Microbiological production of Food and Dairy product

Introduction – it is estimated that food and beverage biotechnology occupies a prominent

place world over. There are records that there was mass making of bread wine curd etc as
early as 4000. B.C. These processes collectively referred to as traditional or old
biotechnology mostly employed for the preparation of food and beverages were based on
the natural capabilities of microbes although their existence was known at that time.
With the advances made in microbiology and recently biotechnology, food and
beverages production are major industries. Food biotechnology is also concerned with the
improved quality, nutrition, consistency, colour, safety preservation of food besides making
them available round the year in addition modern biotechnological processes also takes into
the account the health aspects of the people.

Fermented foods

The production of fermented foods is variable. This depends on geographical regions,

availability of raw materials, traditions and food habits of the people.

Advantage of Fermented foods

1. Enhance nutritive values

2. Increase digestibility
3. Improved flavors and texture
4. Serve as a supplements in preparing several dishes
Vitamins, amino acids, enzymes, biopolymers, SCPs and mushrooms are example of many
compounds that are directly or indirectly used as food supplements.

YOGHURT

Yoghurt is prepared on a small scale under controlled fermentation i.e. “controlled d, batch,
dual fermentation”. It is a fermented milk which derives its name from the Turkish word
“Sugurt”
It is most widely available fermented milk in the western world, and it gained
popularity due to its flavor and versatility than from it s keeping properties
All fermented milk products including yoghurt are prepare by allowing lactic acid
bacteria present in milk t o develop, providing natural conditions for their growth, by adding
known amounts of known culture or by adding previously cultured buttermilk of good quality
to the fresh milk.

Sources of milk

Milk of cow goat sheep skim milk or fortified milk can be used as milk sources for
yoghurt preparation the milk used should be free of anti microbials. The solid content of milk
may be supplemented to enhance the final texture of the product during yoghurt preparation
the solids not fat (SNF) content is increased to 10 – 15%. When compared to 8.5% in fresh
milk. This is achieved by adding skim or whole milk powder depending on the fat
requirement. The properties of the product maybe improved and stabilized by adding small
amount of natural and modified gums which bind water and thicken the milk
 If left to stand, the milk fat would segregate out to form a cream layer to prevent this
the milk is homogenized by passing through small orifice under pressure at 50-60°C. To
reduce size of the fat globules to below 2-4mm. This improves product stability the milk
viscosity and also makes I appear whiter (As the no. of light reflecting centers is increased).

Heat Treatment

This is done to reduce the micro flora present in milk I n order to avid competition to
the tarter culture for this purpose the milk is heated to 80-90°C. for 30 min. this heat
treatment also improves the quality of milk as the growth medium for the starter and
improves the interaction between the casein protein and whey. (serum proteins) this
interaction of casein proteins and whey improves the viscously of yoghurt.

Starter culture

Equal proportion of the 2 cultures are used a starter culture which include
1- Streptococcus salivarius thermophilus
2- Lactobacillus delbruckii bulgaricus
The starter culture is added at the rate OF 2 % v/v which corresponds to 106 - 107
microlitres/ml of the starter in the milk (homogenized).

Relationship between the 2 starter culture microbes.

Relationship is termed as protocol-operation. In this 2 bacteria get benefited from one

and another but are not completely independent on each other they exist in mutually
favorable interaction L.b slightly proteolytic uses the casein protein and produces free
amino acids like valine which inturn stimulates the growth of S.s.t .It produces formate,
pyruvate and CO2 all of which stimulate the growth of lactobacillus

Culture development

Both the microbes are isolated from milk or any other natural source and then
repeatedly subculture till pure culture of the same is isolated they are preserved as master
culture and the subculture of the same are used to build up the inoculum in a sequential
manner

Fermentation Process

The milk ix consisting of 11-15% of SNF and whole or skimmed milk powder and
natural gum Arabic solution is heated to 80 -90 °C. For 30 min. this renders proper
pasteurization of the milk mixture killing all he contaminants which can prevent
contamination of milk and prevent contamination of starter culture

