Food Preservation with

Biotechnology

Lesson 9
Objectives:
 define the term "biotechnology"
 describe the beneficial role that microorganisms have in the
production of fermented foods and of food ingredients or
additives
 outline the sequence of microbiological, chemical and physical
changes that are involved in the conversion of milk to cheese
 compare and contrast the terms "biotechnology-derived foods"
and "genetically modified foods", and describe the position of
the Canadian government on these foods
 articulate a personal set of values pertaining to use of
biotechnology in foods
 Definition of biotechnology

"application of science and engineering in direct or indirect

use of living organisms or their parts in their natural or
modified forms”
(Canadian Environmental Protection Act)

◼Broad spectrum of tools and techniques:

◼ Fermentation (use of microorganisms, enzymes)
◼ Plant and Animal Breeding
◼ Cell and Tissue Culture
◼ Genetic Engineering
 desirable products of biotechnology:
FOOD PRODUCTS INGREDIENTS AND ADDITIVES
obtained by biotechnology obtained by biotechnology
fermented beverages amino acids (e.g. methionine,
(eg. wine, beer, sake) glutamic acid)

fermented dairy products biopolymers (eg. xanthan gum,

(eg. yogurt, specialty cheeses, alginates)
cheddar cheese)

enzymes (eg. chymosin or rennin)

fermented meat products
(eg. salami, bologna, prosciuto)
Vitamins (B-vitamins)
traditional fermented foods
(eg. sauerkraut, soy sauce, tempeh,
idli, yakult, miso)
 Is food fermentation the same as
biotechnology?
 the word biotechnology -new term,
 practiced in the agriculture/food industries for 1000s yrs
◼ yogurt production in India, bread in Egypt.

Food fermentation vs food biotechnology

“a new name for an old process…”
 Modern Food biotechnology is far more than just food
fermentation
◼ probiotics, genetically modified organisms for
production of ingredients,……
 Preservation principle:
◼ Microbial Antagonism
⚫ Suppress growth & metabolic activities of spoilage-
causing microflora
◼ Metabolites: End products of fermentation
⚫ antimicrobial compounds (eg. acids, alcohols)
⚫ Acids (formed in situ) can also lower pH of food

may need additional methods for preservation

• eg. refrigeration or pasteurization or vacuum packaging
 Benefits of fermentation

❖ Increased Nutritional value & availability

⚫ more nutritious than the starting materials
⚫ Mould fermented foods (B-vitamins)
⚫ Microbial hydrolysis of cellulose materials that are indigestible by
humans
⚫ Liberate nutrients from plants (otherwise indigestible in human GI
Tract)

❖ Add Variety to our food supply/diet (Table 9.1)

See Table 9.1

Microorganism Food commodity Fermented product

Lactic acid bacteria Cucumbers Dill pickles, sour pickles

Olives Green olives, ripe olives
Cabbage Sauerkraut
Coffee cherries Coffee beans
Vanilla beans Vanilla
Meat Meat sausages (salami)
Dairy Sour cream, yogurt

Malt Beer, ale, stout

Yeasts
Grapes (Fruit) Wine, vermouth
Wines Brandy
Rice Saké
Bread dough Bread

Mould Soybeans Miso, soy sauce , some cheese

 Table 9.2 Microbial-induced changes
raw material → fermented product
Agent Raw material Product

Lactic acid bacteria Lactose, glucose Lactic acid

Yeasts Glucose, other Ethanol

fermentable
carbohydrates
Acetic acid bacteria Ethanol Acetic acid

Microbial lipases Lipids Free fatty acids, aldehydes,

ketones
Microbial proteinases Proteins Polypeptides, peptides, free
amino acids, amines
(cause textural and flavour
changes in food)
Mould cellulases Cellulose Oligosaccharides, glucose
 Factors affecting fermentation

◼ Starter culture
details in Lesson 9

◼ Formation of metabolites

◼ Temperature

◼ Oxygen

◼ Salt
 Starter cultures
◼ Specifically selected microbial cultures

◼ Desirable traits for the particular fermentation

◼ metabolism leads to desired products
 eg. acids, alcohol, flavour compounds

◼ dominant microorganism
Starter cultures
1. as a “pure” culture (eg. Yeast in bread)
2. from a previous batch of the
fermented food containing that culture
(eg. sourdough or yogurt culture)
3. part of the normal microflora (eg.
Sauerkraut, olives)
 Temperature

◼ starter cultures will only grow at their optimum temp.

◼ eg. Sauerkraut flavour- proper succession of lactic acid

bacteria
 Oxygen

◼ Fermenting MOs have different O2 requirements

◼ baker's yeast (Saccharomyces cerevisae)- grows better under

aerobic conditions

◼ however, the yeast ferments sugars more rapidly under

anaerobic conditions
 Salt
◼ Favours growth of Lactic acid-producing bacteria

◼ Inhibits growth of spoilage /disease-causing MOs

◼ Synergistic effect between acid and salt
 more inhibitory to disease- and spoilage-
causing MOs
 Example of Technology of Fermentation
or Biotechnology of cheese…
◼ Cheese making Process (Figure 9.1)
Video on cheese processing (6 minutes) and
notes in Lesson 9

◼ Why is the milk pasteurized before starter culture is added?

◼ In addition to starter culture, what else is added to coagulate
the milk → curd?
◼ Note the following terms:
◼ curd, matting, Cheddaring, milling, pressing
◼ How is the whey removed from the curd?
◼ What are the roles of heat, pressure, salt, ripening (aging) in
the production of cheese?
Milk (10 kg)
(Fat, casein, lactose, minerals)

Pasteurization

Starter

Enzyme

Fermentation

Curd formation
 Curd formation

Curd Whey

Matting , cheddaring curd pcs. fuse together form

rubbery slab
salting

Pressing

Ripening

Packaging (1 kg)
Critical Thinking Question
 Why is the milk pasteurized before starter
culture is added?

