Kefir Full

Food processing technology Kefir
Content
1. Raw materials ...........................................................................................................................................4

1.1. Milk................................................................................................................................................4
1.1.1. Milk composition ..................................................................................................................4
1.1.2. Criteria for milk ...................................................................................................................6
1.2. Kefir grains ...................................................................................................................................8
1.2.1. Microbiology of kefir grains ................................................................................................9
1.2.2. Structure of grains .............................................................................................................11
1.2.3. Growth cycle of kefir grains ..............................................................................................12
1.2.4. Criteria for kefir grains selection......................................................................................13
1.2.5 Criteria for freeze-dried kefir starter ...............................................................................13
1.3. Materials for milk standardized ....................................................................................................14
1.3.1 Anhydrous milkfat ....................................................................................................................14
1.3.2 Skim milk powder .....................................................................................................................15
1.4. Food Additives ............................................................................................................................16
1.4.1 Gelatine ......................................................................................................................................16
1.4.2. Food additive E.471 ..................................................................................................................17
2. Processing flow chart of kefir 1 .........................................................................................................18
2.1 Fat standardisation ....................................................................................................................19
2.2 Deaeration and evaporator ........................................................................................................22
2.3 Homogenization ..........................................................................................................................24
2.4 Pasteurization .............................................................................................................................25
2.5 Propagation .................................................................................................................................27
2.5.1 Raw milk .................................................................................................................................27
2.5.2 Pasteurization: ....................................................................................................................27
2.5.3 Inoculation and Propagation:............................................................................................28
2.5.4 Filtration: ............................................................................................................................31
2.5.5 Washing and storing of grains: .........................................................................................32
2.5.6 Storing of the filtrate: ........................................................................................................32
2.6 Inoculation ..................................................................................................................................32
2.7 Incubation ...................................................................................................................................33
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2.8 Cooling ........................................................................................................................................39

2.9 Packaging ....................................................................................................................................40
3. Processing flow chart of kefir 2 .........................................................................................................42
3.1 Fat standardization: .........................................................................................................................43
3.2 Homogenization: ...............................................................................................................................43
3.3 Freeze-dried kefir starter ................................................................................................................44
Freeze-dried kefir starter ..........................................................................................................................44
3.4 Cooling: .............................................................................................................................................45
4. Comparison of two production lines: ...............................................................................................46
5. Criteria for evaluation of product quality .......................................................................................47
5.1 Chemical characteristic: ............................................................................................................47
5.2 Physico-chemical characteristic ......................................................................................................49
5.3 Microbiological characteristic .........................................................................................................50
5.4 Sensorial characteristic ...................................................................................................................50
List of figures
Figure 1. 1 Kefir grains ..................................................................................................................................9
Figure 1. 2 Electron micrograph of a kefir grains ........................................................................................11
Figure1. 3 Kefir health benefits....................................................................................................................12
Figure1. 4 Structural of E471 .......................................................................................................................17
Figure 2. 1 Centrifugal separation ................................................................................................................20

Figure 2. 2 Principle for direct in-line standardization of cream and milk. .................................................20
Figure 2. 3 Direct in-line standardization system.........................................................................................21
Figure 2. 4 Deaeration equipment ................................................................................................................23
Figure 2. 5 Single-stage homogenisation .....................................................................................................25
Figure 2. 6 Plate heat exchanger ..................................................................................................................27
Figure 2. 7 The bulk starter tanks .................................................................................................................31
Figure 2. 8 Fermentation tank ......................................................................................................................33
Figure 2. 9 Pathway used by bifidobacteria for lactose metabolism ............................................................35
Figure 2. 10 Heterofermentative pathway for lactose metabolism among lactic-acid bacteria ...................36
Figure 2. 11 Gate agitator .............................................................................................................................38
Figure 2. 12 Typical cone-bottom fermentation tanks used in a kefir plant.................................................39
Figure 2. 13 Filling machine ........................................................................................................................41
Figure 3. 1 Two-stage homogenisation head. ..............................................................................................44

Figure 3. 2 Freeze-dried kefir starter ............................................................................................................44
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Figure 3. 3 Bulk starter tank .........................................................................................................................45

Figure 3. 4 The tubular heat exchanger ........................................................................................................46
List of tables
Table 1 1 Milk composition ...........................................................................................................................6
Table 1 2 Milk chemistry composition ..........................................................................................................7
Table 1 3 Biochemistry criteria .....................................................................................................................8
Table 1 4 Bacteria, yeast and mold isolated from ........................................................................................10
Table 1 5 Culture characterization for the Codex Standard for Fermented Milk. Source: Codex
Alimentarius (2004). ....................................................................................................................................11
Table 1 6 Kefir chemistry criteria ................................................................................................................13
Table 1 7 The chemical criteria ....................................................................................................................13
Table 1 8 The content of microorganism in freeze-dried kefir starter .........................................................14
Table 1 9 Value composition of skim milk powder .....................................................................................15
Table 1 10 The chemical composition values of skim milk powder ............................................................16
Table 2. 1 Microorganisms used in starter cultures for cultured in kefir .....................................................29

Table 2. 2 Characteristics of mesophilic starter bacteria used for cultured kefir. ........................................30
Table 4. 1 Comparison of two production lines ...........................................................................................46
Table 5. 1 The chemical composition and nutritional values of kefir ..........................................................48

Table 5. 2 Chemical composition of various kefirs......................................................................................49
Table 5. 3 Suggested microbiological standards for dairy product ..............................................................50
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Chapter 1: INTRODUCTION
Kefir
Kefir is the fermented dairy product that is described in many reference texts and has long been
of academic interest to microbiologists, but was one for which few U.S students (or consumers for
that matter) had any first-hand knowledge. Although several brands of kefir are now available
throughout the United States, it is still a most unknown product. This is despite the popularity of
this product throughout a large part of the world. In the Middle East, Eastern Europe, and Central
Asia, especially Turkey, Russia, Ukraine, Poland, and Czech Republic, kefir is one of the most
widely consumed cultured dairy products, accounting for as much as 70% of all cultured milk
products consumed. It is interesting that yogurt manufacturers have begun to introduce fluid or
pourable yogurt-like products that are only slightly different than traditional kefir.
Kefir originated in the Caucasus Mountain in Russia. The traditional kefir manufacturing
process, which is still widely practiced, relies on a mixed assortment of bacteria and yeast to initiate
the fermentation. The kefir fermentation is unique among all other dairy fermentations in that the
culture organism are add to the milk in the form of insoluble particles call kefir grains. Moreover,
once the fermentation is complete, the kefir grains can be retrieved from the fermented milk by
filtration and reused again and again.
The flavor of plain kefir is primarily due to lactic and acetic acids, diacetyl, and acetaldehyde,
produced by homo-fermentative and hetero-fermentative lactic acid bacteria. However, because
kefir grains also content yeast, in addition to lactic acid bacteria, other end-products are formed
that make the finished product quite different from other cultured dairy products. This is because
ethanol is produced when the yeast ferment lactose, such that kefir can contain as much as 2%
ethanol (accounting perhaps, for its appeal). In the United States, this much ethanol would trigger
action from the regulatory agencies. Thus, if yeast is present in the culture (most kefir products
made in the United States claim yeasts on their labels), they must be low or non-ethanol producers.
Kefir grains can also contain acetic acid-producing bacteria, such as Acetobacter aceti.
1. Raw materials
1.1.Milk
Kefir is made using milk of different species of mammals (cow, sheep or goat). In the
United States, kefir is usually made with low-fat or whole milk. However, in this article, we just
discuss about bovine whole milk as the major ingredient.
1.1.1. Milk composition
Chemistry speaking, milk is a complex fluid in which more than 100,000 separate
molecules and chemical entities have been found, the levels of which vary with the species. In term
of physical chemistry, milk is the opaque, white heterogeneous fluid in which various constituents
are held in multi-dispersed phases of emulsion, colloidal suspension, or solution.
The composition of milk is generally described in terms of its commercially important
constituents, milk fat, and nonfat solids (NFS) or milk solids-not-fat (MSNF). The MSNF consists
of protein, lactose, and minerals. These solids are also referred to as “serum solids”, the term “total
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solids” refers to the serum solid + the milk fat. Factors affecting composition of milk: individuality
of cow, milking (stages, intervals, completeness, frequency, irregularity, portion), lactation period,
yield of cow, season, feed, environment, administration of drugs and hormones…
Physical structure: milk have well-defined physical equilibrium between various
constituents that exits mainly in three forms, namely, emulsion and colloidal and true solution.
Milk lipids are present as an “oil in water type”, of emulsion in the form of microscopic globules
varying from 0.1 to 22 µm in diameter. The colloidal phase contains casein micelles, calcium
phosphates and globular proteins. Whey proteins are in colloidal solution and the casein is in
colloidal suspension. Lactose, vitamins, acids, enzymes and some inorganic salts are present as
true solutions.
Major constituents:
 Water: 85.4 – 87.7%
 Fat: (3.4 – 5.1%) depending on the breed of the cow, occurs as microscopic globular
emulsion of lipid fat in aqueous phase of milk plasma (average size varies from 3.4
to 4.5 µm), consist chiefly of triglycerides of fatty acids (95-97% of milk fat), along
with fat-soluble vitamin A, D, E and K, essential fatty acids (EPA, DHA, ARA…),
cholesterol, phospholipids (approximately 36 mg/100mL)
 Proteins: contains hundreds of proteins and most of them occur in trace amounts.
The major proteins of milk are broadly classified as caseins (make up 80% of total
protein) and whey proteins (make up 20% remaining).
 Lactose: the major carbohydrate of milk, ranges from 4.8% to 5.2%, relatively
constant. It is a reducing sugar and extensive heating of milk results in Maillard
reaction between lactose and proteins creating brown pigments and brownish color
of milk. The role of lactose in yogurt and fermented milks is extremely important
because the culture nutritionally requires it as a substrate for growth. It is the source
of carbon and after fermentation about 30% of the lactose content is converted to
lactic acid.
 Minerals: The ash content is not quite equivalent to salt level in milk, because some
salts like chlorides and organic salts are volatilized or destroy as a result of high-
temperature exposure.
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Table 1 1 Milk composition
1.1.2. Criteria for milk

