Dr AMOR L.
II. Use of Microorganisms in Food Fermentation
II.1. Milk
Milk is a white, opaque liquid with a slightly sweet taste, constituting a complete and balanced
food, secreted by the mammary glands of women and female mammals for the nutrition of the
young. Raw milk is milk that has not undergone any preservation treatment except refrigeration at
the farm. The sales deadline corresponds to the day after milking. Raw milk must be brought to a
boil before consumption (because it contains pathogenic germs).
II.1.1. Types of milk according to fat content
a) Whole milk: is heat-treated milk which, regarding its fat content, meets one of the following
formulas:
Standardized whole milk: milk whose fat content is at least 3.50% m/m. However, Member
States may provide for an additional category of whole milk whose fat content is greater than or
equal to 4.00% (m/m).
Non-standardized whole milk: a milk whose fat content has not been modified since the milking
stage, neither by addition or removal of milk fats, nor by mixing with milk whose natural fat
content has been modified. However, the fat content cannot be less than 3.50% (m/m).
It is customary for the packaging color to be predominantly red.
b) Semi-skimmed milk is heat-treated milk whose fat content has been reduced to a rate that
amounts to at least 1.50% (m/m) and at most 1.80% (m/m).
It is customary for the packaging color to be predominantly blue.
c) Skimmed milk is heat-treated milk whose fat content cannot exceed 0.50% (m/m).
It is customary for the packaging color to be predominantly green.
II.1.2. Milk according to heat treatment applied
a) Pasteurized milk
The designation "pasteurized milk" is reserved for milk:
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1. Obtained by a treatment using a high temperature for a short time span (at least 72°C for
fifteen seconds or any equivalent combination) or by a pasteurization process using
different combinations of time and temperature to obtain an equivalent effect;
2. Immediately cooled after pasteurization to be brought, as quickly as possible, to a
temperature not exceeding 6°C.
B. UHT sterilized milk
The ultra-high temperature process is also a long-term preservation process that allows shortening
the heating time: the taste qualities of milk are better preserved than with simple sterilization. It
involves rapidly bringing the milk to a temperature of minimum 135°C for 2 to 4 seconds, then
packaging it in a sterile environment.
UHT milk can be whole, semi-skimmed, or skimmed. It is found in commerce under the name
"UHT sterilized milk". It can be stored at room temperature as long as the packaging has not been
opened.
II.1.3. Milk composition
Milk is an important source of very high-quality proteins, rich in essential amino acids,
particularly lysine which is par excellence the amino acid of growth. Its lipids, characterized in
relation to other dietary fats by a high proportion of short-chain fatty acids, are much richer in
saturated fatty acids than in unsaturated fatty acids. They also carry appreciable quantities of
cholesterol and vitamin A as well as small quantities of vitamins D and E.
The average composition of cow's milk is represented in the table.
Table: Average composition of cow's milk (g/l).
Milk Components Content (g/l)
Mineral Components
Water 902
Mineral salts 6,9
Dissolved gases 0,1
Organic Components
Organic salts 1,7
Lactose 49
Fat 38
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Proteins or Protein Nitrogen Components
Casein 32
Soluble proteins 26
Non-protein nitrogen components 6
Other components 1,5
II.2. Cheese making
II.2.1. Microorganisms
In the field of cheese making, multiple useful microorganisms are involved such as bacteria,
molds, yeasts.
a) Bacteria
−Lactic acid bacteria: These are bacteria producing lactic acid through fermentation of simple
carbohydrates (lactic fermentation). Their essential roles are to acidify milk and curd, participate in
flavor formation (proteolysis, aroma production), texture, and the opening of dairy products. These
bacteria are now widely used in the form of selected starters.
−Propionic bacteria: These are bacteria fermenting lactates to produce acetic and propionic acid,
as well as CO2 (propionic fermentation). They participate in flavor formation and the opening of
cooked pressed cheese (Emmental, Comté, Gruyère).
