
prevent a disease or class of diseases
“Use as part of your diet when taking insulin to
help maintain a healthy blood sugar level”
Has a role in the body’s response to a disease or a vector
of disease
“Supports the body’s ability to resist infection”
Product treats, prevents, or mitigates adverse events
associated with a therapy for a disease and manifested by
a characteristic set of signs or symptoms
“Help maintain intestinal flora for individuals on
antibiotics”

If a functional food is promoted using statements that FDA considers to be disease claims,
it is subject to regulation as a drug. In contrast, drug regulation can be avoided through the
use of general health and nutrition-related claims that do not implicate disease, or through
claims specifcally authorized as outside of the drug category (e.g., FDA-approved health
claims).
2. Implications of classification
In general, products regulated as “drugs” are subject to greater regulatory scrutiny than
food. New drugs must undergo a lengthy approval process before marketing, must be
demonstrated to be safe and effective for their intended use (generally through one or more
well-controlled clinical trials), and may bear only FDA-approved labeling. All drugs must
be produced in accordance with prescriptive good manufacturing practice requirements
Application of Biotechnology for Functional Foods
45

for pharmaceuticals. In contrast, food, including food ingredients, is subject to pre-market
review and clearances in more limited circumstances, and generally enjoys greater fexibility
in labeling and production.
C. Conventional Food
1. scope
In the absence of an expressed or implied intent to diagnose, cure, mitigate, treat, or
prevent disease, a product is likely to be regarded as a conventional food if the product
is in the form of an “ordinary” food or beverage, and is consumed primarily for taste,
aroma, or nutritive value. Although not a legally defned term, “conventional food” as
used here denotes products in a traditional food form that are intended for consumption
by the general population, including products produced through modern biotechnology.
Food in traditional form includes fats and oils used in food preparation, vegetables, grains,
beverages, dairy products, nuts, legumes, meat, poultry, or any combination of the above.
Such products are subject to all the requirements applicable to food products generally,
including safety and labeling requirements enforced by FDA.
2. Implications of classification
Safety. Of the food safety standards set out in the FFDCA, two are of central importance.
First, all food, regardless of the regulatory category into which it may fall, must not
bear or contain any added poisonous or deleterious substances that may render the food
injurious to health. (FFDCA § 402(a)(1), 21 U.S.C. § 342(a)(1))
10
Historically, the statutory
prohibition against added poisonous or deleterious substances has served as the primary
tool FDA has used to ensure the safety of all foods, including plant-based products of
modern biotechnology.
Second, food ingredients must, as a general rule, be regulated “food additives” or be
generally recognized as safe (GRAS) for their intended use or otherwise they and food
products containing them are adulterated under the law.
11
If a substance is a “food
additive,” it requires pre-market clearances from FDA on the basis of safety data and other
information submitted in a food additive petition. A legal term of art, “food additive” is
defned in pertinent part to mean—
any substance the intended use of which results or may reasonably be
expected to result, directly or indirectly, in its becoming a component of
or otherwise affecting the characteristic of any food (FFDCA § 201(s), 21
U.S.C. § 321(s))
The term does not include color additives and certain other substances such as pesticides.
Food additives include preservatives such as BHA and BHT, nutrients such as folic
acid, sweeteners such as sucralose, and the kanamycin marker gene product, the only
recombinant DNA product to be regulated as such (See 21 C.F.R. § 173.170).
10 If a poisonous or deleterious substance occurs naturally in a product, and is not increased to abnormal
levels due to mishandling or other intervening acts, the substance must not be “ordinarily injurious” to
health, a less stringent standard. Id.
11 For a discussion of FDA’s authority to require pre-market clearances for food components, including the
important distinction between food components and whole foods, see Edward L. Korwek, FDA Regulation
of Biotechnology as a New Method of Manufacture, 37 Food Drug Cosm. L.J. 289 (1982).
Application of Biotechnology for Functional Foods


Food substances that are GRAS are exempt from the defnition of “food additive” and do
not require pre-market clearances from FDA. In order to achieve GRAS status, however,
there must be evidence supporting the safe use of the ingredient that is generally available
(i.e., published) and accepted by experts qualifed by scientifc training and experience to
evaluate food safety. Under the GRAS standard, a determination of safety may be based on
scientifc procedures or, for substances used prior to January 1, 1958, through experience
based on common use in food. GRAS determinations can be made independently, without
FDA input.
12
For purposes of both GRAS determinations and food additive clearances,
“safety” means that “there is a reasonable certainty in the minds of competent scientists
that the substance is not harmful under the intended conditions of use” (21 C.F.R. §
170.3(i)).
Functional food derived from modern biotechnology is subject to the same FDA safety
framework as other food, meaning that such products similarly must contain only
components that are cleared food additives or GRAS. In guidance, FDA has advised that it
is the transferred genetic material and the intended expression product that are subject to
the food additive and GRAS standards (57 Fed. Reg. 22984, 22990 (May 29, 1992)). Thus,
if a plant were modifed to provide a particular nutrient or phytochemical, the nutrient
or phytochemical would need to be either an FDA-cleared food additive or GRAS for its
intended use.
FDA has cleared relatively few “functional” food ingredients as “food additives,” so many—
if not most—of these ingredients are currently used in food on the basis of self-determined
GRAS positions. Those that are regulated food additives include Vitamin D, folic acid, fsh
protein isolate, and amino acids. In a letter to industry, FDA broadly questioned the legal
basis for using “botanicals and other novel ingredients” in conventional food, asserting that
many of “these added ingredients are not being used in accordance with an applicable food
additive regulation and may not be GRAS for their intended use” (CSFAN 2001).
The need for FDA clearance as a food additive or for the establishment of GRAS status
therefore represents an area of caution for many functional food ingredients, including
those produced through methods of modern biotechnology. Indeed, the safety of functional
food ingredients has been a target of critics of the current regulatory framework. GAO
recommended that Congress amend the FFDCA to require companies marketing functional
food to submit advance notifcation to FDA of ingredients determined to be safe (GAO
2000). CSPI has urged FDA to implement GAO’s proposal based on existing statutory
authority (CSPI 2002).
Fortifcation. Conventional food is subject to FDA’s fortifcation policy, a set of principles
intended to serve as “a model for the rational addition of nutrients” to food (21 C.F.R.
Part 104). The policy, which is nonbinding in most circumstances, recommends a relatively
narrow set of conditions under which FDA believes it is appropriate to add nutrients to
12 FDA has established a voluntary procedure for notifying the agency of independent GRAS determinations,
affording the agency an opportunity to object to, or “question” the status of a substance claimed to
be GRAS. See 62 Fed. Reg. 18937 (1997). FDA has accepted GRAS notifcations for many functional
ingredients, including lutein, fsh oil, phytosterols, and inulin. GRAS notifcations are not yet accepted
for animal feed ingredients, although FDA’s Center for Veterinary Medicine (CVM) has traditionally
addressed the safety of feed ingredients through the issuance of so-called “no objection” letters. See
infra section IV.C.
Application of Biotechnology for Functional Foods
47

food. For example, the policy suggests that vitamins and minerals may be added to food to
correct a recognized dietary defciency or to restore nutrients lost in processing. The policy
does not address nontraditional substances that are not vitamins or minerals. Application
of the policy to functional food is uncertain, especially with respect to non-traditional
nutrients.
Labeling Generally. Nutrition and health-related claims for conventional food are
subject to prescriptive requirements under the FFDCA, as amended by the NLEA. These
requirements, which address “health claims,” “structure/function claims” and “nutrient
content claims,” apply to product labels and to “labeling.” The reach of these requirements
is broad, as “labeling” is generously defned to include “all labels and all other written,
printed, or graphic matter (1) upon any article or any of its containers or wrappers, or
(2) accompanying such article” (FFDCA § 201(m), 21 U.S.C. § 321(m))
13
FDA considers a
product to be “accompanied” by written, printed, or graphic material when the material
supplements the product label or otherwise explains the product.
14
Thus, the agency has
characterized brochures, pamphlets, point-of-purchase signage, internet pages, press
releases, and in certain circumstances, even books and journal articles, as labeling.
Health Claims. Conventional food is permitted to bear statements regarding the
relationship between a food and disease only if such statements are an FDA-authorized
“health claim.” A “health claim” is defned as—
any claim that expressly or by implication characterizes the relationship
between a food or a substance therein and a disease or health-related
condition (FFDCA § 403(r), 21 U.S.C. § 343(r); 21 C.F.R. § 101.14).
FDA may authorize health claims by issuing a health claim regulation, or by accepting
a health claim “notifcation” based upon an authoritative statement of a scientifc body
with responsibility “for public health protection or research directly relating to human
nutrition.” To date, FDA has approved 12 health claims by regulation, and has authorized
additional health claims through the statutory notifcation process.
15

13 The same defnition of “labeling” applies for all categories of FDA-regulated products.
14 See, e.g., Kordel v. United States, 335 U.S. 345 (1948) (holding that the phrase “accompanying such article”
is not restricted to materials that are physically on or in an article or package, and reasoning that it is the
textual relationship between a material and a product, not physical attachment, that is dispositive to a
fnding that the material is “labeling”).
15 The health claim notifcation process was created in 1997 to streamline the authorization of claims that
are based upon fndings (specifcally, “authoritative statements”) of certain scientifc bodies, such as the
National Academy of Sciences (NAS), with responsibility “for public health protection or research directly
relating to human nutrition.” A party that wishes to base a claim on an “authoritative statement” must
notify FDA of the claim at least 120 days before food bearing the claim is frst introduced into interstate
commerce. The claim will be permitted so long as the agency does not object to its use, which it may do
by issuing a regulation prohibiting or modifying the claim, or by fnding that the applicable statutory
conditions have not been satisfed.
Application of Biotechnology for Functional Foods
48

taBle III-3 select relationships that may Be
the subject of Fda-approved Health claims
Calcium and osteoporosis
Dietary lipids and cancer
Sodium and hypertension
Dietary saturated fat and cholesterol and coronary heart disease
Fiber-containing grain products, fruits, and vegetables and cancer
Folate and neural tube defects
Soy protein and coronary heart disease
Plant sterol/stanol esters and coronary heart disease
Whole grains and coronary heart disease and cancer
FDA has advised that two elements must be present before a statement will be classifed as
a “health claim” (58 Fed. Reg. 2478, 2479-80, 1993). First, the statement must be reasonably
understood to refer to a food (e.g., specifc fruits and vegetables, such as apples) or a specifc
substance in a food (e.g., fber). Second, the statement must refer to a “disease or health-
related condition.” FDA has defned “disease or health-related condition” to mean “damage
to an organ, part, structure, or system of the body such that it does not function properly
(e.g., cardiovascular disease), or a state of health leading to such dysfunctioning (e.g.,
hypertension) (21 C.F.R. § 101.14(a)(5)).”
If a statement does not identify both a specifc food or substance and a disease or health-
related condition, it is not a health claim. For example, the statement “5-a-Day for Better
Health” as applied to fruits and vegetables is not a health claim because it does not
reference a specifc disease or health-related condition; the statement “diets rich in fruits
and vegetables may reduce the risk of cancer” is not a health claim because it does not
reference a specifc food. FDA considers such claims to be in the nature of general dietary
guidance, not health claims. General dietary guidance claims are permitted so long as they
are not false or misleading in any particular.
Several important limitations restrict the circumstances under which health claims may
be used. First, FDA traditionally has required that health claims be based on “signifcant
scientifc agreement.” The “signifcant scientifc agreement” standard requires a fnding that
“based on the totality of publicly available scientifc evidence … there is signifcant scientifc
agreement, among experts qualifed by scientifc training and experience to evaluate such
claims, that the claim is supported by such evidence” (FFDCA § 403(r)(3)(B), 21 U.S.C.
§ 343(r)(3)(B)). Although “signifcant scientifc agreement” does not require a consensus or
incontrovertible evidence, on a continuum between emerging science and consensus, it lies
closer to consensus, according to FDA.
Second, for each health claim that is currently permitted, detailed criteria concerning the
conditions for using the claim are spelled out in the relevant authorizing regulation or
notifcation. These criteria address the elements that must be present in each claim and
Application of Biotechnology for Functional Foods


the nature of the food that may bear the claim. For example, the authorized health claim
concerning saturated fat, cholesterol, and heart disease must state that heart disease risk
depends upon many factors, and may appear only on food products that meet FDA’s
defnitions for “low fat,” “low saturated fat,” and “low cholesterol” (21 C.F.R. § 101.75).
In all cases, the claimed health beneft must be described as a potential reduction in
disease risk that may occur when the relevant food or substance is consumed as part of an
appropriate diet. FDA has interpreted the health claim provisions not to permit claims that
a food or substance may treat, cure, or mitigate disease.
Third, in addition to the claim-specifc criteria, all health claims are subject to certain
general principles set forth in FDA’s regulations, in 21 C.F.R. § 101.14. Perhaps the most
signifcant of these is the “disqualifying” nutrient concept. Specifcally, food products
that contain fat, saturated fat, cholesterol, or sodium at levels that FDA has deemed to
be of nutritional concern are generally “disqualifed” from bearing a health claim. The
disqualifying levels for fat, saturated fat, cholesterol, and sodium are 13 grams, 4 grams, 60
milligrams, and 480 milligrams, respectively.
In its call for reform of FDA’s regulation of functional food, IFT recommended that FDA
streamline the agency’s review of health claims by adopting a notifcation procedure for
claims that are “generally recognized as effcacious” (GRAE) (IFT 2005). As proposed by
IFT, companies wishing to use a health claim could convene an expert panel, described by
IFT as a GRAE panel, to evaluate the science supporting the claim. If the panel found the
claim to be GRAE, the claim could be notifed to FDA and used within 90 days if the agency
did not object. FDA, however, does not believe it has the legal authority to adopt such a
system (71 Fed. Reg. 62400, 62404, Oct. 25, 2006).
Qualifed Health Claims. In a 2003 policy change, FDA adopted an interim process that
cleared the way for use of certain “qualifed” health claims that are not supported by
signifcant scientifc agreement. The new policy was prompted by a series of court decisions
that held FDA’s implementation of the “signifcant scientifc agreement” standard for
health claims to violate First Amendment principles.
16
As a result, FDA is not permitted
to automatically reject health claims simply because the claims are not supported by
signifcant scientifc agreement; rather, the agency must frst consider whether a disclaimer
could be used to convey the relative strength of the science and eliminate potential
deception.
Under its interim policy, FDA will entertain requests for the use of claims not supported by
signifcant scientifc agreement if the following conditions are met:
nThe claim is the subject of a health claim petition that is fled for comprehensive review
with FDA;
nCredible scientifc evidence supports the claim;
nThe claim is appropriately qualifed to refect the extent of the scientifc evidence; and
nThe claim otherwise meets the general regulatory requirements for health claims.
For claims that meet the criteria in the new health claim policy, FDA will allow the claim
through the exercise of “enforcement discretion” (i.e., an agreement to not take action
against the claim). An example of a qualifed health claim that FDA has allowed for
conventional food and dietary supplements containing certain omega-3 fatty acids is
16 See, e.g., Pearson v. Shalala, 164 F.3d 650 (D.C. Cir. 1999).
Application of Biotechnology for Functional Foods
50

“Supportive but not conclusive research shows that consumption of EPA and DHA omega-
3 fatty acids may reduce the risk of coronary heart disease. One serving of [name of food]
provides __ gram of EPA and DHA omega-3 fatty acids. [See nutrition information for total
fat, saturated fat, and cholesterol content.]”
At the time of its adoption, the qualifed health claims process was both praised and
criticized for increasing the fexibility of health claim regulation and making possible new
claims based on emerging evidence. FDA’s implementation of the process, however, has
raised questions about its utility. For instance, several “approved” claims (i.e., claims for
which FDA agreed to exercise enforcement discretion) characterize the subject diet-disease
relationship in a negative manner:
“Three studies, including a large clinical trial, do not show that calcium
supplements reduce the risk of preeclampsia during pregnancy. However,
two other studies suggest that calcium supplements may reduce the risk.
Based on these studies, FDA concludes that it is highly unlikely that
calcium supplements reduce the risk of preeclampsia” (FDA 2005a).
“One weak and limited study does not show that drinking green tea
reduces the risk of prostate cancer, but another weak and limited study
suggests that drinking green tea may reduce this risk. Based on these
studies, FDA concludes that it is highly unlikely that green tea reduces the
risk of prostate cancer” (FDA 2005b).
In addition, consumer perception studies have called into question whether consumers
are able to understand disclaimers intended to convey the strength of scientifc evidence
supporting a claim.
17
Structure/function claims. A food may bear, without FDA pre-market review or approval,
substantiated claims identifying the effect of the food on a bodily structure or function. As
explained previously, the legal basis for structure/function claims is the “drug” defnition
in section 201(g)(1)(C) of the FFDCA, which defnes “drug” in pertinent part to mean
“articles (other than food) intended to affect the structure or any function of the body of
man or other animals.” By exempting food promoted for structure/function effects from the
defnition of “drug,” Congress in effect exempted food bearing structure/function claims
from the FFDCA’s stringent requirements applicable to new drugs.
To avoid “drug” status based on disease claims, structure/function claims must be written
in a manner that presumes the intended audience is healthy and seeks solely to maintain,
promote, or otherwise support that state of good health. An example of a “structure/
function” claim is “conjugated linoleic acid supports a healthy immune system.” Structure/
function claims for conventional food do not require FDA approval or review, nor has the
agency identifed specifc criteria (e.g., disqualifying criteria) for their use. FDA has issued,
however, a draft guidance to address the scientifc data and information that industry
should have to substantiate structure/function claims as not false or misleading in any
particular (FDA 2004).
18
17 See Brenda M. Derby and Alan S. Levy, Working Paper: Effects of Strength of Science Disclaimers on
Communication Impacts of Health Claims (Sept. 2005); International Food Information Counsel (IFIC),
Qualifed Health Claims Consumer Research Project Executive Summary (Mar. 2005) (available at http://
www.ifc.org) (accessed Oct. 2005).
18 Although the draft guidance applies on its face only to dietary supplements, the principles set out in the
guidance apply equally to conventional foods.
Application of Biotechnology for Functional Foods
51

For conventional food, such as cranberries or cranberry juice suggested to promote urinary
tract health, FDA has taken the position that structure/function claims must be based upon
nutritive value. FDA’s reasoning for this position is based on the statutory exemption from
“drug” regulation for food (i.e., the “other than food” language quoted above): because
the statute exempts food intended to affect bodily structures/functions from regulation as
a drug, and a food can affect a bodily structure or function only through nutritive value, a
structure/function claim should be tied to such nutritive value, and not a pharmacological
effect. FDA, however, has not clearly explained the meaning of nutritive value in this
context, and appears to have moved away from a strict application of this standard in recent
years. In the agency’s view, FDA has “provided signifcant fexibility in determining whether
a substance possesses nutritive value” (71 Fed. Reg. 62400, 62405 (Oct. 25, 2006)).
19
For dietary supplements, DSHEA specifcally authorizes the use of structure/function
claims (FFDCA § 403(r)(6), 21 U.S.C. § 343(r)(6)). Accordingly, dietary supplements may
bear structure/function claims without regard to nutritive value. As a result of DSHEA and
FDA’s policy on structure/function claims for conventional food, the “disease” provisions of
the drug defnition are of greater practical importance in assessing potential drug status.
Because structure/function claims can be made with relative ease (assuming adequate
substantiation exists), they are commonly used on functional food products, and have been
the target of critics of functional food regulation, who suggest that more stringent controls
are needed. The GAO has recommended that the FFDCA be amended to require marketers
of functional food in conventional food form to follow the same procedures for structure/
function claims as dietary supplements (GAO 2000). In other words, GAO suggests
that conventional food marketers should be required to notify FDA regarding the use of
structure/function claims, and to provide a disclaimer that claims made in labeling have
not been reviewed by FDA. The GAO has also recommended that FDA develop regulations
or other guidance concerning the scientifc evidence needed to support structure/function
claims. These recommendations have been supported by groups such as CSPI (CSPI 2002).
20
Nutrient content claims. A claim that characterizes the level of a nutrient in a food, such
as “low fat” or “high in calcium,” is a “nutrient content claim.” A statement is deemed to
“characterize” the level of a nutrient in food if it states or implies that the level has some
nutritional signifcance (i.e., suggests that there is “a lot or a little” of the nutrient relative
to a dietary recommendation). Like health claims, nutrient content claims may be expressed
or implied, and cannot be used unless authorized by FDA in a regulation or a nutrient
content claim notifcation (FFDCA § 403(r), 21 U.S.C. § 343(r); 21 C.F.R. § 101.13). Express
nutrient content claims may be phrased in absolute or relative terms (i.e., by comparing the
nutrient value of one food to another).
Examples of FDA-approved nutrient content claims are provided in Table III-4. As a general
rule, a product may use these terms in food labeling only if the product meets the FDA
defnition and criteria for the term.
19 The IFT has asked FDA to replace the “nutritive value” concept with a more inclusive defnition, allowing
structure/function claims based on either nutritive value or any “physical or physiological effect that
has been scientifcally documented or for which a substantial body of evidence exists for a plausible
mechanism.” IFT Report, at 51.
20 See also CSPI Reports, Functional Foods: Public Health Boon or 21
st
Century Quackery? (March, 1999).
Application of Biotechnology for Functional Foods
52

taBle III-4 select Fda-approved nutrient content claims
Good source, Contains, Provides
High in, Excellent Source of
More, Fortified, Enriched, Added, Extra, Plus
Antioxidant
Light or lite
Calorie free, low calorie, reduced calorie
Sodium free, very low sodium, low sodium, reduced sodium
Sugar Free, No Added Sugar
Lean
Of particular importance to many functional food products, a nutrient generally may be
the subject of a nutrient content claim only if the nutrient has an established reference
intake, either in the form of a daily value that may be used in nutrition labeling, or a
Dietary Reference Intake adopted by the Institute of Medicine or another authoritative
body. Thus, because FDA has established a daily value for vitamin E, a peanut that is
enhanced to provide “more” vitamin E than conventional peanuts may be promoted on
this basis if the increased vitamin E satisfes FDA’s regulation for the use of “more” claims.
In contrast, spinach that is enhanced to provide increased lutein, for which there is no
reference value, cannot be claimed to be a “good source” of lutein, or to contain “more
lutein than conventional spinach,” no matter how much lutein the fnished food contains.
The only nutrient content claim currently available to such a food would be a factual
statement concerning the amount of lutein in the product (e.g., 2 mg of lutein per serving).
Labeling—Other Considerations. In addition to the requirements for nutrition and health-
related claims detailed above, two additional labeling considerations may have relevance to
functional foods prepared using modern biotechnology methods: the requirement for food
products to bear appropriate statements of identity; and the mandate for labels to reveal
“material” facts about a food.
Labels of conventional food products must bear appropriate statements of product identity.
By regulation, these statements should refect (1) the name required by a federal regulation,
if any, (2) the common or usual name of the food, or if none, (3) an appropriately
descriptive term, in that order of preference (21 C.F.R. § 101.3). If a bioengineered food is
signifcantly different from its conventional counterpart such that the traditional name no
longer adequately describes the new food, then a new product identity must be developed.
Accordingly, depending upon the nature of the modifcation, functional and other food
produced through modern biotechnology may require unique product identity statements.
An example is oil that is altered to contain signifcantly higher levels of unsaturated fats, in
which case the product identity would highlight the predominant fats that are present, such
as “high oleic acid soybean oil.” Where terms such as “high” are necessary to describe the
basic properties of a food, they are not subject to regulation as nutrient content claims.
Application of Biotechnology for Functional Foods
53

In addition, under 201(n) of the FFDCA, a food label must reveal all facts that may be
material in light of representations made in labeling or in light of consequences that may
result from its suggested or customary conditions of use. Consequences that are deemed
material typically relate to substantial issues of safety or usage to which consumers should
be alerted. For example, if a modifed fruit includes an allergen at a level that could trigger
an adverse reaction, and consumers would not expect the allergen to be present based on
the name of the food, the presence of that allergen must be disclosed (57 Fed. Reg.
at 22991).
D. Dietary Supplements
1. scope
In 1994, Congress amended the FFDCA to create a new regulatory category for “dietary
supplements.” A dietary supplement is defned as—
[A] product (other than tobacco) intended to supplement the diet that
bears or contains one or more of the following dietary ingredients:
(A) a vitamin;
(B) a mineral;
(C) an herb or other botanical;
(D) an amino acid;
(E) a dietary substance for use by man to supplement the diet by increasing
the total dietary intake; or
(F) a concentrate, metabolite, constituent, extract, or combination of
any ingredient described in clause (A), (B), (C), (D), or (E) … (FFDCA §
201(ff), 21 U.S.C. § 321(ff))
In addition to containing one or more dietary ingredients, a dietary supplement must
be intended for ingestion (as opposed to topical application, inhalation, or other routes
of application that do not involve the gastrointestinal tract), must not have been frst
approved as a “new drug” or as an “investigational new drug” for which substantial public
investigations were conducted, must be expressly identifed as a “dietary supplement,” and
must not be represented for use as a conventional food or meal replacement.
21
The restriction on representations for conventional food uses means that a dietary
supplement cannot be promoted as a product to be consumed primarily for taste or
aroma (e.g., “Delicious and refreshing beverage”) or for conventional food uses (e.g., as a
salad dressing). FDA has challenged several “functional food” products that attempted to
combine a dietary supplement identity with attributes closely aligned with conventional
food. For example, FDA rejected arguments that a spread fortifed with phytosterols was
a dietary supplement, where promotional materials for the product emphasized taste and
depicted the product being used with toast and other conventional food. At the same time,
many dietary supplements have been marketed in the form of a bar or liquid beverage-type
product without challenge, demonstrating possible overlap between the dietary supplement
and conventional food categories.
21 A dietary supplement also must not have been frst certifed as an antibiotic or licensed as a biologic,
or authorized for investigation as a new antibiotic or biologic for which substantial public clinical
investigations have been conducted.
Application of Biotechnology for Functional Foods
54