Physical:
Optical properties
Color: the color and appearance of milk has significance because the producers perceive it
as parameter of quality. The opaque, white or turbid color of milk is due to scattering of light by
the dispersed phase of fat globules, casein micelles, and the colloidal calcium phosphate. The
intensity of color is directly proportional to the size and the number of these particles. The creamy
color of whole milk is due to its β-carotene and xanthophyll content. The homogenization increases
the number and total volume of fat globules, the result is whiter color of homogenized products.
Flavor: taste and aroma are critical to the assessment of milk. Flavor constitutes a critical
criterion of quality for the producers. It is a sensory property in which odor and taste interact. The
sweet taste of lactose is balanced against the salty taste of chloride and both somewhat moderated
by proteins. Although milk has a clean, pleasantly sweet flavor, it is quite bland and therefore, any
off-flavors are readily discernible. Off flavors result when the balance of flavor compounds is
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altered due to the microbiological activity or processing treatments, or chemical or bio-chemical

reactions, or absorb aromatic odors.
Density and viscosity:
Density of milk (mass/volume) is the sum total of the densities of its constituents, their
concentration, and state at a particular temperature. Milk density at 20oC range from 1.027 to
1.033 with an average of 1.030 g/cm-3 . Accordingly, the weight of 1L of milk would range from
1.027 to 1.033kg. The lower the density, the higher the fat content, which is the higher the risk of
fat separated during the production line.
Viscosity: depend on the temperature and on concentration and state of casein micelles and fat
globules, representative values at 20oC is 1.9 cP. It has bearing on fat separation/creaming, rate of
heat transfer and flow conditions during processing of milk.
Thermal properties
Thermal expansion: when warmer the volume increases which affects the design considerations
for flow rates through processing treatments. The coefficient of thermal expansion of fresh milk
is approximately 0.335 cm3/kg at temperature range of 5-400C.
-Heat capacity (0.93 cal/g/oC) and specific heat are important in processing for determining the
amount of heat or refrigeration necessary to change the temperature of milk.
Chemistry
The pH of normal, fresh, sweet milk usually varies between 6.6 and 6.8. Higher pH values for fresh
milk indicate udder infection and lower values, bacterial action.
 No antibiotics or drugs
 No cleansing chemicals from equipment or milk containers
 Chemistry composition require
Table 1 2 Milk chemistry composition
Biochemistry
There are microorganisms naturally present in milk, but out of producers ’s desire. The permitted
number of microorganism are in the table below:
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Table 1 3 Biochemistry criteria
Raw milk naturally contains low levels of endogenous enzymes, including alkaline
phosphatase, plasmin and lipase – which are important from processing point of view. The absence
of alkaline phosphatase after pasteurization indicates effective pasteurization was accomplished.
Plasmin, which is stable at pasteurization temperature, hydrolyzes both β- and α- CN in milk.
Lipase are associated with fat metabolism and with quality deterioration in kefir product. Other
enzymes like lysozyme attacks the Gram-positive bacterial cell walls.
 To be summed up, a sample will be taken from each bulk tank of milk shipped. This sample
may be tested for:
- Inhibitors: milk must be free from substances that inhibit bacterial growth in raw milk.
These substances are normally veterinary drug residues. Penalties are applied if
inhibitors of any type are in penalty range in the bulk tank milk sample. Losses and cost
may also be charged to an offending producer when a tanker-truck is contaminated.
- Abnormal freezing point: milk must have a freezing point that is less than -0.506oC. A
penalty is applied if the official Cryoscope result is greater than the specified level for
abnormal freezing point (AFP) of -0.507oC.
- Bacteria: Milk must contain less than 122,000 individual bacteria cells (IBC) per mL.
A milk sample that contains greater than 121,000 IBC is in the penalty range.
- Somatic cell count (SCC): milk must contain less than 400,000 individual cells (IC) per
mL. A milk sample with a test result greater than 399,000 IC/mL is in the penalty range.
1.2.Kefir grains
Kefir grains resemble small cauliflower florets: they measure 1-3 cm in length, are
lobed, irregularly shaped, white to yellow-white in color, and have a slimy but firm texture. They
are insoluble in water and most solvents. Grains are kept viable by transferring them daily into
fresh milk and allowing them to grow for approximately 20 hours; during this time, the grains will
have increased their mass by 25%. Grain must be replicated in this way to retain their viability,
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since old and dried kefir grains have little or no ability to replicate. Kefir grains replicated in milk
“at home with daily changes of milk” and stored for three months either at room temperature or at
4oC had microbiological profiles that were different to those of fresh grains. In addition, washing
grains in water also reduced viability. It has been recommended that in a commercial operation
using grains to produced kefir, grain should be kept viable through daily transfers and should only
be replaced if their ability to ferment milk becomes impaired. Low temperature storage appears to
be the best way to maintain kefir grains for long periods. Some show that storage of kefir grains at
-80 or -20 for 120 days did not change their fermentation properties compared to grains that had
not been stored; however, grains stored at -4oC did not produce acceptable kefir after thawing.
Kefir grains replicated in soy milk have been reported. There have been no reports of successful
production of kefir grains from pure cultures.
While early studies of kefir grains employed light microscopy, later investigations used
electron microscopy to describe the complex microbial community of which they were comprised.
Figure 1. 1 Kefir grains
1.2.1. Microbiology of kefir grains

To microbial ecologists, kefir grains represent an amazingly diverse ecosystem. A single
grain can harbor more than a million cells, with perhaps dozens of different species present. The
grains are essentially a natural immobilized cell bioreactor, with the kefir grain serving as an inert
support material for the cells.
Scientists stated that kefir bacteria and yeasts, when separated as pure cultures, either do
not grow in milk or have a decreased biochemical activity, which further complicates the study of
the microbial population of kefir grains. Several media have been proposed for the isolation and
identification of bacteria in kefir grains.
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Table 1 4 Bacteria, yeast and mold isolated from
Yeasts has been recognized its important role in the preparation of fermented dairy
products, where they can provide essential growth nutrients such as amino acids and vitamins, alter
the the pH, secrete ethanol and produce CO2. The yeast in kefir have been less well studied than
kefir bacteria; although it is obvious that the yeast provide an environment for the growth of kefir
bacteria, producing metabolites that contribute to the flavor and mouthfeel of kefir.
The predominant organisms in kefir grains are lactic acid bacteria, with Lactobacillus
species accounting for up to 80% of the total. Other studies have shown homo-fermentative species,
including Lactobacillus kefirgranum and Lactobacillus kefirranofaciens, were less common. Other
lactic acid bacteria that have been identified include species of Lactococcus, Streptococcus,
Leuconostoc. Yeasts are also well-represented, and include lactose-fermenting (Saccharomyces
kefir) and non-fermenting strains.
It was reported recently that kefir grains likely contain organisms that cannot easily be
cultured and that are instead only viable within the confines of the grain environment. Collectively,
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the kefir microflora has considerable metabolic diversity, which explains, in part, the incredible
stability and viability of the grains, as well as the wide spectrum of products produced during the
fermentation.
Table 1 5 Culture characterization for the Codex Standard for Fermented Milk. Source: Codex
Alimentarius (2004).
1.2.2. Structure of grains
Figure 1. 2 Electron micrograph of a kefir grains

