Department of Dairy processing technology

Prepared by: Garo Kabeto IDNO: TTMR/305/16

Individual Assignment- 1

Title: Biochemistry of Yoghurt, Cheese, Kefir, Buttermilk, Sour

Cream Dahi AND Koumis
Table of Contents
...................................................................................................... 1
1. Introduction ............................................................................. 3
1.1 Yoghurt ................................................................................ 4
1.2 Cheese ..................................................................................5
1.3 Kefir ....................................................................................6
Fig. 1.1 .....................................................................................7
Figure 1.2 ................................................................................ 8
1.4 Buttermilk ............................................................................9
1.6 Sour Cream .......................................................................... 9
1.7 Koumiss ............................................................................. 10
Fig 1.3 ....................................................................................12
Reference ...............................................................................14

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 1. Introduction
Milk has always soured spontaneously but at some point in human history, artisans deliberately
caused milk to sour or ferment. Fermentation is one of the oldest methods for preserving milk
and probably dates back ~10,000 years to the Middle East where the first evidence of organized
food cultivation and production is known to have occurred. Fermentation improves shelf life,
increases microbiological safety, adds flavor, and enhances palatability and organoleptic
qualities.

The fermentation process involves a series of complex reactions carried out by microorganisms,
which transform milk constituents rendering new molecules of enhanced nutritive value and
digestibility. Moreover, fermentation generates metabolites that can be major contributors of a
daily healthful diet.

Traditional fermented milk products have been developed in dependently worldwide and were,
and continue to be, especially important in areas where transportation, pasteurization and
refrigeration facilities are inadequate. Nowadays, the primary function of fermenting milk is to
extend shelf life, to improve taste, to enhance digestibility and to manufacture a wide range of
dairy-based products.

If removed aseptically from a healthy udder, milk is essentially sterile but in practice, milk
becomes contaminated by various bacteria, including lactic acid bacteria

(LAB) during milking. During storage, these contaminants grow at rates dependent on the
temperature. LAB probably dominate the microflora of uncooled milk expressed by hand. Since
LAB are well suited for growth in milk, they grow rapidly at ambient temperature, metabolizing
lactose to lactic acid and reducing the pH of the milk to the isoelectric point of caseins (~pH
4.6), at which they form a gel under quiescent conditions, thus producing cultured milks.
Traditionally, and until relatively recently, fermentation was caused by the indigenous
microflora or a “slop-back” culture (some of today’s product is used to inoculate fresh milk).
The production of fermented milks no longer depends on acid production by the indigenous
microflora. Instead, the milk is inoculated with a carefully selected culture of LAB and for some
products with LAB plus lactose-fermenting yeasts. Unlike cheese manufacture, the whey phase
is retained within the coagulum of fermented milk products. As a result, fermented milks are
high-moisture products (>80 %). Most fermented milks have a low pH (~pH 4.0), too low for
most spoilage bacteria and potential pathogens to grow.

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1.1 Yoghurt

Yoghurt is the best known of the fermented milk products and is consumed worldwide. The
consistency, flavor and aroma of yoghurt vary between countries from being a highly viscous
liquid to a softer gel-like product. Yoghurt may also be produced in frozen form as a dessert or
drink. Broadly, yoghurt can be classify as follows:

1. Set type, incubated and cooled in its package

2. Stirred type, incubated in tanks and cooled before packaging

3. Drinking type, similar to the stirred type but the coagulum is broken before packaging

4. Frozen type, incubated in tanks and frozen like ice cream

5. Concentrated yoghurt, which is incubated in tanks, concentrated and cooled before

packaging—also called strained yoghurt, labneh or labaneh.

The yoghurt fermentation is essentially homo fermentative, using a mixed culture of Lb.
delbreuckii and Str. thermophiles.

Depending on the product, the milk used may be full-fat, partially skimmed or fully skimmed. If
it contains fat, the milk is homogenized at 10–20 MPa to prevent creaming during fermentation.
For yoghurt, the milk is usually supplemented with skim milk powder to improve gel
characteristics. Acid milk gels are quite stable if left undisturbed but if stirred or shaken, they
synerese, expressing whey, which is undesirable. The tendency to synerese is reduced by
heating the milk at, e.g., 90 °C × 10 min or 120 °C × 2 min; heating causes’ denaturation of
whey proteins, especially β-lacto globulin, and their interaction with the casein micelles via κ-
casein.