Functional food products and dietary supplements are often treated as distinct categories.
As suggested above, there is no bright line test for distinguishing functional food from
dietary supplements, nor is there an absolute prohibition against marketing what might
be a functional food as a dietary supplement. Accordingly, there may be circumstances
under which a “functional food” might be marketed as either a conventional food or a
dietary supplement. For certain products, such as products that can only be consumed
with conventional food (e.g., spreads), a dietary supplement position may not be feasible;
moreover, it may not be desirable to refrain from marketing a food on the basis of its taste
or favor. For other products, such as those in bar and certain liquid forms, manufacturers
have arguable discretion to position the products as either conventional food or dietary
supplements.
2. Implications of classification
Safety. Safety requirements for dietary supplements differ substantially from those for
conventional food. In contrast to conventional food ingredients, which must be determined
to present a “reasonable certainty of no harm,” the safety standard for dietary supplements
provides that dietary supplements must not present “a signifcant or unreasonable risk of
illness or injury” under the conditions of use (FFDCA § 402(f)(1), 21 U.S.C. § 342(f)(1)).
The safety standards for dietary supplements are widely considered to be less stringent than
conventional food standards.
Application of the “signifcant or unreasonable risk of illness or injury” standard requires
a case-by-case assessment. In its 2004 rulemaking to ban ephedrine alkaloid supplements,
FDA determined a supplement to present an “unreasonable” risk if the risks posed by the
product outweigh the substantiated benefts. Although FDA’s interpretation was initially
rejected in a legal challenge, the agency’s approach was ultimately upheld on appeal.
22
In
upholding FDA’s interpretation, the appellate court found the statute to impose two distinct
safety standards that may cause a supplement to be adulterated: a “signifcant risk” of
illness or injury, which the court found to imply a “great danger; and an “unreasonable”
risk, which the court found to require an accounting of whether the claimed benefts justify
the risk.
23
As for pre-market review, dietary ingredients are exempt from the defnition of “food
additive,” so pre-market clearance is not required, nor are such ingredients subject to the
GRAS standard. If a dietary ingredient is “new,” however, FDA must be notifed of the basis
for its safe use before it may be marketed (FFDCA § 413, 21 U.S.C. § 350b). A new dietary
ingredient notifcation is required for any dietary ingredient that was not marketed in the
United States before October 15, 1994, unless the new dietary ingredient has been present in
the food supply as an article used for food in a chemically unaltered form.
Labeling. Labeling requirements for dietary supplements are similar to requirements for
conventional food in many respects. Like conventional food, dietary supplements may bear
nutrient content claims and health claims only if such claims are expressly authorized by
FDA. Dietary supplements also may bear structure/function claims, although FDA must
be notifed of the use of the claims, and the claims must be accompanied by a disclaimer
stating that—
22 Nutraceutical Corp. v. Von Eschenbach, 459 F.3d. 1033, 1038 (10
th
Cir. 2006)(holding that Congress
“unambiguously” required FDA to conduct a risk-beneft analysis when assessing whether a supplement
presents an unreasonable risk).
23 Id. at 1040.
Application of Biotechnology for Functional Foods
55

This statement has not been evaluated by the Food and Drug
Administration. This product is not intended to diagnose, treat, cure, or
prevent any disease (Id. § 403(r)(6)(C), 21 U.S.C. § 343(r)(6)(C)).
Dietary supplement manufacturers are required to notify FDA of their intended use of
structure/function claims. If FDA objects to the notifed claims, it responds to the manu-
facturer in a so-called “courtesy letter” that explains the basis for the agency’s objection.
Statements that have been the subject of courtesy letters include ”helps maintain healthy
cardiac risk ratios,” “cholesterol protection formula,” “support normal body function
during the cold season,” “treatment for anxiety, anorexia, and depression,” and “helps
maintain healthy blood sugar levels,” among numerous other claims deemed objectionable
by FDA.
E. Food for Special Dietary Uses
1. scope
Perhaps the most ambiguous category of all, “special dietary uses” is defned to mean—
[P]articular (as distinguished from general) uses of food, as follows:
(i) Uses for supplying particular dietary needs which exist by reason of a
physical, physiological, pathological or other condition, including but not
limited to the conditions of diseases, convalescence, pregnancy, lactation,
allergic hypersensitivity to food, underweight, and overweight;
(ii) Uses for supplying particular dietary needs which exist by reason of
age, including but not limited to the ages of infancy and childhood;
(iii) Uses for supplementing or fortifying the ordinary or usual diet with
any vitamin, mineral, or other dietary property. Any such particular use
of a food is a special dietary use, regardless of whether such food also
purports to be or is represented for general use (21 C.F.R. § 105.3(a)(1)).
Although this language could, on its face, be broadly interpreted to cover numerous food
products—including, perhaps, all functional food—its reach has been gradually restricted
over the years. Two FDA interpretations have played a particularly important role in
shaping the special dietary use category.
First, FDA has taken the position that needs relating to general dietary guidelines are
not “special dietary uses.” Thus, FDA has determined that most statements concerning
nutrients such as sodium and sugars are most appropriately regulated under the nutrient
content claim provisions of the FFDCA, since advice concerning restrictions of these
nutrients are applicable to the general public. This determination demonstrates a unique
aspect of special dietary use regulation—a food can lose its special dietary usefulness once
a need related to a particular condition has broad applicability to the public at large. Needs
relating to the prevention of diet-related chronic diseases of broad concern seem most apt
to fall into this category, and thus to be considered of general usefulness. Among the food
products retaining “special” usefulness are products relating to food allergy and sensitivities
(e.g., gluten intolerance, peanut allergy), products relating to intake problems common
to several diseases or conditions (e.g., dysphagia, or diffculty swallowing), and products
intended for weight loss or weight management programs.
Application of Biotechnology for Functional Foods


Second, FDA has provided by regulation that fortifed food is deemed to be for a special
dietary use if vitamins or minerals added to the food provide 50% or more of the applicable
daily values in a single serving (Id. § 101.9(a)(4)). / This determination appears to eliminate
many fortifed products from the special dietary food category. Because the interpretation
applies only to nutrients with a reference value, however, it does not address many
functional food products, which may contain phytochemicals and other substances for
which a reference intake has not been established.
2. Implications of classification
Safety. Like ingredients of conventional food, ingredients used in food for special dietary
use must generally be regulated food additives or GRAS for their intended use.
Labeling Generally. Food for special dietary use is subject to several unique labeling
requirements. The central requirement provides that a food for special dietary use is deemed
misbranded unless its label bears information adequate to inform purchasers of its value
for—
such information concerning its vitamin, mineral, or other dietary
properties as the Secretary determines to be, and by regulations prescribes
as, necessary in order fully to inform purchasers as to its value for such
uses (FFDCA § 403(j), 21 U.S.C. § 343(j)).
In a few instances, FDA has issued regulations addressing the labeling information that
must be provided for food for special dietary use. For example, food produced through
modern biotechnology and intended for use as a hypoallergenic food, an infant food,
or a weight loss product would be subject to specifc labeling requirements as provided
for in FDA’s special dietary use regulations. In all other cases, the informational labeling
requirement for special dietary uses is subject to interpretation on a case-by-case basis.
In addition, FDA has advised that statements that appear in labeling pursuant to a special
dietary use regulation will not be subject to the agency’s requirements for health claims
and nutrient content claims. The agency has suggested that the exemptions apply only
if the claim is made in compliance with a “specifc provision” of the special dietary use
regulations, and is made solely to note the special dietary usefulness of the product. The
reach and practical application of these exemptions, however, is unclear. Following NLEA,
FDA suggested that it would initiate rulemaking to clarify the relationship between special
dietary use claims and health and nutrient content claims, but the agency has not done so to
date.
Labeling—Hypoallergenic Food. There is some promise that modern biotechnology may
be used to produce food, such as peanuts, without allergenic proteins. If promoted as such,
these products would be subject to FDA’s regulations for food represented to be for a special
dietary use by reason of “the decrease or absence of any allergenic property” (21 C.F.R.
§ 105.62). The following labeling information technically would be required under FDA’s
regulation for hypoallergenic claims:
nThe common or usual name and quantity or proportion of each ingredient;
nA qualifcation of the name of the food, or the name of each ingredient thereof, to reveal
the specifc plant or animal that is the source of each ingredient; and
nAn “informative statement of the nature and effect of any treatment or processing of
the food or any ingredient thereof,” if the changed allergenic property results from
processing.
Application of Biotechnology for Functional Foods
57

Adopted many years ago, these requirements, particularly the frst two, seem more naturally
suited to multiple-ingredient, processed food than single-ingredient commodities produced
through modern biotechnology. Their application to food produced using modern biotech-
nology would need to be assessed on a case-by-case basis.
F. Medical Food
1.scope
The term “medical food” has been defned to mean—
[F]ood which is formulated to be consumed or administered enterally
under the supervision of a physician and which is intended for the specifc
dietary management of a disease or condition for which distinctive
nutritional requirements, based on recognized scientifc principles, are
established by medical evaluation (Orphan Drug Amendments of 1988,
Pub. L. No. 100-290 (amending 21 U.S.C. 360ee); 21 C.F.R. § 101.9(j)(8)). /
As noted previously, the frst product recognized as a “medical food” by FDA was the
infant formula Lofenalac®, which is intended for the dietary management of PKU. Initially
regulated as a drug, Lofenalac® was recognized by FDA to meet distinctive nutritional
requirements—namely, an impaired ability to metabolize the essential amino acid
phenylalanine—and thus was reclassifed as a distinct type of food for special dietary use.
The impetus for this reclassifcation was a practical one: FDA recognized the value that
specialty formulas and other highly specialized products bring to a small but vulnerable
population, and did not wish to discourage the development or marketing of such products
by subjecting them to stringent requirements for drug products.
Now a distinct category from food for special dietary use, medical food is defned in an
extremely narrow manner. Specifcally, FDA has provided by regulation that a product can
be classifed as a medical food only if the product is—
nA specially formulated and processed product, as opposed to a naturally occurring food
in its natural state;
nA product intended for the partial or exclusive feeding of a patient by means of oral
intake or enteral feeding by tube;
nIntended for the dietary management of a patient who, because of therapeutic or
chronic medical needs, has limited or impaired capacity to ingest, digest, absorb, or
metabolize ordinary food or certain nutrients, or who has special medically determined
nutrient requirements that cannot be achieved via modifcation of the diet alone;
nIntended to provide nutritional support for the management of unique nutrient needs
that result from a specifc disease or condition, as determined by medical evaluation;
nIntended for use under medical supervision; and
nIntended only for patients receiving active and ongoing medical supervision.
An example of a product positioned as a medical food is a moderate protein, low
electrolyte, low fuid, high calorie formula intended to provide balanced nutrition for
dialyzed patients with chronic or acute renal failure.
1. Implications of classification
Safety. Like ingredients of conventional food, ingredients of medical food must generally
be regulated food additives or GRAS for their intended use. For example, in providing for
the use of the food additive folic acid in food, FDA specifcally authorized its use in medical
food products.
Application of Biotechnology for Functional Foods
58

Labeling. In recognition of their unique status, medical food products are expressly
exempt from FDA requirements for nutrition labeling, nutrient content claims, and health
claims. Although these exemptions make a medical food positioning extremely attractive,
the criteria detailed above reveal that FDA has interpreted the category in a very narrow
manner. As a practical matter, few functional food products will be in a position to be
marketed as a medical food in substantial compliance with the FFDCA.
IV. Food suBject to addItIonal or unIQue reQuIrements:
meat and poultry, sHell eggs and egg products,
and anImal Feed, IncludIng pet Food
Although the FFDCA provides a broad framework that is of central importance to most
functional food products, certain products are subject to additional or unique requirements
under the FFDCA and other laws. A comprehensive discussion of such additional or unique
requirements is beyond the scope of this report; a brief overview of three regulatory schemes
of particular interest to functional food products is provided below: meat and poultry
products, shell eggs and egg products, and non-human animal food, including pet food.
A. “Meat Food Products” or “Poultry Products”
Products that are “meat food products” or “poultry products” are regulated by the Food
Safety and Inspection Service (FSIS) of USDA pursuant to the Federal Meat Inspection Act
(FMIA) and Poultry Products Inspection Act (PPIA), respectively. A “meat food product”
includes any product capable for use as human food made from the carcass of cattle, sheep,
swine, or goats. “Poultry products” include human food products made from the carcass
of any domesticated bird. A functional food would be subject to regulation by FSIS if it is
meat or poultry, or it is a product that contains a meat or poultry ingredient, at more than
a de minimis level (generally, greater than 2 to 3%), and in a form of the type generally
considered to be a product of the meat or poultry industries. As is the case with other food
under the FFDCA, meat and poultry products cannot be adulterated or misbranded under
the FMIA and PPIA.
Although FDA and FSIS requirements are similar in many respects, FSIS generally takes the
position that it must specifcally approve the use of all meat and poultry product ingredients
and labels. Thus, FSIS requires that all ingredients used in meat and poultry products be
specifcally approved as safe and suitable, and that most labels (and claims) be reviewed
and approved before use. Complicating the situation, FSIS has approved the use of many
nutrient content claims by regulation (e.g., “low fat,” “lean”), but has issued no regulations
or formal policies concerning health claims or structure/function claims. As a general rule,
FSIS has evaluated labels containing proposed health claims and structure/function claims
on a case-by-case basis, and has permitted such claims under limited circumstances. For
example, FSIS has allowed the labels of appropriate products to bear the American Heart
Association Heart Check program logo and the FDA-approved health claim concerning
dietary saturated fat and cholesterol and risk of coronary heart disease.
Of particular relevance to functional food, FSIS has historically taken the position that
meat or poultry, per se, is not suitable for fortifcation. It has permitted certain meat and
poultry products to contain FDA-regulated ingredients that are fortifed in accordance with
FDA requirements (e.g., enriched pasta), if the actual fortifcation occurs outside of a FSIS-
inspected meat or poultry plant. FSIS’ limits on fortifcation would generally be expected to
limit the marketing of “functional” meat or poultry products, including products to which
functional ingredients are added through genetic means or product formulation.
Application of Biotechnology for Functional Foods


FSIS has not yet taken a public position concerning the status of transgenic animals
that are intended to produce functional food in the form of meat or poultry for human
consumption. If presented with a transgenic functional food, such as a ground beef product
promoted as containing a favorable fatty acid profle due to genetic enhancement, FSIS
would be expected to follow its usual pre-market approval approach. Thus, the agency
would likely engage in a review of the suitability of such enhancements (e.g., the effect
on the characteristics and cooking properties of the resulting meat, as well as a review of
whether the enhancements constitute impermissible “fortifcation”), the product’s safety
(regarding which USDA would defer to FDA, but would likely require a specifc statement
from the agency), and the product’s labeling, including substantiation for any claims made.
B. Shell Eggs and Egg Products
Jurisdiction over shell eggs and egg products is shared by FDA and USDA. FDA bears primary
responsibility for the safety and labeling of shell eggs for consumer use under the FFDCA
and the Public Health Service Act. Processed egg products, such as liquid eggs, are similarly
subject to regulation by FDA; the production and labeling of such products, however, is also
subject to USDA oversight pursuant to the Egg Products Inspection Act (EPIA).
Shell eggs are subject to the FFDCA provisions concerning safety and labeling, and thus
must meet the same basic requirements as all other FDA-regulated food. For example,
a shell egg that contains a functional component, such as the omega-3 fatty acid,
docosahexaenoic acid (DHA), at elevated levels, must be safe under the FFDCA. FDA
would likely take the position that DHA, the targeted and desirable component that
differentiates such products from other eggs, must be either GRAS or specifcally cleared
by FDA as a food additive for its intended use.
24
Additionally, the labeling of such a product
is subject to FDA’s rules for health claims, nutrient content claims, and other labeling
statements. Because eggs are most reasonably classifed as conventional food, shell eggs
enhanced in a particular way would be regulated as either conventional food or food for
special dietary use, depending on the types of alterations and claims that are made.
An “egg product” is defned to mean “any dried, frozen, or liquid eggs, without or without
added ingredients, excepting products which contain eggs only in a relatively small
proportion or historically have not been … considered by consumers as products of the
egg food industry” (9 C.F.R. § 590.5)
25
Like shell eggs, egg products must comply with the
safety and labeling requirements of the FFDCA. At the same time, pursuant to the EPIA, egg
products must be produced under continuous inspection by FSIS, and must use labels and
formulations that are reviewed and approved by that agency, with possible consultation with
FDA. The EPIA scheme for egg products results in greater oversight of this industry segment.
24 See, e.g., FDA, Letter Regarding Eggs with Enhanced Omega-3 Fatty Acid Content and a Balanced 1:1
Ratio of Omega-3/Omega 6 Fatty Acids and Reduced Risk of Heart Disease and Sudden Fatal Heart
Attack (Docket No. 2004Q-0072, Apr. 5, 2005) (assessing the petitioner’s assertion that eggs enriched with
omega-3 and omega-6 fatty acids were GRAS, though not reaching a conclusion on the issue, due to denial
of the petition on other grounds). In a recent letter announcing the agency’s intent to exercise enforcement
discretion and permit a health claim concerning the omega-3 fatty acids DHA and EPA, FDA concluded
that certain uses of DHA and EPA are safe and lawful, provided that daily intakes of DHA and EPA from
conventional food and dietary supplements do not exceed 3.0 g per person per day. As discussed below,
if DHA content is enhanced through the addition of DHA ingredients to animal feed, FDA’s Center for
Veterinary Medicine (CVM) would likely assert that a separate basis for use of DHA-enriched feed would
need to be established and provided for the review of that Center.
25 By regulation, FSIS has exempted several foods from regulation as “egg products,” including egg substitutes
and “dietary foods.” The scope of the FSIS exemption for “dietary foods” is unclear.
Application of Biotechnology for Functional Foods
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C. Animal Feed, Including Pet Food
All food, including pet food, falls within the defnition of “food” under the FFDCA,
although in common usage often non-human animal food is distinguished as “animal
feed.” Animal feed, however, is subject to quite different regulatory requirements. Some
of the differences are statutory: for example, the NLEA scheme for health and nutrient
content claims does not apply to animal feed, nor does the DSHEA framework for dietary
supplements.
26
Other differences can be explained in large part by the different Centers
within FDA responsible for the regulation of human and other animal food products and
their interpretation of the law: human food is regulated by FDA’s Center for Food Safety
and Applied Nutrition (CFSAN), while other animal food is regulated by FDA’s Center for
Veterinary Medicine (CVM). Oversight of feed by state feed control offcials provides yet
another explanation for the disparate regulation. Most, if not all, states require such food,
including pet food, to be manufactured or distributed within their borders pursuant to a
registration or license, and subject such feed to oversight under state laws similar to the
FFDCA but that require registration or licensing of feed.
Regarding safety, the FFDCA and state feed laws require that substances added to food
either be, as a general rule, regulated food additives or GRAS for their intended use. In
the human food industry, self-determinations of GRAS status are common, and are the
basis upon which many food ingredients are used. In contrast, CVM, as well as state feed
control offcials, routinely take the position that a substance cannot be used in feed unless
it has been formally or informally evaluated by FDA. Since very few ingredients are cleared
as feed additives and since CVM does not have a GRAS notifcation approach,
27
feed
ingredients that have been evaluated by CVM are typically the subject of so-called “no-
objection” letters. Once an ingredient is favorably reviewed by CVM, it is generally allowed
for use in feed, including pet food, under state law. As a result of CVM and state oversight,
the regulatory requirements for using functional ingredients is typically greater for feed
products than for human food products because new feed ingredients must be proven
to be safe and to have utility to FDA’s satisfaction before a “no objection” letter can be
obtained. Examples of a functional feed are those formulations that contain phytase. CVM
has allowed limited claims for such feed after having issued no objection letters for a few
phytases that are produced through the use of recombinant DNA methods.
Another important difference between CFSAN and CVM regulation concerns the
application of the “drug” defnition, especially the “structure/function” exception for
food. CVM treats most structure/function claims as drug claims; thus, “production” claims
such as increased milk production, increased leanness, improved growth, and effciency of
weight gain are viewed as drug claims and feed ingredients with such claims are subject
to regulation as unapproved new animal drugs (CVM 1998). Accordingly, feed containing
such unapproved new drug ingredients cannot be legally marketed. CVM has permitted
structure/function feed claims in limited instances, but only after review and approval.
Examples include “urinary tract health” and “dental health” claims on cat food products
(CVM 2004).
26 See 61 Fed. Reg. 17706 (1996). CVM has indicated that it would not object to the marketing of nutritional
supplements for companion animals in limited circumstances. FDA Compliance Policy Guides, CPG
7126.04 (sec. 690.100)(Rev. 03/1995). CVM has issued no regulations to govern special dietary uses for feed
products, and there is no formal “medical food” category for feed products.
27 See supra note 21.
Application of Biotechnology for Functional Foods
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In summary, CFSAN and CVM approach functional food, including functional ingredients,
in a different manner, resulting in distinct safety and labeling requirements for feed
products. CVM’s prescriptive approach to functional feed, coupled with the lack of a feed-
specifc framework for nutrient content claims, health claims, and dietary supplements,
limits the circumstances under which functional feed ingredients may be marketed without
their being treated as new veterinary drugs.
Application of Biotechnology for Functional Foods
Application of Biotechnology for Functional Foods
ó3

Summary
The application of biotechnology to foods expressly to improve nutritional and
health characteristics holds great potential, as demonstrated by many research
accomplishments. Food enhancements cover a wide range, including improved
fatty acid profles for more heart healthy food oils, improved protein content and
quality for better human and animal nutrition, increased vitamin and mineral levels
to overcome widespread nutrient defciencies throughout the world, and reduction
in anti-nutritional substances that diminish food quality and can be toxic. On
the horizon are efforts to increase the concentration of various antioxidants and
functional substances such as phytosterols and probiotic bacteria. Improvements in
the digestibility of animal feed through the reduction of phytic acid, gossypol and
glycoalkaloids also have potential to enhance the safety of human foods. Although
biotechnology has made some advances in reducing allergenic proteins in some
foods, the complexity of allergenic responses, differences in sensitivity among
people, and the presence of multiple allergens in a single food make this challenge
particularly daunting. It appears that food oils with improved fatty acid profles
are the closest to reaching commercialization, once regulatory clearance has been
obtained.
Just what form those regulatory clearances may take is not entirely certain. There is
no regulatory scheme for functional food, per se, but functional food products are
clearly subject to federal regulation. If a functional food product is marketed for
a therapeutic purpose (e.g., to treat a disease), it will be subject to regulation as a
“drug.” If a product is subject to regulation as a “food,” it may be further classifed
as a conventional food, dietary supplement, food for special dietary use (including
infant formula), or medical food, again depending upon its intended use and other
factors.
How a product is categorized has substantial implications for how it is marketed,
the safety and labeling standards it must meet, and what requirements it must
address for regulatory review and clearance or approval. A conventional food may
be freely marketed on the basis of taste, and enjoys some fexibility regarding
certain types of claims, such as structure/function claims, which need not be
presubmitted to FDA nor accompanied by a disclaimer. Conventional food,
including food ingredients, however, must meet safety standards requiring a
Application of Biotechnology for Functional Foods
ó4

“reasonable certainty of no harm.” The safety standard for dietary supplements
is less stringent, but supplements are restricted to marketing in certain forms
(e.g., as a tablet or powder, and in some circumstances, as a bar or liquid), and
can make structure/function claims only if such claims are submitted to FDA and
accompanied by the DSHEA disclaimer.
Food for special dietary use enjoys some fexibility as to nutrient content and
health claim requirements, but the extent of this fexibility, and of the special
dietary use category itself, is unclear and probably very fact-specifc. Medical food
is entitled to the most fexibility of all, but is permitted in extremely narrow and
carefully defned circumstances. Certain food products, including meat and poultry,
egg products, and animal feed, including pet food, are theoretically eligible for
marketing in a functional food form. Such products are, however, subject to distinct
regulatory requirements and oversight that may limit functional food opportunities
as a practical matter. Animal feed, in particular, has been regulated by CVM in
a manner that restricts the types of ingredients and promotional claims that may
be used, despite the statutory classifcation of animal feed as “food.” The use of
modern biotechnology to enhance human and other animal food will likely not
change these regulatory paradigms, but may challenge the boundaries of some of
the regulatory classifcations.
Application of Biotechnology for Functional Foods
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Application of Biotechnology for Functional Foods
Application of Biotechnology for Functional Foods
Application of Biotechnology for Functional Foods
Application of Biotechnology for Functional Foods

© 2007 Pew Initiative on Food and Biotechnology. All rights reserved. No portion of this paper may be reproduced by any means, electronic or mechanical, without permission in writing from the publisher. This report was supported by a grant from The Pew Charitable Trusts to the University of Richmond. The opinions expressed in this report are those of the authors and do not necessarily reflect the views of The Pew Charitable Trusts or the University of Richmond.