The grains consist of proteins, polysaccharides. Attachment to the grains is mediated via
one of several different types of polysaccharide-like material (kefiran being prototype) that are
produced by the resident organisms. Microbial cells were not bound to one another but
encapsulated within a muco-polysaccharide believed to be produced by the encapsulated
microorganisms, call kefiran (constitutes between 24-25% of dry weight of kefir grain, and is a
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matrix of fibrillary amorphous material that consists largely of polysaccharides, soluble in water,
holds the grains together). The kefir organisms do not appear to be randomly distributed; rather,
some are contained mainly within the core of the grains, whereas others are located primarily in
the exterior. Symbiotic and synergistic relationships likely exits between different organisms, but
these relationships are difficult to establish in such a complex system.
Figure1. 3 Kefir health benefits
Although the precise chemical composition of kefir grains varies depending on the source,
they generally contain about 80% to 90% water, 2% to 5% protein, and 8% to 10% carbohydrate
casein, whereas most of the carbohydrate portion is polysaccharide. The irregular, cauliflower
shaped grains range in size from 0.2 cm to more than 2cm in diameter. In their dry form, they are
stable for many months. Although kefir grains are available commercially, many modern
manufacturers mow use pure lyophylized cultures containing many of the strains ordinarily found
in the grains.
1.2.3. Growth cycle of kefir grains
External surface area of each grain, may vary from smooth areas of diverse irregularity,
containing an array of irregular small rounded protrusions randomly scattered over the surface.
Some grain’s surface texture may be smooth, while the others may exhibit a greater proportion of
surface area covered with multiple irregular rounded protrusions. Other grains may exhibit a
mixture of both surface-textures.
The grains form from irregular flat sheaths. These sheaths grow in an irregular fashion, to
form multiple and irregular lobes (refer to baby-grains). These lobes have a natural tendency to
form as self-enclosed lobular bio-structures, with a growth-signature unique to each baby-grain.
Each of these lobules are conjoined at a common midsection, radiating outwardly while attached
to a center-point of a mother grain. On appearance, the growth pattern of baby-grains share self-
similarities with mother-grains. After a period of time, and possibly due to external stress or trauma,
lobular sections break free from the mother grain. These smaller bodies or baby-grains, eventually
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propagate into mother-grains, by increasing in overall size. This growth-cycle simply repeats, to
continue the process in a similar fashion (self-propagation).
1.2.4. Criteria for kefir grains selection.
Chemistry criteria
Table 1 6 Kefir chemistry criteria
Sensory criteria
-The state: resemble small cauliflower florets, each grain is 3-20mm in diameter, soft,
gelatinous biological mass, not contaminated by the impurities in the surface of grains or
contaminated by the strange compounds.
-The color: white to yellow, swell inside water
Biological criteria
 Microorganism in kefir don’t have the toxic synthesis ability
 The symbiotic relationship between the organisms in kefir grains
 High activity of the kefir grain’s microorganism
 The stability of microbes and yeast’s activity
 Easily storage in a long time
1.2.5 Criteria for freeze-dried kefir starter
Chemical criteria
Table 1 7 The chemical criteria
Chemical composition Content (%)

Water 3.5
Lipid 4.4
Ash 12.1
Muco-polysaccharides 45.7 45.7
34.3 (insoluble protein 27%, soluble protein
Total protein
1.6% and amino acid 5.6%)
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Biological criteria
Table 1 8 The content of microorganism in freeze-dried kefir starter
The microorganism Content (%)

Homofermentative streptococci 75
Citric-fermenting streptococci 24
Lactobacilli 0.5
Yeasts 0.1
In kefir production, the manufacturers always care about some follow criteria of kefir starter:
 Don‟t contain any toxic compounds.
 High activity of the kefir starter ‟s microorganism.
 The stability of microbes and yeast ‟s activity.
 Easily storage in a long time.
Sensory criteria
The state: the starter contains two parts: the grains particles are the kind of yellowish chunks, the
whiter powder is milk, the grains particles are bigger than milk particles. The starter do not
contaminated by the impurities or contaminated by the strange compounds.
The color: the milk powder is white, the grains‟ particles are yellow.
The odour: smell like cheese, do not have the strange smell.
1.3. Materials for milk standardized
1.3.1 Anhydrous milkfat
Anhydrous milk fat and butteroil are products consisting of more or less pure milk fat.
Although they are modern industrial products, they have ancient traditional roots in some cultures.
Defined by Codex Alimentarius standard CODEX STAN 280-1973, Anhydrous milkfat,

milkfat, anhydrous butteroil and butteroil are fatty products derived exclusively from milk and/or
products obtained from milk by means of processes which result in almost total removal of water
and non-fat solids. Anhydrous milk fat, butter oil, can be manufactured from either butter or from
cream. For the manufacture from butter, non-salted butter from sweet cream is normally used, and
the process works better if the butter is at least a few weeks old. Melted butter is passed through a
centrifuge, to concentrate the fat to 99.5% of greater. This oil is heated again to 90-95oC and
vacuum cooled before packaging
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 Anhydrous milk fat must contain at least 99.8 % milk fat and be made from fresh
cream or butter. No additives are allowed, e.g. for neutralization of free fatty acids.
 Anhydrous butteroil must contain at least 99.8 % milk fat, but can be made from
cream or butter of different ages. Use of alkali to neutralize free fatty acids is
permitted.
 Butteroil must contain 99.3 % milkfat. Raw material and processing specifications
are the same as for anhydrous butteroil.
Butter has been the traditional form of storage for milk fat, but in some cases AMF is more
preferable, because it requires less storage space.
Butter is regarded as a fresh product, although it can usually be stored at +4°C for up to six
weeks. If it is stored for a longer period of time, say up to 10 – 12 months, a storage temperature
of at least –25°C is mandatory.
AMF is typically packed in 200-litre drums nitrogen (N2) headspace, and can be stored for
several months at +4°C. AMF is liquid at temperatures above 36°C and solid below 16 – 17°C.
AMF is convenient to use in liquid form because it is easy to mix with, and meter into other
products. Thus, AMF is used for recombination of various dairy products, but it is also used in the
chocolate and ice cream manufacturing industries.
Demand for butter is decreasing, partly due to the increased use of AMF. One field of
application where the use of AMF will increase is in “blends” of different fat contents and with
mixtures of butter and vegetable oils, to make products with different functional properties.
1.3.2 Skim milk powder
Nonfat dry milk is a product that is created when water is removed from pasteurized nonfat
milk. By depriving microorganisms of the water which they require in order to develop, drying
allows skim milk powder to have a shelf life of up to 3 years if stored properly. It contains lactose,
milk proteins and minerals in the same relative proportions as the fresh milk from which it was
made. Skim milk powder is by far the most common type of milk powder available.
Table 1 9 Value composition of skim milk powder
The powder should be stored under cool, dry conditions and should be protected from
contact with water during storage. With its low moisture content, chemical reactions in skim milk
powder stored at room temperature take place so slowly that the nutritive value is not affected,
even after years of storage.
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Skim milk powder (SMP) must contain no less than 95 percent milk solids and must not
exceed 4 percent moisture or 1.5 percent of fat, unless otherwise indicated. In the case of
instantized SMP, Vitamins A and D are added. Regular and instantized SMP may also contain an
anti-foaming agent.
Table 1 10 The chemical composition values of skim milk powder
Strict microbiological requirements are placed on the quality of the raw milk that is used in
the manufacture of milk powders. Consequently, SMP should be free from bacteria of the
genus Salmonella. One gram of powder must not contain more than 50 000 mesophilic bacteria,
10 coliforms and 100 Staphylococcus aureus. These counts correspond respectively to about 5000,
1 and 10 per litre of reconstituted product, provided that contamination does not occur after
manufacture.
Characteristics
 Appearance: Dry milk powder must be reasonably uniform in composition, white- or

cream-coloured, free from the brown or yellow colour (typical of overheated product) or
any other unnatural colour and must also be substantially free from brown specks. Its
flavour and odour must be sweet, clean and free from rancid, thallowy, fishy, cheesy,
soapy or other objectionable flavours and odours.
 Spray-drying: is the principal method used for drying milk in the dairy industry. Most
skim milk powders used as ingredients are spray dried and are manufactured in two basic
varieties: instant (agglomerated) and non-instant (regular). The instantized spray-drying
process gives the powder a larger grain size than regular spray dried powder and renders it
instantly soluble when reconstituted in cold water. Furthermore, the heat treatment
applied during the manufacture of specific spray dried skim milk powders will, to a
certain extent, affect the denaturation of the milk proteins, and consequently, their
solubility and functionality as milk ingredients. The degree of denaturation is normally
expressed by the Whey Protein Nitrogen Index (WPNI) as milligrams of undenatured
whey protein (u.w.p.) per gram of powder.
1.4. Food Additives