The whey protein-coated micelles form a fi near (smaller whey pockets) gel then that formed
from unheated or HTST pasteurized milk, with less tendency to synerese.

In some countries, it is common practice to add sucrose to the milk for yoghurt, production to
reduce the acid taste. It is also very common practice to add fruit pulp, fruit essence or other
flavoring, e.g., chocolate, to yoghurt, either to the milk (set yoghurt) or to the yoghurt after
fermentation (stirred yoghurt).

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 Concentrated Fermented Milk Products

Throughout the Middle East, concentrated fermented milk products are produced, probably the
best known of which is Labneh for which the fermented milk is concentrated by removing part
of the serum (whey). This was done traditionally by stirring the yoghurt and transferring it to
muslin bags to partially drain. The typical composition of Labneh is: ~25 % total solids, 9–11 %
protein, and ~ 10 % fat and ~0.85 % ash (its protein content is similar to that of fresh, acid-curd
cheese).

 Novel Yoghurt Products

Since the late twentieth century, a number of yoghurt-based products have been introduced,
focused mainly on children: frozen (ice cream) yoghurt, dried yoghurt

(For long-term storage, intended to be rehydrated and to set on rehydration but the quality of the
gel is poor), and yogurt-based desserts (mousse).

 Rheology of Yoghurt

Fermented milk products exhibit thixotropic rheological properties, i.e., the viscosity (resistance
to flow) decreases as the rate of shear increases. The rheological properties are major
parameters of quality and are controlled by varying the total solids content of the milk, heat
treatment and homogenization of the milk or by the use of hydrocolloids, e.g., gelatin or
carrageenan, or including an exocellular polysaccharide-producing strain in the culture.

 Exocellular Polysaccharides

Many strains of all species of starter LAB produce exopolysaccharides (EPS) which are
responsible for the thickening of yoghurt and give a ropy property to the product; such products
include several Scandinavian fermented milk products, e.g., Taette, Skyr and Villi.

1.2 Cheese
Animal skins and inflated internal organs, particularly the rumen, have provided storage vessels
for a range of foodstuffs since ancient times. Hence, we can presume that cheese making was
discovered accidentally when storing milk in ruminant stomachs, which resulted in milk
curdling by the residual gastric rennin. Most modern cheeses are manufactured from pasteurized
milk coagulated in a vat by recombinant enzymes or proteases of vegetable origin with added
Lactococcus, Lactobacillus and/or Leuconostoc as starters. However, traditional raw milk
cheeses naturally fermented by its

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Indigenous microbiota are still produced in some Mediterranean countries. For specific cheeses
like blue or soft cheeses, bacteria of the genera Brevibacterium and Propionibacterium and
molds of the genus Penicillium are added to develop their characteristic organoleptical properties.
Raw milk can contain over 400 bacterial species. This microbial biodiversity decreases in the
cheese core usually dominated by few species of LAB but persists on the cheese surface with
high numbers of species of bacteria, yeasts and molds. It is commonly accepted that cheese
flavor develops as a result of the overall microbial metabolism beginning during clotting,
progressing further during cheese ripening.

Semi-hard cheeses typically contain non-starter lactobacilli (NSLAB), which can reach up to
107-108 cfu/g for long periods of time during production and storage. Propionibacterium
freudenreichii, a ripening culture in Swiss-type cheese, produces conjugated linoleic acid (CLA)
and may have bifidogenic and immunomodulatory properties.

1.3 Kefir
Kefir and Koumiss contain ~1 and ~6 % ethanol, respectively, which is produced by lactose-
fermenting yeasts, usually Kluyveromyces marxianus . The ethanol modifies the flavor of the
products and the CO 2 produced in the fermentation affects both their flavor and texture. Kefir,
which originated in Northern Caucasus Mountains, is most popular in northern and eastern
Europe. It is produced mainly from cows’ milk but the milk of goats and sheep, or mixtures of
the three, are also used.

There are two methods for preparing kefir, (1) using kefir grains and sub- culturing the resultant
fermentation or, (2) inoculating milk directly with starter cultures.

The traditional culture, “kefir grains”, contains a blend of lactic acid bacteria m(80–90 %),
lactose-fermenting yeast (10–15 %), acetic acid bacteria ( Acetobacter spp.) And possibly mould
( Geotricum candidum ) which are bound together by exopolysaccharides. Several species of
LAB are present, including Lactococcus spp. (especially L. lactis ssp. lactis ), Lactobacillus spp.,
S. thermophilus and Leuconostoc spp. Yeasts include Kluyveromyces marxianus var. lactis ,

Saccharomyces cerevisiae and Candida spp. A symbiotic relationship exists between the yeasts
and bacteria in kefir grains; yeasts produce vitamins, amino acids and other growth factors which

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are essential to maintain the integrity and viability of the microflora, while bacterial end products
are used as energy sources by yeasts

(Farnworth and Mainville 2003). The grains are up to 2 cm in diameter and contain

10–16 % dry matter, ~3 % protein, 0.3 % fat and ~6 % non-protein nitrogen.

Fig. 1.1 Production of kefir

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The culture for kefir is prepared by inoculating heated (95 °C × 30 min) milk at 20 °C with kefir
grains, incubating for ~20 h (to ~0.8 % lactic acid) and ripening at ~10 °C for ~8 h to facilitate
the growth of yeast. The grains are then strained off and the “filtrate” used to inoculate fresh
milk, at 1–3 %, and incubated to produce kefir or a bulk starter for large operations.

Figure 1.2 Production of kefir using commercial direct-to-vat cultures.

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1.4 Buttermilk
Originally, buttermilk was a by-product of butter production from ripened (sour) cream acidified
by adventitious mesophilic LAB; a similar product is now produced from cream ripened by a
culture of mesophilic LAB. However, cultured buttermilk is also produced from skimmed or
low-fat milk inoculated with a mesophilic LAB culture; this product is produced mainly in
English-speaking countries (USA,

Canada, UK, Australia), where most butter is produced from sweet cream. It is primarily a
drinking product and is also used in the production of soda bread. Basically similar products,
some including an extra-cellular polysaccharide-producing strain of LAB, which increases the
viscosity of the product making it ropy, are produced throughout North European countries Such
products include Tatmjolk, Surmjolk, Filbunke, Skyr, Langfi l, Villi (which contains Geotricum
spp.), Filmjolk and Ymer (concentrated, 3.5 % fat, 5.6 % protein) (see Tamine 2006 ). The
characteristic fl avour of cultured buttermilk is due mainly to diacetyl which is produced from
citrate by Lactococccus lactis ssp. lactis biovar. diacetylactis , which is included in the culture
for this product.

1.6 Sour Cream

Cultured cream is produced using a culture containing L. lactis ssp. lactis, L. Lactis ssp. cremoris
L. lactis ssp. lactis var. diacetylactis and Leu. mesenteroides ssp.

Cremoris; the former two are mainly responsible for acid production and the latter two for aroma
production (diacetyl). The typical fat content is 10–12 % but may be as high as 30 %.; the pH is
about 4.5 but it tastes less acidic than buttermilk or yoghurt, owing to the mellowing effect of the
fat. The inoculated cream may be distributed in cartons before fermentation at 22–24 °C until the
pH reaches 4.5 in about 20 h and is cooled in the package (set type), or it may stirred during
fermentation and then packaged; the former is very viscous. The cream for stirred cultured cream
is homogenized at 10–20 MPa. A long-life version of stirred cultured cream can be produced by
heat-treating the fermented product at 85–90 °C for a few seconds followed by packaging
aseptically. Cultured cream is used in many dishes, e.g., sauces, soups and dressings; it is
popular on baked potato.

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1.7 Koumiss
Koumiss (Kumys) is a traditional fermented product made from equine milk in

Central Asia, Russia, Mongolia, Kazakhstan, etc., and is widely consumed in these regions,
primarily for its therapeutic value. Russians, in particular, have long advocated the use of
koumiss for a wide variety of illnesses but the variable microbiology of the product has made it
difficult to confirm any theoretical basis for the claims (Tamime and Robinson 1999). In
Mongolia, koumiss is the national drink (Airag) and a high-alcoholic drink made by distilling
koumiss, called Arkhi, is also produced (Kanbe 1992 ). Per caput consumption of koumiss in
Mongolia is estimated to be about 50 L per annum .

The oldest method for the production of koumiss was by fermentation of lactose by adventitious
bacteria and yeasts to lactic acid and ethanol, respectively.

Horses were hand-milked with the foal in close proximity. Traditional koumiss (from fresh raw
milk) was usually prepared by seeding milk with a mixture of bacteria and yeasts using part of
the previous day’s product as an inoculum (‘slop-back culture’). The milk was held in a leather
sack called, a ‘turdusk’ (also called a ‘saba’ or ‘burduk’) which was made from smoked
horsehide taken from the thigh of a horse, i.e., it has a broad bottom and long narrow sleeve, with
a capacity of 25–30 L. Fermentation took from 3 to 8 h with a mixed microbial population which
consists mainly of Lb. delbrueckii subsp. bulgaricus , Lb. casei , L. lactis subsp. lactis ,
Kluveromyces fragilis and Saccharomyces unisporus . During the agitation and maturation stages
of production, more equine milk is added frequently to control the acidity and alcohol level. The
whole process was poorly controlled and often resulted in a product with an unpleasant taste, due
to the presence of too much yeast or excess acidifi cation. Turdusks, often containing caprine
milk from the previous season, were stored in a cool place over winter and the starter culture was
reactivated in Spring by gradually fi lling the turdusk with equine milk over about 5 days.

Koumiss is still manufactured in remote areas of Mongolia by traditional methods but with
increased demand elsewhere it is now produced under more controlled and regulated conditions.

A standardized protocol for koumiss production is of considerable interest for increasing the
market for, and consumption of, equine milk products in countries where it has not normally
been consumed. As well as using pasteurized equine milk, pure cultures of lactobacilli, such as

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Lb. delbrueckii subsp. bulgaricus , and yeasts are used for koumiss manufacture. Saccharomyces
lactis is considered best for the production of ethanol and S. cartilaginosus is sometimes used for
its antibiotic activity against Mycobacterium tuberculosis. Other microorganisms such as
Candida spp., Torula spp., Lb. acidophilus and Lb. lactis may also be used in koumiss
production. A schematic of the manufacture of commercial koumiss is shown in which outlines
the three stages of production: mother culture preparation, bulk starter preparation and koumiss
manufacture. The inoculation level of equine milk with bulk starter at 30 % is probably the
highest used in the manufacture of any fermented milk. Agitation is crucial for aeration of the
mix which promotes the growth of the yeast. The characteristics of good koumiss are optimal
when the lactic and alcoholic fermentations proceed simultaneously so that the products of
fermentation occur in definite proportions. As well as lactic acid, ethanol and CO2, volatile acids
and other compounds are formed which are important for aroma and taste and ~10 % of the milk
proteins are hydrolyzed. Products with varying amounts of lactic acid and ethanol are produced
and generally three categories of koumiss are recognized: mild, medium and strong. Koumiss
contains about 90 % water, 2–2.5 % protein (1.2 % casein and 0.9 % whey proteins), 4.5–5.5 %
lactose, 1–1.3 % fat and 0.4–0.7 % ash. Viable counts of ~4.97 × 10 7cfu /ml −1 and ~1.43 × 10
7cfu /ml −1 for bacteria and yeast, respectively, have been reported in koumiss.

Lactic acid in koumiss may occur in either the L (+) and D (−) isomer, depending on the type of
LAB used (Table 13.6). Both L (+) and D (−) isomers are absorbed from the gastrointestinal tract
but differ in the proportions converted to glucose or glycogen in the body. The L (+) isomer is
rapidly and completely converted to glycogen whereas Koumiss is thought to be more effective
than raw equine milk in the treatment of various illnesses due to the additional peptides and
bactericidal substances from microbial metabolism. Nowadays, the main interest in fermented
foods such as koumiss is their apparent ability to positively promote functions of human
digestion, i.e., to have a probiotic effect.

The low lactose content of koumiss compared to raw equine milk is favorable for those suffering
lactose intolerance; ~88 % of Mongolians are lactose intolerant but consume koumiss without ill-
effects, probably due to intra-intestinal digestion of lactose by microbial β-galactosidase in
koumiss, an enzyme that is not denatured in the acidic environment of the stomach. Furthermore,
koumiss is thought to be more effective than raw equine milk in disease treatment due to the

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presence of additional bioactive peptides and bactericidal substances produced during microbial
metabolism while retaining the high levels of lysozyme and lactoferrin of the original milk,
which have proven antibacterial activity.

Fig 1.3 Schematic for the production of koumiss (adapted from Berlin 1962).