Application of Biotechnology for Functional Foods

Contents
Preface...........................................................................................................................................................................5 Part.1:.Applications.of.Biotechnology.for.Functional.Foods..................................................................7 . Part.2:.Legal.and.Regulatory.Considerations.Under.Federal.Law......................................................37 Summary....................................................................................................................................................................63 . Selected.References..............................................................................................................................................65 

Application of Biotechnology for Functional Foods

Application of Biotechnology for Functional Foods .

sometimes collectively referred to as output traits. genetic modifications that make insect. virus and weed control easier or more efficient. the long-awaited promise of genetically engineered food with more direct consumer benefits moves closer to reality. but generally refers to foods that provide health benefits beyond basic nutrition. have been a longer time in development. but is rather a snapshot in time to give readers a sense of the kinds of products that may one day be available. also known as “functional foods.  Application of Biotechnology for Functional Foods . and we learn more about the genes and biochemical pathways that control those attributes that could offer more direct consumer benefits.S. This report looks at the potential to develop functional foods through the application of modern biotechnology. As the technology advances. and the second section analyzes the legal authorities that could govern the use of biotechnology-derived functional foods. scientists have envisioned harnessing the power of genetic engineering to enhance nutritional and other properties of foods for consumer benefit.Preface S ince the earliest days of agricultural biotechnology development. Agricultural biotechnology innovations aimed directly towards consumers. One category of potential products aimed at consumers is those products with added health benefits. to cassava with increased protein content to help fight malnutrition in developing nations. were more geared towards so-called input traits. The range of work being done on functional foods described in this report—from oils that product no trans fats or contain heart healthy omega-3 fatty acids. This report is not intended to be an exhaustive catalog. farmers.” The term functional food means different things to different people. however. These first products have been rapidly adopted by U. The first generation of agricultural biotechnology products to be commercialized. The first section describes some recent scientific advances that could lead to functional foods on grocery store shelves. cotton and corn grown in the United States. however. to foods with enhanced levels of antioxidants—is impressive. and now account for the majority of soybeans.

turns again to look at a category of new products on the horizon. The use of modern biotechnology to produce functional foods will not likely fundamentally challenge existing regulatory structures. economic or other reasons. food manufacturers and consumers. The analysis of relevant statutory authorities suggests that there is ample legal authority to cover the kinds of functional foods currently being explored in laboratories. but that different authorities may come into play for different kinds of foods and that the application of different authorities can have significant consequences for product developers. Michael Fernandez Executive Director April 2007  Application of Biotechnology for Functional Foods . provided a broad overview of what could be the “next generation” of genetically engineered agricultural products. and may never make its way into consumer products for technical. The Pew Initiative on Food and Biotechnology’s first report. Different authorities impose different safety and labeling standards. We would like to acknowledge the contributions of Joyce A. have different requirements for regulatory review and clearance or approval. It is fitting that this. Korwek. for the review of regulatory authorities that could govern future functional foods.It should also be noted that much of the work described here is still in preliminary stages. Nettleton. who created the scientific review used in the development of this paper. Harvest on the Horizon (2001). and could result in different levels of transparency to the public. but may challenge the boundaries of some regulatory classifications. the last of the Initiative’s reports. and of Edward L.

uses genetically engineered chymosin. “any modified food or food ingredient that may provide a health benefit beyond that of the traditional nutrients it contains” (Food and Nutrition Board 1994). the enzyme chymosin. More than 270 foods have FOSHU status in Japan. It is estimated that today 70% or more of cheese made in the U.” Bioactive components of functional foods may be increased or added to traditional foods through genetic engineering techniques. Foods qualify as “functional foods” because they contain non-essential substances with potential health benefits.  B.S. Background Applications of Modern Biotechnology to Functional Food Applications of Biotechnology for Functional Foods A.S. The first genetically engineered food. Applications of Biotechnology in Food Crops In 1990. in 1994. An example would be the high lycopene tomato. soy foods (isoflavones). The broad classification of functional foods carries some irony. The original concept of functional foods originated in Japan from its development of a special seal to denote Foods for Specified Health Use (FOSHU). “It is unlikely that a non-functional food exists. used in cheesemaking. Examples of the diverse foods and their bioactive substances that are considered “functional foods” are: psyllium seeds (soluble fiber). was approved for human consumption in the U.PART 1 I.S. Functional Foods A relatively recent concept in the U. This report focuses on biotechnology applications in functional and improved foods. Application of Biotechnology for Functional Foods . the FlavrSavr™ tomato. cranberry juice (proanthocyanidins). and green tea (catechins). using the National Academy of Sciences definition as a guideline. to describe the broad healthfulness of foods is the term “functional foods. purple grape juice (resveratrol).” These foods are defined as foods that provide health benefits beyond basic nutrition (International Food Information Council 2004). as John Milner.S. which has potent antioxidant capabilities.Food and Drug Administration (FDA) approved the first genetically engineered food ingredient for human consumption. The Food and Nutrition Board of the National Academy of Sciences described a functional food as. a genetically modified tomato with delayed ripening characteristics that is high in lycopene. the U. Chief of the Nutrition Science Research Group at the National Cancer Institute noted. tomatoes (lycopene).

transgenic virus-resistant papaya and squash are also cultivated. Although genetically engineered crops with enhanced health. and South Africa. the absolute growth in GM crop area between 2002 and 2003 was almost the same in developing countries (4.4 billion. Cultivation of genetically engineered crops globally has expanded more than 10% per year for the past seven years. or other agricultural technologies and in these instances in particular. Thus. biotechnology has proven to be an effective agronomic tool. ISAAA highlighted its key findings this way: In 2003. India. traditional breeding.S. and cold. such as herbicide tolerance and insect resistance. such as resistance to rootworm in maize. reduce the need for pesticides. South Africa. in spite of continuing controversy. Biotechnology traits developed and commercialized to date have largely focused on pest control (primarily Bt crops) or herbicide resistance. Brazil. soil salinity. ISAAA suggested that five new Bt and novel gene products for insect resistance in maize could be introduced. living on six continents in the North and the South: Asia. Europe and Oceania…. Canada. according to the International Service for the Acquisition of Agri-biotech Applications (ISAAA. In its 2003 report. China. the technology continues to be adopted by farmers worldwide. Herbicide resistance allows farmers to control weeds with chemicals that would otherwise damage the crop itself. Argentina. and North America. and increase the ability of the plant to survive adverse growing conditions such as drought. maize (corn).  While the improvement of agronomic characteristics in major crops has been highly successful. are expected to increase.S. and canola. a renewed emphasis on developing agricultural biotechnology applications more relevant to consumers has accompanied continuing efforts to develop crops with improved agronomic traits. cotton. In the U. functional. Many plant pests have proven either difficult or uneconomical to control with chemical treatment.6 million hectares) … the three most populous countries in Asia—China. Transgenic Acreage Expands Steadily Seven million farmers in 18 countries now grow genetically engineered crops. Such an expansion rate amounts to a 40-fold increase in the global area of transgenic crops from 1996 to 2003. Agronomic Traits Prevail Research in plant biotechnology has focused primarily on agronomic traits—characteristics that improve resistance to pests. Recently. and the combinations of traits with similar functions. Brazil and Mexico. and Indonesia.S. These applications Application of Biotechnology for Functional Foods .. and consumer benefits have lagged behind agronomic applications. The addition of new traits. are all officially growing genetically engineered crops. nutrition. James 2004). have been introduced in cotton and corn. The leading genetically engineered crops globally and in the U. D. are soy.C. the three major economies of Latin America—Argentina. such as two genes for resistance to lepidopteran pests in maize. research on many such products is in the advanced stages of development. few products genetically engineered to meet the specific needs of either food processors or consumers have yet been commercialized. GM crops were grown in 18 countries with a combined population of 3. Varieties combining two different traits. Africa and Latin America. and the largest economy in Africa. Leading countries are the U.4 million hectares) and industrial countries (4. however..

To reduce the saturated fatty acid content of foods. and flavor to foods. doughnuts. One example is canola oil with 6% to 7% total saturated  Fats and Fatty Acids – Like Oil for Water Fats are slippery substances that usually do not dissolve in water. Substantial amounts also hide in foods such as cheese. Fatty acids with just one double bond are called “monounsaturated” and the amount in a food appears on the nutrition label. increase the risk of heart disease and other chronic diseases when consumed in excess. From a nutritional perspective. So-called “good fats” are rich in unsaturated fatty acids. Quantity and Quality of Food Oils Food oils have both nutritional and functional qualities. Application of Biotechnology for Functional Foods . and create ingredients with superior properties for food manufacturing and processing. such as saturated fatty acids. peanut butter. Each fat contains three fatty acids. and chips. texture. mayonnaise. II. fats and oils contribute more energy (calories) than any other nutrient category. At the same time. Olive oil and high oleic sunflower oil contain mainly monounsaturated fatty acids. about nine calories per gram. Food applIcatIons For Human HealtH A.and polyunsaturated fatty acids. Research over the past several decades has shown that some categories of fatty acids. salad and cooking oils.could improve human and livestock nutrition and health. the kind rich in saturated fatty acids. These fats or oils are usually liquid at room temperature. This compares with about four calories per gram from carbohydrates and protein. the more unsaturated the fatty acid is. so they are not simple to replace. The more double bonds there are. We see them in foods in marbled meat. Butter. specific fatty acids that comprise most of what we call “fat” can affect a person’s risk of developing certain chronic diseases such as heart disease. The amount of polyunsaturated fat is also listed on the nutrition label. Heart healthy foods are those having a majority of mono. For example. which may be a combination of three different types. plant breeders and food manufacturers increased their use of vegetable oils rich in polyunsaturated fatty acids and developed food oils low in saturated fatty acids. and meats are rich in saturated fatty acids. Fatty acids also influence how foods behave during manufacturing and processing. thereby increasing the risk of heart disease. These warnings relate to the ability of most saturated fatty acids to raise blood cholesterol levels. People have been warned for years to limit their intake of saturated fat. Other vegetable and fish oils are abundant in polyunsaturated fatty acids with two to six double bonds. and spreads such as margarine and butter. cheese and other dairy foods. What distinguishes fats from one another is their fatty acids. the nutritional quality of food animals for human consumption. saturated fatty acids add stability. Unsaturation refers to the presence of “double bonds” in the fatty acid.

5%). These results illustrate that alterations in one type of fatty acid may affect the levels of others. chocolate. To date. Laurical™. transgenic palm oil enriched in oleic and stearic acids is under development (Parveez et al. 10 Application of Biotechnology for Functional Foods . scientists reported the development of transgenic mustard greens (Brassica juncea) containing 1% to 2% saturated fatty acids. 2000). genetically engineered palm is not anticipated for another two decades (Parveez et al. 2000). As a result.3%). The transgenic plants also contained slightly higher amounts of oleic acid. one functional food oil created with the tools of biotechnology has been commercialized. a monounsaturated fatty acid. is used in confectionary products. The following section describes research to date focused on developing crop varieties with other unique oil profiles. hydrogenated fats. Palm oil low in saturated fatty acids is currently in development. with the recent success of biotechnology techniques in palm. Where traditional plant breeding reaches its limits. suggesting that combined strategies or genetic transformations may be necessary to achieve specific fatty acid profiles. This tropical oil contains about half saturated fatty acids (49. Because of the long life cycle of palm and the time required to regenerate the plants in tissue culture. which were subsequently associated with adverse health effects. FDA approved its use in foods in 1995 (FDA 1995). Strategic Aims of Altered Fatty Acid Profile Improving the healthfulness and functionality of food oils can be accomplished in several ways.g. a level significantly less than in the control plants (Yao et al. Achievements in Altered Fatty Acid Profile Reduced saturated fatty acid content: Genetically modified soybeans have been developed that contain about 11% saturates compared with 14% in conventional soybeans (Table 1).. food manufacturers developed partially hydrogenated oils. 43. biotechnology may be used to: n n n n n n Reduce saturated fatty acid content for “heart-healthy” oils Increase saturated fatty acids for greater stability in processing and frying Increase oleic acid in food oils for food manufacturing Reduce alpha-linolenic acid for improved stability in food processing Introduce various omega-3 polyunsaturated fatty acids including long-chain forms Enhance the availability of novel fatty acids. primarily palmitic acid (16:0.fatty acids. Calgene’s high lauric acid canola. but created trans fatty acids. and higher levels of the polyunsaturates. However. Food manufacturers are developing other ways to reduce undesirable saturated fat content while maintaining stability such as using short chain saturated fatty acids and monounsaturated fatty acids. Laurical™ is a substitute for coconut and palm oils. Conventional canola oil does not contain lauric acid. B. In May 2003. gamma-linoleic acid C. The process of hydrogenation reduced the polyunsaturated fatty acid content and increased oil stability. e. linoleic and alpha-linolenic acids than the control plants. containing 38% lauric acid. and non-food items such as shampoo. 2003). To improve the stability of vegetable oils rich in polyunsaturated fatty acids. are now being eliminated from foods. the main source of dietary trans fatty acids.

One of these genes led to the accumulation of up to 22% stearate in transgenic canola seed oil. Others have developed canola oil with more than 80% oleic acid (Wong et al. their use in cooking and baking is essential. usually at the expense of polyunsaturated fatty acids. These fatty acids are absent in conventional canola oil. canola oil moderately high in oleic acid was developed using traditional plant breeding techniques. Conventional canola oil contains no lauric acid. 1991. 1996). was developed using the enzyme acyl-ACP thioesterase isolated from the California Bay Laurel (Umbellularia californica). saturated fatty acids. Biotechnology offers a means to increase the oleic acid content of vegetable oils. Scarth and McVetty 1999). Calgene’s high lauric acid canola. the predominant monounsaturated fatty acid in seed oils. Researchers at Calgene. chocolate. with lower linoleic acid (18:2) than the conventional oil. While traditional plant breeding allowed a modest increase in oleic acid. Like saturated fatty acids.100% over conventional varieties (Hawkins and Kridl 1998). myristic (14:0). high oleic acid oils are useful in food processing and manufacturing for maintaining functionality and stability during baking and frying. Davis. palmitic (16:0). CA. genetic engineering techniques have been used in canola and soy to develop oils with more short chain saturated fatty acids—12 to 18 carbons long—mainly lauric (12:0).. depending on the particular transformations used. For example. however. The stearic acid-rich oil shown in Table 1 had 28% stearic and 20% oleic acids. Unlike saturated fatty acids. they do not raise blood cholesterol concentrations and are therefore considered more healthful. avocado (65%). a saturated fatty acid that scientists believe does not raise serum cholesterol levels. Using a palmitoyl-acyl carrier protein thioesterase gene from a Mexican shrub. biotechnology has been necessary to achieve the high levels desired. To avoid the use of animal fats and hydrogenated vegetable oils with trans fatty acids. Laurical™. and non-food items such as shampoo as a substitute for coconut and palm oils. Increased oleic acid content: The most recent approach to developing more healthful food oils is increased oleic acid content. 11 Application of Biotechnology for Functional Foods . a tropical tree that stores up to 56% of its seed oil as stearate. oleic acid content increased to 75% (Corbett 2002). Dehesh and colleagues developed lines of canola with as much as 75% caprylic (8:0) and capric acids (10:0). The concomitant reduction in polyunsaturated fatty acids has the added advantage of increasing the stability of the oil and ultimately the processed food. and only about 6% short chain saturated fatty acids. High oleic acid oils are lower in saturated and polyunsaturated fatty acids compared with conventional oil. FDA approved its use in 1995.Increased saturated fatty acid content: Because saturated fatty acids confer certain functional properties to food fats and oils and are more stable to heat and processing than unsaturated fatty acids. these water-soluble fatty acids are mainly oxidized for energy. Gene transfer technology also boosted the stearic acid content of canola (Hawkins and Kridl 1998). but not in others. those with eight and ten carbons. Soybeans have been genetically modified to produce oil enriched in stearic acid (18:0). and stearic (18:0) acids. cloned three thioesterase genes from mangosteen. This was the first transgenic oilseed crop produced commercially. Inc. is abundant in olive (72%). containing 37% lauric acid. When consumed. Oleic acid. For example. With the application of biotechnology. High laurate canola is used in confectionary products. Enrichment of canola with even shorter chain saturated fatty acids. Cupea hookeriana. an increase of more than 1. has also been accomplished (Dehesh et al. and canola (56%) oils. and sometimes.

Humans can convert alpha-linolenic acid to the more biologically active longchain forms. Like Buhr and colleagues. oleic acid content increased to 85%. Gene silencing has also been used to produce high oleic and high stearic acid cottonseed oils (Liu et al.More recently. the level of oleic acid increased from 18% in the wild-type seed to 57% in the transgenic seed. Such oils would be desirable for the commercial uses mentioned. Cottonseed oil is high in palmitic acid. and frying and increases their susceptibility to oxidation or rancidity. higher plants lack the enzymes to make 20. because they are associated with many health benefits. rainbow trout. Cotton was also genetically modified to produce high stearic oil having 40% stearic and 39% linoleic acids. especially fatty species such as salmon. Oils with appreciable amounts of alpha-linolenic acid such as canola and soybean. these scientists were able to turn off the production of the enzyme that converts oleic acid to polyunsaturated fatty acids. Omega-3 fatty acids: There is extensive interest in increasing Americans’ consumption of omega-3 fatty acids. with 15% palmitic acid. Examples of genetically modified high oleic acid oils compared with their conventional counterparts are shown in Table 1. plant foods with alpha-linolenic acid may be insufficient to supply the need for long-chain omega-3 fatty acids. processing. alpha-linolenic acid. with saturated fatty acids reduced to 6%. Marketed under the brand TREUS™ the company claims that the oil eliminates the need for hydrogenation in food processing. soybean. Cosuppression occurs when the presence of a gene silences or turns off the expression of a related gene. a DuPont company. Reduced alpha-linolenic acid: Several genetic transformations designed to increase oleic or stearic acid content do so at the expense of the polyunsaturated fatty acids alpha-linolenic and linoleic acids. However. Vistive™. A combination was also developed to have 40% stearic. scientists at DuPont used the technique of cosuppression to reduce the production of polyunsaturated fatty acids in soybeans. Buhr and colleagues at the University of Nebraska used genetic engineering to increase oleic acid levels in soybeans by inhibiting the ability of the plant to convert oleic acid to polyunsaturated fatty acid (Buhr et al. 2001). respectively. These fatty acids have desirable nutritional characteristics. with palmitic acid reduced from 26% to 15%. Some plants—mainly canola. When two gene transformations were applied. especially during pregnancy and lactation (Pawlosky et al. respectively. but their presence reduces the stability of oils for baking. The result was greatly increased production of oleic acid and reduced production of polyunsaturated fatty acids. very high in linoleic acid. A similar product from Monsanto. When the conversion enzyme was inhibited. Using a different approach. herring. and sardines. When gene silencing was used to transform cotton. 2002). mackerel. Thus. 2002). In 2002. the National Academy of Sciences’ Institute of Medicine recognized that omega-3 fatty acids are essential in the diet and established an estimated adequate intake for them (Institute of Medicine 2002). Conventional cottonseed oil has about 13% oleic acid. The main food sources of the long-chain omega-3 fatty acids are fish. with about 10% and 8% alpha-linolenic acid. 12 Application of Biotechnology for Functional Foods . and free of alpha-linolenic acid. offers a similar level of reduction in alpha-linolenic acid. but are consumed only in small amounts. These examples illustrate the power and specificity of this technology to develop tailored seed oils. 37% oleic and only 6% linoleic and 14% palmitic acid. but they do so very inefficiently. have been genetically modified to reduce this fatty acid. developed low alphalinolenic acid soybean seeds through conventional breeding techniques with less than 3% alpha-linolenic acid in its oil. Pioneer Hi-Bred. the resulting oil had 78% oleic and only 4% linoleic acids.and 22-carbon polyunsaturated fatty acids needed by mammals. and flax oils—provide the 18-carbon omega-3 fatty acid.

especially plants. and cottonseed oils. increasing worldwide demand has put severe pressure on wild aquatic resources and limited seafood availability. corn. Ironically. It is now recognized that diets high in omega-6 fatty acids and low in omega-3 fatty acids may exacerbate several chronic diseases (Simopoulos et al. Thus. although product developers are working to combine high omega-3 and low alpha-linolenic traits in one product. reducing the level of alpha-linolenic acid in soy and canola oils used in food processing. 2000). Such foods would also be useful for animal and fish feed.Western diets contain predominately omega-6 polyunsaturated fatty acids found in soybean. it would be desirable to increase the availability of these fatty acids or their precursors in a variety of other foods. canola. Because of the many health benefits associated with the regular consumption of omega-3 fatty acids. several health organizations. One limitation to boosting consumption is that they occur naturally mainly in fatty fish and some seeds. oIl canola Conventional High oleic High oleic Low linolenic Low linolenic P6 High myristate/palmitate High laurate (37%) sunFlower Conventional High oleic Mid oleic soyBean Conventional soy Low linolenic High palmitic (17%) High stearic (28%) High oleic soybean otHers Conventional safflower High oleic safflower Conventional corn High oleic corn Olive Avocado oleic (18:1) 60 75 84 65 78 34 34 20 82 56 23 23 17 20 83 14 75 24 70 75 65 linoleic (18:2) 20 14 5 22 11–13 15 12 65 10 33 51 60 55 35 2 75 14 58 8 15 alpha-linolenic (18:3) 10 3 3 4 2–3 4 7 <1 <1 <1 7 2 8 7 3 0 0 <1 <1 1 total saturates 7 <7 5 7 N/A 43 45 10 8 9 14 15 20 35 12 6 6 13 14 14 Application of Biotechnology for Functional Foods . including the American Heart Association and the 2005 Dietary Guidelines for Americans. Although aquaculture has increased the availability of some fish and shellfish species. have called for increased consumption of these substances. 1 taBle 1. may actually reduce consumption of this fatty acid. selected fatty acid content of vegetable oils with modified fatty acid profiles compared with the commodity oil. sunflower.

a landmark paper announced the production of long-chain polyunsaturated fatty acids—both omega-6 and omega-3 types—in Arabidopsis thaliana. Then the body’s enzymes convert stearidonic acid to 20-carbon polyunsaturated fatty acids. 14 Application of Biotechnology for Functional Foods . Although the study used supplements.One strategy to increase the availability of long-chain omega-3 fatty acids is to develop oilseed crops such as canola and soybean that contain stearidonic acid (18:4n-3). the findings suggest that plants with stearidonic acid would have potential to provide EPA. Stearidonic acid is the first product formed when alpha-linolenic acid is converted to eicosapentaenoic acid (EPA). This work marks an important advance in the development of plant-based sources of long-chain omega-3 fatty acids that could be consumed directly or incorporated into food products. 2004). possibly from vitamin E. because stearidonic acid contains four double bonds. but increasing the level of stearidonic acid helps overcome this limitation. a type of cress widely used as a model plant in biotechnology research. United Kingdom. None of the fatty acids consumed had any effect on cell DHA levels. In contrast. a desirable long-chain omega-3 fatty acid. This is the first demonstration of the incorporation into edible plants of a biologically potent source of long-chain omega-3 fatty acids. respectively (Qi et al. a level similar to conventional canola. personal communication 2004). The engineered plants accumulated up to 23% stearidonic acid in the seed oil with a reduction in oleic acid content from about 60% to about 22%. By breeding the transgenic lines with various lines of canola the investigators were able to develop a line of canola containing more than 55% of alpha-linolenic acid and stearidonic acid. 2003). The implications of Calgene’s work with stearidonic acid are substantial. One can imagine that transgenic canola and soybean could be developed using additional traits to boost antioxidant protection. Usually. and accounted for 43% of the total 20-carbon polyunsaturated fatty acids. Total omega-6 fatty acids remained about 22%. another long-chain omega-3 fatty acid associated with health benefits. scientists at Calgene. this first step limits the amount of EPA produced. Dr. The additional genes were necessary to provide the enzymes to make these long-chain fatty acids. have successfully transformed canola so that it makes stearidonic acid. In addition to the production of EPA and arachidonic acid. This omega-3 fatty acid occurs naturally in only a few plants such as black currant seed oil and echium. This achievement was also remarkable because it used a pathway seldom found in plants. This genetic engineering feat required two genes from the fungus Mortierella alpina and one from canola for the three enzymes needed to produce sufficient stearidonic acid (Ursin 2003). However. when the study volunteers consumed alpha-linolenic acid. This finding meant that the stearidonic acid was converted to EPA and appeared in red cells just as readily as the preformed EPA. Virginia Ursin and colleagues at Calgene studied the metabolism of stearidonic acid in people (James et al. not stearidonic acid from transgenic plants. Toward this end. Baoxiu Qi and colleagues at the University of Bristol. Her studies showed that when either stearidonic acid or EPA was consumed the amount of EPA in red blood cells increased significantly. Yields of EPA (13%) and arachidonic acid (29%) in leaves were significantly higher than in conventional cress. the concentration of alpha-linolenic acid was reduced from 48% to 14%. it is vulnerable to oxidation and would require antioxidant protection. there was no change in their red cell EPA content. Dr. transferred to Arabidopsis thaliana three genes encoding for different enzymes in the metabolic pathway from linoleic and alphalinolenic acids to arachidonic and eicosapentaenoic acids. Calgene scientists have also developed soybean that contains stearidonic acid (Ursin. In May 2004. which usually does not produce these fatty acids.