1.4.1 Gelatine
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The primary purpose of using a stabilizer in yogurt is to produce smoothness in body and
texture, impart gel structure, and reduce syneresis. The stabilizer increase shelf-life and provides a
reasonable degree of uniformity of product. Stabilizer function through their ability for form gel
structure in water, thereby leaving less free water for syneresis.
Gelatin is derived by irreversible hydrolysis of the proteins collagen and ossein. It is used
at the level of 0.3-0.5% to get a smooth shiny appearance in refrigerated yogurt. Gelatin is a good
stabilizer for frozen yogurt. The term “Bloom” refers to the gel strength as determined by a Bloom
gelometer under standard conditions. Gelatin of a Bloom strength of 225 to 250 is commonly used.
The gelatin level should be geared to he consistency standards for yogurt. At the temperature below
10oC, the yogurt acquires a pudding-like consistency. Gelatin tends to degrade during processing
at ultrahigh temperature dependent. Consequently, the yogurt gel is considerable weakened by a
rise in temperature.
1.4.2. Food additive E.471
Mono- and diglycerides of fatty acids (E471) is a food additive used as an emulsifier. These
synthetic fats are produced from glycerol and natural fatty acids, from either plant or animal origin.
E471 is generally a mixture of several products, and its composition is similar to partially digested
natural fat. White or cream coloured hard fats of waxy appearance, plastic products or viscous
liquids. Using as thickener, stabilizer, emulsifier.
Figure1. 4 Structural of E471

E471 is mainly produced from vegetable oils, although animal fats are sometimes used and
cannot be completely excluded as being present in the product. The fatty acids from each source
are chemically identical. The Vegan Society, which discourages eating animal-based foods, flags
E471 as a "thing to look out for" that "can be animal or non-animal based"
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Chapter 2: PRODUCTION LINE

2. Processing flow chart of kefir 1
Raw milk/ skim

milk/reconstitut
ed milk
Raw milk Pasteurization Inoculum

Raw milk/
skim
Inoculation milk/reconstit
uted milk
Fat standardization
Propagation Washing
Gas Deaeration
Additives Homogenizatio filtration

n
Kefir
Pasteurization grains
Gelatin
Inoculum for
Inoculation production
Incubation
Cooling
Package Packaging
ss
Kefir ( cool
store)
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Information of each operation in the production line

2.1 Fat standardisation
Objective: improvement operation of product
This operation used for standardizing the fat content in milk to satisfy the requirement for
the fat content in kefir product. Normally, the fat content of kefir varies between 5% and 6%.
Standardisation of fat content involves adjustment of the fat content of milk by addition of
cream or skim milk as appropriate to obtain given fat content. The cover mixtures can be mixture
of whole milk with skim milk, cream with whole milk, cream with skim milk and skim milk
with anhydrous milk fat (AMF).
Transformation of raw material:
Physical transformation: Some physical criteria can change such as density, thermal conductivity,
coeffiective,…
Chemical transformation: The fat content in the milk can be decrease or increase so the chemical
composition of milk changes and dry matter changes,…
Equipment:
The fat content in the raw milk is not stable but quite varies, can be lower or higher than the fat
content in the milk standardization.
 If the incoming milk has the higher fat content than the standardized milk, we separate the
fat from milk by using the equipment for lower-fat-content standardized milk.
 If the incoming milk has the lower fat content than the standardized milk, we add AMF to
the milk by using the equipment for higher-fat-content standardized milk.
 Centrifugal separation
Principle of Centrifugal separation:
In the separator the milk is supplied to the separator bowl from an inlet, normally in the top,
through a stationary axial inlet tube.
When the milk enters the ribbed distributor (1), it is accelerated to the speed of rotation of
the bowl before it continues into the separation chan-nels in the disc stack (2). The centrifugal force
throws the milk outwards to form a ring with a cylindrical inner surface. This is in contact with air
at at-mospheric pressure, which means that the pressure of the milk at the sur-face is also
atmospheric. The pressure increases progressively with increas-ing distance from the axis of
rotation to a maximum at the periphery of the bowl.
The heavier solid particles settle outwards and are deposited in the sedi-ment space. Cream
moves inwards towards the axis of rotation and passes through channels to the cream paring
chamber (3). The skimmilk leaves the disc stack at the outer edge and passes between the top disc
and the bowl hood to the skimmilk paring chamber (4).
19
Figure 2. 1 Centrifugal separation

With:
1 Distributor
2 Disc stack
3 Cream paring chamber
4 Skimmilk paring chamber
In standardization milk system:

People use the automatic standardization control system to carry out the fat standardization.
The direct in-line standardization system is used with the centrifugal separator.
Figure 2. 2 Principle for direct in-line standardization of cream and milk.
20
The direct in-line standardization system:

In modern milk processing plants with a diversified product range, direct in-line
standardization is usually combined with separation. Previously, standardization was done
manually, but, along with increased volumes to process, the need for fast, accurate standardization
methods, independent of seasonal fluctuations of the raw milk fat content, has increased. Control
valves, flow and density meters and a computerized control loop are used to adjust the fat content
of milk and cream to desired values. This equipment is usually assembled in units.
The pressure in the skim milk outlet must be kept constant in order to enable accurate
standardization. This pressure must be maintained, regardless of variations in flow or pressure
drop caused by the equipment after separation, and this is done with a constant-pressure valve
located close to the skim milk outlet.
For precision in the process, it is necessary to measure variable parameters such as:
 Fluctuations in the fat content of the incoming milk

 Fluctuations in throughput
 Fluctuations in pre-heating temperature
Figure 2. 3 Direct in-line standardization system
21
Usually, whole milk is heated to 55 – 65 °C in the pasteurizer, after that the system will
separate warm milk into cream and skim milk by the centrifugal separator, then automatically and
continuously recombine the two streams in the proportion required to obtain the desired skim milk:
cream ratio to have the standardized milk.
Technological parameter:
The fat content of standardization milk between 0.5% and 6%, frequently specified at 2.5% to
3.5%.
2.2 Deaeration and evaporator

Objective:
Deaeration preparation operation for homogenization and the other is improvement.
Deaeration make gel structure of kefir is smooth ang prevent forming in product.
Removes air from the medium to produce a more conducive medium for microaerophilic
lactic cultures to grow. The air content of the milk used to make fermented milk products should
be as low as possible.
The advantages gained through deaeration are:

 Improved stability and viscosity of the yoghurt
 Shortened fermentation time
 Improved working conditions for the homogenizer
 Less risk of fouling during heat treatment
 Removal of volatile off-flavors (deodorization)
Objective evaporator is adjust sodil content in raw milk → improvement operation.
Biological : inhibited aerobic bacteria
Chemical: removed air from raw material, therefore reduced various chemicals are available which
react with the oxygen in milk to produce chemical forms that are not harmful ( ex: oxidation lipid
reaction,…) and happen reaction Maillard between reducing group of lactose with amin group and
peptide in raw milk.
Physical: prevent foaming during production, density and viscosity of milk will be changes in
heating.
Equipment:
22
In kefir production, evaporation is utilised to concentrate milk from 11% to 13% or 16.25%
solids content respectively. This treatment simultaneously deaerates the product and rids it of off-
flavours.
Equipment consist of: heating and vaccum deaeration, barometer.
Milk will be heating in plate exchanger equipment, then go into the vaccum deaeration
cylinder equipment, gas will be evaporate and remove on the top. The gas into barometer
equipment. Air and other noncondensable gases are extracted from the condenser by a vacuum
pump. The product eventually loses velocity and falls to the inwardly curved bottom, where it is
discharged.
With: 1- plate exchanger equipment
2- vaccum deaerator
Figure 2. 4 Deaeration equipment

Technogical parameter: the temperature of milk in plate exchanger do not overcome 800C,
concentration of raw milk is about from 11% to 13%.
After the deaeretion stage, food additives E471 was put into the balance tank while mixing,
amount above 0.35% tend to give milk.
23
2.3 Homogenization
Objective: improvement, preservation and preparation operation for pasteurization.
The homogenization distribution of dispersed pacticlis in continuous phase helps to

stabilize the fat emulsion against gravity separation, primarily causes disruption of fat globules
into much smaller ones then diminishes creaming and the tendency of globules to clump or
coalesce, so that homogenization makes the product become homogeneous, limited separation
phase during preservation, increase coeffective heat transfer, made structured of product is
smoother than.
The effect of homogenization on the physical structure of milk has many advantages:
 Smaller fat globules leading to less cream-line formation
 Whiter and more appetizing color
 Reduced sensitivity to fat oxidation
 More full-bodied flavor, and better mouthfeel
 Better stability of cultured milk products
Physical: The sizes of fat globules will reduce, the temperature of milk will increase slightly
because of the pressure’s effect. When the pressure for homogenization increases 40 atm, the
temperature of milk will rise 10C.
Physico-chemical: The surface rea between two phases oil and water in the emulsion will rise,
then the surface tension of fat globules will increase, too.
Equipment:
Homogenization have two styles:
 A single stage homogenization valve
 Multiple stages homogenization valve
High-pressure single-stage homogeniser is selected because the fat content in kefir is not too much.
24
Figure 2. 5 Single-stage homogenisation