This heating also improves the milk mix as the growth medium for the starter culture
by eliminating immunoglobulin resulting in the expulsion of O2. to produce microaerophillic
environment and by releasing stimulating levels of sulphahydryl groups

The heating also promote the interaction between whey and casein which inturn
increases the yoghurt viscosity stabilizes the gel and limits the separation of whey

The heat treated milk is cooled through 40 – 60 °C. the optimum temperature for
fermentation the starter cultures are added and fermentation carried out for 4 hours at a pH
of 6.3-6.5. The fermentation can be carried in retail packs to produce a firm continuous
coagulum known as set yoghurt or in bulk tanks to produce stirred yoghurt where the gel is
broken by mixing in other ingredient and then come into packs

The lactose present in the milk mix is converted into lactic acid which decreases he
pH of the medium from 6.3-6.5 to 4.6-4.7. This is the isoelctric point of casein which once
reached the lactic acid solubilizes calcium and phosphate ions which inturn destabilizes the
complex casein micelles and denature whey proteins

The micelles aggregate together to produce a continuous gel in which all the
components are entrapped with little or no wheying off.

During fermentation the growth of one is fastest in the early stages but as the pH
drops below 5.5 the growth of one slows down and the 2. Predominates at the end of the
process the product has a total activity of 0.5-0.9 %. And the organisms in excess of 108 cfu
per ml

The fermentation is stopped by cooling the product to 20°C. and maintained at 4°C
for 3 weeks so that at 4°C the total activity of bacteria will be stopped

During fermentation 2 major flavoring products are formed which are

acetaldehyde (most important flavoring agent) and Diacetyl. Yoghurt should contain 22-
41mg/kg of acetaldehyde which accumulates due to lack of enzyme alcohol dehydrogenase
in the organism otherwise acetaldehyde converts into alcohol

Diacetyl at the range of approx. 0.5mg/kg of yoghurt is preferred to be present in

yoghurt if its content increases beyond 0.5 mg/kg of yoghurt the diacetyl impart bitter taste
to yoghurt
 CHEESE

Introduction- Cheese production is the largest dairy industry in the world there are around
100 type of different cheese irrespective of the type of the cheese all of them are invariably
made from the casein of milk i.e. produced after separating the whey. Milk from different
animals can be used ex. - sheep cow goat buffalo
Cheese contains milk fat casein and small amounts of salt. Most of the lactose and
soluble milk components remain in the whey the manufacture of all cheese depends upon
the activity and type of microbes used as starter culture

Types of cheese

Cheeses are broadly categorized into 2 types ripened cheese and unripened cheese.
In general cheese may be classified in several ways.
1. On the basis of hardness
1. Hard cheese – Swiss cheese, cheddar cheese
2. Semi Hard cheese – rockeforte cheese, Danish blue cheese
3. Soft cheese – limburger cheese, camemberg
2.On the basis of principle ripening agent
1.Bacterial cheese- by propionibacterium sp.
2. Mould cheese- eg- camemburg cheese – ripened by penicillium camemberitii
3.On the basis of method of coagulation
1.Rennet Cheese – Limberger cheese
2. Acid curd cheese – Cottage cheese

Production of cheese

Most of the cheese is made either from whole or skimmed milk of goats cows mares
and other animals this is carried out by a process of dehydration where in casein and fats
are concentrated 5 -15 time cheese production is very complicated and broadly involves 4
stages
1- Inoculation and acidification of milk

The milk is inoculated with starter culture depending on the temperature of milk if the
milk is held at 38°C the inoculum includes Streptococcus lactis or S. Cremoris.
Similarly if the milk is held at 50°C then the inoculum includes Lactobacillus lactis and
L.bulgarius.Once he tarter culture is added this culture converts the sugar in the milk
that is lactose to lactic acid this lowers the pH of the milk to around 4-6 thus acidifying
the milk.
2- Coagulum formation