 Technological Developments in Koumiss Manufacture

Blends of microorganisms in starter cultures have been developed that enhance flavor
development and extend the shelf-life up to 14 days. The presence of a high level of thermo-
stable lysozyme in equine milk may interfere with the activity of some starter cultures in the
production of fermented products. Equine milk heated to 90 °C for 3 min to inactivate lysozyme
has been reported to produce an acceptable fermented milk. In sensory tests, fermented

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unmodified equine milk has an unacceptable viscosity and scores very low in comparison to
fortified products for appearance, consistency and taste. In an attempt to improve the rheological
and sensory properties, fortification with sodium caseinate (1.5 g per 100 g), pectin (0.25 g per
100 g) and threonine (0.08 g per 100 g) has been investigated; the resultant products are reported
to have good microbiological, rheological and sensory characteristics even after 45 days at 4 °C.
Addition of sucrose and sodium caseinate has a positive effect on the rheological properties of
the product due to strengthening of the protein network.

 Koumiss-Like Products from Non-equine Milk

Koumiss-like products are produced in several areas, e.g., Mongolia, the former

USSR, Southern Europe and North Africa from camel milk (shubat), donkey milk (koumiss),
goat milk (tarag), ewe’s milk (arak or arsa) or buffalo milk (katyk). The

Physio-chemical and microbiological properties of asinine milk, such as low microbiological

load and high lysozyme content, make it a good substrate for the production of fermented
products with probiotic Lactobacillus strains. Asinine milk has been fermented with the probiotic
bacteria, Lb. rhamnosus (AT 194, GTI/1, and GT 1/3) which is unaffected by the high lysozyme
content of the milk and was viable after 15 days at 4 °C and pH 3.7–3.8. Lb. rhamnosus inhibits
the growth of most harmful bacteria in the intestine and acts as a natural preservative in yoghurt-
type products, considerably extending shelf-life. Fermented asinine milk produced using a mixed
culture of Lb. rhamnosus (AT 194, CLT 2.2) or Lb. casei (LC 88) had a high viable bacteria
count after storage for 30 days. Some sensory differences have been reported for fermented
asinine drinks and those made with the Lb. case strain developed a better and balanced aroma
than the boiled vegetable/acidic taste and aroma of the product made with Lb. rhamnosus alone.

Due to shortages of equine milk and the cost, when it is available, research has been
undertaken to produce koumiss-like products from bovine milk, which must be modified to
make it suitable for koumiss production. Koumiss of a reasonable quality has been produced
from whole or skimmed bovine milk containing added sucrose using a mixture of Lb.
acidophilus, Lb. delbrueckii ssp. bulgaricus and Kluyveromyces marxianus var. marxianus
or Kluyveromyces marxianus var. lactis as starter culture. Koumiss has also been made from
diluted bovine milk with added lactose and, more successfully, from bovine milk mixed

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 with concentrated whey using a starter culture of Kluyveromyces lactis (AT CC 56498), Lb.
delbrueckii subsp. bulgaricus and Lb. acidophilus . Starter cultures for koumiss manufacture
from bovine milk may also include Saccharomyces lactis (high antimicrobial activity
against Mycobacterium tuberculosis) in order to retain the ‘anti-tuberculosis image’ of
equine milk.

1.8 Curd (dahi)

Milk, liquid secreted by the mammary glands of female mammals to nourish their young for a
period beginning immediately after birth. The milk of domesticated animals is also an important
food source for humans, either as a fresh fluid or processed into a number of dairy products such
as butter and cheese . Milk is a nutritious choice as it provides nine essential nutrients our body
needs. Milk contains essential nutrients like high-quality protein, calcium, vitamin D and more.
These nutrients help our bodies function properly. For example: Protein helps build and repair
muscle tissue Calcium and vitamin D helps build and maintain strong bones and teeth Milk also
contains B vitamins, which can help your body convert food into energy.

India is leading milk producing country in the world, accounting for 19 percent of the global
market share and expected to grow at compound annual growth rate (CAGR) of 14.8% between
FY 2018 – 2023. As per fiscal year 2019, milk production in India amounted to about 187
million metric tons. As per FY – 2018, around 81% of the Indian dairy and milk processing
market comes under unorganized sector, where milk is processed in unhygienic infrastructure,
which affects the overall quality of milk and milk-based products. Consumption patterns of
liquid milk at the farm level and less infrastructure for processing is the main reason for low
value addition of milk. The demand for value added products especially traditional dairy
products is increasing day by day and the dairy industry of the country is trying to meet the
present demand.

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