A high-pressure homogenizer is a pump with a homogenization device. A homogenizer is
generally needed when high-efficiency homogenization is required.
The product enters the pump block and is pressurized by the piston pump. The pressure that
is achieved is determined by the back-pressure given by the distance between the forcer and seat
in the homogenization device.
When the liquid passes the narrow gap, the flow velocity increases. The speed will increase
until the static pressure is so low that the liquid starts to boil. The maximum speed depends mainly
on the inlet (homogenization) pressure. When the liquid leaves the gap, the speed decreases and
the pressure increases again.
Technological parameters :
The milk should be homogenized at 20 – 25 MPa and 65 – 70 °C after the process the fat
globule’s size is about 0.5 – 1 µm.
After the homogenization step, gelatin was put into the balance tank while mixing. Amounts
above 0.35% tend to give kefir a curdy appearance upon stirring.
2.4 Pasteurization
Objective : preparation operation for fermentation and preservation operation.
The heat treatment:
 Produces a relatively a sterile medium for the exclusive growth of the starter.
 Effectsthermal breakdown of milk constituents, especially proteins, releasing peptones and
sulfhydryl groups, which provide nutrition and anaerobic conditionsfor yogurt culture.
25
 Denatures and coagulateswhey proteins of milk, thereby enhancing the viscosity, leading
to a custard-like consistency in the product.
The intense heat treatment during yogurt processing destroys all the pathogenic flora and most
vegetative cells of all microorganisms con-tained therein. In addition, milk enzymes inherently
present are inactivated. Consequently, the shelf life of yogurt is assured. From a microbiological
stand-point, destruction of competitive organisms produces conditions conducive to the growth of
desir-able yogurt bacteria. Furthermore, expulsion ofoxygen, creation of reducing conditions
(sulfhydryl generation), and production of protein-cleaved nitrogenous compounds as a result of
heat processing enhance the nutritional status of the medium for growth of the yogurt culture.
Physical: changes in the protein as a result of heat treatment have a profound effect on the viscosity
and some physical criteria can change such as density, thermal conductivity,..
Chemical: high temperature will impulse the disadvantageous chemical reactions happen easier.
Some of them are the Maillard reaction between reducing sugar (lactose) and amino acids as well
as peptides in milk, the reaction of vitamin disintegration, the oxidation of fat, … Depending on
the temperature and time for pasteurization, the change levels will be different.
Physico-chemical: The increase and decrease of temperature in operation can change the phase
status of water and volatile substances. Some proteins can be denatured by heat.
Biological: Pasteurization can inactivate many microorganisms and pathogenic.
Bio-chemical: milk enzymes inherently present are inactivated.
Equipment:
In processing, a plate heat exchanger pasteurization system is commonly used.
The equipment must be made stainless steels, four corners have four holes as channels two
for milk, two four heating agents.
The frame may contain several separate plate packs – sections – in which different stages
of treatment such as preheating, final heating and cooling take place. The heating medium is hot
water, and the cooling medium cold water, ice-water or propyl glycol, depending on the required
product outlet temperature. Most plate heat exchangers contain a section that is used for
regeneration. Regeneration is a process that uses the heat of a hot liquid to preheat the cold
incoming liquid . The cold liquid also cools the hot liquid, saving on water and energy.
Regeneration efficiencies of up to 95% can be achieved in efficient plate heat exchangers.
The liquids enter and leave the channels through holes in the corners of the plates. The
indirect heat exchanger between the milk and the medium will be carried out through the thin
plates. Gaskets round the edges of the plates and round the holes form the boundaries of the
channels and prevent external leakage and internal mixing.
26
Figure 2. 6 Plate heat exchanger

Technological parameters:
Control the temperature for the heat treatment at 90 – 950C about 3 - 5 minutes, temperature
cooling about 22 – 250C.
2.5 Propagation
2.5.1 Raw milk
Skim milk is the medium most frequently used for starter production, but reconstituted skim milk
with 9 – 12% dry matter (DM), made from top-grade skim milk powder, is another alternative.
2.5.2 Pasteurization:
Ojective: preparation for propagation
This heat treatment improves the properties of the medium through:
 Destruction of bacteriophages
 Elimination of inhibitory substances
 Some decomposition of protein
 Expulsion of dissolved oxygen
 Destruction of original living micro-organisms
Transformation of material and equipment similar 2.4 Pasteurization
Technologycal parameter;
We have to pasteurize the medium at 90-950 C for about 30-45 minutes. The medium
will be cooled into 22-240 C to prepare for inoculation.
27
2.5.3 Inoculation and Propagation:

Ojective: preparation for inoculation and exploitation, significant increase bacteria quantity in
cultural.
Transformation of material:
Physical:
During the propagation, because of the growth of microorganism, a part of energy will be
released to the medium in heat energy – heat radiation. This phenomenon will increase the medium
temperature.
The microbial population grows by about 10% per week during propagation, this will
increase the weight of kefir grains. Therefore the grains must be weighed and the surplus removed
before the batch is reused.
Chemical:
The products of the metabolism of the microorganism present in the medium.
The pH value decrease during propagation.
Biological:
During propagation, the microorganism cells will carry out the metabolism and produce
more new cells.
The microbiological profile change during propagation (decreased levels of
heterofermentative lactic acid streptococci and yeasts).
28
Table 2. 1 Microorganisms used in starter cultures for cultured in kefir
29
Table 2. 2 Characteristics of mesophilic starter bacteria used for cultured kefir.
Equipment:
The normal practice is to use two tanks in rotation for manufacture of bulk starter. One
contains ready-made starter for use in the day’s production,while the starter for the following day
is being prepared in the other one.
The tanks should be of aseptic design, i.e. hermetically sealed and triple jacketed. They
should also be capable of withstanding negative pressures up to 30 kPa (0.3 bar) and positive
pressures up to 100 kPa (1 bar). The agitators should be double-sealed and powered by two-speed
motors. In addition, they should be fitted with HEPA (High Effiency Particle Air) filters (4) to
exclude airborne infec-tion from the air that is drawn in when the tank is cooled after cleaning and
when the heated medium is cooled to incubation temperature.
30
The bulk starter tank can be equipped with a stationary integrated pH meter (7) designed to
withstand the great temperature differences that occur during cleaning and heat treatment of the
medium.
Figure 2. 7 The bulk starter tanks

Technologycal parameter:
Incubation takes place at about 23°C and the proportion of grains is about 5% (1 part grains
to 20 parts substrate) or 3.5% (1 part grains to 30 parts milk).
The incubation time is about 20 hours; as the grains tend to sink to the bottom, intermit-
tent stirring for about 10 – 15 minutes every 2 –5 hours is recommended. When the desired pH
value (say 4.5) has been reached, the culture is stirred.
2.5.4 Filtration:
Ojective: preparation culture for fementation.
Transformation of material: material do not change the chemical composition of the culture.
Physical: it’s phase seperated during operation
Equipment: culture is pumped into the tank containing filler basket.
Technology parameter:
31
The culture is stirred before the grains are strained off from the mother culture, now also
called filtrate. The strainer has holes with a diameter of about 1 mm.
2.5.5 Washing and storing of grains:
The grains are washed in the strainer with boiled and cooled water (6-100C) to remove
impurities at the surface of the kefir grains.
Sometime we can use sterile skim milk to wash the grains. The kefir grains after washing,
will be stored in chill water or solution sodium cloride 0.9%. This grains can be reused for next
inoculation.
2.5.6 Storing of the filtrate:
The filtrate can be cooled to about 10°C if it has to be stored for a few hours before being
used.
If large quantities of kefir are going to be produced, the filtrate can be immediately inoculated
into the pretreated milk intended as the sub-strate for the bulk starter. The dosage is 3 – 5% of the
volume of the substrate. After incubation at 23°C for about 20 hours, the bulk starter is ready for
inoculation into the kefir milk.
2.6 Inoculation
Objective:
Preparation: inculation preparation for incubation. It supply bacteria for incubation
operation.
Transformation of material:
The addition of inoculum into the medium do not change the chemical composition of the
medium significantly.
Equipment:
This operation is carried out in vertical cylindrical tanks (we call fermentation tanks,
because the next operation is still carried out in these tanks). The tanks are with an insulated layer
and have a stainless steel double jacket outside, agitator with motor inside set at the top of tank
and drain butterfly valve, supported on 4 adjustable. The surface finish of the parts in contact with
the milk must be smooth, no bacteria traps, easily and thoroughly cleaned.
The temperature of medium during the operation can be adjusted by the jacket
surrounding the tanks or by heating/cooling agent kinky path system inside the cylindrical body of
the tank.
Nowadays, the modern tank have an electronic control system for temperature, pH
indication, control of the cooling/heating unit, agitator… and technological parameters control
system (pH, temperature…) are carried out by software programs and pump controller. By this
system, we can adjust the suitable condition easily.
32
The inlet and outlet door for product are set at the bottom of the equipment. Beside that, we
have the observation door, the sample tanking valve...
There are a kinky path system inside the cylindrical body of the tank, set between the double
jacket, to enable rapid heating or rapid cooling temperature, without mixing. The insulated layer
on the shell to maintain optimum operation and processing temperature.
Figure 2. 8 Fermentation tank