−Micrococci, non-pathogenic staphylococci: flavor formation in cheeses, particularly in washed-
rind, bloomy rind, or mixed rind cheeses (Munster, Camembert, Pont l'Evêque, etc....). These are
bacteria constituting the surface flora of ripened cheeses. They play an essential role in cheese
ripening.
b) Yeasts
Yeasts are found more significantly (on average 100 times more) on cheese surfaces (particularly
soft cheeses) than inside. They intervene in the des-acidification of the paste at the beginning of
ripening, thus allowing the subsequent implantation of an acid flora and also intervene in flavor
formation.
c) Molds
Penicillium camemberti is present on the surface of soft-ripened cheeses with bloomy rind
like Camembert or goat cheeses.
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Penicillium roqueforti is the internal mold of blue cheeses like Bleu d'Auvergne (cow's
milk) or Roquefort (sheep's milk).
Mucor is the dominant mold on the surface of Tomme de Savoie and is also present on the
surface of farmhouse Saint Nectaire.
Through their biochemical aptitudes, molds play a determining role in the formation of sensory
characteristics of cheeses.
II.2.2. Main steps of cheese making
Cheese making can be considered as an agglomeration phenomenon, corresponding to a syneresis,
associated with a flow phenomenon. It involves the agglomeration of milk protein elements,
mainly casein, more or less modified, which trap other constituents and, subsequently, the
agglomeration of molded curd pieces. This agglomeration phenomenon is associated with that of a
flow of the liquid phase, composed of milk water and soluble elements trapped in pores, then
released.
Usually, cheese making includes three steps: The formation of a casein gel, which is milk
coagulation; partial dehydration of the gel, which is draining that leads to a curd and salting. These
steps concern fresh cheeses. The rest of the cheeses undergo an additional ripening step, these are
ripened cheeses (Camembert, Roquefort, Gouda, Tulum,...).
a) Milk coagulation
Milk coagulation results from the association of casein micelles more or less modified. This
agglomeration leads to the formation of a coagulum whose volume is equal to that of the milk
used. These physicochemical modifications of caseins are induced either by acidification or by
action of coagulating enzymes.
Milk acidification can be obtained by fermentation products of acidifying bacteria or by chemical
compounds with direct or indirect acidifying action. The concomitant decrease in pH has the effect
of reducing the ionization of casein acid functions, inducing the progressive displacement of
calcium and inorganic phosphate from the micelle to the aqueous phase. This induces the
disorganization of micelles and reorganization of micellar subunits.
Microbial acidification of milk is a progressive, slow, and uniform process. It is characterized by
difficulties related to controlling microbial development (multiplication kinetics, physiological
state, growth factors, metabolic products, and others). The built coagulum is a set of casein flakes
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fitted into each other. The rate and importance of acidification influence the gel texture by
controlling its demineralization rate. The obtained acid gel is friable, smooth, and homogeneous.
In enzymatic coagulation, several proteolytic enzymes of animal origin (calf, young bulls, pork,
and chickens), vegetable (artichoke, thistle), and microbial (Kluyvermyces, Mucormiehi,
Mucorpusills, and Endothiaparasitica) are used.
b) Draining
Draining is a dynamic phenomenon characterized by the quantity of whey eliminated during time.
Indeed, it sets the physical characteristics (pH and aw) and chemical characteristics of the curd and
consequently the cheese ripening. The draining process is linked to direct factors corresponding to
mechanical and thermal treatments, indirect factors (acidification and enzymatic coagulation), and
factors related to the raw material (richness in milk casein, soluble proteins, and fat).
c) Salting
In cheese making, salting is an essential phase in the production of ripened products. The salt
content of cheeses varies according to the type of cheese, on average it is 0.5-2 g/100 g in most
cheeses, in some cases (blue cheeses and some goat cheeses), it can rise to 3-4 g/100g. However,
some oriental cheeses preserved in brine have quite high levels (8-15 g/100 g). The salting
methods are by brining (Emmental and Camembert), dry salting, and mass salting. Mass salting is
used in traditional manufacturing of some typical Mediterranean basin cheeses. It allows milk
preservation, prolongs the coagulation and draining phases of cheese.