2004). with significant reduction in the amounts of the 18-carbon precursors linoleic and alphalinolenic acid compared with conventional plants. Once the genes for these enzymes were identified and cloned they could be incorporated into model organisms to see whether DHA would be produced. Several research groups have examined many algae and identified the specific enzymes for this conversion (Sayanova and Napier 2004. Gamma-linolenic acid is naturally present in appreciable amounts in few plants. In late 2004. The team then went on to develop transgenic flax.S. 2003). improved liver function in patients with liver cancer. Germany. the question of making Docahexenoic Acid (DHA). Amine Abbadi at the University of Hamburg. A recent study reported that gamma-linolenic acid content in transgenic canola ranged from 22% to 45% (Wainright et al. Gamma-linolenic acid: This fatty acid is the first step in the conversion of linoleic acid to arachidonic acid in the omega-6 fatty acid pathway. but the enzymes to do so are not present in mammals. Davis. In mammals. but also the pathways for achieving the desired ends. One is that it demonstrates the feasibility of developing plants capable of synthesizing long-chain polyunsaturated fatty acids. black currant oil (Ribes nigrum) and echium (Echium plantagineum). For that and other reasons.This work is important in several regards. The ability to increase the production of gamma-linolenic acid in tobacco plants by transferring the gene for the delta-6 desaturase enzyme from various sources was first shown in 1996 by Reddy and Thomas at Texas A&M University. Several steps remain before long-chain polyunsaturated fatty acids will be available in commercial crops. a 22-carbon polyunsaturated omega-3 fatty acid important in retina and brain function and other body systems remained unsolved. Another is the relatively high efficiency of conversion of the precursor fatty acids to the long-chain forms. 2003). an anti-estrogenic medication used to prevent the recurrence of breast cancer.K. Meyer et al. and with anti-cancer effects in cell culture studies. Sayanova et al.. It is possible. notably borage (Borago officinalis). and the U. it is poorly converted to arachidonic acid. reported the successful transformation of yeast that yielded small amounts of DHA (Abbadi et al. it may have potential benefit in cardiovascular disease (Fan and Chapkin 1998). Gamma-linolenic acid has been associated with improved skin conditions in human subjects. It was also shown to enhance the effectiveness of tamoxifen. This accomplishment required four gene transformations. Application of Biotechnology for Functional Foods 1 . 1997). has also reported transgenic safflower plants with 65% gamma-linolenic acid in the oil. in theory. When consumed in evening primrose or borage oils. Arcadia Biosciences. the demonstration that plants can be modified to make these important nutrients means that many of the scientific hurdles have been conquered. A likely next step will be to apply this technology to seed oil crops such as canola and soybean to see if the long-chain polyunsaturated fatty acids will accumulate in the seed. Yet another is the demonstration that plants can be engineered not only with respect to the outcome of final products. the conversion of EPA to DHA is inefficient and requires several steps. It is believed to suppress the production of estrogen receptors in cells. Although production of EPA in plants represents an enormous scientific achievement. working with colleagues in the U. to perform this conversion in a direct manner. A third advantage is the improved balance of omega-6 and omega-3 fatty acids. However. a plant with abundant alpha-linolenic acid for conversion to long-chain fatty acids. CA. Evening primrose has also been genetically modified for enhanced gamma-linolenic acid content (Wainright et al. 2004). This work gives a large boost to the potential for plants to be an important dietary source of these fatty acids. evening primrose (Oenothera biennis). and by others in 1997 (Reddy and Thomas 1996.

This example illustrates the variety of fatty acids that can be developed in seed oils using a combination of genetic engineering and traditional plant breeding techniques. at the expense of the non-essential amino acids. The gene is for a storage protein rich in nutritionally essential amino acids. staple foods in several parts of South America and Africa. an essential amino acid. III. The amino acids most often present in inadequate amounts are lysine. When the gene was expressed in cassava. The experiment demonstrated the feasibility of increasing the quantity and quality of protein in cassava. One is to increase the total amount of protein produced by selecting germplasm with an altered balance of seed proteins. William Folk and his team at the University of Missouri. Columbia. asparagine and glutamic acid. tryptophan. both of which are consumed in human diets. and methionine. Wu et al. Their strategy was to substitute more desirable and scarce amino acids for more abundant ones in certain seed proteins (Chen et al. Another approach is to introduce genes from other sources for proteins that have a favorable balance of essential amino acids. Foods such as potato and cassava. An example is the introduction into potato of a gene for seed albumin protein from amaranth. In 2003. 2003). They applied this concept to rice by increasing the production of lysine. A third approach seeks to increase the production of specific amino acids such as lysine. pioneered another approach to improve seed protein quality. This approach was used in the development of Quality Protein Maize. the precursor of vitamin A. This may be done by traditional cross breeding or genetic engineering. QuantIty and QualIty oF plant proteIn Efforts to improve the protein content and quality of staple foods have been underway for decades. Cassava: A staple food for some 500 million people in tropical and sub-tropical parts of the world. Application of Biotechnology for Functional Foods 1 . For example. cassava (Manihot esculenta Crantz). Improvements in protein quality benefit both human and animal nutrition and increase the feed efficiency of crops fed to food animals. There are various ways of improving protein quantity and quality. 1998. where nutrient shortfalls are widespread and dietary diversity limited. However. MO. Zhang and colleagues reported using a synthetic gene to increase the protein content in cassava (Zhang et al. glutamine. also known as yucca or manioc. The main focus is crops grown in developing countries. Corn with higher levels of these amino acids would significantly improve feed efficiency and lower input costs to farmers. Improved corn varieties consumed by humans would also have nutritional benefits.Ursin’s study of transgenic canola enriched in stearidonic acid discussed above also reported that a cross between the transgenic line of canola producing stearidonic acid and a canola line high in gamma-linolenic acid yielded a canola containing about 11% gammalinolenic acid and about 14% stearidonic acid (Ursin 2003). and parts of Asia and South America. discussed below. 2003). the leaves are also consumed and these are a good source of beta-carotene. It is widely consumed in Africa. have less than one percent protein. Cassava root has less than 1% protein and poor nutritional value. corn is widely fed to cattle but it is limiting in lysine and methionine. transformed plants expressed the gene in roots and leaves. Efforts to improve protein quality strive to increase the amount of limiting essential amino acids provided by the protein in the food. thrives in marginal lands having little rain and nutrient-poor soils.

Researchers at the Max Planck Institute. In 2004. researchers at Monsanto. Africa. currently in the preliminary stage. and South America. 2004). Efforts to develop cassava varieties low in these toxicants is a high research priority. CIMMYT researchers succeeded in developing hardy corn varieties that contained twice the lysine and tryptophan content as traditional varieties. work with the opaque-2 gene continued using both traditional breeding and molecular methods. Louis. potatoes contain about 2% protein and 0. Increased production of other proteins in the corn led to higher levels of lysine. Africa. it was discovered that corn bearing a gene known as opaque-2 contained increased concentrations of lysine and tryptophan and had significantly improved nutritional quality (Food and Agriculture Organization 1992). Potato: Potato (Solanum tuberosum) is a dietary staple throughout parts of Asia. sun-drying.1% fat. researchers at the National Application of Biotechnology for Functional Foods 1 . another limiting amino acid. had nearly twice the content of protein and oil as conventional corn (Young et al. one of which usually dies. Typically. These kernels contained more protein and oil than conventional corn. In 2004. opaque-2 corn proved to have low yields. 2004). Riverside. Recent researchers at CIMMYT reported the development of transgenic corn with multiple copies of the gene from amaranth (Amaranthus hypochondriacus) that encodes for the seed storage protein amarantin (Rascon-Cruz et al. At the International Maize and Wheat Improvement Center (CIMMYT) in Mexico. Total protein in the transgenic corn was increased by 32% and some essential amino acids were elevated 8% to 44%. used genetic engineering techniques to reduce the amount of zein storage proteins. corn ears develop flowers in pairs. MO. and Asia. Pursuing a different strategy to improve protein quality. This work. cytokinin. have focused on methionine. Normally.Cassava also contains cyanogenic glucosides that can produce chronic toxicity if not eliminated or reduced by grating. increased susceptibility to diseases and pests. and methionine (Huang et al. They elucidated several key steps in methionine metabolism in plants. Although some varieties may contain appreciable quantities of protein. a team of researchers at the University of California. reported that transgenic corn with increased production of the plant regulating hormone. However. Germany. Quality Protein Maize varieties have been adapted to and released in over 40 countries in Latin America. These storage proteins constitute over half the protein in corn and are deficient in lysine and tryptophan. St. 2004). In 1964. It was reported in 2000 that. as in cassava. and inferior functional characteristics. After at least 12 years’ work. Scientists Surinder K. Under the influence of the additional cytokinin. Vasal and Evangelina Villegas of CIMMYT were awarded the World Food Prize in 2000 for their work developing ‘Quality Protein Maize’. its quality is poor because of low lysine and tryptophan content. both flowers developed but yielded only a single kernel. could pave the way for using genetic engineering techniques to improve the methionine content of plants (Hesse and Hoefgen 2003). but were disease-resistant and high-yielding. 2000). tryptophan. Corn: Corn (Zea mays) is the predominant staple food in much of Latin America and Africa. The agronomic and nutritional properties of these lines are currently being evaluated. This development resulted from an unusual change in the way the plant developed. or fermenting. transfer of the gene for seed albumin protein from Amaranthus hypochondriacus to potato resulted in a “striking” increase in protein content of the transgenic potatoes (Chakraborty et al.

Momma and colleagues at Kyoto University. In July 2004. is a primary center for rice research and development of improved varieties. Commodity rice contains about 7% protein. The most limiting amino acid in rice is lysine. Although details are sparse. which enhance the quality of the additional protein (Council for Biotechnology Information 2004). 1998).).0% protein and an improved essential amino acid profile compared with 6. Dr. The lead scientist in the golden rice project. 2003).5% protein in the control rice. it has been possible to genetically engineer potatoes that contain fat (triglycerides). at least once a day (IRRI undated). In February 2004. Jesse Jaynes. Dubbed the “protato. It is the primary source of energy and nutrition for millions. In 1999. They did so by modifying the process of protein synthesis. now retired from the Swiss Federal Institute of Technology. Although lysine content remained below optimum levels. The gene. Transgenic plants containing a soybean gene for the protein glycinin contained 8. Dr. 1999). Klaus and colleagues at the Max Planck Institute of Molecular Plant Physiology demonstrated increased fatty acid synthesis in potatoes (Klaus et al. the scientists suggested that additional transformations and modifications could further boost lysine levels. notably black rice. named Asp-1. Thus. Perhaps the most famous genetic transformations in rice are those in “Golden Rice” involving the vitamin A precursor. particularly its children. Precedent for the expression of a synthetic gene in rice grown in cell culture suggests that Jaynes’ approach is viable (Huang et al. rather than by gene transfer or the expression of new proteins.” the protein-rich potato had significant increases in lysine and methionine. reported a genetically engineered rice having about 20% greater protein content compared with control rice (Momma et al. Rice: Almost half the world’s population eats rice (Oryza sativa L. this potato was reported “approaching release” to farming communities. Ingo Potrykus. They achieved an overall 6% increase in lysine content in the grain (Chen et al. India. 2002). was also involved in applying biotechnology for the improvement of rice protein. William Folk and his team genetically modified rice to increase its content of the amino acid lysine (Wu et al. Dr. especially in Asia and parts of Africa and South America. but some varieties. Japan. beta-carotene. Rice is the staple food among the world’s poor. and iron. who synthesized a synthetic gene coding for an ideal high-quality storage protein with a balanced mixture of amino acids. The International Rice Research Institute (IRRI).5% (Food and Agriculture Organization 2004). contain as much as 8. Efforts to increase the nutritional value of rice target protein content and quality along with key nutrients often deficient in rice-eating populations. Philippines. It should be noted that while potatoes are known for their high starch content.Centre for Plant Genome Research. The transgenic rice plants accumulated the Asp-1 protein in their endosperm in a range of concentrations and provided essential amino acids but data are not yet available on the concentrations achieved or their nutritional relevance. 2004). such as vitamin A and iron. Potrykus described the work of Dr. improving the nutritional quality of rice could potentially improve the nutritional status of nearly half the world’s population. reported the development of a nutritionally improved potato line with 25% higher yields of tubers and 35%–45% greater protein content (ISAAA 2004). 1 Application of Biotechnology for Functional Foods . was transferred to rice with the appropriate genetic instructions for its production in the endosperm or starchy part of the rice grain. As mentioned briefly above.

branching. 2002). clarity. determines what ratio of amylose to amylopectin is most suitable. Many lines of corn have been developed with different characteristics derived from modified starch ratios and increased amylose content. Members of the potato family—potato. Starch Starch from cereals. sweet potato. an enzyme involved in the plant’s energy metabolism and starch production. grains. However. and bulking agent. Besides providing energy. cassava—have 17% to 20% amylose. a fructan found in Jerusalem artichokes. and degree of polymerization. Considerable publicity was given to potatoes engineered by Monsanto in the early 1990s to have increased starch content. and tubers contribute a substantial share of dietary calories and in many poor countries. These characteristics depend on the proportion of amylose and amylopectin. corn (28%). gel stability. modIFIed carBoHydrate A. The resulting transgenic potatoes had substantially increased adenylates and a 60% increase in starch compared with wild-type plants (Regierer et al. 2004). gelatinization. Inulin comprised 5% of the dry weight of the transgenic tubers and did not influence sucrose concentration. The full spectrum of inulin molecules present in artichokes was expressed in the transgenic potatoes. They are an example of the type of starch modification that may have secondary health benefits as a consequence of how they are used. Fructan Fructans are polymers (repeating units) of the sugar fructose. smaller granules. etc. Potatoes were genetically altered to increase the activity of adenylate kinase. High amylose starches include high amylose corn (70%). and increased reducing sugars (Hofvander et al. Unexpectedly. cosmetics.IV. They may also stimulate the growth of desirable colonic bacteria. Application of Biotechnology for Functional Foods . the main components of starch. provide the majority of food energy. wheat (26%) and sago (26%). solubility. Transgenic high amylose potatoes developed by inhibiting two branching enzymes were reported to yield more tubers and have lower starch content. How a particular starch will be used in foods. Biotechnology has also been directed to increasing starch content (Geigenberger et al. source of dietary fiber. They serve in food products as a low-calorie sweetener. These were touted as more desirable for French fries because they would absorb less fat during frying. Starch is also important for feed and industrial purposes. Amylose and amylopectin differ from each other in chain length. 2001. and tuber yield increased. 2000). Amylose is linear and amylopectin is highly branched. Its use as paste goes back at least 4000 years BCE to the Egyptians who cemented strips of papyrus stems together with starch paste for writing paper. starch content was reduced. 2002). waxy rice and waxy sorghum contain no amylose. starch confers functional characteristics to foods: texture. Inulin. viscosity. Regierer et al. such as bifida. and detergents. In contrast. was successfully synthesized in potatoes following the transfer of two genes from globe artichokes (Cynara scolymus) (Hellwege et al.to 4-fold. 1 B. Fructans have environmentally friendly non-food applications in the manufacture of biodegradable plastics. but agronomic shortcomings in growing these crops have limited their use. Fructans are naturally occurring in Jerusalem artichokes (sunchokes) and chicory. the concentrations of several amino acids were increased 2.

particularly rice. Research to date includes the isolation of several genes for fructosyl transferase enzymes involved in the production of fructans. It is the precursor of vitamin A and can be converted to the active vitamin during digestion. contain no beta-carotene or its precursor carotenoids. Many staple foods. the Potrykus group reported the application of three transgenes in the development of rice expressing the entire pathway for the production of beta-carotene (Ye et al. is the rate-limiting step in generating carotenoids (Cunningham 2002). severe infections. However. the European Commission funded multidisciplinary research programs in agriculture and fisheries. in 118 countries. it was reported in 2004 that genes encoding for fructosyl transferases in onion were isolated and transferred to sugar beet. and linoleic and alpha-linolenic acids were reduced compared with non-transgenic seeds. Following the transfer of the genes. oleic acid content increased. 1997). the first product in the pathway for carotenoid synthesis. V. could express phytoene. was a landmark achievement in the application of biotechnology to nutrition and public health. It is clear that production of phytoene. a key intermediate in the synthesis of beta-carotene (Burkhardt et al. and fieldtesting the new lines for nutritional value and agronomic performance. The development of “Golden Rice.” so named because of its yellow color conferred by the presence of beta-carotene.In a program called the Agriculture and Fisheries Programme. 20 Application of Biotechnology for Functional Foods . was the first to show that transgenic rice. Humans cannot synthesize carotenoids and therefore depend on foods to supply them. Diets lacking other food sources of vitamin A or beta-carotene are associated with vitamin A deficiency which can result in blindness. The development of transgenic plants able to produce a variety of carotenoids is an active area of research. 2002). Overcoming this nutrient deficiency is an urgent global health challenge. increases the synthesis of carotenoids substantially. IRRI is currently cross-breeding the nutrient-enhanced transgenic rice with local rice varieties from Asia and Africa. Using gene transfer technology to increase the expression of phytoene synthase. in canola (Brassica napa). The deficiency affects some 134 million people. vitamin E levels decreased significantly. Other carotenoids do not have potential vitamin A activity. Beyer et al. For example. 2000. including a project on fructans for food and non-food uses. mainly alpha and beta-carotene. or FAIR. Increased VItamIn content In plants A. According to the World Health Organization. The feasibility of using these enzymes has been demonstrated in model plants and target crops such as sugar beet (Anonymous 2000). Varieties of Golden Rice are not expected to be ready for farmers for several more years. Additional work with Golden Rice included the insertion of a gene to increase the iron content (Potrykus 2003). Subsequently. In addition. Shewmaker and colleagues (1999) from Monsanto reported an increase up to 50-fold in carotenoids. 2004). Peter Burkhardt. the enzyme that makes phytoene. particularly children. These other changes would have to be modified or evaluated to determine whether they might have meaningful nutrition implications. vitamin A deficiency is the leading cause of preventable blindness worldwide. Beta-carotene and Other Carotenoids Beta-carotene belongs to the family of carotenoids and is abundant in plants of orange color. onion-type fructans were produced from sucrose without loss in storage carbohydrate. working with Ingo Potrykus and colleagues in Switzerland. a plant that does not normally synthesize fructans (Weyens et al. and sometimes death. carrying a gene from daffodil.

whereas only trace amounts were found in the control plants.to 130-fold. Bramley’s group transferred the gene that increases carotenoid synthesis from a bacterium to tomatoes and measured total and specific carotenoids in the transgenic fruits (Fraser et al. 2002). transformed tomatoes using a bacterial gene encoding for an enzyme that converts phytoene to lycopene. Kristal 2004). An additional consequence of this transgenic modification was a severalfold increase in lycopene content. Dr.7-fold. 2003). They also reported a 43-fold average increase in beta-carotene (Lindgren et al. 2002). but zeaxanthin was only increased by a factor of 1. United Kingdom.6-fold increase in total pigment and a 13-fold increase in these three carotenoids (Stalberg et al. 21 Application of Biotechnology for Functional Foods . though definitive data are lacking (Etminan 2004. 2003). 2000). but increased the ratio of beta to alpha-carotene from 2:1 to 3:1 (Ravanello et al. Lutein and zeaxanthin are nutritionally important carotenoids for protection of the retina and reduced risk of age-related macular degeneration (Krinsky et al. were able to use biotechnology to block the conversion of zeaxanthin to another carotenoid and thereby increase its content in potatoes (Romer et al.In a separate study on canola. Lutein is found in dark green leafy vegetables such as spinach and collards. 2002). an enzyme involved in the ripening process (Mehta et al. Romer and colleagues at Universitat Konstanz. the Monsanto group transferred to canola three genes from bacteria that affect the phytoene synthesis pathway. 2003. corn. Alphatocopherol (vitamin E) was increased 2. the precursor of beta-carotene. Fine-tuning these alterations has the potential to significantly enhance the nutritional value of potatoes. but in some. Total carotenoids were increased by 5. Stalberg and colleagues in Sweden also studied the effects of phytoene synthase on carotenoid synthesis in transgenic Arabidopsis thaliana. Tomatoes with delayed ripening were produced as a result of inserting a gene encoding for S-adenosylmethionine decarboxylase. but tomatoes with increased lycopene content may have enhanced nutritional value. 2003). They also observed substantial levels of lycopene and alpha-carotene in the seeds. Lutein. Others have examined the effect of transgenes affecting phytoene metabolism on carotenoid synthesis. In another study.1. but total carotenoids were not increased (Romer et al. Peter Bramley’s group at the University of London. phytoene desaturase and lycopene cyclase—the resulting transgenic canola seeds maintained the same amount of total carotenoids.4-fold. and zeaxanthin occurs in yellow foods such as mangoes. another nutritionally important carotenoid. Lycopene is normally converted to beta-carotene.8. However. lutein content was decreased. transformed seeds had a 4. 2003). The altered carotene content did not affect plant growth and development. Tomatoes carrying the bacterial gene had about a 3-fold increase in beta-carotene content. Gale et al. When they included a triple construct— genes for three different enzymes. Lycopene consumption has been linked to reduced risk and spread of prostate cancer.to 3-fold in the transgenic potatoes. germination was delayed in proportion to the increased levels of carotenoids. and lutein of ranging from 1. The latter is not particularly abundant in Western diets. phytoene synthase. with increases in phytoene.to 2. Germany. and peaches. They examined three ketocarotenoids. lycopene. Total carotenoids were 2. beta-carotene. was significantly increased. With this approach they obtained increased levels of zeaxanthin in potatoes ranging from 4.to 4-fold higher in transgenic fruits than in nontransformed plants.

22 In soybeans. Louis. because vitamin E is an anti-oxidant. and delta forms are more widespread in plants. 2000). Efforts to increase the content of vitamin E in food plants. Lonn et al. Dr.The carotenoid astaxanthin is synthesized by algae and plants and is responsible for the pink color in shrimp and salmon. The most potent form of the vitamin is alpha-tocopherol. Transgenic tobacco plants produced astaxanthin and changed color (Mann et al. however. In September 2003. Israel. Vitamin E There is strong interest in vitamin E because of studies linking it to decreased occurrence of several degenerative diseases and cancers. Also. Application of Biotechnology for Functional Foods . Edgar Cahoon of the U. beta. this alteration may increase its oxidative stability after harvest. Besides enhancing the potential therapeutic and nutritional value of corn. researchers at The Hebrew University of Jerusalem. announced the development of transgenic corn with increased levels of vitamin E. Insertion of a gene from barley into corn increased the conversion of vitamin E precursors to vitamin E itself (Cahoon et al. 1999). 2003). have sought to increase the amount of precursor substances by overexpressing the genes for various enzymes involved in the biosynthetic pathway. 2003). particularly cereals and grains. Some recent trials with vitamin E supplementation reported no protection against cardiovascular disease or cancer and some chance of increased risk of heart failure (Eidelman et al. Tocotrienols. The result was a 5-fold increase in vitamin E activity (Van Eenennaam et al. 2003). Department of Agriculture (USDA) and co-researchers at the Donald Danforth Plant Science Center. Vitamin E biosynthesis involves complicated pathways so that multiple genetic manipulations are required. Vitamin E is found mainly in vegetable oils. MO. St. 2005. was increased up to 6-fold. transferred the gene into tobacco (Nicotiana tabacum). are poorly absorbed in humans. they may have cholesterol lowering properties (Theriault et al. much of the antioxidant produced was tocotrienols rather than vitamin E (Aijawi and Shintani 2004). a closely related substance. It is less certain whether it is an antioxidant in human health. This ability to manipulate pigmentation in fruits and flowers may have commercial potential and possible implications for increasing the availability of carotenoids for human health. wheat germ. it is useful in foods and oils to provide oxidative stability. which may have low amounts. B. This work paves the way for the development of vitamin E-rich oils and plants with potential health benefits. Astaxanthin is used commercially in feed for cultured salmon and trout. Astaxanthin is of interest because of its strong antioxidant properties in vitro. Van Eenennaam and colleagues developed transgenic plants that were able to increase the conversion of the weaker forms of tocopherol typically present in soybeans to the more potent alpha-tocopherol (vitamin E) form. but absorption is increased in the presence of fat (Mercke Odeberg et al. although potent antioxidants in vitro. Using a gene from the alga Haematococcus pluvialis. although efficacy remains unproven and data are inconsistent (71. Miller et al. 72). 2005). and a few other foods not widely consumed. Production of astaxanthin in flowers and fruits has also been accomplished with the techniques of biotechnology. but the less potent gamma. Humans absorb astaxanthin poorly. However.S. The content of vitamin E and tocotrienol. 2004.

They accomplished this increase by transferring a gene from wheat for an enzyme that recycles vitamin C and prevents its breakdown. several foods are fortified with folic acid. Folates are available in small amounts in a variety of fruits and vegetables. 2 D. Other investigators have developed transgenic tomatoes. Spain (Agius et al.to 4-fold increase in vitamin C content in the kernel. Folic Acid Inadequate intake of the vitamin folic acid. Antioxidants Vitamins C and E function as antioxidants in the body. that had twice the amount of folate as control fruit (Diaz de la Garza et al. In the U.to 4-fold increase in folates and pterins. birth defects. This group was able to boost folate content 10-fold by including a second gene transformation to increase the content of another substance. also enriched in the same gene. it cannot be synthesized by humans nor stored to any appreciable extent. many other substances widely distributed in foods in small amounts also provide protection against potentially damaging breakdown products arising from oxidation during normal metabolism. In 2004. These studies provide good evidence of the potential to increase the availability of this vitamin in widely consumed foods. 2004). Moreover. For this reason. Thus. is abundant in citrus and other fruits such as strawberry and kiwi. This study demonstrated the feasibility of using this enzyme to raise the level of vitamin C in plants. or ascorbic acid. However. MA. 2003). but is very low in cereals and grains. and free Application of Biotechnology for Functional Foods . one of a family of folates. In 2003. para-aminobenzoate. strategies to increase the vitamin C in cereals and grains hold considerable potential to improve health. impaired cognitive development. Oxidation breakdown products. needed for folate synthesis. In areas of the world where foods containing vitamin C are not widely consumed. such as reactive oxygen species. Vitamin C Vitamin C. announced the successful transformation of corn and tobacco that resulted in a 20% increase in vitamin C (Chen et al. Gallie and colleagues at the University of California. is associated with megaloblastic anemia. The ability to increase the vitamin C content of strawberries.C. 2003). Application of this approach to other food plants remains to be developed and evaluated. a good source of the vitamin. the precursors of folates. in transgenic Arabidopsis thaliana modified by the incorporation of the transgene for the first step in the synthesis of folic acid (Hossain et al. Using a gene for the enzyme D-galacturonate reductase transferred to strawberry (Fragaria spp) this group showed that vitamin C content in the modified strawberry fruit increased with the expression of the transgene. it would be desirable to increase the concentration of folates in dietary staples and foods widely consumed. oxidized lipids. Medford. Hossain and colleagues at Tufts University. was reported in 2003 by a research team at the University of Málaga. but intakes tend to be low. 2004). S. Increased expression of this enzyme in transgenic corn and transgenic tobacco resulted in a 2. E. we depend on regular dietary consumption to meet our vitamin C needs. and some chronic diseases. Riverside. and the Donald Danforth Plant Science Center reported a 2.

