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Chapter 5

Fermented foods

5.1. Introduction
5.2. Microbial ecology of fermentation
5.3. Fermented food products
1. Introduction
• The term fermented foods is used to describe a
special class of food products characterized by
various kinds of carbohydrate breakdown.
• Fermented foods may be defined as foods which
are processed through the activities of MOs but
in which the weight of the microorganisms in
the food is usually small.
• The influence of microbial activity on the nature
of the food, especially in terms of flavor and
other organoleptic properties, is profound.
• Fermentation is a technology that utilizes the
growth and metabolic activities of
microorganisms for the preservation and
transformation of food materials.
Cont’d…
• During food fermentation, the growth of spoilage and
pathogenic organisms is inhibited by the metabolites
generated by the fermenting organisms, thereby
extending the shelf life of perishable produce.
• For instance, during lactic acid fermentation, lactic acid
bacteria synthesize metabolites such as lactic acid, acetic
acid, CO2, ethanol, H2O2, bacteriocins, and antimicrobial
peptides, which synergistically suppress the survival and
growth of pathogenic and spoilage microorganisms.
• Besides preservation, fermentation imparts characteristic
aroma, flavor, texture, and nutritional profile into food.
• Thus, although ancient civilizations developed
fermentation primarily as a way of preserving perishable
agricultural produce, the technology has evolved beyond
preservation into a tool for creating desirable
organoleptic profiles in foods and improving their
palatability.
Cont’d…
• Bread is a classic example for this case, where the
primary function of dough fermentation is to
create the characteristic structure, texture, and
organoleptic profile of bread after the baking
process.
• Fermentation also helps to remove anti-nutritional
factors and toxins in food materials and improve
their nutritional profile.
• For instance, fermentation of soybean into
products such as tempeh, natto, and soy sauce
leads to reduction of anti-nutritional factors such
as phytic acid and trypsin inhibitors and results in
the hydrolysis of complex soy proteins into more
digestible and bioavailable peptides and amino
acids.
Table Examples of fermented foods
Cont’d… food fermentation processes can be
• Traditional
broadly classified into:
lactic acid fermentation,
fungal fermentation, and
alkaline fermentation.
• Examples of lactic acid fermented products, i.e.,
products primarily fermented by lactic acid
bacteria, include yoghurt, sausages, cheese,
sauerkraut, and kimchi (fermented and spiced Napa
cabbage from Korea).
• Yeast spp. are also involved in the fermentation of
many of the lactic acid-fermented products,
including:
Kefir: a slightly alcoholic dairy beverage from the
Caucasus,
Kombucha: a fermented sweetened tea from China.
Cont’d…
• Most of the well-known soy-based fermented foods
from Asia such as tempeh and soy sauce are
produced by fungal fermentation, except natto,
which is produced by alkaline fermentation.
• Industrial fermentation processes use either
submerged or solid-state bioreactors that are
operated in batch or continuous mode.
• Most food fermentation processes from sauerkraut
and kimchi to miso and tempeh use SSF processes
operated in batch mode, where microorganisms are
cultivated on the surface of a water-insoluble
substrate.
• SmF processes are used in the production of
yoghurt and other dairy-based beverages, alcoholic
beverages, and food condiments such as vinegar.
Cont…
• Most fermented foods contain a complex mixture of
carbohydrates, proteins, fats, and so on, undergoing
modification simultaneously, under the action of a
variety of microorganisms.
• This creates the need for additional terms to
distinguish between major types of change.
• Those reactions involving carbohydrates and
carbohydrate-like materials (true fermentations) are
referred to as "fermentative."
• Changes in proteinaceous materials are designated
proteolytic or putrefactive.
• Breakdowns of fatty substances are described as
lipolytic.
• When complex foods are "fermented" under natural
conditions, they invariably undergo different degrees
of change.
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Cont…
• Fermented foods are influenced mainly by
the:
nature of the substrate and the MOs involved,
length of the fermentation and
treatment of the food during the processing.
• In specific food fermentations, control of the
types of microorganisms and environmental
conditions to produce desired product
characteristics is necessary.
• Fermented foods have been known from the
earliest period of human existence, and exist
in all societies.
ble: History and origins of some fermented foods

Food Approximate year Region


of introduction

Mushrooms 4000 BC China


Soy sauce 3000 BC China, Korea, Japan
Wine 3000 BC North Africa, Europe
Fermented milk 3000 BC Middle East
Cheese 2000 BC Middle East
Beer 2000 BC North Africa, China
Bread 1500 BC Egypt, Europe
Fermented Meats 1500 BC Middle East
Sourdough bread 1000 BC Europe
Fish sauce 1000 BC Southeast Asia, North Africa
Pickled vegetables 1000 BC China, Europe
Tea 200 BC China
Fermented foods comprise about one-third of the
world wide consumption of food and 20-40% (by
wt.) of individual diets.
ble: Worldwide production of some fermented foods
Food Quantity (t) Beverage Quantity (hl)