When the acidified milk is treated with rennet the enzyme chymosin of animal r fungal
origin casein gets coagulated.Casein mainly consists of 3 components insoluble
alpha casein, insoluble beta casein, kappa casein that keeps them in a soluble state
By he action 0of chymosin kappa casein is degraded consequently alpha and
beta casein and the degraded products of kappa casein combine to form a coagulum
i.e. curd this process of coagulation is dependent on the Ca + ions
3- Separation of curd from whey and processing of curd

When the temperature of coagulum I raised to around 40°C the curd and the whey
liquid gets separated. This separation of curd from the whey can also be done by
driving or processing the curd followed by heating to reduce the moisture
Texture of the cheese depends on the manner of separation of curd from the
whey i.e.
• On during the cheese is soft
• On pressing the cheese is hard
Whey has 93% water ,5% lactose, and the remaining components like vitamins salts
and minerals and some amount of fat
The curd separated is subjected to a process of salting which can be done in any of
the following 2 ways
a- the curd is floated on a strong brine solution
b- hard cheese can be rubbed with NaCl salt directly

4- Ripening of Cheese

Their flavour of raw cheese is very bland, therefore to enhance the flavour, to make
changes in its texture, the cheese should be ripened either by microbes or by
chemical or by injecting CO2 gas
Depending on the type of cheese to be prepared the procedures adopted for
ripening or maturation are highly variable. During ripening, degradation of the lactose
left in the cheese, protein, fats is brought about
The block of curd separated are subjected to the action of proteases and on
lipases alternatively as in the case rockeforte and Danish blue cheese curd ids
inoculated with penicillium rockeforte spores after salting process. In Swiss cheese
the milk is inoculated with lactobacilli, streptococci and propiobacterium even before
the curdling process occurs. In limburger cheese the surface of cheese is allowed to
be contaminated with brevibacterium linens. Hard cheeses are generally layered by
lactic acid bacteria. Propionibacterium ferments lactose and produces CO2 and lactic
acid in Swiss cheese this CO2 is responsible for the formation of holes or eyes on the
surface
Lipases present in milk are reduced by bacteria degrade the fats in curd to produce
fatty acids which impart flavour to cheese
 BREAD

Bread making from the dough by employing microbes is one of the oldest examples of
fermentation process known to mankind. There is evidence that prepared in Egypt in 3000
B.C. Primarily bread is a fermented product of cereal flour such as wheat and rye. The
cereal flour is mixed with water salt sugar fat and other ingredients ( as desired for
enrichment of bread is subjected to fermentation by yeast Saccharomyces crevisiae – Top
fermenting strain. The main reaction that occurs during bread formation is the fermentation
of hexoses to CO2 and ethanol

C6H12O6 ------------- 2C2H5OH + 2CO2

The ethanol produced either gets evaporate or forms esters. The CO2 gets entrapped in the
dough resulting in its expansion the expansion and stretching of the dough particularly with
heat is during to the unique elastic protein namely gluten. It is mainly responsible for
retaining the shape of breadBesides yeast enzymes ex- amylases of other microbes also
help in fermentation and baking of bread
The texture of bread is influenced by fats and emulsifiers added top the dough.
Breadmaking is carried out with 3 objectives
1- Good leavening due to CO2 formation
2- Flavour development
3- Good texture
The yeast fermented bread has the above characteristics in contrast to the bread produce
by baking powder which also produces CO2 but this does not have the same flavour and
texture as that produced by yeast. Thus yeast is approx. refers to as BAKERS Yeast . IT is
a package of enzymes to give a desired product
In recent years some workers have reported the development of genetically
engineered strains of saccharomyces cerevisiae with improved fermentation properties.
such organisms when used in baking industry are believed to further enhance the quality of
bread with regard to flavour, texture etc.