Potatoes are a source of antioxidant flavanoids and vitamin C. Phenolic compounds are the most widespread antioxidants in foods. potato. Mineral-efficient plants are also more drought resistant and require fewer chemical inputs (Bouis 2002). 2005). Transgenic plants exhibited significantly increased levels of phenolic acids and anthocyanins. wine. Such plants are able to unbind minerals in the soil and make them available to the plant. they can act as prooxidants and may be harmful. They have become familiar to consumers because they are touted in foods as diverse as berries. Several examples of the applications of biotechnology for enhanced antioxidant capacity in foods are described below. Resveratrol is usually associated with grapes and wines where it is abundant. accumulates in some crops and is found in apples. Further. they will not only improve their nutrient intake. If they have access to and consume improved crop varieties.radicals have been associated with chronic diseases. an enzyme in the pathway for resveratrol synthesis. starch and glucose levels were decreased. 2004). vitamin C and glutathione. In this study. The transgenic tomatoes also showed improved antioxidant capacity. Yet another transformation in tomatoes was recently reported to result in the synthesis of resveratrol. In 2004. tocopherols.g. eggplant). and many others. 2004). Caution should be sounded. These findings point to complex relationships between antioxidant content and other compounds. because in small amounts many of these substances are protective. however. they may improve their crop yields and consequently their economic wellbeing. VI. They include such substances as flavanols. 24 Application of Biotechnology for Functional Foods . tea. green coffee beans. olive oil. suggesting that such enhanced tomatoes might provide additional antioxidants. quercetin. trace mIneral content and BIoaVaIlaBIlIty Improving human nutrition by increasing the availability of trace minerals in crops is potentially highly efficient and effective. This strategy may reach more people in developing countries than fortification of foods. chlorogenic acid. Another phenolic antioxidant. but indicate that antioxidant levels in potatoes can be altered using biotechnology. were also increased. an antioxidant not normally found in tomatoes.. had a resveratrol content of 53 mg/g fresh tomato upon ripening (Giovinazzo et al. However. The contents of two other antioxidants. To enhance the antioxidant content of potatoes. because many subsistence farmers grow their own food and are outside the market system. Lukaszewicz and colleagues conducted a series of transformations using one or multiple genes encoding enzymes in the bioflavanoid synthesis pathway (Lukaszewicz et al. This is because trace minerals are essential to the plant’s ability to resist disease and other environmental stresses (Bouis 2002). tomatoes incorporating the gene for stilbene synthase. and tea. tomato. resveratrol. thus making use of an abundant resource in the soil that is otherwise unavailable. so there has been considerable interest in the availability of antioxidants. plants with improved ability to take up minerals from the soil will not deplete nutrient-poor soils. it was reported that transgenic tomatoes carrying the gene for this enzyme accumulated higher levels of chlorogenic acid with no side effects on levels of other phenolics (Niggeweg et al. in high doses. It is synthesized by the enzyme hydroxycinnamoyl transferase in solanaceous plants (e. tomatoes. plus improved antioxidant capacity. and many others.

was used in the transformation of rice to increase iron content (Goto et al. the enrichment of staple foods. Iron Iron deficiency anemia is one of the most widespread nutritional deficiencies in the world. much is stored in the husk and subsequently lost during milling and polishing. manganese. A different source of ferritin genes. 2000). A significant breakthrough in improving the iron content of cereals was achieved by Ingo Potrykus and colleagues. 1996). iron was significantly increased in vegetative tissues but not in seeds (Drakakaki et al. This is because iron has low solubility and is toxic in excess. commonly used in iron supplements. the latter two can be toxic. iron deficiency anemia is caused by insufficient dietary iron. Thus. Phaseolus vulgaris. This gene encoded for the iron storage protein. For this reason. Japan. mineral transport and storage in the plant. strategies to increase the iron content of cereals and grains face the challenge of targeting iron storage in a form and location in the plant where it will be bioavailable when consumed. transferred the gene for ferritin from soybean into rice and confirmed the stable incorporation of the ferritin subunit in the rice seed. Iron transport and uptake in plants is carefully regulated. soybean. and in rice just 5% of the iron in leaves is transported to the grain. The problem for women and children is more severe because of their greater need for iron. For example. and other conditions have substantial effects on the outcome of genetic engineering experiments to increase mineral content. absorption inhibitors such as phytate. type of promotor used. Hanover. This finding introduces the possibility of engineering plants with the ability to take up desirable minerals while excluding toxic and undesirable ones. the experimental conditions. Thus.A. Although much is known about the uptake of iron and zinc in roots and transport of minerals to and from vegetative parts of the plant. NH. Iron content in the transgenic rice seeds was up to threefold greater than in non-transgenic control plants. and when expressed in the transgenic rice increased the iron content twofold (46). Recent studies have examined the function of iron transporter proteins in transgenic plants. IRT1. The bioavailability of iron in transgenic rice varieties containing ferritin was shown to be as good as ferrous sulphate. 2002). have shown that slight changes in the amino acid composition of the transporter protein affects the selectivity of metals transported into the plant (Rogers et al. In cereals. These proteins have been shown to increase iron uptake into roots when iron is deficient (Eide et al. One of the genes transferred to Golden Rice came from the common bean. and cadmium. ferritin. is one of the top priorities in international agricultural and nutrition research. In rice-eating populations. lead. The United Nations estimates that over three billion people in developing countries are iron-deficient (Administrative Committee 2000). as reflected in biochemical indices of iron status in iron-deficient laboratory rats (Murray-Kolb et al. and lack of enhancing factors for iron absorption such as ascorbic acid. especially those consumed in poor countries. 2000). in wheat only 20%. first isolated from Arabidopsis thaliana. 2 Application of Biotechnology for Functional Foods . Mary Lou Guerinot at Dartmouth College. 2002). also transports other metals such as zinc. The iron transporter protein. Researchers in the laboratory of Dr. 1999). Others have used recombinant soybean ferritin under a different promotor to increase the iron content in wheat and rice. Researchers at the Central Research Institute Electric Power Industry. some plants accumulate very little trace minerals in the grain (Holm et al. In this case.

2001). Zinc is also an important constituent of more than 100 enzymes. This observation suggests ways in which selective genetic transformations might be used to enhance the uptake of some minerals while excluding others. anorexia. immune disorders. Unlike the IRT1 transporter. 2 Increasing the zinc content of cereals and grains.A second iron transporter protein. in screening over 1. Ramesh and colleagues (2004) in Australia. higher zinc levels were obtained as well. a nutritional staple in many countries. may be an effective way to improve human nutrition and at the same time increase plant yields. zinc uptake in the transgenic lines was higher in the short term compared with control plants. found that iron content averaged about 55 mg/ g iron. Aromatic rice tends to have the highest iron levels and several varieties have been successfully crossed with elite rice lines having excellent agronomic characteristics and grain qualities. IRT2. Multiple transgenic lines exhibited higher zinc and iron contents in their seeds compared with control plants (Ramesh et al. especially where soils are low in zinc. For example. Zinc Zinc is one of several trace minerals that can be deficient in human diets. uptake of zinc decreased Application of Biotechnology for Functional Foods . studied the effect on seed zinc content in barley (Hordeum vulgare cv Golden Promise) in plants transformed to increase the expression of zinc transporter enzymes. B. 2004). Another approach to improving the availability of iron for infants was reported by Suzuki and colleagues (2003) at the University of California. Their findings suggest that recombinant lactoferrin grown in plant culture may be a functional alternative to human lactoferrin in infant formula and provide another way to improve iron availability during infancy. High iron rice developed from traditional breeding is currently being tested for iron bioavailability and effects on iron nutrition status in young women in the Philippines (World Bank 2000). These observations suggest that genetic modification of selected varieties to further increase iron content might boost zinc levels too. a milk protein that binds iron. and a variety of other symptoms. These micronutrient traits were shown to be stable across different growing environments and could be crossed with high yielding varieties to improve the nutrient density. Nigeria. however.000 varieties of common beans. but some varieties from Peru averaged more than 100 mg/g iron (Gregorio 2002). As in beans. iron uptake was increased (Vert et al. Zinc content. When grown under zinc deficient conditions. although the average content of each differs. has also been identified in the roots of Arabidopsis. These investigators developed transgenic rice in cell culture that expressed the gene for human lactoferrin. iron and zinc concentrations differ across varieties of rice.and zinc-deficient plants. When zinc content was restored. averaged 35 mg/g. When they compared the recombinant lactoferrin with native human lactoferrin both proteins retained functional activity after mild heat treatment. It should be noted that iron and zinc levels tend to be present together in many plants. When the gene for IRT2 was incorporated into iron. Results are not yet available. Absorption of zinc from cereals and grains can be impaired or blocked by the presence of some substances such as phytate. and in vitro digestion. IRT2 did not transport manganese and cadmium when it was expressed in yeast. scientists at the International Institute of Tropical Agriculture. Zinc deficiency is associated with impaired growth and reproduction. When varieties were selected for their iron content. especially where meat is not consumed. high acidity. Davis.

C. 2 VII. Areas where soils are deficient in selenium are well known and low to deficient intakes have been observed among human and animal populations in these regions. resveratrol—to name a few. a study from the University of California. Berkeley reported that genetically engineered Arabidopsis thaliana and Indian mustard (Brassica juncea) were able to incorporate more selenium from soil and convert it to non-toxic methylselenocysteine than wild type plants (LeDuc et al. improve immune function. This study suggests that increasing the production zinc transporter proteins may be one approach to increasing the zinc content of cereals. Because they are widely distributed in fruits and vegetables. colo-rectum and prostate (El-Bayoumy and Sinha. Combs 2004). Genetic engineering technology offers considerable potential for increasing the uptake of selenium from soils and incorporating the mineral into non-toxic compounds in the edible parts of plants. Thus. data appear promising. However. isoflavones. and isothyocyanates. pHytonutrIents and noVel suBstances Intense research activity is being devoted to the identification and study of phytonutrients in plants with an eye to their ability to protect health. polyphenols. these various categories of substances appear to hold significant potential health benefits when consumed in modest amounts. 2004). diets rich in these foods are likely to furnish generous amounts of many of these phytonutrients. suggesting that the transporter proteins may be degraded when zinc is adequate. These studies demonstrate the feasibility of developing crop plants with improved ability to take up selenium from the soil and store it in a non-toxic form. Isoflavones: Isoflavones are a type of phytoestrogen. However. However. 2004). West Lafayette. Consumption of selenium has been linked to reduced risk of all cancers. the enhancement of foods with particular phytonutrients usually lacks sufficient scientific grounding to justify the development of foods with enhanced levels. the biological effects of isoflavones differ markedly from estrogen and many are non-hormonal. Examples of phytonutrients include: phytoestrogens. can be transformed to do so (Ellis et al. For these reasons. For example. and reduce the risk of chronic diseases ranging from heart disease and cancer to age-related macular degeneration. but are not consistent or conclusive. IN. selenium is toxic at levels only a little greater than those required in a healthy diet. Plants genetically modified to absorb above average quantities of minerals could be used to improve human or animal nutrition. so caution is warranted with supplementation and increased intakes. Selenium is also important for specific enzymes and proteins in the brain and is necessary for proper immune function. Researchers at Purdue University. but particularly those of the lung. Selenium Selenium is an essential trace mineral incorporated into plants from soil. Application of Biotechnology for Functional Foods . Extensive media and manufacturer publicity about many of these compounds generated expectations exceeding scientific justification.in both transgenic and control plants. For many substances—lycopene. so named because they bind to estrogen receptors and mimic some of the effects of the hormone estrogen. 2004. selenium at appropriate concentrations would be safe for the plant and for human consumption. a few examples can be cited. Unfortunately. also showed that plants not normally accumulating selenium. In spite of their cumbersome technical names. there is insufficient scientific data from carefully controlled studies that adequately demonstrate safety and efficacy of substances with potential promise. such as Arabidopsis.

In 2003.5-fold vs. Yu and colleagues (2003) at the DuPont Company. Increases were greatest for phytoene and beta-carotene. they are effective in reducing blood cholesterol levels. it is possible to increase the level of isoflavones in soybeans. Several novel phytostanols were obtained as well. total seed sterol levels increased by 2. researchers at Unilever Research. and glycitein. Netherlands. Wilmington. reported the application of genetic engineering techniques to increase the isoflavone content of soybeans. and possibly reducing the risk of prostate cancer. By activating genes in the phenylpropanoid pathway. Thus. DE.4-fold increase in phytosterols. but isoflavones occur in other legumes such as broadbeans. Currently. The two transgenes increased total seed sterol content more than with either gene expressed singly (2. diadzein. Their concentration in legumes is greatly affected by growing conditions and climate. Seeds from both types of plants exhibited conversion of the major phytosterols to phytostanols and no other functionalities were affected. yet still confer cholesterollowering benefits. Because these substances are hydrogenated they would be expected to be more stable during food processing. Application of Biotechnology for Functional Foods . This group also developed transgenic tobacco carrying two genes involved in regulating carbon flux through the sterol biosynthesis pathway (Holmberg et al. are plant sterols found in small quantities in vegetable oils. improving bone health.6-fold). 2003). 2003). Such an alteration in a widely consumed food would potentially increase the consumption of these substances in a large share of the population.4-fold. By blocking the anthocyanins branch of this synthetic pathway. 2 Most recently. 2003). so genetic modifications to increase isoflavone content might not be expected to be a high priority. Plants were transformed with a gene from yeast for the enzyme 3-hydroxysteroid oxidase (Venkatramesh et al. 1. Sufficient data of their efficacy and safety exist that the FDA has permitting food manufacturers to claim a role for plant sterols or stanols or their esters in reducing the risk of coronary heart disease (see section on health claims below for a further description). Researchers at Monsanto reported the application of genetic engineering to modify the ratio of phytosterols to phytostanols in rapeseed (Brassic napus) and soybean (Glycine max). and these variables could potentially override genetic modifications. reported the generation of transgenic tobacco seeds with enhanced total seed sterol level (Harker et al. Following the insertion of the gene for a key enzyme in sterol biosynthesis. They have been linked with easing menopausal symptoms. When consumed in sufficient amount. the investigators obtained higher concentrations of isoflavones. and in many vegetarian (“meatless”) foods made from soy products (USDA 2002). However.Soybeans are the richest food source of isoflavones. reducing cardiovascular risk. The main soy isoflavones are genistein. (2005) reported the development of transgenic tomatoes with a 2. diadzein levels increased and genistein levels fell. isoflavones are abundant and readily available. Enfissi et al. Phytosterols and Phytostanols: Phytosterols and their saturated derivatives. phytostanols. The additional sterol was present as fatty acid esters and several intermediate sterols accumulated.

For example. many benefit human health and plants. In foods. Further. or may be toxic if consumed in sufficient quantity. biotechnology. heat treatment. Examples of anti-nutritional substances are shown in Table 2. 2 VIII. Genetic engineering should make it possible to strengthen the effects of existing bacterial strains and create new ones (Steidler 2003). immunomodulation. Probiotics have been used to treat inflammatory bowel disease by creating more hostfriendly gut flora. Many of these substances are important in plant metabolism and provide resistance to environmental stress and pests. production of antimicrobial metabolites and nutraceuticals. dietary fiber improves colon function and reduces plasma cholesterol levels. Probiotic microorganisms may be concentrated and added directly to a food or added to a milk product in small amounts and allowed to grow. but others are resistant to digestion. genetically engineered bacteria have been used to improve the flavor and stability. and other means requires careful consideration of the complexities. Genetically engineered probiotic bacteria have been used to overcome problems associated with more traditional technologies for developing such bacteria. lactis subsp. polyphenols and other substances have anti-carcinogenic properties.Probiotics: The term “probiotics” refers to live microorganisms that have a health benefit when consumed in adequate amounts.S. 2002). including resistance to infectious diseases. They are usually bacteria selected from species found in the intestinal tract. Some strains of Lactobacilli have prevented the development of colitis in genetically susceptible mice. reduce the ability to break down complex molecules such as proteins and starch.S. Application of Biotechnology for Functional Foods . or other forms of processing. and others (Ahmed 2003). Foods containing probiotic bacteria are abundant in Japan and parts of Europe. 2002). prevention of vaginitis. Other genetically engineered bacteria have been used to secrete the anti-inflammatory cytokine IL-10 (Guarner et al. Probiotics have been used as dietary supplements and oral agents for intestinal disorders. They may interfere with intestinal cell function. diacetyllactis is used as a buttermilk starter culture in the U. Selective use of probiotic bacteria can create an environment where stimulation of the immune system is restrained and intestinal inflammation reduced (Guarner et al. Some of these substances are rendered harmless with cooking or processing. (Renault 2002). Although anti-nutrient substances reduce nutrient availability. but are less developed in the U. antI-nutrItIonal suBstances Many plants contain substances that inhibit nutrient uptake in various ways or are toxic themselves. Many health benefits have been attributed to probiotics. The most common foods having probiotic organisms are fermented dairy products such as yogurt containing Lactobacillus acidophilus. and this suggests that humans may be better able to adapt to these substances than was once thought (Welch 2002). but few products have reached application in the U. or to block the formation of unwanted flavors. The question of improving micronutrient availability by reducing anti-nutritional substances through plant breeding.S. A genetically engineered strain of L. Considerable research is being devoted to the identification and effects of probiotic bacteria and their use as therapeutic agents. some of the adverse effects on nutrient availability may diminish over time or with different levels of consumption.

peanuts. lectins Cyanogens or glycoalkaloids Cassava. cadmium. coffee. mercury. beans Glucosinolates Saponins Goitrogens Heavy metals. wheat. magnesium.g. e..g. soy. e. Phytate: Phytic acid is a phosphorus storage compound found in the seeds of many edible crops. other crops 0 There are several ways of reducing anti-nutritional substances. calcium.. can be toxic Inhibit acetylcholinesterase activity which impairs nerve transmission. Hemaglutinins. e. copper that reduces mineral absorption Interfere with cells lining the gastrointestinal tract causing acute symptoms. wheat. or impair health antI-nutrIent Phytic acid (phytate) Fiber. anti-nutrients in plant foods that reduce nutrient bioavailability. e. peas. rice Legumes.g. lignin Trypsin inhibitor eFFect Binds minerals. fumonisin Cabbage. cereal grains Whole cereal grains. can bind metals and some vitamins. Mg.g. toxic to humans and animals.. zinc. can be toxic Binds calcium to prevent its absorption Can be toxic. e. and genetic engineering. can damage cell membranes May adversely affect thyroid activity May irritate the gastrointestinal tract and interfere with nutrient absorption Suppress thyroid function May have toxic effects. heat.g. affects gastrointestinal and nervous systems Toxins produced by certain molds. potatoes Tea. barley. sorghum.g. aflatoxin. garlic Contaminated leafy vegetables Cottonseed Spinach leaves. including plant breeding. corn. corn. Examples of the various anti-nutritional substances that have been reduced by the application of biotechnology in different plants are described below. When Application of Biotechnology for Functional Foods . Phytic acidcontaining foods bind minerals in the intestinal tract rendering them unavailable.g. cellulose. e. solanine Mycotoxins. processing. and other minerals that cannot be absorbed. rice.g.. linseed. e. may decrease vitamin & mineral absorption Reduces the activity of the enzyme trypsin and other closely related enzymes that help digest protein Form complexes with iron. tannins. broccoli. beans. e. Fe. Zn Decreases fat and protein digestibility. cereals. e. fermentation and drying.g. high levels of Hg impair fetal brain developmnet May harm kidney function and reduce sperm counts. broccoli Soybeans. peanuts.taBle 2. lead Gossypol Oxalic acid Phytotoxins. can be carcionogenic dIetary source Whole legume seeds... zinc. Legumes Polyphenolics.g. rhubarb Green parts of potato Grain. e. K.. hemicellulose. Phytic acid forms salts (phytates) of potassium. Ca. iron. sugar beets Brassica and Allium foods.

using animal feed engineered to produce phytase addresses many of the problems associated with high phytate levels in animal feed. 2003). a drawback of such a transformation for human consumption is that the enzyme is inactivated above 60°C and would be destroyed by cooking (Holm et al. which was then cooked. the investigators were able to increase the enzyme activity above that of the original yeast gene. Similarly. This is a particularly important consideration in the diets of women and children where legumes and cereals are staple foods. Feeding the transgenic soybeans resulted in an 11% greater reduction in phosphorus excretion than feeding with conventional soybeans to which the enzyme is added. 2002). 2002). Consequently. An additional consequence is the production of high phosphorus animal waste with adverse environmental effects. It is possible that more heat-stable forms of the enzyme could be used. barley. However. the presence of phytic acid can contribute to mineral deficiencies.these minerals are limiting in the diet. 2003). However. Reduction in phytate in the range of 50% to 66% has been achieved with these mutant lines. Zinc absorption from the 80% phytate-reduced corn was three times greater than from the wild-type corn. Transgenic rice expressing phytase derived from modified yeast genes has also been reported (Hamada et al. 2005). (2004) who fed corn tortillas to six healthy adults. only 8% of the activity remained (Holm et al. In soybeans and corn. several hybrids developed with the mutant strains exhibited lower yields. In a study of broiler chickens. phosphorus may have to be supplied in a more available form to overcome the loss due to binding with phytic acid. 2002). especially for poultry and swine. 4. Proof that zinc absorption from low phytate corn compared with wild-type varieties was significantly greater was reported by Hambridge et al. consumption of transgenic soybeans containing phytase led to a 50% reduction in phosphorus excretion compared with a diet supplemented with an intermediate level of nonphytate phosphorus (Denbow et al. 1998).9 mg/day compared with 1. It has now been shown that low phytate mutant corn is linked to the reduced expression of the enzyme myoinositol phosphate synthase.5 mg/day. 80% reduction has been achieved. Lines of corn. By selectively modifying the genes. the first enzyme in the synthesis pathway for phytic acid (Shukla et al. and soybean with slightly different phytic acid characteristics have been used to develop varieties with reduced seed phytic acid (Raboy 2002). biotechnology has been used to express the enzyme phytase in plants (Chier et al. but when more heatstable enzymes were incorporated into rice. Holm et al. This enzyme allows animals to metabolize phytic acid and eliminates the need to supplement feed with phosphorous. Although genetically engineered low phytate crops have not been commercialized. 1 Application of Biotechnology for Functional Foods . 2004). 2004). rice. Transgenic phytase-containing wheat was developed by Holm and colleagues at the Danish Institute of Agricultural Sciences. low-phytate corn and barley fed to broiler chicks resulted in 33% lower phosphorus excretion compared with wild-type grain diets and reduced the need for supplemental phosphorus (Jang et al. Denmark (Brinch-Pedersen 2000. particularly in the case of iron and zinc. Phytase has been successfully incorporated into soybean and wheat and is biologically active when the plants are used as animal feed (Brinch-Pedersen et al. 2000). This finding was confirmed recently by Italian researchers who developed a mutant corn with 90% less phytic acid and a 10-fold increase in seed-free phosphate (Pilu et al. Transgenic plants exhibited up to 4-fold higher phytase activity compared with wild-type seeds. In animal nutrition.

2001). 2003). copper and calcium did not differ between the two formulas. These findings suggest that use of dephytinized soy protein improves zinc absorption and can be recommended for infant foods. In this case. this group reported that transgenic cassava roots expressing a different gene contained significantly less acetone cyanohydrin levels compared with wild-type plants (Siritunga et al. fermenting.6% compared with 16. 22. 2004). Absorption of iron. Transformed cotton plants had up to 70% less gossypol in their seeds compared with non-transformed plants (Martin et al. lotaustralin. soy protein used in infant nutrition may limit mineral absorption because of its phytate content. These pigs expressed the enzyme phytase in their saliva and exhibited complete phytate digestion. but are sufficiently abundant in “bitter” varieties to require removal (Padmaja 1996). To investigate this question.Reducing the phytate content of plants. This substance is found in the green tissue of potato just under the skin. grating or maceration. Davidsson and colleagues at the Swiss Federal Institute of Technology compared regular and dephytinized soy formula in nine infants 69 to 191 days old. long known to be toxic to humans and animals. The investigators reported that zinc absorption was significantly greater from dephytinized formula compared with regular formula. they reduce food quality and safety. 2002. created a transgenic cotton (Gossypium vitifolium) using the antisense gene technology for a key enzyme in the synthesis of gossypol. 2 Application of Biotechnology for Functional Foods . These findings suggest that biotechnology can be used to reduce the gossypol levels to render the seed oil more suitable for feed and food. Cyanogens: Cassava (Manihot esculenta Crantz) produces various cyanogenic glycosides such as. TX. 2004 ). a characteristic of countries where cassava is a staple. Canada. manganese. but those with higher levels require. gossypol also appears to have anti-cancer properties toward several human prostate and breast cancer cell lines (Liu et al. Drs. Another approach to solving the phosphorous uptake problem in animal production was undertaken by researchers at the University of Guelph. required no dietary inorganic phosphorus and excreted 75% less phosphorus. Regular and dephytinized formula contained 300 and 6 mg phytic acid/kg liquid. Columbus. respectively. For example. reported the development of transgenic cassava with a 99% reduction in linamarin in the roots and between 60% and 94% reduction in leaves (Siritunga and Sayre 2003). and acetone cyanohydrin in its roots and leaves. and sundrying to reduce cyanogens adequately. College Station. linamarin. has direct implications for human nutrition. 2004). OH. These accomplishments open the door to the development of cassava truly safe for human and animal consumption. While these compounds protect the plant from pests. Steroidal glycoalkaloids: Potatoes (Solanum tuberosum spp) contain potentially toxic steroidal glycoalkaloids. Gossypol: Cottonseed contains the polyphenolic compound gossypol. These plants were transformed by inhibiting the expression of two genes that catalyze the first step in the synthesis of linamarin. swine were engineered to produce the enzyme phytase (Golovan et al. The following year.7% absorption (Davidsson et al. Interestingly. These potentially toxic substances are only present in small amounts in “sweet” varieties of cassava. soaking. Boiling and drying reduce these substances to safe levels in low cyanogen varieties. Siritunga and Sayre at the Ohio State University. Jiang et al. the best known of which is solanine. In 2003. The toxicity of these cyanogens is exacerbated by low protein intakes. Glycoalkaloids are synthesized from cholesterol. Martin and colleagues at Texas A&M University. particularly soybean.