Cheese Beer 1000 million


15 million
Yoghurt Wine 350 million
3 million
Mushrooms 1.5 million
Fish sauce 300 000
Dried stockfish 250 000
Benefits of fermentation
• In general, fermented foods have several
advantages. Fermentation:
help to preserve the food: Many foods such as
ripened cheeses, pickles, sauerkraut, and
fermented sausages are preserved products in
that their shelf life is extended considerably
over that of the raw materials from which they
are made.
improve aroma and flavour characteristics:
Organoleptic properties of fermented foods are
improved in comparison with the raw
materials from which they are prepared;
increase vitamin content or digestibility of the
food compared to the raw materials;
Cont’d…
reduces the toxicity of some foods.
Fermentation sometimes removes unwanted or
harmful components in the raw material. For
example, fermentation:
removes flatulence factors in soybeans, and
reduces the poisonous cyanide content of cassava
during garri preparation.
The nutritive content of the food is improved in many
items by the presence of the microorganisms; thus
the lactic acid bacteria and yeasts in garri and the
yeasts in bread add to the nutritive quality of these
foods;
Fermentation often reduces the cooking time of the
food as in the case of fermented soy bean products, or
ogi the weaning West African food produced from
fermented maize;
Cont’d…
• In addition to the roles of fermentation in preservation
and providing variety to the diet, there are further
important benefits of fermentation.
• Several of the end products of food fermentation,
particularly acids and alcohols, are inhibitory to the
common pathogenic microorganisms that may find
their way into foods.
• The inability of Clostridium botulinum to grow and
produce toxin at pH values of 4.6 & below has already
been cited.
• Increasing the acidity of foods by fermentation is very
common.
• Foods as diverse as yogurt, hard sausages, and
sauerkraut all contain acid as a result of fermentation.
Cont…
• When microorganisms ferment food constituents, they
derive energy in the process and increase in numbers.
• To the extent that food constituents are oxidized, their
remaining energy potential for humans is decreased.
• Compounds that are completely oxidized by
fermentation to such end products as carbon dioxide
and water retain no further energy value.
• Most controlled food fermentations yield such major
end products as alcohols, organic acids, aldehydes, and
ketones, which are only slightly more oxidized than
their parent substrates, and so still retain much of the
energy potential of the starting materials.
• Fermentation processes are attended by temperature
increases.
Cont…
• The energy dissipated as heat represents a fraction
of the total energy potential of the original food
material no longer recoverable for nutritional
purposes.
• Fermented foods can be more nutritious than their
unfermented counterparts.
• This can come about in at least three different ways:
1) Microorganisms not only are catabolic, but they
also are anabolic and synthesize several complex
vitamins and other growth factors.
• Thus, the industrial production of such materials as
riboflavin, vitamin B12, and the precursor of vitamin
C is largely by special fermentation processes.
Cont…

2)
The second important way in which fermented foods can be
improved nutritionally has to do with the liberation of nutrients
locked into plant structures and cells by indigestible materials.
This is especially true in the case of certain grains and seeds.
•Fermentation, especially by certain molds, breaks down
indigestible coatings and cell walls both chemically and
physically.
•Molds are rich in cellulose-splitting enzymes; in
addition, mold growth penetrates food structures by
way of its mycelia.
•This alters texture and makes the structures more
permeable to the cooking water as well as to human
digestive juices.
•Similar phenomena result from the enzymatic actions of
yeasts and bacteria.
Cont…
3) A third mechanism by which fermentation can enhance
nutritional value, especially of plant materials, involves
enzymatic splitting of cellulose, hemicellulose, and
related polymers that are not digestible by humans
into simpler sugars and sugar derivatives.
• This goes on naturally in the rumen of the cow
through the enzymatic action of protozoa and
bacteria.
• It also occurs in the process of preparing silage
for animal feeding.
• Cellulosic materials in fermented foods similarly
can be nutritionally improved for humans by the
action of microbial enzymes.
Table: Benefits of fermentation
Raw Fermented
Benefit
material food
Preservation Milk Yoghurt, cheese
(Most materials)
Enhancement of safety
Acid production Fruit Vinegar
Acid and alcohol production Barley Beer
Grapes Wine
Production of bacteriocins Meat Salami
Removal of toxic components Cassava Gari, polviho azedo
Soybean Soy sauce