Salt allows reaching the appropriate humidity of cheese. It exerts, according to its concentration, a
selective microbial action and an inhibitory effect on enzyme activity. For example, the growth of
lactic acid bacteria from starters is inhibited at a salt content above 2.5 g/100 g, and is practically
zero above 5 g/100 g. [Link] undergoes spore germination inhibition for rates of 3-6 g/100 g.
The effect of salt on the development of cheese microbial flora can however only is fully
appreciated by taking into account the salt tolerance of microorganisms in the cheese environment
and the salt content of the cheese paste.
d) Cheese ripening
Ripening is the most complex step in the manufacture of matured cheeses which depends on each
physicochemical or microbiological characteristic of the cheese. It is a complex and long
biochemical process that corresponds to an enzymatic digestion phase of curd constituents by
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different agents. The cheese thus becomes the site of different degradations that occur
simultaneously or successively, leading to the release of sapid and odorous substances while
modifying the texture. The cheese is thus compared to a complex bioreactor whose evolution the
practitioner must master to bring it towards the optimal desired characteristics. The ripening
duration varies according to the cheese; it lasts from a few weeks to two years or more at specific
temperatures for different types of cheeses.
II.2.3. the 8 cheese families
There are numerous cheeses in the world, all different from each other, by their textures, their
shape, their flavors... These varieties are divided into eight large families, to better understand
them.
1. Fresh cheeses
Brousse de Provence, Creuset d'Anjou, Faisselle.... These are cheeses that have not been ripened.
The milk under the action of rennet and lactic ferments transforms to become curd. This is then
drained to extract the whey (petit-lait). Some varieties of fresh cheese are then molded. Their
moisture content is above 60%. These are cheeses with an unctuous and melting texture, which are
consumed quickly after their manufacture.
2. Soft-ripened cheeses with bloomy rind
Soft-ripened cheeses have an unctuous, melting, creamy texture. They are made with curdled milk,
then molded and finally drained. Once unmolded, the curdled milk is then salted and dried. The
outer surface is then seeded with a fungus (generally PenicilliumCandidum). During ripening, a
white and downy rind will form, called the "flower". Their moisture content is between 50 and
60% (higher percentage for double or triple creams, for which cream is added to milk).
Among bloomy rind cheeses, we find Camembert, Chaource, Brie...
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3. Soft-ripened cheeses with washed rind
The manufacture of soft-ripened cheeses with washed rind is the same as bloomy rind cheeses
until ripening. During this period, the cheeses will be washed and brushed several times with brine
(salt water), to which alcohol is sometimes added (Marc de Bourgogne for Epoisses, for example).
These cheeses are known for their strong odor and their often orange-colored humid rind.
Examples include Epoisses, Maroilles, Pontl'Evêque...
4. Uncooked pressed cheese
It's the cheese rind that gives all the flavor and aroma to this paste. The milk also, whether from
cow or sheep, imposes its taste on the cheeses. This paste, more or less thick depending on the
ripening duration, is obtained by mechanically pressing the curdled milk to extract the whey
(lactosérum). Its moisture content is between 45 and 50%.
Among the most well-known are Reblochon, Saint Nectaire, Cantal...
5. Cooked pressed cheese
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To preserve these cow's milk cheeses and be able to consume them all year round, the curd was
heated and pressed, hence their name "hard pastes". Cooked pressed pastes were originally alpine
cheeses, made in the mountains in summer, when milk is rich and abundant, then brought down to
the valley during winter. Some come in the form of very large wheels! Currently, they can be
produced in a more industrial way. Their slow and long maturation gives them their fruity flavor
and firm texture. The paste is yellow in color. These are the richest in calcium and proteins.
The most well-known are Emmental, Comté, Abondance, Beaufort...