The potential for genetic engineering strategies to reduce the production of fungal toxins in food crops has been discussed by Duvic (2001) and Munkvold (2003). Mycotoxins in corn: An important risk to the safety of grains (corn. It remains to be seen whether pest resistance in these transgenic potatoes is affected by the reduction in glycoalkaloids. but many remain unknown. groundnut. Specific food proteins can trigger the immune system and provoke an allergic response. It binds several minerals including calcium and prevents their absorption. responses in susceptible people. 2005). Application of Biotechnology for Functional Foods . Certain types of fungi—Aspergillus flavus and Fusarium— are notorious for their deadly products. These studies indicate that substantial improvements in the reduction of steroidal glycoalkaloids in potatoes may be close at hand. 2002) and in transgenic bacillus thuringiensis (Bt) corn (Bakan et al. pistachios) and cottonseed is the production of toxins by fungi. walnuts. Wilson et al. Scientists at the National Centre for Plant Genome Research (NCPGR). Department of Agriculture developed transgenic walnut trees that displayed increased resistance to aflatoxin synthesis (USDA 2004). and by age five about 80% of allergic infants will lose their food allergies. Eight types of foods account for nearly 90% of all food allergies. al. Aflatoxin. tree nuts. 2002). and soy. India. suggesting that many soy proteins have allergenic potential. Risk of such reaction is greatest in the first two years of life. and chick pea. but weather also contributes to their development. peanuts. soybean. People allergic to one type of food are frequently.S. Other investigators used antisense technology to create transgenic potatoes with up to 40% less steroidal glycoalkaloids in the tubers (McCue et al. is sometimes found in peanuts and corn. a particularly dangerous cancer-causing mycotoxin produced by the fungus Aspergillus flavus. Partial resistance to Fusarium disease was reported in transgenic wheat (Okubara et al. Many proteins associated with food allergies in these foods have been identified. known as P34 (Cordle 2004. fish. wheat. allergic to others. but not always. groundnuts (peanuts). allergens Food allergy. can provoke severe. 2003). sometimes fatal. seek to reduce the oxalic acid content in these foods through the transfer of the gene for oxalate decarboxylase (OXDC). Breeding crop varieties with increased resistance to fungi may reduce the production fungal toxins. These are: milk. a type of antibody involved in allergic responses. barley).Recently. tomato. (2003) showed that the inhibition of plant sterol synthesis in transgenic potatoes reduced the synthesis of glycoalkaloids by 41% in leaves and 63% in tubers. Oxalates: Oxalic acid is present in spinach. crustacea. wheat. Agricultural practices and grain storage conditions help to minimize growth of these fungi. Arnqvist et. More than half of soybean allergic reactions are attributable to a single protein. as many as 28 proteins may bind with IgE. Soybean: In people with soy allergy. Genetically engineered tomatoes bearing the OXDC gene have been successfully grown and are currently undergoing field and biosafety tests (ISAAA. The substances produced by these organisms cause disease in plants and potentially serious illness in people and animals consuming the infected crops. eggs. the enzyme that degrades oxalic acid. Researchers at the U. although relatively rare. 2004). tree nuts (almonds. Fumonosins produced by Fusarium fungi are thought to be carcinogenic and harmful to the immune system.  IX.

EuropaBio (2002) reported that several laboratories in Japan are working to develop hypoallergenic rice. Shrimp: Allergic reactions to shrimp and other crustacea (lobster. 2003). known as Pen a 1. is believed to be the most potent of these three (Koppelman et al. 2002). The major allergen(s) in rice have been identified and hypoallergenic varieties of rice developed using antisense genetic engineering techniques to suppress the synthesis of the predominant allergen (Nakamuro and Matsuda 1996). People with hypersensitivity responses to crustacea may also be allergic to mollusks. Nonetheless. Animals were challenged after 2. The major allergens in peanut have been identified as Ar h1. and some arthropods such as house dust mites and cockroaches. and 10 weeks. not all allergenic potential had been eliminated. some people. commented that removing the other proteins should not be difficult because wild species lacking the genes for the other proteins are already known. Samuel Lehrer of Tulane University. This particularly encouraging research suggests that protection against peanut allergy may be in the foreseeable future. crayfish) are among the most common food allergies. Animals given the medium and high doses remained protected for 10 weeks. Extremely sensitive patients were also sensitive to other proteins. Other genetic engineering strategies that alter the composition of the allergenic proteins may render the proteins harmless to sensitive people. Recombinant versions of these allergens produced in bacteria were heat-killed and used in a vaccine (Li et al. These regions contained 15 to 38 amino acids from whose sequence the corresponding gene sequence can be determined. and their genes have been isolated. Dr. LA. are allergic to rice proteins. has located the gene sequence that encodes these epitopes and has suggested that by altering these epitopes by a single amino acid. Treated mice produced no anaphylactic or histamine response when challenged. two other proteins that trigger allergic reactions may have to be removed before soybeans could be sold as hypoallergenic. a Kunitz trypsin inhibitor. However. The ability to eliminate the most hazardous allergens in soy would have substantial benefit for infant formula feeding and for those who are allergic to soy.Scientists at USDA and the Donald Danforth Plant Science Center have succeeded in silencing the gene for P34 and created soybeans without this protein (Herman et al. and Ar h3. 2004). a researcher involved in the project. Ar h2. Dr. Recent analysis of this allergen revealed five IgE-binding regions or epitopes (Ayuso et. 2003). as it is not possible to eliminate all allergens. but efforts have provided only partial success. particularly in Japan. so that a single genetic transformation was insufficient to overcome their allergic responses. Careful plant breeding with the genetically modified soybeans may be able to produce the hypoallergenic soybeans. IgE binding could be halted. This work suggests that safe recombinant allergens could be synthesized for immunotherapy of those who are allergic to shrimp and related substances. Rice: Rice is generally consider a hypoallergenic food. To date there have been no ways to overcome these allergies. The vaccine was given in three different doses to allergensusceptible mice. and for that reason is one of the solid foods first introduced to infants. The protein Ar h2. 6. New Orleans. The major allergen in shrimp is the muscle protein tropomyosin. when the hypoallergenic rice was tested in sensitive patients. Allergen content in the transgenic seeds was reduced from about 300 micrograms/seed to about 60 to 70 micrograms/seed.al. 4 Application of Biotechnology for Functional Foods . Anthony Kinney. Peanut: Allergy to peanut proteins can be fatal. However.

(2005) developed transgenic grapevines using the gene for stilbene synthase. 2003). Transgenic yeast also has potential for modifying the flavor of beer (Vanderhaegen et al. (2004) reported additional progress on modifying the pathways in Arabica and canephora coffee varieties which resulted in caffeine reductions ranging from 30% to 50%. mIscellany A. Beer and Wine Applications of biotechnology are finding their way into brewing and wine-making. as recently described (Dudareva and Negre 2005). an enzyme that leads to the production of resveratrol.) has been increased content of the antioxidant resveratrol. Application of Biotechnology for Functional Foods . 2004). It is also thought to improve plant resistance to fungal disease. This substance is of special interest in human health for the reduction of oxidized lipids. In a different approach. 2003) and sake (Aritomi et al. Genetic engineering also holds considerable potential for the production and regulation of diverse flavors and aromatic substances.X. Ogita et al. Ogita et al.  B. This genetic modification would eliminate the need for decaffeination processing. One application in grapevines (Vitis vinifera L. the most popular coffee grown. Toward this end. The plants increased their production of resveratrol. Giorcelli et al. GonzalezCandelas et al. (2000) also reported increased resveratrol in wine through the use of transgenic yeast. The researchers aim to modify Arabica coffee plants. Caffeine content in the transgenic plants was reduced by 70%. but showed no improved resistance to a leading fungal pathogen. Decaffeinated Coffee Japanese scientists have succeeded in silencing the gene responsible for caffeine production in coffee plants (Jameel 2003.

Application of Biotechnology for Functional Foods .

or other means). A frequent criticism is that all food is. federal requirements for “functional food” apply regardless of how the food is produced. The focus of much attention in recent years. functional food has been the subject of numerous articles. As previously noted. product formulation.1 The difficulties associated with FDA regulation of functional food historically have been related to whether such food is subject to the more onerous drug provisions of the law. the Food and Nutrition Board of the National Academy of Sciences has suggested that a “functional food” is “any modified food or food ingredient that may provide a health benefit beyond the traditional nutrients it contains” (Food and Nutrition Board 1994). IntroductIon Applications of Modern Biotechnology to Functional Food Legal and Regulatory Considerations Under Federal Law “Functional food” is touted as a convenient means for consumers to promote optimal health. is subject to the same regulatory requirements as any other food.  Application of Biotechnology for Functional Foods . including the Animal and Plant Health Inspection Service of the U. such as through mechanical or genetic methods (e. and the Food and Drug Administration (FDA).S. Department of Agriculture (USDA). the Environmental Protection Agency (EPA). including lycopene. Thus.” or a “drug. This means that a functional food may be regulated as “conventional food. 22984 (1992). modern biotechnology techniques.. fruits and vegetables (and components thereof. See 57 Fed. and other phytochemicals). indoles.” a “dietary supplement.g. including calcium-fortified orange juice.” a food for “special dietary use” (including infant formula) a “medical food. This report focuses on safety and labeling requirements enforced by FDA and USDA’s Food Safety and Inspection Service (FSIS) pursuant to certain food laws administered by those agencies. Despite the widespread attention. whole grains. soybeans. including claims made for it. polyphenols. Food that is deemed functional. Significantly.” depending upon its positioning in the marketplace. rice that has been genetically enhanced to provide beta carotene is subject to basically the same statutory and regulatory framework as rice to which beta carotene is added through product formulation. omega-3 fatty acids. regardless of whether the food has been modified in some fashion.PART 2 I. reports. and cocoa. 1 The use of modern biotechnology is subject to regulation by several federal agencies. Food produced through the use of modern biotechnology is handled somewhat differently by FDA in that it is subject to a voluntary consultation process. therefore. there is no separate regulatory category for functional food in the United States. the “functional food” concept eludes precise definition. phytosterols. A considerable array of food has been described as “functional” in one or more respects. Others argue that a functional food is any food promoted or consumed for a specific health effect. and consumer education materials. From a legal perspective. including the prevention of disease. such as a “Functional Food Guide Pyramid”—a modified version of the USDA Food Guide Pyramid that identifies functional food from each major food group. Reg. functional. in some sense.

tHe FFdca Framework—oVerVIew and BrIeF HIstory In the United States. to include “infant foods.” “Drug. 12 (1934)). slenderizing foods.This section of the report describes how functional food is regulated under certain federal laws governing food in general. 73d Cong. § 321(z).” FFDCA § 201(z).3 In addition to providing for the regulation of “food. invalid foods. 21 U. including pet food. mineral.” the FFDCA establishes comprehensive requirements for the marketing of drugs..” the legislative history reveals that such uses were considered. medical devices. A food for special dietary use is deemed mislabeled or misbranded under the law unless its label bears information adequate to inform purchasers of its vitamin.2 To place the regulation of functional food in an appropriate context.S.. II. 2d Sess. the discussion also identifies select areas of criticism and issues concerning functional food regulation in the United States. first in the context of food generally. products with meaningful amounts of meat or poultry (generally 2–3%) are regulated by the U. a “special dietary use” food for which Congress and FDA have established unique and prescriptive requirements. The emergence of functional food as a unique and important category for marketing purposes has led some to question whether the current legal and regulatory framework is adequate.4 2 3 Under applicable law. and then in the context of meat and poultry products.  4 Application of Biotechnology for Functional Foods . mineral. “Functional food” is primarily intended for human use. the FFDCA established requirements for only three product categories of relevance to functional food regulation. No. infancy).S. or other dietary content. Alcoholic beverages are regulated primarily by the Bureau of Alcohol. at that time. In addition to covering “food” and “drugs. as other animal feed is subject to typically different requirements. A. or other dietary property.” the 1938 Act introduced the concept of food for “special dietary uses.” Although this law contained no definition of “special dietary uses. this report begins with an overview of the basic statutory framework pertinent to food. eggs and egg products. age-related nutritional needs (e. the law has evolved—through flexible agency interpretations as well as statutory amendments—to accommodate new products and advances in science. Tobacco and Firearms of the Department of Treasury. This report does not address infant formula. food refers to use for human beings or other animals and includes (non-human) animal feed. 493. Food and other articles regulated by FDA under the FFDCA cannot be adulterated or misbranded. and other dietary [products] intended for special nutritional requirements” (S. The Statutory Foundation: “Food. It then examines the application of this basic framework to functional food.” and Food for “Special Dietary Use” At the time of its enactment in 1938. Department of Agriculture (USDA) pursuant to the Federal Meat Inspection Act (FMIA) or the Poultry Products Inspection Act (PPIA). and animal feed. As discussed more fully below. Drug.” including uses that may arise from disease or certain health-related conditions. “Infant formula” is defined to mean “a food which purports to be or is represented for special dietary use solely as a food for infants by reason of its simulation of human milk or its suitability as a complete or partial substitute for human milk. and Cosmetic Act (FFDCA). Rep. Although the concept of “functional food” was not contemplated at the time Congress enacted the FFDCA. As appropriate.g. Adulteration refers generally to aspects of a product that typically relate to quality or safety and misbranding refers generally to false or misleading labeling.C. food products other than meat and poultry and alcoholic beverages are regulated primarily by the Food and Drug Administration’s (FDA) Center for Food Safety and Applied Nutrition (CFSAN) pursuant to the Federal Food. and cosmetics. FDA later defined “special dietary uses” by regulation to mean “particular (as distinguished from general) uses of food. or a desire to supplement or fortify the diet with any vitamin.

. which had been regulated as a drug. Accommodations of Advancing Science For nearly thirty years after enactment of the FFDCA.” and food for “special dietary use” remained largely unchanged.e. Intended for the dietary management of phenylketonuria (PKU). the claim addresses the ability of the food. the basic categories of “food. 2124.” Underlying this framework was an assumption that food for targeted nutritional purposes (i. to reduce the risk of disease or a health-related condition (as opposed to disease treatment. Second.. In a regulation issued shortly thereafter. “Health claims” are permitted only if approved or otherwise authorized by FDA. “special” uses) was of little or no relevance to the general population. “Health claims” are statements that characterize the relationship between a food (or a substance in the food) and a disease or health-related condition (21 C. Lofenalac® was recognized by FDA to meet distinctive nutritional requirements—namely. as recently as 1971. FDA created the “medical food” category by deciding on its own initiative to reclassify the specialty infant formula Lofenalac®. which establishes a framework for the regulation of “health claims” and “nutrient content claims. Congress passed the NLEA. Paradoxically. In 1972. Three regulatory developments are of particular relevance to the regulation of functional food.” Following NLEA. As discussed more fully in Part III.” “drug. FDA interpreted almost any use of food for a targeted nutritional purpose to be a “special dietary use. or similar concepts). an impaired ability to metabolize the essential amino acid phenylalanine—and therefore more appropriately regulated as a food. an inborn error of metabolism. These reforms ultimately led to new and distinct categories of “food” and new types of claims. NLEA also authorized “nutrient content claims. the special dietary use concept was gradually reformed to accommodate evolving views concerning the relationship between diet and health. such as specialty infant formulas. mitigation. were strictly regulated as drugs. such as “sugar free” and “low sodium. and (3) authorization of a separate regulatory scheme for dietary supplements in the Dietary Supplement Health and Education Act (DSHEA) of 1994. (2) the authorization of certain nutrition and health-related claims for food in the Nutrition Labeling and Education Act (NLEA) of 1990. FDA today treats “medical food” as a wholly unique category from food for special dietary use.e.” Indeed. Reg. and demonstrate the progressive blurring of the legal distinction between “food” and a “drug” since 1938: (1) the advent of “medical food” in 1972.R.14(a)(1)). FDA determined that many nutrition-related claims were of general use to the public. and food purported for use in sodium-restricted diets. the addition of nutrients to food or ingredients thereof). in 1990. § 101. vitamin and mineral supplements.” which are statements characterizing the level of a nutrient in food. as a distinct type of food for special dietary use. and has developed very specific criteria for the marketing of such products. which exempted medical food from nutrition labeling requirements. as part of an appropriate diet. FDA’s framework for the regulation of food for special dietary uses (commonly referred to as “special dietary food” at that time) included requirements addressing all uses of fortification (i. FDA described the new category as “foods represented for use solely under medical supervision in the dietary management of specific diseases and disorders” (38 Fed.”  Application of Biotechnology for Functional Foods . certain products that seemed to more naturally fit the special dietary use category.F. and thus were no longer indicative of “special dietary uses. among other food or uses deemed to be “special. 2126 (1973)). and the food meets certain qualifying criteria established by FDA.” among other requirements. With advances in nutrition and related sciences. During this time period.B.

classIFIcatIon oF FunctIonal Food under tHe Federal Food. 62400. Requests for Reform The emergence of functional food as a unique and commercially important marketing category has led some to question whether the current regulatory framework is adequate.” the claim could be notified to FDA and used within 90 days if the agency did not object. health claims. have criticized FDA’s oversight of ingredient safety and labeling. NLEA. and other stakeholders regarding the regulation of these products” (71 Fed. 2006))5 FDA’s views of the current framework for regulating functional food are described in detail next. Some groups. such as the Institute for Food Technologists (IFT) have recommended increased flexibility and policy changes to better allow for claims based on scientific advances (IFT 2005). to announce that the agency would hold a hearing to allow consumers. they created new regulatory categories and permitted many products to bear claims that would ordinarily have triggered “drug” regulation. the new law creates a unique framework for supplement regulation. including the Government Accountability Office (GAO) and the Center for Science in the Public Interest (CSPI). both are addressed in this report. the functional food concept is often considered to include both categories. labeling. and other interested parties to provide comments on approaches to the regulation of functional food. which establishes comprehensive requirements for dietary supplements. III. and whether 5 FDA did not specifically seek comment regarding the regulation of dietary supplements. Although DSHEA provides that dietary supplements are still “food” for many purposes. and therefore functional food. For example. Application of Biotechnology for Functional Foods . GAO and CSPI have suggested that companies should be required to notify FDA before using certain “functional” or “novel” ingredients and before making certain claims about the effect of such ingredients on the structure or function of the body (GAO. and cosmetIc act 40 The regulatory classification of a product has substantial implications for its marketing and use. as it influences the legal requirements pertaining to safety. CSPI 2002). accordingly. These new categories and claims are described more in Section III and relate to dietary supplements. if the panel found the claim to be “generally recognized as efficacious. Although there are important legal distinctions between conventional food and dietary supplements.Third. These important developments—DSHEA. and FDA’s “medical food” policy— helped to shape FDA’s regulation of “food” and nutritional claims. As proposed by IFT. 62401 (Oct. and nutrient content claims. in October 2006. Reg. Affirming its flexible approach to the statute. we believe that it would be in the best interest of public health to begin a dialog with industry. Particularly. drug. industry. These and other calls for reform led FDA. FDA stated that “[a]lthough we are confident that the existing provisions of the act are adequate to ensure that conventional foods being marketed as ‘functional foods’ are safe and lawful. IFT has proposed that FDA adopt a notification procedure for health claims. Other groups. including safety and labeling requirements. Congress enacted DSHEA. 2000. medical food. C. 25. consumers. in 1994. companies wishing to use a health claim could convene an expert panel to evaluate the science supporting the claim.

the product is subject to regulatory pre-market clearances or approval.g. or mitigate disease Product used primarily for taste. An overview of the basic categories and claims available for each is presented in Table III-1. or nutritive value. careful consideration of the pertinent regulatory categories is required. and in a conventional food form safety standard New drugs must be demonstrated to be safe and effective Components must present “reasonable certainty of no harm”.g. labeled as a dietary supplement. a review of each regulatory category into which a functional food may fall is provided. among other criteria Food for Special Dietary Use Food intended for supplying See conventional food particular dietary needs standard above (e. prevent. be intended for ingestion. not represented for use as a conventional food. and not approved as a drug. underweight. food for special dietary uses (Section E). Accordingly.. structure/function claims must be notified to FDA. must contain one or more “dietary ingredients” (e. other claims must not be false or misleading in any particular Health and nutrient content claims must be FDAauthorized. These regulatory categories are drug (Section B). treat.g. dietary supplement (Section D). or overweight) Formulated food intended for use under the supervision of a physician for the dietary management of disease with distinct nutritional requirements See conventional food standard above Claims of special dietary usefulness (e. to determine how a specific “functional food” is regulated under the FFDCA. Following a brief review of the pivotal definitions of “food” and “drug” (Section A). and medical food (Section F). substances found in the diet). taBle III-1 regulatory categories and general summary category Drug 41 claims Must be approved by FDA scope Product intended to diagnose. new ingredients must be notified to FDA Conventional Food Health and nutrient content claims must be FDAauthorized. other claims must not be false or misleading in any particular Dietary Supplement Product intended to supplement the diet. vitamins. which form the basis for much of the regulatory landscape for functional foods. pregnancy. herbs. those relating to disease.. must be FDA-cleared unless determined to be generally recognized as safe (GRAS) Must not present a “significant or unreasonable risk of illness or injury”. conventional food (Section C). lactation. aroma. usefulness in reducing body weight) Medical Food Claims for the dietary management of disease Application of Biotechnology for Functional Foods ..

or prevent disease. The food definition is based on a product’s actual (as opposed to intended) use. aroma. mitigation. 1983) (emphasis added).’ it means that the statutory definition of ‘food’ includes articles used by people in the ordinary way most people use food—primarily for taste. Today. or nutritive value is unduly restrictive since some products such as coffee or prune juice are undoubtedly food but may be consumed on occasion for reasons other than taste. which are authorized to bear structure/function claims without regard to their nutritive content or positioning as “food. and (D) articles intended for use as a component of any article specified in clause (A). or nutritive value. 713 F. Schweiker.” See infra section III. on the basis of its intended use to diagnose. . FDA asserted a starch blocking product to be a “drug” within the meaning of section 201(g)(1)(C). (2) chewing gum. or nutritive value. Nutrilab. was a drug. that articles used as food are articles used solely for taste. or nutritive value are “food”: When the statute defines ‘food’ as ‘articles used for food. . and was not “food” because it was not consumed primarily for taste. or (C). cure. treatment. but not exclusively. aroma. or prevention of disease in man or other animals.6 FDA actions against weight loss products marketed as starch blockers illustrate the scope of the “food” definition.C. the drug category is based on intended use. mitigate. aroma. treat. Inc. To hold . for taste. Specifically. § 321(f)). It has been interpreted to mean that articles used primarily. Application of Biotechnology for Functional Foods . Schweiker. not by way of its consumption as food. aroma. In other words. the product affected a bodily structure or function. v.7 Unlike food.A. cure.2d 335. The court reasoned that the product was intended to affect digestion. a bodily function. and (3) articles used for components of any such article (FFDCA § 201(f). aroma. The court agreed that the product. which contained substances that were claimed to inhibit enzymes necessary for starch digestion.C. A product is deemed to be a drug. or nutritive value. but through a pharmaceutical mechanism of action. v. or intended use to affect a bodily structure or function. in most relevant part. 6 7 Nutrilab. Inc. starch blocker products are positioned as dietary supplements.S. Key Definitions: “Food” and “Drug” Two key definitions form the basis for much functional food regulation: “food” and “drug. and 42 (C) articles (other than food) intended to affect the structure or any function of the body of man or other animals. “Food” is defined to mean— (1) articles used for food or drink for man or other animals. 21 U.” An understanding of these overarching regulatory categories is particularly important to appreciate the scope of permissible claims. 338 (7th Cir. In one such case. the term “drug” is defined in section 201(g)(1) of the FFDCA to include— (B) articles intended for use in the diagnosis. (B).