Enhancement of nutritional value


Improved digestibility Wheat Bread
Retention of micronutrients Leafy veges. Kimchi, sauerkraut
Increased fibre content Coconut Nata de coco
Synthesis of probiotic compounds Milk Bifidus milk, Yakult,
Acidophilus yoghurt
Improvement of flavour Coffee beans Coffee
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Grapes Wine
Health benefits of fermented foods
1. Lactose intolerance
• Lactose intolerance is the normal state for adult
mammals, including most adult humans.
• Many more groups are intolerant to lactose than
are tolerant.
• When lactose malabsorbers consume certain
quantities of milk, they immediately experience
flatulence and diarrhea.
• The condition is due to the absence or reduced
amounts of intestinal lactase.
• This allows the bacteria in the colon to utilize
lactose with the production of gases.
• Anaerobic and facultative anaerobic bacteria
utilizing the non-absorbed lactose
3.
2. Cholesterol
Cont’d…
Anticancer
control
• It makes the blood vessels
effect
narrow and narrow • Anticancer
• Some studies indicated activities
dietary intake of fermented demonstrated
milks reduce cholesterols.
• Fermented milk contain a
in animal
factor that inhibits synthesis models by
of cholesterol. employing
• In other studies, LAB were yogurt and
reported to remove some LAB
cholesterol or its precursor
from gastrointestinal tract.
• It is possible to take life LAB
5.2. Microbial ecology of fermentation
• The microbial ecology of food fermentations has been
studied for many years, and the activities of the fermenting
organisms are dependent on the intrinsic & extrinsic
parameters of growth.
• For example:
when the natural raw materials are acidic and contain free
sugars, yeasts grow readily, and the alcohol they produce
restricts the activities of most other naturally contaminating
organisms;
If the acidity of a plant product permits good bacterial growth &
at the same time the product is high in simple sugars, lactic acid
bacteria (LAB) may be expected to grow. Low level NaCl addition
will ensure their growth preferential to yeasts (as in sauerkraut
fermentation);
Products that contain polysaccharides but no significant levels of
simple sugars are stable to the activities of yeasts and LAB due to
the lack of amylase in most of these organisms. To effect
fermentation, an exogenous source of saccharifying enzymes
must be supplied.
• Diverse group of bacteria are involved in the
Cont’d…

manufacture of fermented foods.


• These bacteria are subdivided into 3 phyla:
i. Proteobacteria,
ii. Firmicutes, and
iii. Actinobacteria.
• Firmicutes:
the lactic acid bacteria,
a cluster of Gram positive bacteria
the main organisms used in the manufacture of fermented foods.
• The genera Bacillus and Brevibacterium also belongs
to this phylum and contain species used in the
manufacture of few fermented foods.
• Proteobacteria:
contains Gram negative bacteria
involved in the vinegar fermentation.
Cont’d…
• Actinobacteria contains only few genera relevant to
manufacture of fermented foods.
• These include :
i. Bifidobacteriim: do not have a functional role in fermented
foods; rather they are added for nutritional purposes,
ii. Kocuria,
iii. Staphylococcus, and
iv. Micrococcus.
• Kocuria, Staphylococcus and Micrococcus are used in
fermented meat production and to impart the desired
flavor and color.
• Fermented foods may contain many other
microorganisms, whose presence occurs as a result of
inadvertent contamination.
• The section below describes only lactic acid bacteria,
which are the main organisms used in manufacture of
fermented foods.
Lactic acid bacteria (LAB)
• The LAB are generally defined as a cluster of
lactic acid-producing, low %G+C, non-spore-
forming, Gram-positive rods and cocci that share
many biochemical, physiological, and genetic
properties.
• Composed of 13 genera of Gram-positive
bacteria:
─ Carnobacterium ─ Oenococcus
─ Enterococcus ─ Pediococcus
─ Lactococcus ─ Paralactobacillus
─ Lactobacillus ─ Streptococcus
─ Lactosphaera ─ Tetragenococcus
─ Leuconostoc ─ Vagococcus
─ Weissella
Cont’d…
• LAB are distinguished from other lactic acid
producing Gram positive bacteria by virtue of
numerous phenotypic and genotypic differences.
• All members of LAB produces lactic acid from
hexoses.
• As fermenting organisms, they lack functional
heme-linked electron transport systems or
cytochromes.
• They obtain their energy by substrate-level
phosphorylation while oxidizing carbohydrates.
• They do not have a functional Krebs cycle.
• Based on end products of glucose metabolism, LAB
are divided into two groups:
1. Homofermentative
2. Heterofermentative
1. Homofermentative
• Those LAB that produce lactic acid as the major or sole product
of glucose fermentation are designated homofermentative.
• Homolactics are able to extract about twice as much energy
from glucose as are the heterolactics.
• All members of the genera Pediococcus, Streptococcus,
Lactococcus, and Vagococcus are homofermenters, along with
some of the Lactobacilli.
• Homofermentative pattern is observed when glucose is
metabolized.
• But not necessarily when pentoses are metabolized.
• Some homolactics produce acetic and lactic acids using
pentoses.
• Homofermentative character of homolactics may be shifted for
some strains by altering growth conditions.
• The homolactics possess the enzymes aldolase and hexose
isomerase but lack phosphoketolase.
• Use the Embden-Meyerhof-Parnas (EMP) pathway toward their
production of two lactates per glucose molecule.
2. Heterofermentative
• Heterofermentative: are LAB that produce equal
amounts of lactate, acetic acid, CO2 & ethanol from
hexoses.
• Heterofermenters consist of Leuconostoc, Oenococcus,
Weissella, Carnobacterium, Lactosphaera, and some
lactobacilli.
• They are more important than the homolactics in
producing flavor and aroma components such as
acetylaldehyde and diacetyl.
• Heterofermenters and homofermenters can be readily
distinguished in the laboratory in that heterofermenters
produce CO2 in glucose-containing media.
• Heterolactics have phosphoketolase but do not possess
aldolase & hexose isomerase.
• Instead of the EMP pathway for glucose degradation,
they use the hexose monophosphate or pentose pathway.
Cont’d…
• Although LAB are mesophilic, some can
grow below 5oC and some as high as
45oC.
• With respect to growth pH, some can
grow as low as 3.2, some as high as 9.6,
and most grow in the pH range 4.0-4.5.
• LAB are only weakly proteolytic and
lipolytic
• They do not act on lipids and proteins,
no pawl production.
• They depend on simple sugars
• Thermobacterium lactic acid bacteria
Starter culture