Technological parameter: The filtrate (received after propagation operation) is used to inoculate
into a new source of milk intended as the substrate for the inoculum. The dosage is 2 -3% of
the volume of the substrate. The inoculation temperature about 230C.
2.7 Incubation
Objective: processing and preservation operation
Processing:
The incubation change the chemical composition and sensory properties of milk, transform
the milk into product.
In fermentation lactic bacteria will be change lactose into acid lactic, yeast will be used
lactose and production ethanol plus CO2. Furthermore, a majority of the kefir sold contains optional
bacteria that have been shown to possess probiotic properties, including bifidobacteria,
Lactobacillus acidophilus , Lactobacillus casei and other lactobacilli, which are often referred to
as “probiotic bacteria.” Their inclusion in culture is motivated by their unique health-promoting
effects, such as improvement in protein digestibility, alleviation of lactose intolerance,
enhancement of mineral absorption, control of intestinal health, lowering of serum cholesterol,
antihypertensive effects, anticancer properties and immunity enhancement.
Preservation:
33
The incubation catabolism acid lactic made reduced pH ( pH = 4.5 ) inhibited bacteria, yeast
and enzymes, therefore increase sholf-life.
Biological: there are two important transformations during the incubation: the metabolism and
the growth of microorganism. The metabolism is needed to microbes‟ existence.
Chemical and biochemical:
Lactic fermentation: The lactose found in milk is degraded to lactic acid during fermentation; this
causes the pH to drop and the milk to thicken. As much as 30% of the milk lactose is broken down
during fermentation. β-galactosidase is the enzyme that hydrolyses lactose into glucose and
galactose.
The bacteria in kefir are able to break down glucose to lactic acid. Both D-(−)-lactic and L-
(+)-lactic acid are produced from the milk lactose, and in kefir the L+ form tends to predominate.
However, the ratio of L to D depends on the source and microbial composition of the grains.
Lactic-acid production results in coagulation of milk, beginning at pH 5.2–5.3, at the point
where the casein is first destabilized, and continuing until completion at pH 4.6. During lactic-acid
production there is a gradual removal of phosphorus and calcium bound to the stable casein particle
as tricalcium phosphate. The texture, body and acid flavor of kefir owe their origins to lactic acid
produced during fermentation.
34
Figure 2. 9 Pathway used by bifidobacteria for lactose metabolism

With: 1 = β -galactosidase, 2 = glucokinase, 3 = phosphoglucose mutase, 4 = phosphoglucose isomerase, 5 =
transaldolase, 6 = phosphoglucose isomerase, 7 = fructose-6-phosphate phosphoketolase, 8 = acetate kinase, 9 =
transketolase, 10 = xylulose-5-phosphate phosphoketolase, 11 = ribose-5-phosphate isomerase, 12 = ribulose-5-
phosphate-3 epimerase, 13 = xylulose-5-phosphate phosphoketolase, 14 = acetate kinase, 15 = Embden-Meyerhoff
pathway enzymes, 16 = lactate dehydrogenase.
35
Figure 2. 10 Heterofermentative pathway for lactose metabolism among lactic-acid bacteria

With: 1 = β -galactosidase, 2 = galactokinase, 3 = glucose:galactose-1-phosphate uridyl transferase uridine
diphosphate-glucose epimerase, 4 = glucokinase, 5 = phosphoglucomutase, 6 = glucose-6-phosphate dehydrogenase,
7 = 6-phosphogluconate dehydrogenase, 8 = epimerase, 9 = phosphoketolase, 10 = acetate kinase, 11 = triose phosphate
dehydrogenase, 12 = phosphoglycerokinase, 13 = phosphoglyceromutase, 14 = enolase, 15 = pyruvate kinase, 16 =
lactate dehydrogenase, 17 = phosphoacetyl transferase, 18 = acetaldehyde dehydrogenase, 19 = ethanol dehydrogenase.
Ethanol fermentation: The production of ethanol in kefir is complex; both yeasts and
heterofermentative bacteria produce ethanol. The quantity of ethanol produced is dependent on
the fermentation process and the type of container used (tightly capped or not).
Ethanol appears to be produced towards the end of the fermentation process, and its
formation can continue even when the pH has decreased to the point where the lactic acid bacteria
36
in the product are no longer active. The ethanol concentration can be increased in the final product
by increasing the temperature during fermentation.
Adding 0.4 or 0.5% glucose to the starter milk resulted in ethanol production only during
the yeast fermentation stage, yielding a final product with 0.07 or 0.08% ethanol. When 1.0%
glucose was added, the production of ethanol continued in the lactic fermentation and resulted
in an ethanol concentration of 0.24% in the final product.
Another compound such as:
Amino acid: As the fermentation slows and the kefir enters the ripening stage, the
proteolytic activity of other microorganisms such as acetic acid bacteria and yeasts causes more
peptides and free amino acids to be formed in a manner seen in other fermented milk products.
The remarkable compounds are some organic acid such as propionic acid, formic acid, succinic
acid, some volatile component in aldehyde group (aldehyde acetic and diacetyl) and
macromolecule alcohol.
Lactic acid, acetaldehyde, acetone, diacetyl and other carbonyl compounds produced by
fermentation constitute key flavor compounds of kefir. Acetaldehyde content varies from 4 to 60
ppm in kefir. Acetone varies from 1.3 to 4.0 ppm, acetoin from 1.3 to 4.0 ppm and diacetyl from
0.1 to 0.3 ppm. Acetic acid is present in the range of 50–200 ppm. These key compounds are
produced by kefir bacteria. Certain amino acids (threonine, methionine) are known precursors of
acetaldehyde. For example, threonine in the presence of threonine aldolase yields glycine and
acetaldehyde. Acetaldehyde can arise from glucose, via acetyl CoA, or from nucleic acids, via
thymidine of DNA. Diacetyl and acetoin are metabolic products of carbohydrate metabolism in
ST ( Streptococcus thermophilus ). Acetone and butane-2-one may develop in milk during
prefermentation processing.
Minerals:
Kefir is an excellent dietary source of calcium, phosphorus, magnesium and zinc in human
nutrition. Research has shown that the bioavailability of the minerals from yogurt is essentially
equal to that from milk. Since yogurt is a low-pH product compared to milk, most of the calcium
and magnesium occurs in ionic form.
The complete conversion from the colloidal form in milk to the ionic form in yogurt may have
some bearing on the physiological efficiency of utilization of the minerals.
Vitamins
During and after fermentation, bacteria affect the B-vitamin content of Kefir. The
processing parameters and subsequent storage conditions influence the vitamin content at the time
of consumption. Incubation temperature and fermentation time exert significant balance between
vitamin synthesis and utilization of the culture. In general, there is a decrease of vitamin B12 , biotin
and pantothenic acid and an increase of folic acid during kefir production. As a result of
fermentation, folate concentration in kefir increases to 80 ± 20 μg/l, as compared to 40 ± 10
μg/l in milk.
37
Physicochemical: Yeasts and some heterofermentive lactic acid bacteria are responsible for the
production of the CO2 gas in kefir. The CO2 content increases during fermentation as the pH drops.
The gas production leads to “fine flake” coagulum formation and also imparts a sparkling mouth
feel to kefir.
Physical: the milk coagulation during fermentation results from the drop in pH due to the activity
of the production.
Equipment:
In practice, the inoculation and incubation operations are carried out in one equipment (the
fermentation tanks).
The double jacket outside the tank enables cold water to be flushed next to kefir, without
mixing, to enable rapid cooling temperature.
The tank has an agitator with gear motor, stainless steel shell, with or without insulation.
The agitator will stir kefir during cooling to break down the coagulum formation. The rate of
agitation is set to minimize foam development. We use Gate agitators, which ensure a gentle
agitation of the contents, are commonly used in kefir fermentation tank.
Gate agitators: gate agitators are close-clearance impellers that fit the contour of the vessel.
Gate agitator obtain adequate mixing under the laminar flow conditions encountered in high
viscosity applications. These impellers sweep the whole wall surface of the vessel and agitate most
of the fluid batch through physical contact.
Gate type agitators are used in wide shallow tanks and for materials of high viscosity when
low shear is adequate.
Figure 2. 11 Gate agitator
38
Figure 2. 12 Typical cone-bottom fermentation tanks used in a kefir plant.