4 B.g.R. based on labeling claims made for a product. and other materials as appropriate. but a cosmetic bearing a structure/function claim would be regulated as a drug because there is no “cosmetic” exemption. hypertension). the “disease” prong of the “drug” definition is the primary factor determining whether a functional food will be regulated as a “drug. meaning that a product that appears to be a “food” but that is promoted for therapeutic purposes falls within the drug definition and can be regulated as such. because food obviously affects bodily structures and functions through nutrition. cardiovascular disease).” The line between “disease claims” that render a food a “drug” and structure/function claims can be difficult to discern.D. FDA has defined “disease” to mean “damage to an organ. scope Based on the statutory definitions described above.C. a food could bear a structure/function claim without FDA approval.” FFDCA § 201(g)(1)(C) (emphasis added). or state of health leading to such dysfunctioning (e. “Drug” 1. for example.2. Thus. Although this definition was adopted to aid FDA in the regulation of dietary supplement claims. The “food” and “drug” categories are not mutually exclusive. part.g. Examples include phytosterolenhanced rapeseed oil (i. (21 C. structure or system of the body such that it does not function properly (e.2 and III. it is commonly used in evaluating claims for conventional food as well.Intended use is determined objectively.93(g))9 FDA has further identified by regulation eight types of statements that would cause a product—whether positioned in the marketplace as a conventional food or as a supplement—to be regulated as a drug.e. 8 9 The definition specifically provides that the term “drug” includes “articles (other than food) intended to affect the structure or any function of the body of man or other animals.. Application of Biotechnology for Functional Foods ... Thus. instructions for use. canola oil) promoted for cholesterol lowering properties and yogurt promoted for probiotic properties that are suggested to prevent or treat infectious disease.F. § 101. a manufacturer may exercise substantial control over a product’s classification. These so-called “disease claim” categories are featured in Table III-2.g. Significantly. including the exemption for structure/ function claims that appear on food.. Rules for structure/ function claims for conventional food and dietary supplements are explained more fully in Sections III. the structure/function prong of the drug definition provides a broad exception for food.8 In other words. “Calcium builds strong bones”). advertising. To help identify impermissible disease claims that cannot be used on food.” excluding diseases resulting from nutrient deficiencies. by controlling claims made for a product. a food product is not subject to regulation as a “drug” merely because it bears a structure/function claim (e.

mitigate. Implications of classification In general. cure. products regulated as “drugs” are subject to greater regulatory scrutiny than food. 44 2. or through claims specifically authorized as outside of the drug category (e. using scientific or lay terminology Effect on disease through statements regarding product name. In contrast. drug regulation can be avoided through the use of general health and nutrition-related claims that do not implicate disease. treat. or prevent a disease or class of diseases “Herbal Prozac” “Use as part of your diet when taking insulin to help maintain a healthy blood sugar level” Has a role in the body’s response to a disease or a vector “Supports the body’s ability to resist infection” of disease Product treats. mitigate. formulation. and may bear only FDA-approved labeling. prevents. supportive scientific references. or use of the term disease example of Impermissible claim “Reduces the pain and stiffness of arthritis” “Support for Cystic Acne” “Reduces joint pain” Name: “Hepatacure” Formulation: Contains digoxin Scientific references: Serial Coronary Angiographic Evidence That Antioxidant Vitamin Intake Reduces Progression of Coronary Artery Atherosclerosis (if placement in labeling suggests disease use) Symbols: EKG Disease: “Prevent the onset of disease” Product belongs to a class of products that is intended to “Antidepressant” diagnose. All drugs must be produced in accordance with prescriptive good manufacturing practice requirements Application of Biotechnology for Functional Foods . must be demonstrated to be safe and effective for their intended use (generally through one or more well-controlled clinical trials).g. or mitigates adverse events “Help maintain intestinal flora for individuals on associated with a therapy for a disease and manifested by antibiotics” a characteristic set of signs or symptoms If a functional food is promoted using statements that FDA considers to be disease claims. symbols. New drugs must undergo a lengthy approval process before marketing. FDA-approved health claims).taBle III-2 disease claims that may trigger “drug” status type of claim Effect on a specific disease or class of diseases Effect on an abnormal condition associated with a natural state or process. if the abnormal condition is uncommon or can cause significant or permanent harm Effect on the characteristic signs or symptoms of a specific disease or class of diseases. or prevent a disease Product substitutes for a product that is a therapy for a disease Product augments a particular therapy or drug action that is intended to diagnose.. treat. it is subject to regulation as a drug. cure.

dairy products. sweeteners such as sucralose. and is consumed primarily for taste.C. two are of central importance. “food additive” is defined in pertinent part to mean— any substance the intended use of which results or may reasonably be expected to result. First. and is not increased to abnormal levels due to mishandling or other intervening acts.J.” it requires pre-market clearances from FDA on the basis of safety data and other information submitted in a food additive petition. Although not a legally defined term.11 If a substance is a “food additive. aroma. a product is likely to be regarded as a conventional food if the product is in the form of an “ordinary” food or beverage. Such products are subject to all the requirements applicable to food products generally. or any combination of the above. as a general rule. § 321(s)) The term does not include color additives and certain other substances such as pesticides. all food. treat. including plant-based products of modern biotechnology. and the kanamycin marker gene product. 10 If a poisonous or deleterious substance occurs naturally in a product. Application of Biotechnology for Functional Foods . 4 2. 289 (1982). § 342(a)(1))10 Historically. Second. L. directly or indirectly.S. A legal term of art. “conventional food” as used here denotes products in a traditional food form that are intended for consumption by the general population. a less stringent standard. nutrients such as folic acid. grains. or prevent disease. see Edward L. Of the food safety standards set out in the FFDCA. the only recombinant DNA product to be regulated as such (See 21 C. cure. food. Id. mitigate. the substance must not be “ordinarily injurious” to health.R. In contrast. including food ingredients. regardless of the regulatory category into which it may fall. beverages.C. including safety and labeling requirements enforced by FDA. scope In the absence of an expressed or implied intent to diagnose. Food in traditional form includes fats and oils used in food preparation. and generally enjoys greater flexibility in labeling and production. nuts. 21 U. legumes. including products produced through modern biotechnology. food ingredients must. is subject to pre-market review and clearances in more limited circumstances. be regulated “food additives” or be generally recognized as safe (GRAS) for their intended use or otherwise they and food products containing them are adulterated under the law.for pharmaceuticals.170). § 173. 37 Food Drug Cosm. must not bear or contain any added poisonous or deleterious substances that may render the food injurious to health. Implications of classification Safety. poultry. the statutory prohibition against added poisonous or deleterious substances has served as the primary tool FDA has used to ensure the safety of all foods.F. vegetables. Conventional Food 1. C. Food additives include preservatives such as BHA and BHT. Korwek. or nutritive value. (FFDCA § 402(a)(1). meat. 21 U. FDA Regulation of Biotechnology as a New Method of Manufacture. including the important distinction between food components and whole foods. 11 For a discussion of FDA’s authority to require pre-market clearances for food components.S. in its becoming a component of or otherwise affecting the characteristic of any food (FFDCA § 201(s).

§ 170. See 62 Fed. Fortification. Reg.3(i)).R. folic acid. including lutein. FDA broadly questioned the legal basis for using “botanicals and other novel ingredients” in conventional food. In order to achieve GRAS status. 1958. Indeed.Food substances that are GRAS are exempt from the definition of “food additive” and do not require pre-market clearances from FDA..” so many— if not most—of these ingredients are currently used in food on the basis of self-determined GRAS positions. phytosterols. FDA has cleared relatively few “functional” food ingredients as “food additives. for substances used prior to January 1. fish protein isolate. including those produced through methods of modern biotechnology. In guidance. 22984. The need for FDA clearance as a food additive or for the establishment of GRAS status therefore represents an area of caution for many functional food ingredients.F. The policy. there must be evidence supporting the safe use of the ingredient that is generally available (i. the nutrient or phytochemical would need to be either an FDA-cleared food additive or GRAS for its intended use. if a plant were modified to provide a particular nutrient or phytochemical. which is nonbinding in most circumstances. GAO recommended that Congress amend the FFDCA to require companies marketing functional food to submit advance notification to FDA of ingredients determined to be safe (GAO 2000). although FDA’s Center for Veterinary Medicine (CVM) has traditionally addressed the safety of feed ingredients through the issuance of so-called “no objection” letters. GRAS determinations can be made independently. Functional food derived from modern biotechnology is subject to the same FDA safety framework as other food. through experience based on common use in food. 1992)). 18937 (1997). without FDA input. FDA has accepted GRAS notifications for many functional ingredients. affording the agency an opportunity to object to. See infra section IV . CSPI has urged FDA to implement GAO’s proposal based on existing statutory authority (CSPI 2002). Under the GRAS standard. Those that are regulated food additives include Vitamin D.e. and inulin. Part 104). published) and accepted by experts qualified by scientific training and experience to evaluate food safety. however.12 For purposes of both GRAS determinations and food additive clearances. the safety of functional food ingredients has been a target of critics of the current regulatory framework. recommends a relatively narrow set of conditions under which FDA believes it is appropriate to add nutrients to 4 12 FDA has established a voluntary procedure for notifying the agency of independent GRAS determinations. Conventional food is subject to FDA’s fortification policy. GRAS notifications are not yet accepted for animal feed ingredients. FDA has advised that it is the transferred genetic material and the intended expression product that are subject to the food additive and GRAS standards (57 Fed. or “question” the status of a substance claimed to be GRAS. “safety” means that “there is a reasonable certainty in the minds of competent scientists that the substance is not harmful under the intended conditions of use” (21 C.C. Application of Biotechnology for Functional Foods . and amino acids.F. Reg. meaning that such products similarly must contain only components that are cleared food additives or GRAS. Thus. fish oil. asserting that many of “these added ingredients are not being used in accordance with an applicable food additive regulation and may not be GRAS for their intended use” (CSFAN 2001). a set of principles intended to serve as “a model for the rational addition of nutrients” to food (21 C. 22990 (May 29. In a letter to industry.R. a determination of safety may be based on scientific procedures or.

“authoritative statements”) of certain scientific bodies. with responsibility “for public health protection or research directly relating to human nutrition. United States. Labeling Generally. and in certain circumstances. or graphic matter (1) upon any article or any of its containers or wrappers. 335 U. or by accepting a health claim “notification” based upon an authoritative statement of a scientific body with responsibility “for public health protection or research directly relating to human nutrition. 345 (1948) (holding that the phrase “accompanying such article” is not restricted to materials that are physically on or in an article or package. 14 See. § 321(m))13 FDA considers a product to be “accompanied” by written. press releases.14 Thus. such as the National Academy of Sciences (NAS).g. Conventional food is permitted to bear statements regarding the relationship between a food and disease only if such statements are an FDA-authorized “health claim.14). that is dispositive to a finding that the material is “labeling”). or graphic material when the material supplements the product label or otherwise explains the product. or (2) accompanying such article” (FFDCA § 201(m).S.C. Kordel v. The claim will be permitted so long as the agency does not object to its use.S.C. Application of the policy to functional food is uncertain. which address “health claims. point-of-purchase signage. FDA has approved 12 health claims by regulation. Nutrition and health-related claims for conventional food are subject to prescriptive requirements under the FFDCA. the agency has characterized brochures. as “labeling” is generously defined to include “all labels and all other written.food. These requirements.” A party that wishes to base a claim on an “authoritative statement” must notify FDA of the claim at least 120 days before food bearing the claim is first introduced into interstate commerce.” apply to product labels and to “labeling. 15 The health claim notification process was created in 1997 to streamline the authorization of claims that are based upon findings (specifically. 21 U. as amended by the NLEA. printed.” A “health claim” is defined as— any claim that expressly or by implication characterizes the relationship between a food or a substance therein and a disease or health-related condition (FFDCA § 403(r). Health Claims. or by finding that the applicable statutory conditions have not been satisfied. internet pages.S. The policy does not address nontraditional substances that are not vitamins or minerals. the policy suggests that vitamins and minerals may be added to food to correct a recognized dietary deficiency or to restore nutrients lost in processing. even books and journal articles. pamphlets. 21 U.15 4 13 The same definition of “labeling” applies for all categories of FDA-regulated products.” “structure/function claims” and “nutrient content claims. as labeling. which it may do by issuing a regulation prohibiting or modifying the claim..R. and reasoning that it is the textual relationship between a material and a product. e.” The reach of these requirements is broad. For example. printed. not physical attachment. FDA may authorize health claims by issuing a health claim regulation. 21 C.F.” To date. § 343(r). § 101. and has authorized additional health claims through the statutory notification process. especially with respect to non-traditional nutrients. Application of Biotechnology for Functional Foods .

14(a)(5)). it lies closer to consensus.g.” If a statement does not identify both a specific food or substance and a disease or healthrelated condition. specific fruits and vegetables. not health claims.. for each health claim that is currently permitted. among experts qualified by scientific training and experience to evaluate such claims. or system of the body such that it does not function properly (e.R.F.” The “significant scientific agreement” standard requires a finding that “based on the totality of publicly available scientific evidence … there is significant scientific agreement. These criteria address the elements that must be present in each claim and Application of Biotechnology for Functional Foods 4 . § 343(r)(3)(B)).taBle III-3 select relationships that may Be the subject of Fda-approved Health claims Calcium and osteoporosis Dietary lipids and cancer Sodium and hypertension Dietary saturated fat and cholesterol and coronary heart disease Fiber-containing grain products. 1993). FDA considers such claims to be in the nature of general dietary guidance. hypertension) (21 C. that the claim is supported by such evidence” (FFDCA § 403(r)(3)(B).g. fiber). it is not a health claim. 2478. and vegetables and cancer Folate and neural tube defects Soy protein and coronary heart disease Plant sterol/stanol esters and coronary heart disease Whole grains and coronary heart disease and cancer FDA has advised that two elements must be present before a statement will be classified as a “health claim” (58 Fed. or a state of health leading to such dysfunctioning (e. structure. Reg. General dietary guidance claims are permitted so long as they are not false or misleading in any particular.” FDA has defined “disease or health-related condition” to mean “damage to an organ. such as apples) or a specific substance in a food (e. For example. 2479-80. First.C. First.g. § 101. fruits. Although “significant scientific agreement” does not require a consensus or incontrovertible evidence. the statement “diets rich in fruits and vegetables may reduce the risk of cancer” is not a health claim because it does not reference a specific food. Second. part. 21 U. Second. the statement must be reasonably understood to refer to a food (e... FDA traditionally has required that health claims be based on “significant scientific agreement. according to FDA. the statement must refer to a “disease or healthrelated condition. on a continuum between emerging science and consensus. the statement “5-a-Day for Better Health” as applied to fruits and vegetables is not a health claim because it does not reference a specific disease or health-related condition.g.. cardiovascular disease).S. Several important limitations restrict the circumstances under which health claims may be used. detailed criteria concerning the conditions for using the claim are spelled out in the relevant authorizing regulation or notification.

all health claims are subject to certain general principles set forth in FDA’s regulations. cure. cholesterol. The disqualifying levels for fat. and sodium are 13 grams.” “low saturated fat. Third. Oct. FDA. In all cases. companies wishing to use a health claim could convene an expert panel.. Qualified Health Claims.R. the agency must first consider whether a disclaimer could be used to convey the relative strength of the science and eliminate potential deception. § 101. FDA is not permitted to automatically reject health claims simply because the claims are not supported by significant scientific agreement. Application of Biotechnology for Functional Foods . § 101. 2006). the claimed health benefit must be described as a potential reduction in disease risk that may occur when the relevant food or substance is consumed as part of an appropriate diet. Specifically. in 21 C. and 480 milligrams. As proposed by IFT. food products that contain fat. 62400.16 As a result. Shalala. 164 F.e. 25. An example of a qualified health claim that FDA has allowed for conventional food and dietary supplements containing certain omega-3 fatty acids is 16 See. 62404. and heart disease must state that heart disease risk depends upon many factors. the authorized health claim concerning saturated fat. saturated fat. The new policy was prompted by a series of court decisions that held FDA’s implementation of the “significant scientific agreement” standard for health claims to violate First Amendment principles. respectively. 4 grams. FDA adopted an interim process that cleared the way for use of certain “qualified” health claims that are not supported by significant scientific agreement. and claim otherwise meets the general regulatory requirements for health claims.R.” and “low cholesterol” (21 C. cholesterol.3d 650 (D. however. 60 milligrams. Cir. to evaluate the science supporting the claim.F. IFT recommended that FDA streamline the agency’s review of health claims by adopting a notification procedure for claims that are “generally recognized as efficacious” (GRAE) (IFT 2005). cholesterol.the nature of the food that may bear the claim. or sodium at levels that FDA has deemed to be of nutritional concern are generally “disqualified” from bearing a health claim. Under its interim policy. FDA has interpreted the health claim provisions not to permit claims that a food or substance may treat. rather.C. Reg. In its call for reform of FDA’s regulation of functional food. scientific evidence supports the claim. e. In a 2003 policy change. If the panel found the claim to be GRAE. described by IFT as a GRAE panel. Pearson v. Perhaps the most significant of these is the “disqualifying” nutrient concept.F. the claim could be notified to FDA and used within 90 days if the agency did not object. or mitigate disease. 1999). saturated fat. claim is appropriately qualified to reflect the extent of the scientific evidence.75). For example. FDA will allow the claim through the exercise of “enforcement discretion” (i. in addition to the claim-specific criteria. does not believe it has the legal authority to adopt such a system (71 Fed.g.14. n Credible n The n The For claims that meet the criteria in the new health claim policy. an agreement to not take action against the claim).. FDA will entertain requests for the use of claims not supported by significant scientific agreement if the following conditions are met: n The 4 claim is the subject of a health claim petition that is filed for comprehensive review with FDA. and may appear only on food products that meet FDA’s definitions for “low fat.

” Structure/ function claims for conventional food do not require FDA approval or review. two other studies suggest that calcium supplements may reduce the risk. a draft guidance to address the scientific data and information that industry should have to substantiate structure/function claims as not false or misleading in any particular (FDA 2004). Levy.“Supportive but not conclusive research shows that consumption of EPA and DHA omega3 fatty acids may reduce the risk of coronary heart disease. However. For instance. the legal basis for structure/function claims is the “drug” definition in section 201(g)(1)(C) of the FFDCA. A food may bear. One serving of [name of food] provides __ gram of EPA and DHA omega-3 fatty acids.org) (accessed Oct. which defines “drug” in pertinent part to mean “articles (other than food) intended to affect the structure or any function of the body of man or other animals. has raised questions about its utility. Based on these studies. 2005) (available at http:// www. FDA’s implementation of the process. saturated fat. however.” Congress in effect exempted food bearing structure/function claims from the FFDCA’s stringent requirements applicable to new drugs. without FDA pre-market review or approval. or otherwise support that state of good health. the principles set out in the guidance apply equally to conventional foods. International Food Information Counsel (IFIC). Based on these studies.. however. As explained previously. An example of a “structure/ function” claim is “conjugated linoleic acid supports a healthy immune system.e. do not show that calcium supplements reduce the risk of preeclampsia during pregnancy. claims for which FDA agreed to exercise enforcement discretion) characterize the subject diet-disease relationship in a negative manner: “Three studies. structure/function claims must be written in a manner that presumes the intended audience is healthy and seeks solely to maintain. To avoid “drug” status based on disease claims. Derby and Alan S. FDA concludes that it is highly unlikely that green tea reduces the risk of prostate cancer” (FDA 2005b).18 17 See Brenda M.” By exempting food promoted for structure/function effects from the definition of “drug.. substantiated claims identifying the effect of the food on a bodily structure or function. including a large clinical trial. 2005).]” At the time of its adoption. 18 Although the draft guidance applies on its face only to dietary supplements. but another weak and limited study suggests that drinking green tea may reduce this risk.g. 0 Application of Biotechnology for Functional Foods . [See nutrition information for total fat. disqualifying criteria) for their use. nor has the agency identified specific criteria (e. Working Paper: Effects of Strength of Science Disclaimers on Communication Impacts of Health Claims (Sept. 2005).ific. several “approved” claims (i. In addition. promote. and cholesterol content. the qualified health claims process was both praised and criticized for increasing the flexibility of health claim regulation and making possible new claims based on emerging evidence. FDA concludes that it is highly unlikely that calcium supplements reduce the risk of preeclampsia” (FDA 2005a). “One weak and limited study does not show that drinking green tea reduces the risk of prostate cancer. Qualified Health Claims Consumer Research Project Executive Summary (Mar.17 Structure/function claims. consumer perception studies have called into question whether consumers are able to understand disclaimers intended to convey the strength of scientific evidence supporting a claim. FDA has issued.

e. Like health claims. Express nutrient content claims may be phrased in absolute or relative terms (i. the “other than food” language quoted above): because the statute exempts food intended to affect bodily structures/functions from regulation as a drug. a product may use these terms in food labeling only if the product meets the FDA definition and criteria for the term.S. These recommendations have been supported by groups such as CSPI (CSPI 2002). A claim that characterizes the level of a nutrient in a food.R. 20 See also CSPI Reports.S. 2006)).20 Nutrient content claims.19 For dietary supplements. such as “low fat” or “high in calcium. and have been the target of critics of functional food regulation.” is a “nutrient content claim. FDA’s reasoning for this position is based on the statutory exemption from “drug” regulation for food (i. such as cranberries or cranberry juice suggested to promote urinary tract health. nutrient content claims may be expressed or implied. Examples of FDA-approved nutrient content claims are provided in Table III-4. 21 U.. In other words. § 101. GAO suggests that conventional food marketers should be required to notify FDA regarding the use of structure/function claims. 21 U.C. 1 19 The IFT has asked FDA to replace the “nutritive value” concept with a more inclusive definition. and a food can affect a bodily structure or function only through nutritive value. The GAO has recommended that the FFDCA be amended to require marketers of functional food in conventional food form to follow the same procedures for structure/ function claims as dietary supplements (GAO 2000). and not a pharmacological effect.. and cannot be used unless authorized by FDA in a regulation or a nutrient content claim notification (FFDCA § 403(r). Reg.” A statement is deemed to “characterize” the level of a nutrient in food if it states or implies that the level has some nutritional significance (i. allowing structure/function claims based on either nutritive value or any “physical or physiological effect that has been scientifically documented or for which a substantial body of evidence exists for a plausible mechanism.e.” IFT Report. 62405 (Oct. Functional Foods: Public Health Boon or 21st Century Quackery? (March. Because structure/function claims can be made with relative ease (assuming adequate substantiation exists). FDA.For conventional food. § 343(r).e. has not clearly explained the meaning of nutritive value in this context. As a general rule.F. and appears to have moved away from a strict application of this standard in recent years. FDA has taken the position that structure/function claims must be based upon nutritive value.. Accordingly. who suggest that more stringent controls are needed.C. a structure/function claim should be tied to such nutritive value. § 343(r)(6)). 62400.13). they are commonly used on functional food products. 25. however. and to provide a disclaimer that claims made in labeling have not been reviewed by FDA. The GAO has also recommended that FDA develop regulations or other guidance concerning the scientific evidence needed to support structure/function claims. at 51. Application of Biotechnology for Functional Foods . DSHEA specifically authorizes the use of structure/function claims (FFDCA § 403(r)(6). In the agency’s view. FDA has “provided significant flexibility in determining whether a substance possesses nutritive value” (71 Fed. by comparing the nutrient value of one food to another). dietary supplements may bear structure/function claims without regard to nutritive value. suggests that there is “a lot or a little” of the nutrient relative to a dietary recommendation). As a result of DSHEA and FDA’s policy on structure/function claims for conventional food. 21 C. 1999). the “disease” provisions of the drug definition are of greater practical importance in assessing potential drug status.

In contrast. In addition to the requirements for nutrition and healthrelated claims detailed above.R. If a bioengineered food is significantly different from its conventional counterpart such that the traditional name no longer adequately describes the new food. if any. § 101. depending upon the nature of the modification. (2) the common or usual name of the food.. a nutrient generally may be the subject of a nutrient content claim only if the nutrient has an established reference intake. and the mandate for labels to reveal “material” facts about a food. for which there is no reference value. low sodium. An example is oil that is altered to contain significantly higher levels of unsaturated fats. or if none. such as “high oleic acid soybean oil. Contains. or a Dietary Reference Intake adopted by the Institute of Medicine or another authoritative body.3). either in the form of a daily value that may be used in nutrition labeling. in that order of preference (21 C. Fortified. two additional labeling considerations may have relevance to functional foods prepared using modern biotechnology methods: the requirement for food products to bear appropriate statements of identity. low calorie. or to contain “more lutein than conventional spinach. then a new product identity must be developed. spinach that is enhanced to provide increased lutein. 2 mg of lutein per serving). these statements should reflect (1) the name required by a federal regulation. in which case the product identity would highlight the predominant fats that are present.F. functional and other food produced through modern biotechnology may require unique product identity statements. Labels of conventional food products must bear appropriate statements of product identity. very low sodium.” no matter how much lutein the finished food contains. Labeling—Other Considerations. Accordingly. a peanut that is enhanced to provide “more” vitamin E than conventional peanuts may be promoted on this basis if the increased vitamin E satisfies FDA’s regulation for the use of “more” claims. Provides High in. reduced calorie Sodium free. because FDA has established a daily value for vitamin E.taBle III-4 select Fda-approved nutrient content claims Good source. Plus Antioxidant Light or lite Calorie free. 2 Application of Biotechnology for Functional Foods . Excellent Source of More. (3) an appropriately descriptive term. Extra.g. Thus. cannot be claimed to be a “good source” of lutein. By regulation. reduced sodium Sugar Free.” Where terms such as “high” are necessary to describe the basic properties of a food. The only nutrient content claim currently available to such a food would be a factual statement concerning the amount of lutein in the product (e. they are not subject to regulation as nutrient content claims. Added. Enriched. No Added Sugar Lean Of particular importance to many functional food products.

or authorized for investigation as a new antibiotic or biologic for which substantial public clinical investigations have been conducted. a food label must reveal all facts that may be material in light of representations made in labeling or in light of consequences that may result from its suggested or customary conditions of use. FDA has challenged several “functional food” products that attempted to combine a dietary supplement identity with attributes closely aligned with conventional food.” A dietary supplement is defined as— [A] product (other than tobacco) intended to supplement the diet that bears or contains one or more of the following dietary ingredients: (A) a vitamin.In addition. inhalation.  21 A dietary supplement also must not have been first certified as an antibiotic or licensed as a biologic. at 22991). must not have been first approved as a “new drug” or as an “investigational new drug” for which substantial public investigations were conducted. scope In 1994.” and must not be represented for use as a conventional food or meal replacement. (D) an amino acid. many dietary supplements have been marketed in the form of a bar or liquid beverage-type product without challenge. or combination of any ingredient described in clause (A). (C) an herb or other botanical. At the same time. or other routes of application that do not involve the gastrointestinal tract). Dietary Supplements 1. (B) a mineral. “Delicious and refreshing beverage”) or for conventional food uses (e.S. Congress amended the FFDCA to create a new regulatory category for “dietary supplements. as a salad dressing).g. § 321(ff)) In addition to containing one or more dietary ingredients.C. Consequences that are deemed material typically relate to substantial issues of safety or usage to which consumers should be alerted. (B). For example.. For example. (C). where promotional materials for the product emphasized taste and depicted the product being used with toast and other conventional food.. a dietary supplement must be intended for ingestion (as opposed to topical application. extract. metabolite. and consumers would not expect the allergen to be present based on the name of the food.21 The restriction on representations for conventional food uses means that a dietary supplement cannot be promoted as a product to be consumed primarily for taste or aroma (e. D. constituent. (E) a dietary substance for use by man to supplement the diet by increasing the total dietary intake. demonstrating possible overlap between the dietary supplement and conventional food categories.g. the presence of that allergen must be disclosed (57 Fed. Application of Biotechnology for Functional Foods . Reg. (D). must be expressly identified as a “dietary supplement. 21 U. or (E) … (FFDCA § 201(ff). if a modified fruit includes an allergen at a level that could trigger an adverse reaction. under 201(n) of the FFDCA. FDA rejected arguments that a spread fortified with phytosterols was a dietary supplement. or (F) a concentrate.

dietary ingredients are exempt from the definition of “food additive. nor are such ingredients subject to the GRAS standard.g. 2006)(holding that Congress “unambiguously” required FDA to conduct a risk-benefit analysis when assessing whether a supplement presents an unreasonable risk).” so pre-market clearance is not required. § 342(f)(1)). v. such as products that can only be consumed with conventional food (e. FDA determined a supplement to present an “unreasonable” risk if the risks posed by the product outweigh the substantiated benefits. For other products.3d.Functional food products and dietary supplements are often treated as distinct categories. Labeling requirements for dietary supplements are similar to requirements for conventional food in many respects. although FDA must be notified of the use of the claims. Labeling.23 As for pre-market review. Although FDA’s interpretation was initially rejected in a legal challenge. which the court found to require an accounting of whether the claimed benefits justify the risk. FDA must be notified of the basis for its safe use before it may be marketed (FFDCA § 413. For certain products. 459 F. 23 Id. 1994. the appellate court found the statute to impose two distinct safety standards that may cause a supplement to be adulterated: a “significant risk” of illness or injury.. 1038 (10th Cir. 4 Application of Biotechnology for Functional Foods . which must be determined to present a “reasonable certainty of no harm. nor is there an absolute prohibition against marketing what might be a functional food as a dietary supplement. Accordingly. it may not be desirable to refrain from marketing a food on the basis of its taste or flavor. there may be circumstances under which a “functional food” might be marketed as either a conventional food or a dietary supplement. and the claims must be accompanied by a disclaimer stating that— 22 Nutraceutical Corp. 2.S. such as those in bar and certain liquid forms. at 1040. 21 U. Like conventional food. which the court found to imply a “great danger. dietary supplements may bear nutrient content claims and health claims only if such claims are expressly authorized by FDA. Safety requirements for dietary supplements differ substantially from those for conventional food. In its 2004 rulemaking to ban ephedrine alkaloid supplements. 21 U. manufacturers have arguable discretion to position the products as either conventional food or dietary supplements.” however. Von Eschenbach. Dietary supplements also may bear structure/function claims. the agency’s approach was ultimately upheld on appeal.C. moreover. In contrast to conventional food ingredients. If a dietary ingredient is “new. As suggested above. 1033.S. unless the new dietary ingredient has been present in the food supply as an article used for food in a chemically unaltered form. Implications of classification Safety. spreads). A new dietary ingredient notification is required for any dietary ingredient that was not marketed in the United States before October 15. The safety standards for dietary supplements are widely considered to be less stringent than conventional food standards. there is no bright line test for distinguishing functional food from dietary supplements. § 350b).C. a dietary supplement position may not be feasible.” the safety standard for dietary supplements provides that dietary supplements must not present “a significant or unreasonable risk of illness or injury” under the conditions of use (FFDCA § 402(f)(1). and an “unreasonable” risk.22 In upholding FDA’s interpretation. Application of the “significant or unreasonable risk of illness or injury” standard requires a case-by-case assessment.

or prevent any disease (Id. regardless of whether such food also purports to be or is represented for general use (21 C. underweight.S. as follows: (i) Uses for supplying particular dietary needs which exist by reason of a physical. Among the food products retaining “special” usefulness are products relating to food allergy and sensitivities (e. § 105. all functional food—its reach has been gradually restricted over the years. FDA has taken the position that needs relating to general dietary guidelines are not “special dietary uses.  E. Although this language could. and overweight. be broadly interpreted to cover numerous food products—including.” “cholesterol protection formula. pathological or other condition.. 21 U. since advice concerning restrictions of these nutrients are applicable to the general public. it responds to the manufacturer in a so-called “courtesy letter” that explains the basis for the agency’s objection.3(a)(1)). including but not limited to the conditions of diseases. This determination demonstrates a unique aspect of special dietary use regulation—a food can lose its special dietary usefulness once a need related to a particular condition has broad applicability to the public at large. on its face. lactation.g.” and “helps maintain healthy blood sugar levels. and depression.C. dysphagia. and thus to be considered of general usefulness. § 343(r)(6)(C)). allergic hypersensitivity to food.g. and products intended for weight loss or weight management programs. Application of Biotechnology for Functional Foods ..” “support normal body function during the cold season. products relating to intake problems common to several diseases or conditions (e. Dietary supplement manufacturers are required to notify FDA of their intended use of structure/function claims. or difficulty swallowing). treat.R. Food for Special Dietary Uses 1. Two FDA interpretations have played a particularly important role in shaping the special dietary use category. (ii) Uses for supplying particular dietary needs which exist by reason of age. or other dietary property. This product is not intended to diagnose. If FDA objects to the notified claims. Any such particular use of a food is a special dietary use. pregnancy. convalescence.This statement has not been evaluated by the Food and Drug Administration. perhaps. “special dietary uses” is defined to mean— [P]articular (as distinguished from general) uses of food. Needs relating to the prevention of diet-related chronic diseases of broad concern seem most apt to fall into this category. cure. (iii) Uses for supplementing or fortifying the ordinary or usual diet with any vitamin. First.” Thus.” among numerous other claims deemed objectionable by FDA. physiological. including but not limited to the ages of infancy and childhood.” “treatment for anxiety. anorexia. Statements that have been the subject of courtesy letters include ”helps maintain healthy cardiac risk ratios. FDA has determined that most statements concerning nutrients such as sodium and sugars are most appropriately regulated under the nutrient content claim provisions of the FFDCA. § 403(r)(6)(C). scope Perhaps the most ambiguous category of all. mineral. gluten intolerance. peanut allergy).F.

In a few instances. Following NLEA.62). The agency has suggested that the exemptions apply only if the claim is made in compliance with a “specific provision” of the special dietary use regulations. Because the interpretation applies only to nutrients with a reference value. There is some promise that modern biotechnology may be used to produce food. however. necessary in order fully to inform purchasers as to its value for such uses (FFDCA § 403(j). In addition. FDA has provided by regulation that fortified food is deemed to be for a special dietary use if vitamins or minerals added to the food provide 50% or more of the applicable daily values in a single serving (Id. Labeling Generally.R.S. The following labeling information technically would be required under FDA’s regulation for hypoallergenic claims: n The n A  common or usual name and quantity or proportion of each ingredient. or other dietary properties as the Secretary determines to be. Implications of classification Safety. these products would be subject to FDA’s regulations for food represented to be for a special dietary use by reason of “the decrease or absence of any allergenic property” (21 C. it does not address many functional food products.9(a)(4)). Labeling—Hypoallergenic Food. The central requirement provides that a food for special dietary use is deemed misbranded unless its label bears information adequate to inform purchasers of its value for— such information concerning its vitamin. without allergenic proteins. but the agency has not done so to date. such as peanuts. and “informative statement of the nature and effect of any treatment or processing of the food or any ingredient thereof.F. and is made solely to note the special dietary usefulness of the product. The reach and practical application of these exemptions.Second. Food for special dietary use is subject to several unique labeling requirements. FDA suggested that it would initiate rulemaking to clarify the relationship between special dietary use claims and health and nutrient content claims. FDA has advised that statements that appear in labeling pursuant to a special dietary use regulation will not be subject to the agency’s requirements for health claims and nutrient content claims. n An Application of Biotechnology for Functional Foods . an infant food. the informational labeling requirement for special dietary uses is subject to interpretation on a case-by-case basis. § 343(j)). FDA has issued regulations addressing the labeling information that must be provided for food for special dietary use. or the name of each ingredient thereof. food produced through modern biotechnology and intended for use as a hypoallergenic food. In all other cases. If promoted as such.” if the changed allergenic property results from processing. or a weight loss product would be subject to specific labeling requirements as provided for in FDA’s special dietary use regulations. mineral.C. § 101. 21 U. however. to reveal the specific plant or animal that is the source of each ingredient. which may contain phytochemicals and other substances for which a reference intake has not been established. Like ingredients of conventional food. is unclear. / This determination appears to eliminate many fortified products from the special dietary food category. 2. and by regulations prescribes as. § 105. For example. qualification of the name of the food. ingredients used in food for special dietary use must generally be regulated food additives or GRAS for their intended use.

100-290 (amending 21 U. low fluid. or metabolize ordinary food or certain nutrients. has limited or impaired capacity to ingest. and only for patients receiving active and ongoing medical supervision. n A n Intended n Intended n Intended n Intended An example of a product positioned as a medical food is a moderate protein. as opposed to a naturally occurring food in its natural state. Now a distinct category from food for special dietary use. an impaired ability to metabolize the essential amino acid phenylalanine—and thus was reclassified as a distinct type of food for special dietary use.F. Pub. Implications of classification Safety.scope The term “medical food” has been defined to mean— [F]ood which is formulated to be consumed or administered enterally under the supervision of a physician and which is intended for the specific dietary management of a disease or condition for which distinctive nutritional requirements. § 101. The impetus for this reclassification was a practical one: FDA recognized the value that specialty formulas and other highly specialized products bring to a small but vulnerable population.9(j)(8)). in providing for the use of the food additive folic acid in food.S. which is intended for the dietary management of PKU. Initially regulated as a drug. based on recognized scientific principles. Lofenalac® was recognized by FDA to meet distinctive nutritional requirements—namely. because of therapeutic or chronic medical needs. / As noted previously. for the dietary management of a patient who. particularly the first two. L. Medical Food 1. product intended for the partial or exclusive feeding of a patient by means of oral intake or enteral feeding by tube. 21 C. low electrolyte. Their application to food produced using modern biotechnology would need to be assessed on a case-by-case basis. these requirements. or who has special medically determined nutrient requirements that cannot be achieved via modification of the diet alone. ingredients of medical food must generally be regulated food additives or GRAS for their intended use. seem more naturally suited to multiple-ingredient. and did not wish to discourage the development or marketing of such products by subjecting them to stringent requirements for drug products. F. Application of Biotechnology for Functional Foods . Like ingredients of conventional food. high calorie formula intended to provide balanced nutrition for dialyzed patients with chronic or acute renal failure. to provide nutritional support for the management of unique nutrient needs that result from a specific disease or condition.C. absorb. 1. No. FDA specifically authorized its use in medical food products. FDA has provided by regulation that a product can be classified as a medical food only if the product is— n A  specially formulated and processed product. the first product recognized as a “medical food” by FDA was the infant formula Lofenalac®. medical food is defined in an extremely narrow manner. for use under medical supervision. as determined by medical evaluation. are established by medical evaluation (Orphan Drug Amendments of 1988. For example. digest.R. 360ee).Adopted many years ago. processed food than single-ingredient commodities produced through modern biotechnology. Specifically.

“Poultry products” include human food products made from the carcass of any domesticated bird. and in a form of the type generally considered to be a product of the meat or poultry industries. FSIS has approved the use of many nutrient content claims by regulation (e. swine.. FSIS’ limits on fortification would generally be expected to limit the marketing of “functional” meat or poultry products. Food suBject to addItIonal or unIQue reQuIrements: meat and poultry. As a practical matter. A functional food would be subject to regulation by FSIS if it is meat or poultry. FSIS generally takes the position that it must specifically approve the use of all meat and poultry product ingredients and labels. sHell eggs and egg products.g. but has issued no regulations or formal policies concerning health claims or structure/function claims. It has permitted certain meat and poultry products to contain FDA-regulated ingredients that are fortified in accordance with FDA requirements (e.. Although these exemptions make a medical food positioning extremely attractive. medical food products are expressly exempt from FDA requirements for nutrition labeling. including products to which functional ingredients are added through genetic means or product formulation. For example. Although FDA and FSIS requirements are similar in many respects. Application of Biotechnology for Functional Foods  . certain products are subject to additional or unique requirements under the FFDCA and other laws. and health claims. FSIS has historically taken the position that meat or poultry. IV. Complicating the situation. few functional food products will be in a position to be marketed as a medical food in substantial compliance with the FFDCA. if the actual fortification occurs outside of a FSISinspected meat or poultry plant. FSIS has allowed the labels of appropriate products to bear the American Heart Association Heart Check program logo and the FDA-approved health claim concerning dietary saturated fat and cholesterol and risk of coronary heart disease. and non-human animal food. per se. and has permitted such claims under limited circumstances. shell eggs and egg products. enriched pasta). is not suitable for fortification. Of particular relevance to functional food. A. A comprehensive discussion of such additional or unique requirements is beyond the scope of this report. A “meat food product” includes any product capable for use as human food made from the carcass of cattle. FSIS requires that all ingredients used in meat and poultry products be specifically approved as safe and suitable. Thus. “low fat. sheep.Labeling. greater than 2 to 3%). or it is a product that contains a meat or poultry ingredient. As a general rule. IncludIng pet Food Although the FFDCA provides a broad framework that is of central importance to most functional food products. respectively.” “lean”). “Meat Food Products” or “Poultry Products” Products that are “meat food products” or “poultry products” are regulated by the Food Safety and Inspection Service (FSIS) of USDA pursuant to the Federal Meat Inspection Act (FMIA) and Poultry Products Inspection Act (PPIA).g. the criteria detailed above reveal that FDA has interpreted the category in a very narrow manner. As is the case with other food under the FFDCA. nutrient content claims. meat and poultry products cannot be adulterated or misbranded under the FMIA and PPIA. and anImal Feed. and that most labels (and claims) be reviewed and approved before use. FSIS has evaluated labels containing proposed health claims and structure/function claims on a case-by-case basis. including pet food. or goats. In recognition of their unique status. a brief overview of three regulatory schemes of particular interest to functional food products is provided below: meat and poultry products. at more than a de minimis level (generally.

such as the omega-3 fatty acid. FDA concluded that certain uses of DHA and EPA are safe and lawful. the effect on the characteristics and cooking properties of the resulting meat. Thus. at elevated levels. though not reaching a conclusion on the issue. but would likely require a specific statement from the agency). An “egg product” is defined to mean “any dried. Apr. shell eggs enhanced in a particular way would be regulated as either conventional food or food for special dietary use. frozen. if DHA content is enhanced through the addition of DHA ingredients to animal feed.  24 See. For example. and must use labels and formulations that are reviewed and approved by that agency. Shell eggs are subject to the FFDCA provisions concerning safety and labeling.” including egg substitutes and “dietary foods.g.24 Additionally. egg products must be produced under continuous inspection by FSIS. depending on the types of alterations and claims that are made.0 g per person per day. B. § 590. FDA bears primary responsibility for the safety and labeling of shell eggs for consumer use under the FFDCA and the Public Health Service Act. the targeted and desirable component that differentiates such products from other eggs. pursuant to the EPIA. If presented with a transgenic functional food. FDA. with possible consultation with FDA. FSIS would be expected to follow its usual pre-market approval approach. 5. and the product’s labeling. must be safe under the FFDCA. excepting products which contain eggs only in a relatively small proportion or historically have not been … considered by consumers as products of the egg food industry” (9 C.5)25 Like shell eggs. or liquid eggs. At the same time. e. as well as a review of whether the enhancements constitute impermissible “fortification”). provided that daily intakes of DHA and EPA from conventional food and dietary supplements do not exceed 3. due to denial of the petition on other grounds). without or without added ingredients. and thus must meet the same basic requirements as all other FDA-regulated food. the product’s safety (regarding which USDA would defer to FDA.” The scope of the FSIS exemption for “dietary foods” is unclear.. such as liquid eggs. nutrient content claims. the agency would likely engage in a review of the suitability of such enhancements (e.g. docosahexaenoic acid (DHA). FSIS has exempted several foods from regulation as “egg products. egg products must comply with the safety and labeling requirements of the FFDCA. the production and labeling of such products. The EPIA scheme for egg products results in greater oversight of this industry segment.R. a shell egg that contains a functional component. Because eggs are most reasonably classified as conventional food..F. including substantiation for any claims made. Shell Eggs and Egg Products Jurisdiction over shell eggs and egg products is shared by FDA and USDA. however. Application of Biotechnology for Functional Foods . such as a ground beef product promoted as containing a favorable fatty acid profile due to genetic enhancement. In a recent letter announcing the agency’s intent to exercise enforcement discretion and permit a health claim concerning the omega-3 fatty acids DHA and EPA. is also subject to USDA oversight pursuant to the Egg Products Inspection Act (EPIA). are similarly subject to regulation by FDA. the labeling of such a product is subject to FDA’s rules for health claims. Letter Regarding Eggs with Enhanced Omega-3 Fatty Acid Content and a Balanced 1:1 Ratio of Omega-3/Omega 6 Fatty Acids and Reduced Risk of Heart Disease and Sudden Fatal Heart Attack (Docket No. 2004Q-0072. FDA’s Center for Veterinary Medicine (CVM) would likely assert that a separate basis for use of DHA-enriched feed would need to be established and provided for the review of that Center. and other labeling statements. FDA would likely take the position that DHA. 2005) (assessing the petitioner’s assertion that eggs enriched with omega-3 and omega-6 fatty acids were GRAS. must be either GRAS or specifically cleared by FDA as a food additive for its intended use. 25 By regulation. Processed egg products. As discussed below.FSIS has not yet taken a public position concerning the status of transgenic animals that are intended to produce functional food in the form of meat or poultry for human consumption.

but only after review and approval. as a general rule. is subject to quite different regulatory requirements. Animal Feed. nor does the DSHEA framework for dietary supplements. CVM has indicated that it would not object to the marketing of nutritional supplements for companion animals in limited circumstances. Another important difference between CFSAN and CVM regulation concerns the application of the “drug” definition. 17706 (1996). including pet food. “production” claims such as increased milk production. although in common usage often non-human animal food is distinguished as “animal feed.C. including pet food.26 Other differences can be explained in large part by the different Centers within FDA responsible for the regulation of human and other animal food products and their interpretation of the law: human food is regulated by FDA’s Center for Food Safety and Applied Nutrition (CFSAN). while other animal food is regulated by FDA’s Center for Veterinary Medicine (CVM). falls within the definition of “food” under the FFDCA. Reg. it is generally allowed for use in feed.27 feed ingredients that have been evaluated by CVM are typically the subject of so-called “noobjection” letters. states require such food. the regulatory requirements for using functional ingredients is typically greater for feed products than for human food products because new feed ingredients must be proven to be safe and to have utility to FDA’s satisfaction before a “no objection” letter can be obtained. routinely take the position that a substance cannot be used in feed unless it has been formally or informally evaluated by FDA. Some of the differences are statutory: for example. as well as state feed control officials. Most. Regarding safety. regulated food additives or GRAS for their intended use. In contrast. and subject such feed to oversight under state laws similar to the FFDCA but that require registration or licensing of feed. Application of Biotechnology for Functional Foods . Examples of a functional feed are those formulations that contain phytase.” Animal feed. Examples include “urinary tract health” and “dental health” claims on cat food products (CVM 2004). CPG 7126. thus. to be manufactured or distributed within their borders pursuant to a registration or license. 03/1995). In the human food industry. CVM. feed containing such unapproved new drug ingredients cannot be legally marketed. CVM has allowed limited claims for such feed after having issued no objection letters for a few phytases that are produced through the use of recombinant DNA methods. Including Pet Food All food. increased leanness. improved growth. As a result of CVM and state oversight. and there is no formal “medical food” category for feed products. the FFDCA and state feed laws require that substances added to food either be. Accordingly.100)(Rev. CVM has permitted structure/function feed claims in limited instances. CVM treats most structure/function claims as drug claims. if not all. especially the “structure/function” exception for food. however. including pet food. FDA Compliance Policy Guides. 27 See supra note 21. self-determinations of GRAS status are common. and efficiency of weight gain are viewed as drug claims and feed ingredients with such claims are subject to regulation as unapproved new animal drugs (CVM 1998). Oversight of feed by state feed control officials provides yet another explanation for the disparate regulation. Once an ingredient is favorably reviewed by CVM. under state law. 0 26 See 61 Fed. CVM has issued no regulations to govern special dietary uses for feed products. Since very few ingredients are cleared as feed additives and since CVM does not have a GRAS notification approach.04 (sec. the NLEA scheme for health and nutrient content claims does not apply to animal feed. 690. and are the basis upon which many food ingredients are used.

In summary. in a different manner. 1 Application of Biotechnology for Functional Foods . limits the circumstances under which functional feed ingredients may be marketed without their being treated as new veterinary drugs. CVM’s prescriptive approach to functional feed. and dietary supplements. including functional ingredients. resulting in distinct safety and labeling requirements for feed products. coupled with the lack of a feedspecific framework for nutrient content claims. CFSAN and CVM approach functional food. health claims.

Application of Biotechnology for Functional Foods .

the complexity of allergenic responses. per se. differences in sensitivity among people. improved protein content and quality for better human and animal nutrition.Summary The application of biotechnology to foods expressly to improve nutritional and health characteristics holds great potential. Food enhancements cover a wide range. If a functional food product is marketed for a therapeutic purpose (e. dietary supplement. however. the safety and labeling standards it must meet. and reduction in anti-nutritional substances that diminish food quality and can be toxic.” If a product is subject to regulation as a “food. but functional food products are clearly subject to federal regulation. must meet safety standards requiring a Application of Biotechnology for Functional Foods  . Although biotechnology has made some advances in reducing allergenic proteins in some foods. How a product is categorized has substantial implications for how it is marketed. Conventional food.. or medical food. There is no regulatory scheme for functional food. including improved fatty acid profiles for more heart healthy food oils. including food ingredients. food for special dietary use (including infant formula). to treat a disease).” it may be further classified as a conventional food. once regulatory clearance has been obtained. such as structure/function claims. which need not be presubmitted to FDA nor accompanied by a disclaimer. Just what form those regulatory clearances may take is not entirely certain. and what requirements it must address for regulatory review and clearance or approval. It appears that food oils with improved fatty acid profiles are the closest to reaching commercialization. and enjoys some flexibility regarding certain types of claims. A conventional food may be freely marketed on the basis of taste. gossypol and glycoalkaloids also have potential to enhance the safety of human foods.g. increased vitamin and mineral levels to overcome widespread nutrient deficiencies throughout the world. Improvements in the digestibility of animal feed through the reduction of phytic acid. as demonstrated by many research accomplishments. it will be subject to regulation as a “drug. and the presence of multiple allergens in a single food make this challenge particularly daunting. On the horizon are efforts to increase the concentration of various antioxidants and functional substances such as phytosterols and probiotic bacteria. again depending upon its intended use and other factors.

in particular. subject to distinct regulatory requirements and oversight that may limit functional food opportunities as a practical matter. has been regulated by CVM in a manner that restricts the types of ingredients and promotional claims that may be used. Food for special dietary use enjoys some flexibility as to nutrient content and health claim requirements.g. are theoretically eligible for marketing in a functional food form. however. egg products. and animal feed. Certain food products.. 4 Application of Biotechnology for Functional Foods . as a bar or liquid). and of the special dietary use category itself. Animal feed. and in some circumstances. but may challenge the boundaries of some of the regulatory classifications. but the extent of this flexibility.” The safety standard for dietary supplements is less stringent. but is permitted in extremely narrow and carefully defined circumstances. and can make structure/function claims only if such claims are submitted to FDA and accompanied by the DSHEA disclaimer. despite the statutory classification of animal feed as “food.“reasonable certainty of no harm. but supplements are restricted to marketing in certain forms (e. as a tablet or powder. including pet food. including meat and poultry.” The use of modern biotechnology to enhance human and other animal food will likely not change these regulatory paradigms. Medical food is entitled to the most flexibility of all. is unclear and probably very fact-specific. Such products are.

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Application of Biotechnology for Functional Foods

Application of Biotechnology for Functional Foods .

Application of Biotechnology for Functional Foods .

Application of Biotechnology for Functional Foods .