6. Blue-veined cheeses
Or "the blues" such as Roquefort, Fourmed'Ambert, 1924...
They develop bluish or greenish points in the paste. During molding, the curd is inoculated with a
fungus (penicilliumglaucumroqueforti or penicilliumcandidum) to allow molds to develop in the
paste. For several months, the cheeses are ripened in a humid environment and pierced with long
needles to facilitate air circulation in the paste and stimulate mold development.
7. Processed cheeses
This type of cheese is obtained by melting cheese or a mixture of cheeses. Aromatics, spices, or
other types of dairy products can be added to its preparation. These can be creams (like
Cancaillotte), spreadable cheeses, aperitif cheeses,... Most bear the name of a trademark registered
by a company.
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8. Goat milk cheeses
Goat cheeses have very different textures and tastes. Depending on the manufacturing technique,
there are goat cheeses in all paste families. There are many of them, manufactured as fresh or
ripened cheeses, ash-covered or not, in different shapes: crottins, logs, pyramids, corks, pucks...
Among the most renowned, we find Crottin de Chavignol, Rocamadour, Valençay,...
II.3. Yogurt Manufacturing
II.3.1. Definition
The designation yogurt (or yoghourt) is reserved for fermented milk obtained, according to loyal
and constant practices, by the development of only specific thermophilic lactic acid bacteria called
Lactobacillus bulgaricus and Streptococcus thermophilus, which must be inoculated
simultaneously and be found alive in the finished product until the expiration date, at a rate of at
least 10 million bacteria per gram relative to the milk part. Moreover, the quantity of free lactic
acid contained in 100 g of yogurt must not be less than 0.7 g.
A yogurt (or yoghourt) can come with varying fat content, depending on the milk used for its
manufacture and added dairy materials. It can be plain, sweetened or not, or contain ingredients
other than dairy which must not represent more than 30% of the final weight of the yogurt (fruit
pieces, honey, jam, natural or authorized synthetic flavors). These ingredients are pasteurized or
sterilized before being incorporated. The addition of stabilizers, thickeners, or gelling agents other
than those from fruit preparations is prohibited.
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II.3.2. Lacticacidbacteria
Streptococcus thermophilus is one of the thermophilic lactic acid bacteria, widely used as a
starter in the manufacture of certain fermented dairy products such as yogurt (in mixed
culture with Lactobacillus bulgaricus)
Lactobacillus is a main genus of the Lactobacillaceae family, it contains numerous species
that are lactic fermentation agents involved in many industries species that are encountered
as contaminants
The market offers a whole range of products entitled to the designation yogurt (or yoghourt):
plain yogurts (or yoghourt), sweetened or not fruit yogurts (or yoghourt), pulped yogurts (or
yoghourt) or with small cake pieces (less than 30% added elements),
flavored yogurts (or yoghourt).
Yogurts (or yoghourt) can be classified into 3 main consistency families:
"firm" yogurt (or yoghourt): proteins form a gel under the action of lactic acid.
"stirred" yogurt (or yoghourt): stirred yogurt, Bulgarian, has a smooth, more or less fluid texture.
"drinkable" yogurt (or yoghourt): its texture is liquid and foamy, precisely to allow it to be easily
drunk.
II.4. Dairy Creams
The designation cream is reserved for milk containing at least 30 g of fat coming exclusively from
milk per 100 g of total weight. The designation light cream is reserved for milk containing
between 12g included and 30g not included of fat coming exclusively from milk per 100 g of total
weight.
II.4.1. Raw cream
It's a cream that has not undergone any pasteurization or sterilization treatment. Direct result of
skimming, it is cooled and stored at +6°C. With liquid texture during the first days, and mild
flavor, its fat content is generally higher than that of other creams. The mention "raw" is
mandatory on the labeling.
II.4.2. Pasteurized cream
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Fresh cream designates a cream having undergone only pasteurization treatment and packaged at
the production site within 24 hours after it. There exists thick pasteurized fresh cream and fluid
pasteurized fresh cream, called fleurette.