Technogical parameter:
The incubation period is normally divided into two stages, acidulation and ripening.
The acidulation stage
Fermentation stage is carried out at 23-250 C for approximately 12 hours. The fermentation
stops when the acidity of milk reaches 85-1000 Th.
After fermentation stage, we stir the medium to break down the coagulum formation. The
temperature (after this stage) increases to about 300C, so we cool the medium to decrease the
temperature to the suitable temperature for ripening stage (14-160C).
The speed of rotation of agitator ranging between 8 and < 50 rpm.
The ripening stage
The typical slightly yeasty flavour starts to develop during the following 12 – 14 hours at
temperature 14-160C. Final cooling commences when the acidity has reached 110 – 120 °Th (pH
about 4.4).
2.8 Cooling
Objective:
Preservation
Cooling decrease the temperature to 5-80C in order to reduce the metabolism of microbes
and its enzyme activity in kefir, so cooling will help to prolong the shelf life of the final product.
39
Improving: improvement smooth structure and stability yogurt.

Transformation raw material: transformation in operation are similar with the fermentation
operation. However, temperature is slowly decrease so the speeds and transform levels will be more
lower.
Equipment: used plate heat exchanger for cooling operation.
Cooling is to decrease the temperature to 5-80C.
2.9 Packaging
The principal and fundamental functions of packaging are:
- To enable efficient food distribution
- To maintain product hygiene
- To protect nutrients and flavour
- To reduce food spoilage and waste
- To increase food availability
- To convey product information
Objective:
Preservation:
The package outside will protect the food inside not to be contaminated by the harmful
effect of the environment such as dirt or other foreign bodies, microorganisms (bacteria, yeast and
moulds), gases (e.g. oxygen) which can help the yeasts and moulds to grow and spoil the product,
and light which may oxidase the fat … The correct package and suitable condition for storing will
maintain the quality of food, so it will prolong the food shelf-life.
Improvement
The package outside helps products to be transported and stored easily. The beautiful
package can attract the customers, too.
Transformation of raw material: The fast and closed packaging is not cause the significant
transformations that affect tothe quality of the product.
Equipment:
Using a packing machine to pour kefir into plastic bottles or paper box and then, packaging
them. The machines involve volumetric piston filling. The product is sold by weight, and the
machines delivering volumetric measure are standardized accordingly.
40
We can control the filling volume into each package depend on the volume we want for our
products and types machine. The filling machine requires conveyor belt to transform the package
in and out of the machine.
Figure 2. 13 Filling machine

The common package for kefir is paper box, plastic bottle.
Various packaging machines of suitable speeds (up to 400 cups per minute) are available
to package various kinds and sizes of yogurt products.
Kefir is generally packaged in plastic containers that vary in size from 4 to 32 ounces (113–
907 g). Kefir packaged in tubes generally weighs around 2 ounces (60 g) per tube. The machines
involve volumetric piston filling. The product is sold by weight and the machines delivering
volumetric measure are standardized accordingly.
41
3. Processing flow chart of kefir 2
Raw milk/ skim

milk/reconstitute
d milk
Raw milk Pasteurization
Skim
milk Inoculation Kefir grains
Fat standardization
Additives Homogenization Propagation
Pasteurization
Gelatin
Inoculum for
Inoculation production
Incubation
Cooling
Packages Packaging
s
Kefir ( cool
store)
Differences of the second production line compare to the first one:

 Production line 2 do not have deaeration operation, then, skim milk was put to standardized
milk solid content.
 Replace cooling equipment with tubular heat exchanger and change single stage
homogenizer into two-stage homogenizer.
 Replace the inoculum (kefir grains) by using freeze-dried kefir starter.
42
3.1 Fat standardization:

Production line 2 do not have deaeration operation, then, skim milk was put to standardized
milk solid content.
Ojective: similar production line 1.
Transformation of raw material: similar production 1, but the solid content in the milk increase.
Equipment: similar prodcution line 1, but in the standardized milk tank with mixer equipment to
mixing skim milk, food additives E471 with standardized milk.
The fat content of standardization milk between 0.5% and 6%, frequently specified at 2.5%
to 3.5%.
Food additives amount above 0.35% tend to give milk.
3.2 Homogenization:
Objective, transformation similar production line 1.
Equipment: the process is conducted by applying pressure in two stages
In two-stage homogenisation there are two oil systems, each with its own pump. A new
homogenisation pressure is set by changing the oil pressure. The pressure can be read on the high-
pressure gauge.
Homogenisation always takes place in the first stage.The second stage basically serves two
purposes:
• Supplying a constant and controlled back-pressure to the first stage, giving best possible
conditions for homogenisa-tion;
• Breaking up clusters formed directly after homogenisation.
43
Figure 3. 1 Two-stage homogenisation head.

In the first stage, pressure of the order of approximately 14 MPa (2000 psi) reduces the
average diameter of the average milk fat globule from approximately 4 micrometers (μm) (range
0.1 to 16μm) to less than 1 micrometer.
The second stage uses a pressure of 3.5 MPa (500 psi) and is designed to break the clusters
of fat globules apart, with the objective of inhibiting creaming in the milk.
After the homogenization step, gelatin was put into the balance tank while mixing. Amounts
above 0.35% tend to give kefir a curdy appearance upon stirring.
3.3 Freeze-dried kefir starter
Figure 3. 2 Freeze-dried kefir starter
44
Many plants use frozen direct to vat cultures for yogurt production. However, for cost
savings, large yogurt manufacturers prefer to make bulk starters in their own plant from frozen
bulk cultures.
Raw milk: similar production 1
Pasteurization: similar production 1
Inoculation and propagation:
Ojective: to prepare for the enough inoculum for the fermentation.
Transformation of raw material: similar production 1
Equiment : similar equiment bulk starter tank but do not have filter.
Figure 3. 3 Bulk starter tank

Preparation of the pouch holding the culture: wiping the top with 70% alcohol, cutting the
top and into the starter tank.
The dosage used: 1 g of grains to 3 litres of milk to get the inoculum.
3.4 Cooling:
Objective, transformation similar production line 1.
Equipment:
In most case, producers use the tubular heat exchanger to cool kefir. Mechanical agitation
in pumps, pipes and filling machines must therefore be minimized. Air entrainment must also be
45
avoided, as air increases the risk of serum separation in the product, this serum will rise to the kefir
surface to form a serum layer.
In this cooling operation, we use double tube heat exchanger.
The double tube heat exchanger consists of two concentric tubes; one fluid flows through
the inner tube while the other fluid flows in the annulus. The flow arrangement could be cocurrent
or counter-current. Usually, the counter-current flow is more efficient from the heat transfer
point of view.
Kefir after incubation will be pumped to the inner tube, the cooling water will be pumped
to the outer tube to get the required temperature of kefir.
Figure 3. 4 The tubular heat exchanger

Technogical parameter: similar production line 1.
4. Comparison of two production lines:
Table 4. 1 Comparison of two production lines
Production line 1 Production line 2

Deaeration -Production line is complex Don’t require deaeration
-Consume more energy -the production line is more
-Improved working condition for simple than.
the homogenization. - consume less energy
-Prevent foaming during - the homogenization will be
production. more difficult.
-Stabilize solid level in product. - may be foaming during
production.
Homogenization -single-stage homogenization -Require more energy
spend less energy. -high content of soluble
-the homogenization will be more substances
easy than.
46
-break the clusters of fat globular