Microbiology of Bread

Microbes are useful in bread making in 2 ways.

1- They leaven the dough to a desirable range to hold CO2 gas
2- They may produce some flavoring agents to make the bread more palatable and
flavored

Components of bread

Wheat flour, sugar , yeast, invert syrup, preservatives etc. The dough is made by mixing all
the above components with water and then leaven.

Process of bread making

Involves
1- Making the dough
 2- Flavor production
3- Baking process
Making the dough

The prepared dough is subjected to leavening which can be performed by yeast , other
microbes , chemicals or gases

1- leavening by yeast

Primary function of added yeast is to ferment the sugar present in the dough resulting
in gas and alcohol production which in turn add flavour to the bread.

In the first 2 hours of leavening there is little or no growth of yeast after 2-4
hours slight growth of yeasts occurs and after 4-6 hours decline in growth occurs.

Fermentation by yeas begins as soon as the dough is mixed and continues till 6
hours and then a decline in growth occurs after 4-6 hours when decline occur the
fermentation process can be stepped up by increases in the temperature by keeping
in the oven. The oven temperature kills the yeast and inactivates the enzymes during
fermentation by yeasts conditioning of the dough or flour proteins like gluten occurs
upon gluten conditioning the dough gluten becomes spongy and springy. Once it
becomes spongy & springy it can hold more CO2 produced by yeast fermentation
proteolytic enzymes from yeasts fermentation or flour or malt or added enzymes also
act on gluten and condition it apart from the above reduction in pH by means of acid
formation also condition the gluten proteins . The gluten protein conditioned hold the
CO2 in more amounts and so give a good texture and structure of the bread

Dough conditioner added externally are called yeast foods .The ammonium
produced sometime stimulated the yeast growth and now these yeasts produce more
proteolytic enzymes which condition the gluten and therefore it holds more CO2.

The rate of Gas production by yeast can be increased by adding more yeast ,
more sugar and yeast foods within limits. Similarly the rate of gas production can be
decreased by adding more salt, addition of too much yeast foods or increasing or
decreasing of pH. The main objectives of the baker during leavening process are to
have enough gas produced and to have the dough in such a condition that it will hold
the gas at the right time.

Here are 2 ways of bread leavening

a- Sponge method of bread making
Some of the ingredients are mixed at temperatures of 23-24°C and allowed to
ferment after some time the remaining ingredient are mixed and further
fermentation is carried out at the same temperature
b- Continuous or straight method of bread making
All the ingredients are mixed and the fermentation is carried out at 26 –
28°C most of the leavening process is usually held at about 27°C these
temperature are mot favorable for yeast fermentation
 2- Leavening by other microbes

This can be accomplished by gas forming other than yeast like coliform bacteria,
sachrolytic anaerobes and heterofermentative lactic acid bacteria. Bread can also be
leavened y addition small portions of previously fermented dough

3- Leavening by chemicals

This process doesn’t involve only microbes. The CO2 gas may be directly
incorporated into the dough . Baking powders which are combination of chemical
compounds which release CO2 when mixed with sough may also be used for
leavening

Flavor production

Yeast and bacteria during fermentation produce products responsible for desirable
flavour the flavoring substance include alcohol diacetyl aldehydes isoalchols acetic acid
succinic acid and their esters. The ingredients of the dough also add to the flavour

Baking process

Dough once leavened by any of the 3 methods is transferred into oven. This stops the
fermentation and kills the yeast , inactivates the enzymes which produce CO2.A temperature
of 100 - 150°C is maintained in the oven . These temperatures expand the gas present in
dough and raise the dough to give a proper texture and structure to the bread. The baking
also contributes to flavour the heating process removes the alcohol like ethanol produced in
the dough and impart desirable flavour and quality to the bread the bread becomes more
palatable on account of gelatinization of starch. Once gelatinized the starch supports the
structure of the baked bread
Credits to – Mrs Krishna priya maam