• Traditionally the fermenting organisms came


from the natural microflora or a portion of the
previous fermentation.

• In many cases the natural microflora is either


inefficient, uncontrollable, and unpredictable,
or is destroyed during preparation of the
sample prior to fermentation (eg
pasteurisation).

• A starter culture can provide particular


characteristics in a more controlled and
predictable fermentation.
Cont’d…
• Lactic starters always include bacteria that
convert sugars to lactic acid, usually:
Lactococcus lactis subsp. lactis,
Lactococcus lactis subsp. cremoris or
Lactococccus lactis subsp. lactis biovar
diacetylactis.

• Where flavour and aroma compounds such


as diacetyl are desired the lactic acid starter
will include heterofermentative organisms
such as:
Leuconostoc citrovorum or
Leuconostoc dextranicum.
Cont’d…
• The primary function of lactic starters is the
production of lactic acid from sugars.
• Other functions of starter cultures may
include the following:
flavour, aroma, and alcohol production
proteolytic and lipolytic activities
inhibition of undesirable organisms
• A good starter culture will:
Convert most of the sugars to lactic acid
Increase the lactic acid concentration to
0.8 to 1.2 % (Titratable acidity)
Drop the pH to between 4.3 to 4.5
5.3. Fermented food
products
• Microorganisms have long played a major
role in the production of fermented foods.
• Traditional fermented foodstuffs include:
i. Dairy products: particularly cheeses, yoghurt,
sour cream and kefir;
ii. Fish products: such as fish sauce and fish
pastes;
iii. Meat products: fermented sausages which are
produced generally as dry or semidry products;
and
iv. Plant (vegetable) products: notably cereal-based
breads and fermented rice products; along with
fermented fruits, vegetables and legumes.
Cont’d…
• The most economically relevant vegetable products
include:
Sauerkraut: a fermentation product of fresh cabbage,
Pickles: fermentation products of fresh cucumbers and
Olives.
• As raw vegetables have a high microbial load and cannot
be pasteurized without compromising product quality,
most vegetable fermentations occur as a consequence of
providing growth conditions (such as added salt) that
favour the LAB.
• These bacteria are present on fresh vegetables in very
low numbers, accounting for only 0.15-1.5% of the total ¨
population (Buckenhuskes, 1997).
• The starting material for fermented juice is pasteurised
mash or juice and starters of lactobacilli (including Lb.
plantarum, Lb. casei, Lb. acidophilus), L. lactis and Lc.
mesenteroides.
Fig. Examples of traditional and novel fermented foods and
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