 Can unpasteurized (raw) milk be used?

 What is cheddaring?
What aspects of cheese (making) result in
preservation? (longer shelf life compared to milk)

 microbial antagonism
 Lactic acid bacteria → Acidic conditions (metabolites)
 Lactic acid bacteria → microbial antagonism
 Moisture content and Water activity

 Salt
◼ Lowers water activity
◼ Inhibits the growth of proteolytic microorganism
 What aspects of cheese making result in
different varieties of cheese?

Differences between cheeses

◼ Milk - pasteurized or not; cow, goat, sheep ….

◼ Method for curd formation (acid, enzyme, or acid-heat

coagulation).
◼ Specific starter cultures, Time, temp of cooking
(affects moisture content)
◼ Washing, matting etc, Pressure

◼ Ripened (second starter cultures, enzymes) or not

◼ Acidity
Virtual Cheese Tasting
 Examples of different cheeses –
fermentation using different starter cultures

Examples of hard cheeses

⚫ Cheddar cheese – lactic acid bacteria
⚫ Swiss cheese –
⚫ lactic acid bacteria and
⚫ propionic acid bacteria (Propionibacterium shermanii)
⚫ carbon dioxide – “eyes”
⚫ proline – sweet
⚫ propionic acid- nutty flavour
 Examples of different cheeses –
fermentation using different starter cultures

Example of a semi-soft cheese

⚫ Blue veined cheese –

⚫ Blue cheese (U.S.), Stilton (England), Gorgonzola
(Italy): cow’s milk

⚫ Roquefort (France): sheep’s milk

 Examples of different cheeses – fermentation
using different starter cultures

⚫ mould (Penicillium roquefortii) grow throughout the curd

⚫ Curd is needle-inoculated with the mould (making

‘channels’)

⚫ blue colour

⚫ distinct flavours from action of lipase

⚫ Free fatty acids, aldehydes & ketones

⚫ Sharp, peppery odour and flavour

Blue-veined cheese
 Examples of different cheeses –
fermentation using different starter cultures

Examples of soft cheeses

⚫ Camembert and Brie cheese

⚫ Mould-ripened cheese–
⚫ surface inoculated with mould (Penicillium camembertii)
⚫ P. candidum is sometimes used instead
⚫ Curds are not pressed
⚫ Sprayed with mist of mould spores
⚫ mould mycelia – outer layer (surface)
⚫ action of proteases
⚫ Small molecular weight peptides (charac. flavour)
⚫ soft/creamy texture
Camembert cheese
Virtual Cheese Tasting
What to remember
 Type (mould, bacteria, …….) and name of
secondary starters and their metabolites,
 Sensory features
◼ E.g holes, sweet taste, nutty taste in Swiss Cheese
◼ Blue veins, sharp taste in blue cheese
◼ Velvety appearance, soft texture in Camembert
and Brie
Genetically Modified Organisms, Novel
Foods and Biotechnology-Derived Foods
What are GMOs (genetically modified organisms) ?

Plants, animals and MOs -there is a change to the

heritable trait(s) by intentional manipulation:

e.g. modern gene technologies

A foreign piece of DNA is inserted into the genetic

material of the host organisms.
What is the relationship between the terms biotechnology,
genetically modified organisms (GMO) and genetic
engineering (GE)?

 Biotechnology refers to a broad range of

techniques of which GMO is one example.
 Within the term GMO is also more
specifically the technique of GE.
Use of Biotechnology in Cheese Making

Milk (10 kg)

calf or adult cow In cheese production,
rennet (contains genetic engineering may be
Pasteurization used to:
chymosin or rennin)
(a)improve the starter
or microbial rennets cultures
(rennin-like Starter
substitutes) (b)produce the coagulating
enzyme
or microbially Enzyme
produced chymosin

Fermentation

Curd formation
 Biotechnology (GM microorganisms) in
cheese production
 Genetically engineered starter cultures
◼ Cultures with improved resistance to viruses, or improved
enzyme activity

 Recombinant chymosin
◼ MOs genetically engineered to include the gene for producing
the enzyme chymosin or rennin, which is originally from the
fourth stomach of calves or lambs (or cows, sheep, goats)
◼ Approved as a food additive that may be used as a food
enzyme
Food and Drug Regulations (Division 8 – dairy)
Ingredient Common name

1 or more species of bacteria Bacteria culture

1 or more species of mould Mo(u)ld culture

Preparation containing rennin Rennet

Milk coagulating enzymes from Microbial Enzyme

Aspergillus oryzae RET-1 (pBoel777),
Endothia parasitica, Mucor miehei
(Cooney and Emerson) or Mucor
pusillus Lindt
Food and Drug Regulations –
enzymes allowed in cheese
Milk coagulating enzymes derived from Mucor miehei
…., aminopeptidase derived from Lactococcus lactis,

Chymosin-A derived from Escherichia coli … Chymosin

B derived from Aspergillus niger … or Kluyveromyces
marxianus var lactis

e.g. of regulation:
“Lipase and proteases for …. (name of) cheese”

“Lysozyme derived from egg white”

Microbial
enzyme
Rennet and/or
microbial
enzymes
Rennet and/or
pepsin and/or
microbial
enzyme
Terms to remember
 Biotechnology  Curd, whey
 Genetic engineering  Lactic acid bacteria
 Aerobic, anaerobic  Propionibacterium
 Microbial antagonism shermanii
 Antimicrobial agents  Penicillium roquefortii
 Starter cultures  Penicillium
 Rennin, chymosin camembertii
 Rennet  Penicillium candidum
 Cheddaring