apart, inhibiting creaming in the
milk.
Kefir grains - Kefir grains can be reuse many - Powdered starter culture can
of times. be reused for a number of
-Propagation operation used more batches.
equipment → production line -Propagation operation is more
more complex simple → saving money
- A larger number of probiotics - The probiotics in freeze-dried
(about 30) depend on the origin of starter are fewer than in kefir
grains and storage condition. grains.
Cooling -From the standpoint of heat -Coefficient is high
transfer the tubular heat
exchanger is less efficient than a
plate heat exchanger.
-The tubular heat exchanger
can also run longer between
cleanings than the
plate heat exchanger
Chapter 3: PRODUCT
The flavor of plain kefir is primarily due to lactic and acetic acids, diacetyl, and
acetaldehyde, produced by homo-fermentative and hetero-fermentative lactic acid bacteria.
However, because kefir grains also content yeast, in addition to lactic acid bacteria, other end-
products are formed that make the finished product quite different from other cultured dairy
products. This is because ethanol is produced when the yeast ferment lactose, such that kefir can
contain as much as 2% ethanol (accounting perhaps, for its appeal). In the United States, this much
ethanol would trigger action from the regulatory agencies. Thus, if yeast is present in the culture
(most kefir products made in the United States claim yeasts on their labels), they must be low or
non-ethanol producers. Kefir grains can also contain acetic acid-producing bacteria, such as
Acetobacter aceti.
5. Criteria for evaluation of product quality
The flavour, viscosity and microbial/chemical composition of the final kefir product can be
affected by the size of the inoculum added to the milk, the occurrence of any agitation during
fermentation, and the rate, temperature and duration of the cooling and ripening stages following
fermentation.
5.1 Chemical characteristic:
The composition of kefir is variable and not well defined. It depends on the source and the fat
content of milk, the composition of the grains or cultures and the technological process of kefir.
47
The major products formed during fermentation are lactic acid, CO2, alcohol (ethanol) and
aromatic compounds such as diacetyl and acetaldehyde. Kefir is also rich in vitamin B such as B1,
B12, biotin, pantothenic acid, folic acid; vitamin K; minerals such as calcium, magnesium and
some essential amino acids, especially trytophane, which help the body with healing and
maintenance functions. Furthermore, the kefir‟s abundance of beneficial yeast and bacteria provide
lactase, a useful treatment to consume lactose for lactose intolerant individuals.
The amounts of ethanol and CO2 produced during fermentation of kefir depend on the
production conditions used, values of 0.85–1.05 g/l for kefir produced from kefir grains) and 1.7
g/l for kefir produced from purified cultures. Ethanol levels in the finished product is 0.01–0.1%
although kefir with ethanol concentrations as high as 0.25% have been produced from grains in the
laboratory.
The fat content in kefir depends on the type of milk used and the requirement for fat
standardization operation.
Table 5. 1 The chemical composition and nutritional values of kefir
48
Table 5. 2 Chemical composition of various kefirs
5.2 Physico-chemical characteristic

 Fat: The fat content of the kefir did not differ significantly (p > 0:05) from the fat content
of the milk it was made from. However, the fat content was observed to undergo a decrease
by the end of the storage period and it could be related to the growth of moulds, since those
ones are the principal lipolytic agents in fermented milks.
 Dry matter: These values were not significantly different from the dry matter content of the
source milk. Accordingly, as was the case for the fat content, fermentation did not affect
the dry matter content of the source milk used. The last days of storage period, the
dry matter decreased and it is also explained by the growth of moulds.
 Viscosity: The viscosity decreased appreciably over the course of storage. The viscosity
is claimed to be higher for the kefir made using the higher percentage added kefir grain
inoculate.
49
 pH value: The pH of the product is usually 4.3 – 4.4. The pH of the kefir did not
vary during the storage which is possible because at the presence of yeasts.
Researchers found that lactic acid bacteria multiply and produce lactic and acetic
acids more slowly in mixture with yeasts than in pure culture.
5.2 Microbiological characteristic
Table 5. 3 Suggested microbiological standards for dairy product
5.4 Sensorial characteristic

Natural kefir has a refreshing, acid, yeasty taste and a”sparkling” mouth feel.
Kefir should be viscous and homogenous with shiny surface.
The distinctive taste of kefir results from the presence of several flavour compounds which are
produced during fermentation. Acetaldehyde and acetoin have contributed the role in making
flavour; both have been found to increase in concentration during kefir fermentation.
During storage, acetaldehyde increases in concentration and acetoin decreases. Fructose was
found to increase production of several flavour volatiles, but did not increase fermentation time.
The samples used for analysis were taken 24h after inoculation and then after 2, 7, 14 days
of storage at 5 ± 10 C. The content of Kefir grains used is 5%.
The attributes considered were: odour intensity, milky odour, fermented odour, vegetable
odour, mouth odour, viscosity, flavour intensity, dairy taste, sour taste, bitter taste, milky taste,
astringency and acceptability. Each attribute was scored on an increasing scale of from1
(not present) to7 (very intense).
The correlations indicated that the panel was positively influenced by milky taste, milky odour,
viscosity and negatively influenced by astringency, bitter taste, sour taste and fermented odour.
Miscellaneous tastes and odours also adversely affected product acceptability.
50
REFERENCE
1. Prof.Dr Van Viet Man Le, food processing, VietNam National University HCM, 2011, 980-
990p.
2. Bylund G., Dairy processing handbook, Tetra-Pak processing systems AB Publisher, Lund,
1995, 436p.
3. Semih Ot1es and Oz1em Cagindi, Kefir: A Probiotic Dairy-Composition, Nutritional and
Therapeutic Aspects, Pakistan Journal of Nutrition 2 , 2003.
4. Edward R. Farnworth and Isabelle Mainville, Handbook of Fermented Functional
Foods, Second Edition, CRC Press, Boca Raton, 2008, 602p.
5. J. Scott Smith and Y. H. Hui, Food ProcessingPrinciples and Applications, blackwell
publishing, 2004, 510p.
6. Y.H.Hui and associate editors, Handbook of food products manufacturing (volume 2):
health, meat, poultry, seafood, and vegetable, wiley-interscience, A John Wiley & Sons,
inc.,publication, 2007
7. Edward R. Farnworth, Kefir-a complex probiotic, Food research and development centre,
Agriculture and Agri-food Canada
8. Robert W.Hutkins, Microbiology and technology of fermented foods, IFT Press, Blackwell
Publishin, 2006
9. Ramesh C. Chandan, Manufacturing Yogurt and Fermented Milks, Wiley-Blackwell
Publishing, 2006, 478p.
10. A. Y. Tamime and R. K. Robinson, Tamime and Robinson 's Yoghurt Science and
technology Third edition, Published by Woodhead Publishing Limited, Abington Hall,
Abington, 2007, 808p.
51

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Food processing technology Kefir

1. Raw materials ...........................................................................................................................................4

2.8 Cooling ........................................................................................................................................39

Figure 2. 1 Centrifugal separation ................................................................................................................20

Figure 3. 1 Two-stage homogenisation head. ..............................................................................................44

Figure 3. 3 Bulk starter tank .........................................................................................................................45

Table 2. 1 Microorganisms used in starter cultures for cultured in kefir .....................................................29

Table 4. 1 Comparison of two production lines ...........................................................................................46

Table 5. 1 The chemical composition and nutritional values of kefir ..........................................................48

Table 1 1 Milk composition

1.1.2. Criteria for milk

altered due to the microbiological activity or processing treatments, or chemical or bio-chemical

Table 1 3 Biochemistry criteria

Figure 1. 1 Kefir grains

1.2.1. Microbiology of kefir grains

Table 1 4 Bacteria, yeast and mold isolated from

1.2.2. Structure of grains

Figure 1. 2 Electron micrograph of a kefir grains

Figure1. 3 Kefir health benefits

Chemical composition Content (%)

The microorganism Content (%)

Defined by Codex Alimentarius standard CODEX STAN 280-1973, Anhydrous milkfat,

 Appearance: Dry milk powder must be reasonably uniform in composition, white- or

1.4. Food Additives

Figure1. 4 Structural of E471

Chapter 2: PRODUCTION LINE

Raw milk/ skim

Raw milk Pasteurization Inoculum

Additives Homogenizatio filtration

Information of each operation in the production line

Figure 2. 1 Centrifugal separation

In standardization milk system:

Figure 2. 2 Principle for direct in-line standardization of cream and milk.

The direct in-line standardization system:

 Fluctuations in the fat content of the incoming milk

Figure 2. 3 Direct in-line standardization system

2.2 Deaeration and evaporator

The advantages gained through deaeration are:

With: 1- plate exchanger equipment

Figure 2. 4 Deaeration equipment

The homogenization distribution of dispersed pacticlis in continuous phase helps to

Figure 2. 5 Single-stage homogenisation

Figure 2. 6 Plate heat exchanger

2.5.3 Inoculation and Propagation:

Table 2. 1 Microorganisms used in starter cultures for cultured in kefir

Table 2. 2 Characteristics of mesophilic starter bacteria used for cultured kefir.

Figure 2. 7 The bulk starter tanks

Figure 2. 8 Fermentation tank

Figure 2. 9 Pathway used by bifidobacteria for lactose metabolism

Figure 2. 10 Heterofermentative pathway for lactose metabolism among lactic-acid bacteria

Figure 2. 11 Gate agitator

Figure 2. 12 Typical cone-bottom fermentation tanks used in a kefir plant.

Improving: improvement smooth structure and stability yogurt.

Figure 2. 13 Filling machine

3. Processing flow chart of kefir 2

Raw milk/ skim

Raw milk Pasteurization

Additives Homogenization Propagation

Differences of the second production line compare to the first one: