FFST 3121: Dairy Science and Technology

Component: Principles of Dairy Science and

Technology
Academic year: 2021-2022
Lecturer: F. Matsiko
 INTRODUCTION
• Milk is a fluid secreted by the mammary glands of
mammals
• It provides good quality proteins, Ca, and vit A, D,
B2 (riboflavin), niacin and folic acid
• High content of water (87% on average)
• Milk has shorter shelf life
• Shelf stable dairy products: yogurt, cheese and Kefir
• Overview of physical-chemical properties of milk
• Basic quality tests on raw milk
• Discussion of processing technologies of dairy
products
Chap1. Chemical & Physical properties of Milk
1.1Milk Composition and Structure
• Milk = polydisperse system of water with:
– Emulsified fat globules enveloped by a
membrane (Ø 0,1-10 μm)
– Casein micelles (Ø 10-100 nm)
– Lipoprotein particles (Ø 10 nm)
Milk plasma = Milk - fat globules
~ separated milk (skimmed milk)
Milk serum = milk plasma - casein micelles
= milk - fat, caseins and insoluble salts
Dry matter (DM) = all dry milk components
Fat-free DM = dry matter - fat
Milk viewed at different magnifications
Composition and Structure of milk (Approximate average quantities in 1kg of milk
Properties of the main structural elements of milk
 1.2.Different kinds of milk
Some of the properties in comparison with cow’s milk:
Goat's milk
– relatively more proteins and fat
– used for cheese manufacturing
• Sheep's milk
– rich in proteins and fats
– used for cheese manufacturing
• Buffalo milk
– composition comparable to cow's milk
– high fat content (6-7%)
• Horse's milk
– low fat and protein content
– used for yogurt
– low nutritional value
– used in Eastern countries, Russia, Mongolia
 1.2.Different kinds of milk
• Casein-rich milks: cow, sheep, goat, buffalo
– usable for cheese manufacturing; compact
coagulation
– difficult digestion
• Albumin rich milks: humans, horse
– no cheese manufacturing possible; fine
coagulation
-better digestion
Unless specified otherwise, cow’s milk will be
dicussed in this subject.
Composition of human milk with reference of cow’s milk
• Human milk
– specific composition
– more natural defense components (immunoglobulins,
oligosaccharides, Bifidus factors)
– large amounts of lactose (= stimulates absorption of Ca,
Mg, Mn, Cu and Zn)
– Rich in vitamins and Poly-unsaturated fatty acids (n-6/n-
3)
– low in proteins (more whey proteins, fewer caseins)
– Absence of allergens
– Dangers: presence of (fat-soluble) pesticides (DDT,...),
PCB’s, dioxins, Phthalates, Medicines, Nicotin, alcohol
 1.3Factors influencing Composition of milk

• Breed
– a wide diversity
– predominately the result of selection by
people
Various breeds
Component Breeds
s
Ayrshire Brown Guernsey Holstein Jersey Shorthorn
Swiss
Fat 3.95 3.98 4.72 3.54 5.13 3.5
Lactose 3.48 3.64 3.75 3.29 3.97 3.32
Ash 0.72 0.74 0.76 0.72 0.76 0.75
Total Solids 12.77 13.07 14.04 12.16 14.42 12.27

Different breeds of cows and the gross composition of milk

• Season/nutrition
– milk composition is largely influenced by seasons
– winter: silaged feed; summer: fresh grass
- Differences in fat and protein content

-Major Implications on processing

 •Stage of lactation
-Time elapsed after calving influences milk composition
-Colostrum milk = First milk produced after calving;
more salts, proteins (mainly serum proteins) and less
lactose; composition evolves towards its normal
composition within 4 days
- Further changes occur: decrement in protein content,
casein, serum proteins, ashes, fat-free dry matter; at
the end of the lactation, an increment in DM content
occurs
-Lactose content is almost constant during lactation; fat
content is correlated to fat-free dry matter during
lactation
-pH changes: 6,6 6,7 6,9 (at the end)
 Factors affecting the milk composition
• Age of cow
– has a very small but consistent effect on milk composition
fat and fat-free dry matter decrease slightly with each
successive
lactation
• Mastitis
– = inflammation of the udder
– several species of pathogenic bacteria may cause mastitis
– causes a decrease in milk yield and a change in milk
composition
– decrease in fat content, fat-free dry matter, lactose and casein
and
increase of serum proteins and chloride content
– the number of somatic cells increases (> 400.000/ml)
-catalase activity increases
– Cl-lactose ratio increases
Correlations
• Lactose and chlorides are negatively correlated
– the result of the activity of the udder which
maintains milk isotonic
– if lactose-production is restricted by an infected
udder, more chlorides (and thus also sodium) are
added to the milk by the udder
– this phenomenon can be used to detect mastitis
infection: the number of Koestler is a valuable
parameter to differentiate normal from mastitis milk
Normal milk=(100*% Clorides)/% Lactose= 1.5-3.0
Mastititis milk=(100*% Clorides)/% Lactose> 3.0
1.3 Factors influencing the composition
• Method of milking
– during milking the fat content of the milk
leaving the udder increases (e.g. from 1-
10%, marked differences among cows in
this respect); fat-free dry matter remains
constant
– the interval between milking influences fat
content, but not fat-free dry matter; longer
intervals result in a higher fat content
1.3 Factors influencing the composition

• Individual
– variations within breeds are the result of genetic and
environmental factors
- by selection high-productive animals are obtained
• Quarters
– differences in composition in the milk of different quarters
of the udder of one cow mostly are negligible unless a
quarter is or has been affected by mastitis
• Other factors
– contamination and processing
Chap.2 Chemical Constituents of Milk

2.1 Carbohydrates
 2.1.1 Lactose
• Lactose: prominent carbohydrate of milks
of most species virtually unique to milk
having been found elsewhere only in fruits
of certain members of Sapotacea
• The components and breakdown elements
of lactose are found in minor quantities:
glucose and galactose, lactic acid and
butyric acid
• Other carbohydrates are found only in
small quantities
Chemical and Biochemical Properties of Lactose
• Lactose: Disaccharide of D-glucose and D-galactose
joined in a ß-1,4glucosidic linkage
– both moieties occur predominantly in the pyranose ring
form
– lactose = 1,4-β-D-galactopyranosyl- D-glucopyranose
• Reducing sugar
– property important in the analysis of milk
– also causes non-enzymatic browning, called Maillard
reaction
• amino acids and reducing sugars react forming aroma
products (HMF: Hydroxymethyl Furfural,aldehydes and
pyrazines) and brown components; nutritional value of milk
lowers during this reaction
 Lactulose
• Compound found in heated milk products
• Lobry de Bruyn-Alberda van Ekenstein transformation:
• mechanism by which lactulose is formed
• Sweeter than lactose
• Bifidus factor: promotes the growth of Bifidobacterium
bifidum and thus beneficial in the diets of human infants
• Concentrations up to about 1% may occur in commercial
evaporated milk
• A parameter to evaluate the thermal treatment of milk
(sterilized versus UHT milk, limit of 600 ppm)
Lobry de Bruyn-Alberda van Ekenstein
transformation
 Lactose content
• Relatively constant at 4,8 to 5,2 %
• Lower levels occur in colostrum and mastitis milk to
• Lactose comprises about 52 % of milk solids-non-fat,
about 70 % of whey solids and > 90 % of the solids in milk
ultrafiltrate
• Lactose intolerance
– Caused by a shortage of the lactase-enzyme (β-
galactosidase)
– Results in non-hydrolysis of lactose into glucose and
galactose
– Undigested lactose may support growth of undesirable
intestinal
flora, as well as draw water into the intestine causing
diarrhoea
and abdominal cramps
 Lactose sweetness
Lactose is not as sweet as other common sugars:Sucrose, Fructose and glucose
• Relatively sweeter at higher concentration
• β-lactose is sweeter than α-lactose
– this difference is not important since the small
difference between freshly prepared solutions is
eliminated quickly by equilibration of the
anomers
 Lactose mutarotation
• Lactose exists in both α and β forms (interchanging the
OH and H on the reducing group)
• Lactose is optically active because of its asymmetry
– α-form can be distinguished from the β form by its greater
rotation of polarized light
• α and β -forms of lactose exist in solution in a
temperature dependent equilibrium
[β]/[α] = 1,64 - 0,0027.T with T = temp. in °C
• The equilibrium is reached after 24 hours
– example: at 15 °C [β]/[α] = 1,60 or in this solution 38 % of
the lactose is present in the α form, 62 % in the β form
 Mutarotation in Lactose Solutions
A.Course of reaction (% finished ) as function of time
B.Effect of pH on the mutarotation rate constant (K/h)
 Lactose solubility
• α and β lactose differ considerably in solubility and in the
temperature dependence of solubility
– this is influenced by the mutarotation
• Lactose solutions can be supersaturated easily, i.e.
nucleation does not occur easily
– at concentrations over 2,1 times the final solubility,
spontaneous crystallization occurs (homogeneous
nucleation)
– at a relative supersaturation below 1,6 seeding with
lactose crystals usually is needed to induce
crystallization (solution is meta-stable)
 Lactose crystallisation
• αLactose crystallises as a hydrate of equimolar
amt of water and lactose
– Crystallization is of great technological importance
– because it may crystallize in some milk products,
notably
sweetened condensed milk and ice cream
• During evaporation processes, the solubility is
exceeded
– in condensed milk, a directed crystallization in the
α-form is observed, so that crystals don't exceed
10 μm
– crystals of 30 μm give the defect sandy mouthfeel
 Lactose crystallization

• Above 93 °C anhydrous β-lactose

crystallizes
– β-lactose dissolves much faster than α-
lactose hydrate at room
• temperature, as its solubility is about 10
times higher and the
• crystals are usually smaller with a larger
surface area
 Amorphous Lactose
• Is formed when a solution is dried rapidly, as in a
spray drier, or frozen
• Is often called “glass”
• Is a very concentrated solution which dissolves
on addition of water, but then α-lactose may
start to crystallize
• Is very hygroscopic, which is of importance in
milk powder
• Instant milk powder has a better solubility,
caused by
addition of moisture and a directed crystallization
 Lactic acid fermentation
• Lactose can be fermented by bacteria that have a β-
galactosidase (lactase)-system: LAB
• Lactose glucose + galactose 4 x lactic acid
– glycolitic or Embden-Meyerhof pathway
– dependent on conditions and culture, the following
reaction products can be formed: D(-) lactic acid, L(+)
lactic acid, racemic solution
• When 1 % lactic acid is formed, the reaction process is
slowed down due to the pH drop (at this moment 20% of
the lactose is metabolized; yogurt: 40%)
 Lactic acid fermentation

• Important flavor compounds are:

– diacetyl (butter flavor)
– acetaldehyde (yogurt flavor)
– propionic acid can be formed out of lactic
acid by propionic acid bacteria
• 3 CH3CHOHCOOH 2 CH3CH2COOH +
CH3COOH + H2O + CO2
 Alcohol fermentation
• Lactose pyruvic acid acetaldehyde + CO2
ethanol
• This fermentation doesn't occur frequently
• Mostly an undesired fermentation by
yeasts
• In some sour foaming milk drinks it is
desired (kefir, koumiss)
 Butyric acid fermentation

• 2 CH3CHOHCOOH C3H7COOH + 2 CO2

+ 2 H2
• This fermentation is highly undesirable in
cheese because of the gas and off-flavor
• Characteristic for Clostridia bacteria
 Other Carbohydrates

• Lactulose
– a derivative of lactose present in heated milk
– often used as a parameter for heat-treatment of
milk, so that sterilized and UHT milk can be
distinguished
• Free glucose and galactose
– detected readily in fresh milk
– lack of agreement on their concentrations
• No carbohydrates as glycogen or starch
 Other carbohydrates
• Oligosaccharides
– present in small quantities
– composed out of glucose, galactose and fucose
(6-deoxygalactose)
– some are composed of hexosamines: N-
acetylglucosamines
• of great biological importance
• bifidus factor in human milk
• serological activity (anti-hemaglutination)
– some are made of neuraminic derivatives as N-
acetyl neuraminic acid (NANA)
• bounded on k-casein
• anti-bacterial activity
 Lactose separation

• Lactose is water-soluble
• Follows the aqueous phase during
separation processes
• Present in skimmed milk, buttermilk, whey
(often used as source for gaining of
lactose)
 2.2 Lipids

• Milk fat content

• Chemical properties
• Physical properties
• Deterioration of milk fat
• Other lipid components
• Fat globules
 Milk Fat and Composition

• Is variable
• Dependent on a large
• number of factors
– caused by differences between breed and
individuals
– influenced by the season
 Chemical properties

• Global composition of milk fat

– major fraction of neutral lipids, in particular
triglycerides
– small fraction of polar lipids with important
structural functions
• present in the fat globule, in the fat globule
membrane and in the plasma
 Chemical properties
• Milk fat triglycerides
– a very great range of some 250 different fatty acid residues
– a relatively high proportion of short-chain fatty acids (C4-C10)
– butyric acid is specific for milk fat of ruminant species
– a high proportion of saturated fatty acids is high (63% w/w)
– oleic acid is the most abundant of the unsaturated fatty acids
residues
– other unsaturated fatty acids residues are present in a wide
variety of chain length, unsaturation and isomers
– several "odd" fatty acids (uneven, branched, keto, hydroxy)
– fatty acids are stereospecifically distributed: asymmetric
structure, which influences the texture of derived products
 Chemical properties
• Mono- and diglycerides and free fatty acids
also occur in small quantities (hydrolytic
products from lipolysis)
• Fluctuations in the composition occur
– dependent on feed, season and breed
– are of major technological importance as
they influence the physical properties of
the milk fat (harder fat during winter
periods)
Fatty Acid Composition of milk lipids
 Physical Properties
• Melting Point

• Of major importance in the manufacturing of milk

products and milk fat in particular
• Is determined by the fatty acids
– chain length and degree of saturation
– place of the double bond
– the trans-isomers
– the place of the fatty acids within the triglycerides
• Influenced by minor components
– mono- and diglycerides
– free fatty acids
– phospholipds
– water
 Physical Properteis

• Crystallization

• Nucleation
• Crystal growth
• Polymorphism
 Nucleation and crystal growth

• Bulk milk fat

– one catalytic impurity per mg suffices to ensure rapid crystallization
– super cooling of 5 K suffices to induce nucleation
– this concerns the highest melting triglycerides
– as soon as these nuclei have been formed, they act in turn as
catalytic impurities for the nucleation of other triglycerides
– little hysteresis between solidification and melting curves
• Emulsified milk fat
– fat is finely divided: 1 mg of fat is divided among some 108
-globules, in homogenized milk among at least 1011 globules
– in each globule at least one nucleus must be formed: the number
of catalytic impurities may easily become limiting
– super cooling can be more exquisite
– hysteresis can be considerable
 Polymorphism
•Three polymorphic modifications α, β' and β
– each modification is characterized by its crystal lattice type (mode
of packing), not by its geometrical form (habit)
– an important feature is the distance between the chains, the so called
short spacing (X-ray diffraction patterns)
– the melting points increase in the order a, b' and b, as do the
melting heat and the density of the crystals; this implies that
closeness and intricacy of fit of the molecules increase and their
freedom of motion decreases
– generally the a, b' modifications are not stable (meta stable)
– Nucleation of a fat usually occurs in the a modification
• mostly, after a little while, transition to a stabler polymorph occurs
– In milk fat, a crystals can be very persistent
– Conversely, in most fats the a modification has only a short
-lifetime, while b' may persist longer
 Deterioration of milk fat
• Lipolysis
• Oxidation
• Fishy taste
 Lipolysis
• = Hydrolysis of fatty acid esters by the action of lipases
– results in the common flavor defect know as lipolytic or hydrolytic rancidity
and is
distinct from oxidative rancidity
• By endogeneous lipases (mainly lipoprotein lipase) or by bacterial lipases
• The properties of the fat globule membrane are of major importance
– reduced contents of phospholipids or mastitis can increase the sensitivity of
the
fat globule for lipolysis
– other factors that destabilize the fat globule membrane, especially agitation
and
foaming, also promote lipolysis
– lipolysis is promoted in the manufacturing of cheese
• Sensory perception of lipolytic rancidity is strongly affected by the pH of the
product
– at low pH they are more readily tasted
• – in fresh milk threshold values corresponded to acid degree values (ADV)
of 4,1
• to 4,5 mmol per 100 g of fat
 Oxidation
• Proceeds by the well-known auto oxidation reaction in
three stages initiation, propagation and termination
– unsaturated fatty acids are transformed to
hydroperoxides, the primary reaction products
– during propagation, antioxidant compounds such as
tocopherols and ascorbic acid are depleted while
peroxide derivatives of fatty acids accumulate
– peroxides, which have little flavor, undergo further
reactions to form a variety of carbonyl, the secondary
reaction products, which are responsible for the
rancid taste and odour
 Oxidation
Factors affecting oxidation can be categorized into 2 groups

• Intrinsic factors • Extrinsic factors

– metalloproteins such as milk – contamination with metals
peroxidase and xanthin oxidase – temperature of storage
– endogenous ascorbic acid, – oxygen tension
which acts as a cocatalyst with – heat treatment
copper to promote oxidation – agitation
– endogenous copper content – light
– endogenous antioxidants, – acidity
mainly tocopherols
 Oxidation
• Effect of heat treatment
– migration of copper from the plasma to the fat globule
– denaturation of metalloproteins and increase the availability of metals
for oxidation
– high heat treatment stabilizes milk against oxidation
• probably due to exposure of sulfhydryl groups of denaturated proteins
and the releaseof hydrogen sulfide
• Effect of homogenization
– reduces the sensitivity of milkfat to both copper- and light-induced
oxidation, probably because oxidation sensitive membrane lipids are
displaced
• Effect of light
– photooxidation is accompanied by depletion of riboflavin, ascorbic
acid and some amino acids
– so-called sunlight-flavor is caused by oxidation of methionine to
methional
 Fishy taste
• Can be induced by transformation of
Phosphatidylcholine to trimethylamine
 Other lipids
• Phospholipids
• Sterols
• Other lipids
Phospholipids
 Sterols
• Form the largest fraction of the unsaponifiable
lipids
• Consist largely of cholesterol
• A small fraction of the cholesterol is esterified
(saponifiable sterol fraction)
• Sterols can be found in:
– The fat globule
– The fat globule membrane
– The serum phase
 Other lipids
• Pigments, mainly carotenoids (β-carotene =
provitamin A)
– concentration is season-dependent; in summer
double quantities compared to winter
– in sheep, goat and buffalo milk no β-carotene is
present
• Vitamins (mainly A, D and E)
• Antioxidants (mainly tocopherols)
• Polar fatty acids: keto acids en HO acids, important
for lacton formation
• Squalene
 Fat globules
• Milk fat present in milk as a globule that
reduce interfacial tension of lipids-serum
• Fat globule distribution
• Fat globule structure
• Agglutination
 Fat globules distribution
• Thermodynamically, no emulsion is stable
– the stability is a kinetic time-dependent phenomenon
– milk separates or creams spontaneously and rapidly and
many processes involve manipulation of the creaming
phenomenon
– emulsion stability is largely dependent on the size
distribution of the globules
– in raw milk, fat globules range in size from 0,1 to 15 μm
• The milk emulsion contains three distinct populations of fat
globules
– small globules (<1μm) that represent 80% of the total
number of the globules, but only a small fraction of the total
milk fat
– large globules (>12μm) that represent 2-3% of the total milk
fat
– the medium group which represents 95% of the total milk fat
 Fat globules distribution
• The distribution can be characterized by some
parameters
– the average diameter
– the average volume
– the average ratio volume/surface
• A large activity at the level of the membrane: 70
m2 per liter of milk (3,5 % fat)
• The fat globule size distribution can be influenced
by many factors :
– shift to larger globules during lactation
– in fatty rich milk larger globules occur
 Fat globule structure
• Fat globule membrane
-8 to 10 nm in thickness
-reduces the lipid/serum interfacial tension
-composition dependent on the procedure of
isolation (difficult quantification)
 Fat globules structure
• Composition of natural fat globule membranes
– phospholipids and cholesterol
– proteins, especially glycoproteins with properties similar to the
proteose-pepton-fraction of the serum
– enzymes (xanthine oxidase and alkaline phosphatase)
– high melting glycerides, which are probably crystallized on the
membrane during cooling
 Agglutination
• In undisturbed milk, lipid globules rise and
form a cream layer
• The creaming effect can be described by
the law of Stokes:
v= 2r2 (dp - df) g
9ŋ

V = speed of creaming g = gravity acceleration

r = radius ŋ = viscosity
dp, df = density of plasma and fat
 Agglutination
• = Flocculation of fat globules
– speed of spontaneous creaming is in reality much
higher than predicted by the law of Stokes
– adverse temperature effect is found; at lower
temperatures creaming increases
– doesn't often occur in heated milk: a consequence
of denaturation of agglutinins
– in homogenized milk no flocculation occurs, but
clustering
• Destabilization of the emulsion can be performed by
– mechanical movement (free fat in cooled milk)
– enzymes
– churning (butter still contains fat globules; 10-50 %
of the fat
Destabilization processes in
emulsions
 Positive Nutritional Aspects of Milk Fat

• Conjugated linoleic acid (cis-9, trans-11

octadecanoic acid)
• Produced in the rumen by Butyrivibrio fibrisolvens
Up to 100 μmol/g milk fat
• Potent anti-carcinogen (especially colon cancer,
melanoma and breast cancer)
• Antioxidant
• Lowers blood LDL-cholesterol
• Promotes muscle and fat growth, raises rate of
metabolism
• Positive influence on the immune system
 Nutritional aspects of Milk Fat
• Sphengomyeline
• Structural function in membranes
• Positive effects on cholesterol metabolism
• Protection against fumonisin, bacterial toxins
and pathogens
• Anti-proliferative effect on cancer cells (colon
cancer) by influencing cell differentiation, and
programmed cell death
• (apoptosis)
 Other positive nutritional aspects
• Butyric acid
– Unique feature of milk fat (7-13 mol/100 mol
milk fat)
– Results from fermentation of non-absorbed
carbohydrate by colonic microflora
– Is utilized by colonocytes as an important
energy source inducing colonic generation
(importance of dietary fibers!)
– Potent inhibitor of cell proliferation and
inducer of differentiation and apoptosis in a
number of cancer cell lines
– Synergic anti carcinogenic effect with vit. A, D
and E
 Chemical & Physical Properties
• Proteins
 Introduction
 Heterogeneity of milk proteins
 Caseins
 Casein micelles
 Whey proteins
 Milk fat globule membrane proteins
 Non-protein nitrogen
 Enzymatic coagulation of caseins
 Heat-induced coagulation
 Acid-induced coagulation
 Ethanol coagulation
 Age-gelation of sterilized milks
 Denaturation of whey proteins
 Introduction
• Milk proteins: of great importance
– human nutrition: nutritional functionality
– behavior/properties of dairy products: technological
functionality
• Normal bovine milk: 30-35 g protein/litre
• Nitrogen content of milk
– caseins
– whey proteins
– non-protein nitrogen (NPN)
– minor proteins associated with the milk fat globule
membrane
Amino acid composition of the milk proteins
 Caseins
• αs1-Caseins
• αs2-Caseins
• β-Caseins
• k-Caseins
 Casseins
• Original definition:
“Those phosphoproteins which precipitate from
raw skim” milk upon acidification to pH 4.6 at
20°C”
• Account for 76-86% of the total milk protein
• Essentially all occur in micelles
• Most of the caseins can be represented by
four gene
• products: αs1-, αs2-, β- and k-caseins in the
approximate ratio 40:10:35:12
 Caseins
• Nowadays:
classification according to chemical structure, rather than
on the basis of an operational definition
– not all of the caseins contain phosphorus
– some are found in acid whey
• Nomenclature
– a Greek letter with or without a numerical subscript to
identify the family of proteins
– an uppercase Latin letter to indicate the genetic variant
– post-translational modifications such as number of
phosphorylations or formation of sub-fractions are indicated
after the genetic variant
– e.g. as1-CN B-8P
 Casseins
• Caseins
– contain high numbers of proline residues distributed
relatively uniformly throughout the polypetide chains
– proline inhibits formation of an ordered, stable a-helix
 lack of tertiary structure of caseins
 stability of caseins against heat denaturation
 considerable exposure of hydrophobic residues to water
 strong association of caseins
 insoluble in water
• Milk protein genetic variants
– Relationship between genetic polymorphism of milk proteins
and technological properties
– Milk composition is different for various phenotypes
 Caseins
• Contain negatively charged groups such as
phosphates,side-chain carboxyls, terminal
carboxyls, and sulfhydryls
– binding of a number of different cations, such as
calcium, barium, magnesium, potassium, sodium,
etc.
– highly charged segments facilitate electrostatic
interactions
• Calcium
– calcium binding by phosphoseryl residues is of
primary interest
– involved in micelle formation
 αsCasseins
• Calcium-sensitive casein fraction
• Precipitated with 0.4 M CaCl2 at pH 7 and
4°C
• αs1-caseins
• αs2-caseins
 αs1Cassein
• Five known genetic variants: A, B, C, D, E
– the polymorfs are breed specific
– the B variant: predominant in Bos taurus
(European cattle)
• Predominant genetic B variant
– consists of 199 amino acid residues
– a calculated molecular weight of 23,614 D
(dalton)
 αs1Cassein
• Polypeptide chain
– two predominantly hydrophobic regions (1 - 44,
and 90 - 199)
– a highly charged polar zone (45 - 89)
– rather loose and flexible polypeptide
• λ-caseins
– may be present in milk in small amounts
– fragments of αs1-casein as the result of plasmin
proteolysis
 αs2-Caseins
• Four recognized genetic variants: A, B, C, and
D
– the same primary amino acid sequence
– different degrees of posttranslational
phosphorylation
– A and D variants observed in European breeds
(Bos taurus)
• αs2-casein A-11P
– primary structure of 207 amino acid residues
– a calculated molecular weight of 25,230 D
 αs2-Caseins

• More sensitive to precipitation by Ca++

than αs1-casein
• Polypeptide chain
– a remarkable dipolar structure
– negative charges near the N-terminus
– positive charges near the C-terminus
 β-Caseins
• 1 major component with at least seven
genetic variant(A1, A2, A3, B, C, D, and E)
8 minor components: proteolytic fragments of
the major component
• A variants: predominant (nearly 100%)
polymorphs in all species and strains of Bos
 β -caseins
β -casein A2-5P
– a single polypeptide chain of 209 residues
– a calculated molecular weight of 23,983 D
• Polypeptide chain
– a strongly negatively charged N-terminal portion
– rest of the molecule has virtually no net charge
– the β-casein molecule is somewhat similar to an
anionic detergent with a negatively charged
head and an uncharged essentially hydrophobic
tail
 β -caseins
• γ1-, γ2- and γ3-caseins
– present in raw milk
– formed by the action of the plasmin on β-casein
in milk
– carboxyl terminal fragments of b-casein
remaining associated with the casein micelle
and recovered by pH 4.6 precipitation
– corresponding N-terminal fragments
• classified as proteose-peptones
• hydrophilic and appear as heat-stable fractions in
whey
 ΚCaseins
• A mixture of polymers held together by disulfide
bonds
– monomers consist of a major carbohydrate-free
component and at least six minor components
• Minor k -casein components
– same primary amino acid sequence as major
component
– contain various amounts and types of carbohydrate
moieties attached to the polypeptide chain by post-
translational glycosylation
 Κcaseins
• Two genetic variants are known: A and B
– the A variant tends to be the predominant variant in most
breeds
• k-casein B-1P
– consists of 169 amino acid residues
– a calculated molecular weight of 19,007 D
• Possess considerable heterogeneity
– genetic differences
– variation in carbohydrate content and/or phosphate
content
– a possible variation in the para-k-portion
 Κcaseins
• Mostly contain only one phosphoseryl residue
• Do not bind Ca++ strongly and are soluble in the
presence of high calcium concentrations
• k -casein associates with as1- and/or b-casein
 stabilizes them against precipitation by Ca++
 formation of stable colloidal particles similar to
but less stable than native casein micelles
 Casein Micelle
• Casein micelle structure
• Stability of micelles
• Forces
 Casein Micelles
• Micelles
– = coarse colloidal particles
– ≈ 95% of the casein in normal un-cooled milk
– comprised of some 20 - 150,000 casein
molecules
– molecular weights of ≈108 Dalton
– mean diameters of ≈100 nm (range 50 - 500
nm)
 Composition of casein micelles
• Composition of casein micelles
– consist on dry weight base of » 94% protein
and » 6% of small ions principally calcium,
phosphate, magnesium and citrate, referred
to collectively as colloidal calcium phosphate
(CCP)
– milk micelles are highly hydrated, typically ≈ 2
g H2O/g protein
 Casein Micelle
• Important and characteristic macromolecular
assembly of mammalian biology
• Occurring in all milks that have been examined in
detail
• Biological functions
– to form a coagulum in the stomach of the nursling,
allowing the slow release of nutrients down the
digestive tract
– to act as a means of transporting calcium and
phosphate in a readily assimilable form from mother
to young
– to provide a source of amino acids
– to produce biologically active peptides by enzymatic
cleavage of casein polypeptide chains such as the
casomorphins
 Models of Casein micelle structure

• Detailed structure is still not • Walstra , 1999 structure

known!
• Submicelles
– basic units that compose a
micelle
– 10 - 15 nm in diameter
– a porous structure
– varying composition of the four
principal caseins
linked together in the micelles by
colloidal calcium phosphate
(cementing agent) and
hydrophobic bonds
Models of casein micelle structure
David Horne, 2003
Double binding model
 Models of casein micelle structure
• Dalgleish (2004)
– No spherical sub-micelles
– Caseins organized as tubules

Calcium phosphate
nanoclusters

Κ-casein on surface
Gaps big enough available for reaction
which allow proteins with denaturated
to pass trough or bind whey proteins
 Casein micelle structure
• k-casein
– the principal micelle-stabilizing factor
– located predominantly on the surface
– very hydrophilic C-terminal part = flexible
"hairs”
– essential in providing stability against
flocculation of the micelles
– hydrodynamic thickness of the hairy layer
≈ 7 nm
 Stability of casein micelle
• Heat stability
– limited heat treatment has no effect on the micelles
– sterilization can disintegrate micelles into a fibrillar
structure
– stable at the following temperature-time
combinations: 10 minutes at 130 °C and 50 minutes
at 115 °C
• Lowering temperature
– lower temperatures cause a disintegration of micelles
in particular β-casein is sensitive
– important as milk is frequently cooled to below 5°C
– effect is reversible when increasing the temperature
– low temperatures give a finer, more voluminous
precipitate and a weaker gel by acidification
 Stability of casein micelle
• Change in mineral composition
– removal of calcium ions from the micelle causes reversible
dissociation of β- and k-casein from the micelles without real
micelle disintegration
– addition of excess Ca++ favors micellar component
aggregation
– aggregation of casein micelles in concentrated and frozen
milk products seems to be caused by salting out (a
decrease steric repulsion)
• Dehydration
– dehydration leads to micelle-aggregation
– dehydration of k-casein causes presumably a decrease in
voluminosity and steric repulsion
Chemical & Physical
Properties
Whey Proteins
 Whey Proteins
• β-Lactoglobulin
• α-Lactalbumin
• Bovine serum albumin
• Immunoglobulins
• Lactotransferrin
• Enzymes
 Whey Proteins
• Whey proteins are the major nitrogen compounds
remaining in milk after precipitation of the caseins
by acid (pH 4.6) or by rennet (pH ≈6.7) and
represent ≈ 20% of the nitrogen in bovine milk
• Include a characteristic group of globular proteins
• Are synthesized in the mammary gland of the cow
– e.g. β-lactoglobulin and α-lactalbumin
or
• Are derived from the blood
– e.g. bovine serum albumin and immunoglobulins
 Whey Proteins
• Some minor whey proteins have antimicrobial
properties,
e.g. lactotransferrin, lactoperoxidase, and lysozyme
• Some identified polypeptides include the proteose
pepton components which occur in the acid and
rennet whey and the glycomacropeptides present
only in rennet whey
• Milk contains ≈ 30 enzymes, derived mainly from
blood and secretory cell membranes. Some of
these enzymes, especially lipoprotein lipase and
proteinase, are technologically important in milk
and dairy products
 Whey proteins
•β lactoglobulins

• Is a major milk protein

– ≈ 50% of the whey proteins
– ≈12% of total milk proteins
• Monomeric molecular weight is about 18,300 D
• Has a high affinity for retinol
– speculations that its biological function is related to vitamin A
transport
• 8 genetic variants: A, B, C, D, Dr, E, F, G
– A and B, have been found in all breeds of Bos taurus and B.
indicus examined
– some breeds of B. taurus also secrete variants C and D
– milk of the Draughtmaster breed contains a fifth variant Dr
– other variants are found in the milk of B. grunniens (Yak) and B.
javanicus (Bali cattle)
Primary structure of Bos β- lactoglobulin &α
Lactalbumine
 β -Lactoglobulin
• Conformation
– depends on the pH
– between pH 6.7 and 5.2 (its isoelectric point) at
room temperature:
dimerization (MW of 36,700 D)
– between pH 5.2 and 3.5: octamerization of
especially the A variant (MW of 147,000 D);
maximal octamerization from pH 4.4 to 4.7 at 0°C
– below pH 3.5: monomers due to strong electrostatic
repulsive forces
– the B variant (predominant one in Western cattle)
octamerizes to a much smaller extend, possibly due
to increased electro-static repulsion
 β -Lactoglobulin
• Has a free thiol group at Cys119 or Cys121
– of great importance for changes occuring
in milk during heating
– is involved in reactions with other proteins,
notably k-casein and α-lactalbumin
• Binds a variety of hydrophobic molecules
 α-Lactalbumin
• Represents » 20% of the proteins of bovine whey
• Is a small molecule with a molecular weight of about 14
kD
• Iso-electric point is pH 4.8
• 2 to (3) genetic variants, A, B, (C)
– only variant B is found in the milks of western breeds
– milks of African Fulani and Indian Zebu cattle contain
both A and B variants (differing by only one amino
acid replacement)
• Several minor components
– the same amino acid composition as the major
component
– glycosylated forms that contain mannose, galactose,
fucose, Nacetylglucosamine,N-acetylgalactosamine,
 α-Lactalbumin
• Amino acid sequence is similar to that of lysozyme
– α-lactalbumin arose in evolution by gene duplication
of an ancestral gene coding for lysozyme
• Is necessary for the synthesis of lactose
– interacts with galactosyltransferase (an enzyme
which catalyzes the transfer of galactose)
– without α-lactalbumin, glucose is an extremely poor
substrate for galactosyl-transferase
 α-Lactalbumin
• Contains eight cysteine groups, all of which are involved in
disulfide bonds
• Has a highly ordered secondary structure
• Has a compact, spherical tertiary structure
• Undergoes a conformational change at pH < 4.0
– is accompanied by the release of bound calcium
– results in temperature and concentration dependent
aggregation
• Thermal denaturation of a-lactalbumin
– is quite reversible
– is also accompanied by a release of bound calcium
• Is stabilized against heat denaturation and aggregation in
the presence of calcium
 α-Lactalbumin
• Has rather good emulsifying and foaming
properties, although the native molecule
reveals low surface hydrophobicity
• The susceptibility of α-lactalbumin to surface
denaturation at oil-water or air-water
interfaces is involved in its emulsifying and
foaming properties
 Bovine serum albumin
• Is identical to the blood bovine serum albumine
(BSA), the major protein of blood plasma
– milk ≈ 0.4 g BSA/litre
– blood contains 30 - 40 g BSA/litre
• BSA contains 582 amino acids and has a calculated
molecular weight of 66,267 D
• Has no specific function in milk?
– Is presumably a leakage protein
• Is a well-known transport protein for insoluble fatty
acids in the blood circulatory system
– binding of fatty acids stabilizes the protein molecule
against heat denaturation
– this ability to bind fatty acids may promote lipolysis
 Bovine serum albumin
• Has a rather complex tertiary structure
– three about equal-sized major globular
domains dissimilar in hydrophobicity, net
charge, ligand binding sites
– one free thiol
– 17 disulfide linkages
 Bovine serum albumin
• Behavior of BSA in milk and milk products
– little is known
– possible influence on properties of milk and milk
products?
• Heat-induced gelation behaviour at pH 6.5
– initiated by an intermolecular thiol disulfide interchange
• BSA is highly soluble up to 35% (w/w) in pure water at
3°C
• BSA undergoes extensive precipitation in the
temperature range 40-45°C
– this changed solubility above 40°C parallels the
reversible (partial)
• unfolding of BSA
• On acidification to pH 4.0, the BSA undergoes acid
denaturation
 Immunoglobulin(lg)
• Are antibodies synthesized in response to
stimulation by macromolecular antigens foreign
to the animal
• Bovine colostrum contains up to 100 g Ig/litre
– concentration decreases to < 1 g/litre within a
week of parturition
• Their primary function in milk is to provide
passive
• immunity for the neonate
 Immunoglobulin
• Are a very complex, heterogeneous group of proteins
• Five principal classes of Ig are recognized
– IgG (sub-classes IgG1, IgG2, IgG3, IgG4)
– IgA (sub-classes IgA1, IgA2)
– IgM
– IgD
– IgE
• Up to 80% of the immunoglobulin in milk or whey is Ig-G
• The basic sub-unit in each class consists of four
• polypeptide chains linked by disulfide bridges
– two heavy (H, MW ≈ 50 - 70,000) chains
– two light (L, MW = 22,400) chains
Four-peptide chain structure of immunoglobulins
 Transferrins
• = a group of evolutionary related iron-
binding proteins
• Best characterized members
– sero-transferrin (present in blood plasma
and other extracellular fluids, e.g., spinal
fluid, semen)
– lacto-transferrin (milk, pancreatic juice,
tears, leucocytes)
 Lactotransferrin
• A monomeric glycoprotein with a molecular weight of
about 80 kD
• Has the ability to bind reversibly two Fe+++ per molecule
(concomitantly ie together with two HCO3-)
• Suppresses bacterial activity by removing the iron that is
required for bacterial growth
• Concentration
– in bovine colostrums: ≈1 mg/ml
– in milk: 0.02 - 0.35 mg/ml
• Several improved methods for their isolation have been
published
– because of the apparent physiological and nutritional
significance of lacto-transferrins
 Milk fat globule membrane proteins
• MFGM
– a thin membrane that surrounds the fat globules in milk
– contains approximately 50% protein and accounts for about
1% of the total protein of the milk
• Total MFGM protein content
– as observed is dependent upon the past history of the
membrane from its formation to its analysis
– both the temperature and the time of storage before analysis
can alter the membrane composition and physical state
• SDS-PAGE / IEF
Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis
IEF IsoElectroFluoroscence
– most universally accepted method for characterization of
membrane proteins
– milk fat globule membrane proteins are classified based on
their electrophoretic mobilitie
Estimated composition of natural
MFGM
 Non-protein nitrogen
• = a large number of N-containing compounds of low
• molecular weight occur in milk
• 250 - 350 mg of N per litre milk
• Wide variations in concentrations
– probably arise from the fact that many of them are
metabolites of amino acids and nucleic acids and
from the fact that their concentrations in milk
depend on the amounts of those substances
consumed by the cow
Principal NPN components in milk
 Enzymatic coagulation of caseins
• Introduction
• Primary phase of rennet action
• Secondary (non-enzymatic) phase of
coagulation
• Curd formation
• Final stage
• Rennets
 Enzymatic coagulation of caseins
• = clotting of milk by proteolytic enzymes
• Represents one of the oldest operations in
food technology
– is used since antiquity for the manufacture
of cheese
• Is a complex process
 Enzymatic coagulation of caseins
• A primary enzymic phase
– k-casein is altered and loses its ability to stabilize the
remainder of the caseinate complex
• A secondary non-enzymic phase
– aggregation of the altered casein micelles takes place
• A third step: liquid to solid
– the aggregate of casein micelles forms a firm gel structure
• A possibly separate fourth step
– the curd structure tightens and syneresis (the expulsion of
water by the curd arising from structural rearrangements after
formation) occurs. In cheese making, this syneresis enhanced
by cutting
• The enzymic reaction and the subsequent aggregation can
occur simultaneously during the later stages
• Without primary enzyme action, no clotting occurs
 Primary phase of rennet action
• Involves the enzymatic cleavage of the Phe105-Met106
linkage of k-casein
• This results in the formation of
– soluble glycomacropeptide (GMP) which diffuses away from
the micelle and is washed out (106-169)
– para-k-casein = a distinctly hydrophobic peptide that remains
on the micelle (1-105)
• A relatively quick reaction
– turnover: 100 s-1 in milk, pH 6.7, 30°C
– seems independent of micelle size
– decreasing temperature by 10°C reduces the rate by a factor 2
Properties of k-casein and the polypeptides formed
by chymosin action
 Secondary phase of coagulation
• A non-enzymatic phase
= aggregation of the renneted micelles
• Hydrolysis of k-casein reduces the intermicellar repulsive
forces (electrostatic and steric) and the micelle stability
• No aggregation until 80-85 % of k-casein has been destroyed
• During the last 20% of the proteolytic reaction
– concentration of micelles capable of aggregating increases
– the aggregation rate increases rapidly
• Aggregation rate
– is unaffected by the concentration of rennet
– is unaffected by the size of the casein micelles
– is very sensitive to the concentration of calcium ions
– is very sensitive to the temperature
 Secondary phase of coagulation
• Milk will not clot at less than about 15°C
– this is a direct consequence of the slowness of the
aggregation
reaction at low temperatures
• Low temperatures (less than 8°C)
– allow proteolysis of k-casein in the absence of coagulation
– coagulation occurs on subsequent warming
• Temperatures above 45°C
– aggregation is very efficient
– approaches the theoretical maximum particle collision rate
• Addition of CaCl2
– decreases the coagulation time
– at high calcium chloride concentrations (0.4 M), the
coagulation time is retarded severely and only weak curd is
obtained
 Curd formation
• Is characterized by a steady aggregation of the rennet treated
casein micelles
– Chains of micelles are formed at first
– By the rennet clotting time (RCT), the chains have begun to link
into a loose network
– The network extends and becomes more differentiated, with the
chains of micelles aligning together
• The visually-observed rennet clotting time (RCT)
– when the curd starts to form
– the resultant of two reactions: the enzymatic reaction is largely
determinant, as flocculation time is insignificant to splitting time
– at 4°C, the enzymatic reaction may be complete in about 3 h,
while clotting takes a week
 Final stage
• Is not well defined and includes
– syneresis and firming of the curd
– a loss of paracasein micelle identity
– non-specific proteolysis of caseins in the coagulum
• The paracasein micelles fuse into larger units as
CCP rearranges throughout the micellar region
– this may be analogous to binding between
submicelles in a micelle
• Syneresis is enhanced by applying external pressure
to the curd or deforming it (stirring, cheddaring)
 Renneting of milk

S= degree of splitting of k casein

A= Agregation of casein micelle
F= Firmness of the coagulum(gel)
 Rennets
• Traditional rennet
= a crude enzyme extract prepared from calves
stomachs
• used in the milk-clotting-process
• Shortages of this material !
– use of pepsins and acid proteases produced by
other animals, fungi and microorganisms
• Nowadays any milk-clotting enzyme preparation
yielding a relatively stable curd is designated as
rennet
 Animal rennets
• Calf rennet
– generally refers to an enzyme extract obtained from the
fourth stomach (abomasum) of 10- to 30-day-old un-weaned
calves
– used to coagulate milk for cheese production
• Chymosin
– the purified milk-clotting enzyme present in crude rennet
– a molecular weight of 35,600 D
– belongs to the group of aspartic proteases
– the standard against which all other types of milk-clotting
enzymes are compared
– coagulates milk rapidly at its natural pH with little further
degradation of the milk proteins
 Animal rennet
• As the calf ages, chymosin is replaced by pepsin
– in cattle, the chymosin secretion never comes to a complete
stop
– pepsin can also clot milk
• Pepsin’s use as a 100% replacement for calf rennet in
cheese making is limited due to shortcomings
(i) set time for curd formation is extented
(ii) the curd is softer than that obtained with calf rennet
(iii) bitter peptides are formed
(iv) fat loss is excessive
(v) the cheese flavor is generally bland
(vi) above pH 6.5 pepsin activity falls of so rapidly that its use is
limited to the production of only some sweet (e.g. swiss
cheese) and some Italian varieties of cheeses
 Animal rennet
• A worldwide shortage of calf rennet has developed
– largely due to an increase in cheese production
– a greater tendency to slaughter "mature" calves
• Stomachs of other young mammals (lambs)
– obvious sources of rennet substitutes
– they produce acid proteinases that may be expected to
have properties in common with calf rennet
– pepsins and chymosins have also been isolated from the
adult cow, sheep, buffalo, chicken, rabbit, swine, and
goat
• Apparently, humans, dogs, and possibly young pigs do
not secrete chymosin
 Plant Rennet
• Many proteolytic enzymes of vegetable origin are
milk coagulants
– papain from papaya
– ficin from Ficus spp.
– bromelain from pineapple
• Most plant coagulants
– proved to be non-specific in their proteolytic activity
– the excessive proteolysis results in bitter products
• At present there is no commercially available rennet
derived from a higher plant
Plant rennets
 Bacterial rennets
• A number of bacteria are reported to produce milk-
clotting enzymes
• Possible advantages
– high titer of the active preparation
– short generation times
• Commercial production of rennets with bacteria has
not been successful
– invariably strong and nonspecific proteolytic action
– loss of fat and nitrogen in the whey
– reduced yield
– poor quality of the aged cheese
Bacteria reported to produce milk
coagulants
 Fungal rennet
• Many species of filamentous fungi produce a
chymosin like enzyme
• Most fungal coagulants are much too proteolytic
for use as rennets
• The coagulants from three fungal species have
proved
• suitable for large-scale commercialization
– Endothia parasitica (Cryphonectria parasitica)
– Mucor miehei (Rhizomucor miehei)
– Mucor pusillus (Rhizomucor pusillus)
 Recombinant calf chymosin
• Differences still exist between microbial and calf chymosin
(i) the ratio of proteolytic to milk-clotting activity is still a little
higher
or microbial rennets, resulting in greater hydrolysis of cheese
protein during ripening and leading to soft body and texture
(ii) microbial rennets are more thermostable, cannot be
inactivated at
normal pasteurization temperature, and therefore present
problems in processing of cheese whey
• (iii) change in milk-clotting activity from calf to microbial
rennet will
require adjustment of the process parameters such as
temperature, pH, calcium concentration etc.
 Recombinant calf chymosin
• Calf chymosin has been cloned in suitable microbes
• Commercial calf chymosin produced by recombinant DNA
technology
– Aspergillus niger var. awamori : Chymogen® (Chr. Hansen)
– Escherichia coli K12 : Chy-Max® (Pfizer)
– Kluyveromyces lactis : Maxiren® (Gist-Brocades)
• A variety of cheeses have been produced using
recombinant chymosin
– no significant differences could be detected among
cheeses made by rennet and recombinant chymosin
 Heat-induced coagulation
• Bovine milk withstands high processing temperatures
– is essential for many modern milk-processing
operations
• The heat stability of milk is normally far in excess of
that required to withstand all the commercial
processes to which it is subjected.
• Non-casein proteins of milk are relatively heat-labile
• Caseins are remarkably heat-stable
– sodium caseinate in water can withstand heating at
140°C for >60 min at pH 6.7 and typical bulk bovine
milk is stable for »20 min at 140°C
 Heat-induced coagulation
• Most protein denaturation reactions are very pH
dependent
– the pH change caused by heating is primarily
responsible for heat coagulation
– the decrease in pH upon heating is partially due to
changes in the buffer capacity of milk salts and the
release of carbon dioxide
• Heating milk at a pH < 6.5 for 20 to 30 min at 100°C
– it coagulates to form a gel
– casein micelles have denatured whey proteins
attached to the micelle surfaces Heating milk at a pH
> 6.7
– very intensive heat treatment is required for
precipitation
– a gel does not form
 Heat-induced coagulation
• High heat stability of milk casein micelles
– reflects the open, random-coil structure of the caseins
– treatment at elevated temperatures has little effect on
its (absent?) secondary and tertiary structures
– hydrophobic bonds that play a major role in the
association of caseins at low or ambient temperature
are non-existent at temperatures > 100 °C and hence
there is little tendency to aggregate at high
temperatures
– negatively charged phosphate groups of casein may
contribute to its remarkably high heat stability
 Acid induced coagulation
• Caseins are insoluble at their iso-electric points (≈ pH
4.6) at temperatures above 20 °C
• Caseins remain more or less soluble if milk is acidified
to pH 4.6 at temperatures lower than 6°C
– the precipitate formed becomes progressively coarser
and more rubbery as the temperature at precipitation
is increased
– raising the temperature gives an immediate
precipitation
• Although the iso-electric point of the γ-caseins is
about 6, they also precipitate at pH 4.6
• Proteose peptons are soluble at pH 4.6
 Ethanol coagulation
• Milk coagulates on addition of ethanol
(usually to ≈ 40%)
• Ethanol stability
= the ability of milk to withstand treatment
with ethanol without coagulation
– has been used as a not-very-effective
selection test for milk for the manufacture
of sterilized concentrated milk
– is strongly influenced by the Ca++-
concentration and the pH
 Age-gelation of sterilized milk
• In-container sterilized evaporated milk occasionally gels
during storage
• UHT-sterilized (concentrated) milks
– are very prone to gelation during storage
– limits the usefulness of such products
• The cause (s) and mechanism of age-gelation
– have not yet been definitively established
– age-gelation of unconcentrated UHT milk
• plasmin and extracellular proteinases secreted by
psychotrophic bacteria are major causative factors
– age-gelation of UHT-sterilized concentrated milk
• physicochemical changes in the casein micelles appear to be
principally
• responsible for milk sterilized by a direct UHT process
 Age-gelation of sterilized milks
• UHT sterile milk concentrate
– contains casein micelle-denatured whey protein
complexes that are about double the size of native
casein micelles
– these complexes undergo extensive aggregation
during storage
– this eventually causes age thickening
• Age thickening is promoted by
– high milk solids content
– addition of alkali to raise pH
– addition of citrate, phosphate, and other anions that
lower Ca ion activity
• Addition of Ca++ improves stability against age
thickening
 Age thickening of concentrated
milk

Dotted lines show milk with added polyphosphate

Solid lines indicate milk without added polyphosphate
 Denaturation of whey proteins
• Whey proteins
– have typical globular conformations
– are relatively heat labile
– denaturation occurs upon heating
• Susceptibility to heat denaturation is influenced by
– pH
– Ca++-concentration
– protein concentration
– presence of sugars, polyhydric alcohols and protein modifying
agents
• The ranking order of the heat sensitivity
• α-lactalbumin > β-lactoglobulin > BSA > immunoglobulins
 Denaturation of whey proteins
• Heat-induced interaction between b-lactoglobulin
and k-casein
– plays a major role in determing the heat stability
and rennet clotting behaviour of milk
– β-Lactoglobulin precipitates on casein micelles
upon heating
– β-Lactoglobulin is also precipitated by
destabilization of the micelles (β-lactoglobulin -
k-casein interaction)
Denaturation of whey proteins
Chemical & Physical
Properties
Enzymes
 Enzymes
• Milk contains both indigenous and exogenous enzymes,
the latter being mainly bacterial
• Most significant bacterial enzymes occurring in milk are
heat-stable lipases and proteinases elaborated by
psychotrophic bacteria
• Cause several defects in milk but also can be responsible
for some beneficial effects
– desired transformations, e.g. ripening of cheese (natural
proteases)
– undesired degradation processes, e.g. lipolysis of raw milk;
– indicator enzymes for heating of milk
– antimicrobial effects of certain enzymes
– quality parameter
Endogenous Enzymes of bovine milk
Endogenous Enzymes of bovine milk
 Enzymes
• Lactoperoxidase
• Xanthine oxidase
• Catalase
• Lipase
• Phosphatase
• Protease
• Reductase
• Other enzymes
 Lactoperoxidase
• Catalyzes oxidation by H2O2 of a long list of
electron-donor compounds, including
– aromatic amines
– phenols
– aromatic acids
– leuko dyes
– tryosine and tryptophan
– ascorbate
– iodide
– nitrite
– thiocyanate
• Transfer oxygen in its atomic form from hydrogen
peroxide to its substrates
 Lactoperoxidase
• May amount to as much as 1% of the total serum proteins
of milk (i.e. 60 mg.kg-1)
• Its properties are well-known (pH-optimum 6,8; molecular
weight 82.000)
• Is an indicator enzyme for high pasteurization
• Catalyzes oxidation of thiocyanate (SCN-) to a product
(OSCN- ) that inhibits certain bacteria
• Thiocyanate is a natural constituent of milk and
hydrogen peroxide is produced by some bacteria
themselves (self-inhibition)
• In some countries this effect is enhanced by further
addition of hydrogen peroxide
 Xanthine oxidase
• Is very prominent in milk
• Most of it is associated with the fat globule
membrane
• Milk is a source of choice for isolating this
enzyme for investigation
• Different names are used for xanthine
oxidase; e.g
– Schardinger enzyme
– adenine oxidase
– nitrate reducing enzyme
– aldehyde oxidase
 Xanthine oxidase
• Catalyzes the reaction of purine bases (xanthine and
hypoxanthine) to ureic acid & hydrogen peroxide
• Can reduce NO3 (which occurs in milk only in trace
quantities) to NO2 which is a powerful inhibitor of
some bacteria
– put to use in the manufacture of certain types of
cheese, where a little nitrate is added to the milk to
prevent late blowing of the cheese by Clostridia
• Is especially activated by pasteurization and
homogenization
 Catalase
• Catalyzes the decomposition of H2O2 to H20 and O2
• In milk its activity parallels leukocyte count and is
higher in mastitic milk and colostrum than in normal
milk
• Its activity increases with the multiplication of
bacteria
• Is inactivated at 65°C after 30 minutes and is used
as a test to control heat treatment of milk
• The catalase procedure
– H2O2 is added to milk to inactivate spore forming
bacteria
– excess of H2O2 is decomposed by the catalase
 Lipases
• Catalyzes the hydrolysis of triglycerides to glycerol and fatty
acids
• Membrane lipase
– is originally present on the casein micelles, but adsorbs
irreversible to the fat gobule membrane during cooling of milk
– is often called lipoprotein lipase
– is responsible for the rancid flavor of raw milk
• Plasma lipase
– stays in the plasma during the cooling of milk
– can be activated by mechanical movement
• During cool conservation of milk, lipolysis can occur as a
consequence of the combination temperature - mechanical
movement
• Thermo-labile enzymes
 Phosphatase
• Also called phosphomonoesterases
• Of natural origin and responsible for hydrolysis of phosphoesters
• Alkaline phosphatase
– pH-optimum is about 9,6
– is destroyed by pasteurization and an index for the efficiency of
pasteurization
– two major isoenzymes have been identified, α and β
phosphatase,mainly located in the milk plasma and fat globule
membrane
– reactivation can occur
• Acid phosphatase
– pH optimum of 4,5
– extremely heat stable, but is sensitive to light and UV
– is present in low concentrations, in particular in skimmed milk
 Protease
• Plasmin
– the principal milk protease
– belongs to the alkaline serine proteinase class
– is probably identical to the plasmin of blood
– is present on the casein micelles
– has alkaline properties
– survives pasteurization
– is relatively resistant to UHT-treatment; can cause a bitter
flavor (hydrophobic peptides of low molecular weight) and
changes in viscosity and appearance
– is also of importance during ripening of cheese
• A second protease
– with maximal activity at pH 4,0
– is very heat sensitive and hydrolyses preferably α-caseins
– classification not yet clear; an aspartate proteinase?
 Reductase
• Related to the reducing properties of
microorganisms and leukocytes
• Can be used as quality-parameter
• Causes discoloration of methylene blue
and other dyes
• Has lost its importance as a result of better
cooling conditions, which has changed the
bacterial flora in milk
 Other enzymes
• Amylases
– catalyze the hydrolysis of starch to dextrin and maltose,
dependent on its nature (α- or β-amylases)
– the α-amylase is inactivated by heating 55°C during 30
minutes,
– the β-amylase keeps its activity after 30 minutes at 65°C.
• Lysozyme
– is quantitatively an important fraction of the whey proteins of
human milk (400 mg.l-1)
– less important in cow's milk
– is a powerful bactericide as it attacks polysaccharides of the
bacterial cell wall, causing lysis of the bacteria
– it releases bifidus factors
• Lactase
– normally doesn't occur, and if so only in small quantities
Chemical and Physical
Properties of Milk
Minerals and Vitamins
 Minerals and ionic equilibra
• Ash content
• Composition of mineral fraction
• Equilibra
• Trace elements
 Ash content
• ≈ 0,70 - 0,85 %
• < actual present salt content
• Comprises
– oxides of Na, K, Ca, Mg, Fe, P and S
– some chlorides
– S and parts of P and Fe of organic origin
Some minor components in milk
Some minor components in milk
 The composition of mineral fraction
• Three families of salt constituents
1. Na, K and Ca, which exist almost entirely as free ions in milk
and
are readily diffusible (present in milk ultrafiltrate); the
concentrations of these 3 ions are negatively correlated to lactose,
as required to maintain osmotic equilibrium of milk with blood
2. colloidal Ca, Mg, inorganic P and citrate; total concentrations of
Ca, Mg, P and citrate in milk plasma are 30,3; 5,2; 21,4 and 9,5
mM.
3. salts, whose concentrations are affected by the natural pH of
milk,
namely, diffusible Ca, diffusible Mg, diffusible citrate, Ca2+ and
HPO4-2
 Equilibra
• = the form in which the mineral occurs,
whether as ions, whether in colloidal form
• Of importance for physicochemical
properties of milk
-heat stability
-gelling
-clotting
• Ca/P-equilibrum
• Factors influencing equilibra
 Ca/P-equilibrum
• Forms of calcium
1. 20 % as organic Ca: Ca-caseinate
2. 80 % as inorganic Ca
50 % as inorganic colloidal tricalciumphosphate
20 % as not ionized Ca salt : Ca citrate and phosphate
10 % ionic Ca
• Forms of phosphorus
1. 30 % as organic P
10 % in phopsholipids and other esters
20 % in caseins as phophoserine
2. 70 % as inorganic P
35 % colloidal tricalciumphosphate
35 % soluble phosphate
 Ca/P-equilibrium
• Dialysis of milk doesn't result in removal of all Ca
and P,
• which means that these elements are partly bound
• A part of the calciumphosphate is present in an
insoluble form, the colloidal calciumphosphate
• Milk is supersaturated in calcium phosphate; this is
precipitated on the micelles
• On the casein, cations are bound as counter-ion for
the negative charge of the protein
 Factors influencing equilibrium
• Acidification
– During acidification more Ca will be present in its ionic form; the
colloidal calcium phosphate will be more soluble
– By acidification micelles are destabilized; the negative charge of
the micelles decreases, which causes a higher concentration of
ionic Ca
• Heating
– During heating a part of the calcium phosphate will become
insoluble and precipitates on the micelles; this can be
accompanied by a slight pH-drop
– Heating above 100°C causes splitting of phosphate from the
caseins and creates acids out of lactose; the result is a pH-drop
that influences the equilibrium
 Factors influencing equilibra
• Water removal
– During evaporation processes, the
concentration of solubilized components
increases, which can affect the equilibria
• Additions
– Addition of CaCl2 causes an increase of Ca-
activity and a decrease in the stability of the
micelles
– Addition of sodium citrate and certain
phosphates enhances the stability of the
micelles
 Trace elements
• Content of trace elements
• Role of Cu and Fe
 Content of trace elements
• Some of these trace elements have an
important role in the activity of enzymes
• Manganese is known to play an important
role in the development of lactic acid
bacteria
Trace elements
 Role of Cu and Fe
• Cu and Fe have a technological significance as catalyst
of oxidation reactions
• Cu is present in fat globule, where it can catalyze oxidation of
phospholipids
•Cu can enter the milk via dust, apparatus and water; this
Cu is distributed between membrane and serum.
• Technological processes influence the distribution of Cu ions:
– acidification causes a migration of Cu to the fat globule
membrane
– during cooling Cu migrates from the membrane to the plasma
– during heating Cu migrates to the membrane, but high
pasteurization hinders this transport; the distribution of Cu, during
decreaming, depends on the history of milk (heating, cooling)
Chemical and Physical
Properties of milk
Vitamins
 Vitamins
• Milk is a good source of diverse vitamins
• There is often a difference upon their
solubility in fat or water
 Fat Soluble Vitamins
• Vitamin A
• Vitamin D
• Vitamin E
• Vitamin K
 Vitamin A
• Vitamin A and its analogous are important for eye functioning
• β-carotene
– also called pro-vitamin A
– is also present in milk
– is responsible for the yellow color of butter
• Content
– whole milk : 0,4 mg per kg
– skimmed milk : 20 μg per kg
– butter: 7,5 mg per kg.
• During summer, milk contains 50% more vitamin A than
during winter months
• Pasteurization, sterilization and drying have nearly no
effect on vitamin A or β-carotene content
 Vitamin D
• Is important for the skeleton
• Content of milk: about 2 μg per kg
– is very dependent on seasonal variations
– summer milk contains 2 to 3 times more
vitamin D than winter milk
• Is very stable and its content isn't affected by
pasteurization or sterilization
 Vitamin E
• Vitamin E (tocopherol) deficiency causes sterility in male
and female animals
• Content of human milk is about ten times that of cow's milk
• The need for vitamin E increases as the amount of
unsaturated fatty acids in food increases
• Content
– milk: 1 mg per liter
– butter: 23 mg per kg
– principal content is higher during summer than during winter,
caused by the higher tocopherol content of fresh grass
• Heat or other processes don't cause a significant decrease
in vitamin E content
 Vitamin K
• Only in traces in milk, if at all
• Human needs for this vitamin are supplied
from consumption of plant materials
containing it and by microbial synthesis in
the digestive tract
 Water soluble Vitamins in Milk
• Vitamin B1
• Vitamin B2
• Niacin
• Vitamin B6
• Pantothenic acid
• Biotin
• Folic acid
• Vitamin B12
• Vitamin C
 Vitamin B1
• = Thiamine
• Is essential for growth and metabolism of all animals
but also of many plants and microorganisms
• In milk not only present in its free form, but also
phosphorylated (18-45%) and as a protein complex (5-
17%)
• Mean concentration in milk: about 0,43 mg per liter
• Daily requirement of an adult person: about 1,0 to 1,4
mg
• More or less destroyed by heat, dependent on time –
temperature combination
– loss in pasteurized milk and UHT milk: about 3-4%
– loss in sterilized milk: can mount up to 45%
 Vitamin B2
• = Riboflavin
• Very important in
– hydrogen transport
– carbohydrate and protein metabolism
– transformations of sugar and proteins into fatty acids
– eye-functioning
• Mostly present in its free form in cow's milk
• Content of whole, skimmed and butter milk : about 1,7
mg per liter
• Daily requirement of an adult person: 1,5 to 1,7 mg
• In the absence of oxygen very persistent during heat
treatment.
– the effect of pasteurization and sterilization is small
 Niacin
• Is a component of co-enzymes that are
involved in the citric acid cycle, metabolism of
fatty acids and the glycol acid cycle
• Content of milk: about 0,9 mg per liter
• In skimmed and butter milk, this content
decreases
• Daily requirement of a grown person: up to 13
to 18 mg
 Vitamin B6
• Is very important in protein metabolism
• Main content of milk: about 0,6 mg per liter
• Daily requirement: about 1 to 4 mg per day
• Pasteurization or homogenization give no
significant losses of vitamin B6 content
• Great losses are caused by sterilization (up
to 50%)
• Vitamin B6 is also very sensitive to light
 Pantothenic acid
• Is involved in the metabolism of carbohydrates,
fats and amino acids
• Mean content of milk: about 3,4 mg per liter
• Daily requirement of a grown person: about 10
to 15 mg
• Very stable during a number of processes (e.g.
manufacture of evaporated milk, sterilized milk
and milk powder)
 Biotin
• Is involved in carbohydrate and fatty acids metabolism
• Milk is a good source of biotin
• Mean content
– whole milk: 0,030 mg per liter
– skimmed milk: 0,016 mg per liter
– buttermilk: 0,011 mg per liter
• Daily requirement of an adult person is only 30 μg
• Light and heat have only a small effect
– loss in pasteurized and UHT: less than 10 %
– loss in sterilized milk: up to 10 to 15 %
 Folic acid
• Is necessary for the normal development of
red blood cells
• Few data about the contents in milk are
available
– in raw milk about 0,06 mg per kg
– this value is also reached in cheese
• Is very sensitive to heat, oxygen and light
– during sterilization, losses of 50 % can
occur
 Vitamin B12
• Is the only vitamin that contains a metal
(cobalt)
• Deficiencies of this vitamin can cause anemia
• Is only present in animal products and
vegetarians may have these deficiencies
• Mean content of milk: about 4,2 μg per liter
• Daily requirement of a grown person: only 1 μg
• Is sensitive to heat and oxygen
– in strongly heated products, the losses can
reach 100 %
 Vitamin C
• Is involved in the formation of intercellular material
(collagen)
• Content
– fresh raw milk: nearly 20 mg per liter
– skimmed milk: 16 mg per liter
– butter milk: 12 mg per liter
• Daily requirement of an adult person 55 to 60 mg
• Is very sensitive to light, heat and metals (Cu, Fe).
– during pasteurization 20 % is destroyed
– during evaporation 75 % is destroyed
– during sterilization 50 to 100 % is destroyed
Chemical & Physical
Properties of milk
Other Compounds
 Other compounds

• Organic acids
• Gasses
• Biological factors
 Organic acid

• Citric acid
– is very important for the stability of milk
– is also a substrate for microorganisms (used as starter
culture to make dairy products) and is transformed to
aromas
• Besides citric acid, also lactic acid, hippuric acid
(benzoic acid + glycocol) and orotinic acid (precursor
of nucleotides) are found
 Gases
• After milking 8 % (volume) gases, especially
CO2 (6%),are present
• During processing (heating) and storage, CO2
content decreases, while O2 and N2 content
increase
• Raw commercial milk contains at room
temperature about 1,3 % N2 and 0,5 % O2 by
volume, or about 15 and 6 mg.kg-1
 Biological Factors
• Bifidus factors
• Streptogenines
– peptides formed out of casein
– growth factors for lactic acid bacteria
• Lactenines
– active against lactic acid bacteria
– L1, L3 agglutinines
– L2 lactoperoxidase
Physical and Chemical Properties of
Milk

Physical Properties
 Physical Properties
• Density
• Dry matter
• Freezing point
• Acidity
• Redox potential
 Density
• Can be expressed as mass density or volumetric mass
and is designated as ρ
• Is temperature-dependent
• Recknagel effect
– with a change in temperature, a slow stabilization of
the density has to be considered
– density is dependent on the physical properties of fat
and the hydration of proteins can take time
• The density of milk
– at 20°C is on average about 1030 kg.m-3
– normally varies within the range of 1027 to 1033 kg.m-
3
 Density
• The density of milk
– is dependent on composition
– can be calculated from the density and mass fraction
of individual components
– at 20°C the densities of water, milk fat, protein, lactose
and other components are 998.2, 918, 1400, 1780
and 1850 respectively
• Fat content decreases the density of milk, while
solubilized components increase its density
– as a consequence, the density of skimmed milk is
higher than that of whole milk.
– addition of water causes a decrease of the milk density
 Dry matter
• Is in essence a chemical property
• Has a major influence on physical properties
• Its mean value in milk is 12,50 %
• Dry matter is determining weight loss during heating
• The dry matter can also be calculated from some
characteristics by the formula:
DM=1.23 F +2.6 *100 (ρ20-0.9982)/ ρ20
with ρ20 = volume mass at 20°C
D.M. = dry matter (%)
F = fat content (%)
• Fat-free dry matter (F.F.D.M.) is defined as
F.F.D.M. = D.M. - F
 Freezing point
• Is a colligative property that is determined by
the molarity of solutes rather than by the
percentage by weight or volume
• Is a constant that is used to determine addition
of water, which has a warmer freezing point as
a result
• Is determined by some components in solution:
lactose and salts
 Freezing point
• Freezing point determinations may be done by
the Horvet procedure
• For scaling, solutions of sugar or NaCl are used
– 7 % sucrose = -0,422°H (6,879 g NaCl diluted
to 1000 ml with water)
– 10 % sucrose = -0,621 °H (10,175 g NaCl
diluted to 1000 ml with water)
– Conversion formulas are given
m°H = 1,0356 m°C
m°C = 0,9656 m°H
 Freezing point
• The freezing point of milk is usually in the range of -
0,512 to -0,550°C with an average of -0,522°C
• If the freezing point of unwatered milk is known, the
relationship between added water and freezing point
depression is given by
W= (C-D)(100-S)/C
• with W = % (w/w) extraneous water in suspect milk
C = actual or reference freezing point of genuine milk
D = freezing point of suspect milk
S = the percentage (w/w) of total solids in the suspected
milk
 Freezing point
• Soured or fermented milk is not suitable for added
water testing
– because the freezing point is lowered by lactic acid
and increased concentrations of soluble minerals
• Several reports suggest that heat treatment of milk,
including UHT and retort sterilization causes little
permanent effect on freezing points
• It has also been suggested that freezing points are
not a reliable index for added water in processed milk
 Acidity
• An important quality parameter, but is also a characteristic
for fermentation processes
• Fresh milk normally has a pH of 6,6 to 6,8.
– is not the result of lactic acid, but a result of dry matter
• The acidity can be expressed in:
– D°(Dornic): number of 0,1 ml NaOH N/9 needed to neutralize
10 ml of milk
– N°(Normal grades): number of 0,1 ml NaOH 0,1N to neutralize
10 ml of milk
– S.H. (Soxhlet-Henkel): number of 0,1 ml NaOH 0,25N to
neutralize 10 ml of milk
– % lactic acid
• Earlier, triation of milk at reception was based on the acidity:
alizarol test, bromocresol test, alcohol test, ...
 Redox potential
• The redox potential of milk is in the range of +0.2 to +0.3 V
and is mainly determined by dissolved oxygen
• Milk is essentially oxygen free when secreted but about
0,3 mM O2 is present after equilibrium with air is established
• Removal of oxygen by nitrogen lowers the Eh of milk to
about -0,12 V
• Decreased oxygen tension by bacterial respiration is the
basis of the methylene blue reduction test for milk bacterial
quality
• The other redox systems of significance in milk are
ascorbate and riboflavin
 Redox potential
• Ascorbate in freshly drawn milk is all in the reduced
form, but oxidizes during refrigerated storage; this
process also produces singlet oxygen which is
involved in lipid oxidation
• Riboflavin is important for photooxidation reactions
in milk; methionine is transformed into methional,
which is the principal component of "sunlight" flavor
in milk
• Heat treatment is well known to increase the
reducing capacity, mainly due to activation of
protein thiol groups and products of Maillard
browning reactions
– activated thiol groups cause cooked flavor which
decreases as cysteine bonds reform on standing
Technology of Milk and Milk
Products
Primary Treatments
 Primary treatments
• Reception
• Centrifugation
• Standardization
• Homogenization
 Raw milk quality
• Fresh raw milk collected aseptically from the udder:
500-1000 bacteria / cm3
• Milk produced under normal hygienic conditions:
< 10.000 bacteria / cm3
• High counts are indicative of a breakdown in hygiene
– contamination of the cow's udder
– contamination from inadequately cleaned and
sterilized equipment
– the result of mastitic infection
• Raw milk should be maintained at less than 4 °C
– between milking and processing
– to prevent spoilage and pathogenic organisms
 Reception
• At delivery, a sample is taken per tanker
• After a quick control, the tanker is unloaded
• The milk is cooled over a plate heat exchanger and
pumped into storage tanks
• After each collection, the tanker is cleaned and disinfected
by a CIP (clean-in place) system
• In some cases the milk is collected via a collecting station
and transferred to the processing industry
• For a temporary stabilization, a thermisation (60-65°C, 10-
20s) can be applied
• Some regular tests that are performed include freezing
point depression, antibiotics, fat content
 Centrifugation
• Aim
• (1) to obtain skimmed milk and cream
these phases can afterwards be mixed together
to obtain the desired fat content
• (2) to remove the visible dirt
• (3) to remove bacterial spores (bactofugation)
 Spontaneous creaming
• Because of the difference in density between
– fat globules (df = 920 kg.m-3 at room temperature)
– plasma (dp = 1030 kg.m-3 at room temperature)
• Phenomenon we often want to prevent
• Is much enhanced when the globules have been
aggregated into floccules, clusters or granules
• Stokes Law:
V= 2r2 (dp- df )g/9η
v = creaming velocity (m/s); r = radius of fat globules (m)
dp = density plasma (kg/m3); df = density fat (kg/m3)
g = gravity acceleration (m/s2); η = viscosity (Pa.s)
 Spontaneous decreaming
• Example
• r = 1mm
• η = 1,79.10-3 Pa.s at 20 °C
• dp-df = 103,4 kg/m3 at 20 °C
v = 0,45 mm/h
 Centrifugal decreaming
• A considerable increase in the velocity of rising or
settling can be obtained in a centrifugal field
• The following formula can be used:
• 2r2 (dp- df )g*Z/9η
Z=Rn2/894

• Z = centrifugal constant
• R = radius of the orbital path (m)
• n = number of revolutions per minute (rpm)
• Same example of spontaneous creaming
• R = 0,12 m
• n = 6000 rpm
v = 0,60 mm/s
Operation of Centrifuge
 Operation of Centrifuge
• Cream separator
– bowl base and lid
– separating discs (conical)
– feed inlet and outlet
– sludge chamber
• Separators have up to 120 discs
– one above another
– angle of inclination is 45°to 60°
– outer diameter 200 to 300 mm
– made of stainless steel with a wall thickness of 0,4 mm
– space between the discs varies between 0,4 and 2 mm
– laminar flow through the narrow gaps ensuring that separation is
not adversely affected by turbulence
– larger gaps are necessary if there is danger of clogging
– holes forming a channel for the ascending liquids
 Types of Centrifuge
• Tubular bowl centrifuge
– long, narrow, cylindrical
bowl rotating at high speed
in an outer stationary
casing.
– The feed is introduced
through a stationary pipe to
the bottom of the bowl and
quickly accelerated to bowl
speed by means of vanes
– The two liquids are
removed from the annular
layers formed through a
circular weir system and
discharged into stationary
covers.
 Types of Centrifuge
• Disc-bowl centrifuge
– a relatively shallow, wide cylindrical bowl rotates at
moderate speed in a stationary casing
– the bowl is usually bottom driven
– the feed is normally introduced to the bottom of the
bowl through a centrally located feed pipe from above
– the bowl contains a number of closely spaced metal
cones, called discs, which rotate with the bowl and are
located one above the other with a fixed clearance
between them
– the discs have one or more sets of matching holes
which form
channels through which the feed material flows
– the separated liquids are removed by means of a weir
system
 Types of Centrifuge
• Semi open design
– The centrifugal force throws the milk outwards to form a ring
with a cylindrical inner surface.
– This is in contact with air at atmospheric pressure, which means
that the pressure of the milk at the surface is also atmospheric.
– The pressure increases progressively with increasing distance
from the axis of rotation to a maximum at the periphery of the
bowl.
– Cream moves inwards towards the axis of rotation and passes
through channels to the cream paring chamber (paring disks)
– The skim milk leaves the disc stack at the outer edge and
passes between the top disc and the bowl hood to the skim milk
paring chamber.
– The rims of the stationary paring discs dip into the rotating
columns of liquid, continuously paring out a certain amount.
The kinetic energy of the rotating liquid is converted into
pressure in the paring disc.
 Types of Centrifuge

Semi open design

Paring Discks
 Types of Centrifuge
• Hermetic separator
– The bowl of a hermetic
separator is completely filled
with milk during operation.
There is no air in the centre.
– The pressure generated by the
external product pump is
sufficient to overcome the flow
resistance through the
separator to the discharge
pump at the outlets for cream
and skimmilk.
– With this type of separator it is
possible to obtain a high
concentrated cream (>75%),
which is not the case with a
semi open separator.
 Types of Centrifuge
• Nozzle-discharge (self-cleaning) centrifuge
– = a self-opening centrifuge.
– is of the disc-bowl type but the bowl is usually biconal in shape
– a number of ports are spaced around the bowl usually at its
largest diameter
– these centrifuges are used to clarify milk from visible dust
 Process Parameters
• De-creaming of milk is largely dependent on
temperature
• Usually de-creaming is carried out at elevated
temperatures
– viscosity decreases
– difference dp-df increases
• The optimal decreaming temperature is 45 to
55°C
• Elevated temperature has also defects
– e.g. an increased risk of precipitation on the
apparatus
 Performance of a Separator
• Is judged by the residual fat content of the separated
milk or by the degree of cream separation from the
whole milk
• The degree of cream separation is defined as:
E= total fat in cream/total fat in the whole milk should
be as high as possible
Furthermore: mw = mc + ms
and: mw.fw = mc.fc + ms.fs
mw = mass of the whole milk fw = fat content of the
whole milk
mc = mass of the cream fc = fat content of the
cream
ms = mass of the separated milk fs = fat content of the
sep. milk
 Performance of a separator

• E therefore becomes:

• E=mc.fc / mw.fw= fc(fw-fs)/fw(fc-fs)

• As fs << fc, and fc ≈ fc-fs :
E=1-fs/fw
• E is determined by different factors:
– functioning of the separator
– temperature
– presence of gasses
– the season
– quality of the raw milk, especially the amount of free fatty acids
 Standardization
• Continue standardization can be achieved by means of
pipe and valve connections
• A part of the cream flow is led into the separated milk,
so that the desired fat content is obtained
• Mass balance equation:
mC.fC + mS.fS = (mC + mS).fM
mC : mass flow cream fC : fat content cream
mS : mass flow skimmed milk fS : fat content skimmed
milk
mM : mass flow stand. milk fM : fat content stand. milk
• After some transformations:
mc/ms= fM-fs/fc-fM
 Standardization
• Direct automated standardization of milk
– The pressure control system at the skim milk outlet maintains a
constant pressure, regardless of fluctuations in the pressure
drop over downstream equipment.
– The cream regulating system maintains a constant fat content
in the cream discharged from the separator by adjusting the
flow of cream discharged. This adjustment is independent of
variations in the
throughput or in the fat content of the incoming whole milk.
– Finally, the ratio controller mixes cream of constant fat content
with skim milk in the necessary proportions to give
standardized milk of a specified fat content.
– The standard deviation, based on repeatability, should be less
than 0.03% for milk and 0.2 – 0.3% for cream.
 Homogenization
• Is used principally to prevent or delay the formation of a
cream layer in full cream milk
• A reduction in the size of the fat globules
• As a result the stability of the emulsion is improved
• Raw and homogenized milk at 350 atm
 Principle
• Homogenization divides globules into smaller ones with
diameters down to < 1 µm, depending on the pressure
• Done by forcing all of the milk at high pressures through a
narrow slit, which is only slightly larger than the diameter of
the globules themselves
• The velocity in the narrowest slit can be 100 to 250 m/s
• This causes high shearing stress and microturbulence
• Fat globules become deformed, then become wavy and
then break up
 Principle of Homogenization
• Scheme of Homogenizer
 Homogenization Parameters
• Pressure is the most important parameter
• Pressure influence on homogenization degree :
log dva = const - 0,6 log P
dva = average diameter
P = homogenization pressure
• The temperature has different effects
– by lowering the viscosity, turbulence increases
– the fluidity of the fat plays an important role
• Normally, homogenization is executed at 50 to
70°C.
 Homogenization parameters
• The ratio fat to surface active material
determines also the degree of homogenization
– for the homogenization of whole milk, there is
normally enough protein present
– for fat-rich products, the amount of proteins is a
limiting factor; as a result, the fat globules can
aggregate to clusters
 Influence of Homogenization
• Homogenization causes a change in following
properties :
– fat globule diameter
– clustering is directly proportional to pressure and fat
content, and inversely proportional to fat-free dry
matter
– creaming is strongly reduced
– the thickness of the membrane increases with an
increasing homogenization pressure (± 15 nm in
homogenized milk)
– heat stability decreases with increasing
homogenization
 Effect on the product
• The fat globules are surrounded by a membrane
-5 to 10 nm thickness
-Is approximately composed of 1/3 phosphatides
(Phospholipids) and 2/3 proteins
-has emulsifying properties and keeps emulsion, milk
stable
-Phosphatides mainly lecithins form the inner part of
membrane
-The non polar groups of this monomolecular layer are
oriented inside
 Effect on product

• During homogenization, the original membrane is

destroyed
• The first result is rise in interfacial tension
• Surface active components form a new membrane by
adsorption and interfacial tension soon falls again
• The new emulsion therefore remains stable again
after homogenization
• The new membrane mainly consists of casein, the
proportion of phosphatides having decreased
Technology of Milk and Milk
Products
Heat Treatment
 Introduction
• Heat treatments: on a large scale in the dairy
industry
• Intermediate products and end products
• Destruction of all or part of the microorganisms in
milk
– pasteurization: destroys pathogenic microorganisms
and partly spoilage microorganisms, inactivates
some enzymes
– sterilization: destroys all microorganisms and
inactivates the majority of enzymes
• Optimized heating methods (e.g. UHT): to retain as
much as possible the good organoleptic and
nutritive properties
 Pasteurization
• Batch heating
– 62 to 65°C for 30 minutes
– rarely applied (in developed countries)
• HTST
– short time heating: 72 to 76°C for 15 to 40 seconds
– continuously in plate heat exchangers
• Flash heating
– high temperature heating: 85 to 90°C for 1 to 4 seco nds
– continuously in plate heat exchangers
– used for cream and yogurt milk
– peroxidase enzyme inactivated
• Ultra pasteurisation
– 125-138°C for 2s
– Extended shelf life products (ESL)
– 30-40 days shelf life
– Refrigeration remains necessary
Time/Temperature Combination Innactivation Curves
 Sterilization
• Destruction of the whole
microflora including spores
(except heat resistant spores,
e.g. B. stearothermophilus)
• Complete enzyme inactivation
(except some heat resistant
lipases and proteases derived
from Pseudomonas strains)
• 109 to 115°C for 40 to 20
minutes
• Side effects
– non-enzymatic browning
(Maillard)
– cooked or caramelized flavour
due to the decomposition of
lactose
 Ultra High Temperature Treatment

• Less discoloration and flavor changes compared

to classical sterilization
• 135 - 150 °C for 6 - 2 seconds
• Direct UHT
– steam injection followed by vacuum cooling
– less defects
• Indirect UHT
– heat exchangers
– more defects
Apparatus and process line
Plate Heat Exchanger
Apparatus and process line

Tubular Heat Exchanger

 Apparatus and process line

Scraped Heat Exchanger

 Apparatus and process line

Direct UHT with steam injection

Apparatus and process line

Indirect UHT
 Effect on product
• Proteins • Endogenous milk enzymes
– whey proteins • Bacterial enzymes
– caseins • Rennet coagulation
• Lactose • Acid coagulation
• Milkfat globules • Nutritional quality
• Milk salts • Sensory quality and flavor
 Whey Proteins
• β-lactoglobulin, α-lactalbumin, bovine serum
albumine (BSA) and immuno-globulins
• Well developed secondary and tertiary structures
• Susceptible to thermal denaturation: unfolding
– tends to enhance intermolecular interactions
– frequently leads to a loss of protein solubility
• α-lactalbumine:
– lowest denaturation temperature Td
– able to renature after moderate heating: β-lac
more affected
 Whey proteins
• Mildest heat treatments: thermisation and pasteurization
– no significant effect on the whey proteins
• More severe pasteurization (> 72°C and/or > 15 sec)
– may partly denature whey protein
– effect on further processing operations (e.g. increased rennet
coagulation time), or on sensory qualities of the finished product
(e.g. cooked flavour development in highly pasteurized milk)
• More severe heating conditions
– complex formation of β-lactoglobulin with k-casein
– reduced rennetability of heated milk
– applied in yogurt manufacturing process (90 °C, 5 minutes)
– incorporation of whey proteins in fresh cheeses
 Caseins
• Heterogeneous group phospho-proteins (80 % of
proteins)
• Four types: αs1-, αs2-, β-, k-cas (38%, 10%, 36%,
13%)
• Random coil proteins
• Casein in micellar form
– exceptionally heat stable
– withstands 20 minutes heating at 140 °C, pH 6,7
– heat stability of milk decreases abruptly at pH < 6,4
• Interaction between β-lactoglobulin and k-casein
– reaction between thiol groups
 Lactose
• Milk products: very sensitive to thermally
induced non-enzymatic sugar degradation
reactions
• Maillard browning reactions
– involves lactose and lysine rich milk proteins
– may significantly compromise the nutritional
value, especially in the case of sterilized milk:
• destruction of essential amino acids
• destruction of vitamins
• limitation of the bioavailability of other amino
acids
 Milk Fat globules
• Heat-induced changes
– chemical (reactions of fatty acid residues)
– physical (creaming, flocculation, coalescence,
disruption)
• Unheated natural milk fat globules
– quite stable against flocculation and coalescence
– cold agglutination (precipitation of immunoglobulins)
• The effect of heating
– inactivation of immunoglobulines (no cold
agglutination)
– homogenization: heat stability¯
– no changes in size distribution (except direct UHT-
heating)
– no coagulation of the fat globules
 Milk Salts
• Dynamic equilibrium shifts
– concentrations of Ca and P in the serum phase
decrease with increasing heating temperature
– lesser effects appear on Mg and citrate
– reversible when moderate heating temperature (70 -
80 °C)
• Irreversible changes
– irreversible breakdown of lactose to lactic and formic
acid: permanent decrease in pH after cooling
– dephosphorylation of casein: progressively more
important as heating temperature and time increase
 Endogenous Milk Enzymes

• The activity and stability of endogenous milk

enzymes
– over 60 in raw milk
– influenced by temperature, pH, thermal
conductivity and the availability of substrates,
activators or inhibitors
• Thermal inactivation of some milk enzymes is
taken as a control of heat treatments
– example: the universal alkaline phosphatase test
as an index of adequate pasteurization
 Bacterial enzymes
• Heat stable extracellular enzymes of many psychrotrophic
bacteria (Pseudomonas spp.), stable in UHT-range
– proteinases:
• may cause gelation of UHT-sterilized milks
• responsible for taste and textural changes
– lipases:
• influence taste: hydrolytic rancidity
– phospholipases:
• are responsible for bitterness of milk or cream
• Heat-stable enzyme secretion in cold-stored raw milk is
dependent on the initial microbial quality of the raw milk
(thermisation)
 Rennet coagulation
• Severe heating of cheese milk (> pasteurization
temperatures) impairs its renneting properties:
– β-lactoglobulin - k-casein complex: -S-S- bridge
– sterically hinders the aggregation of rennet-altered
micelles
– reversion (if not too severe conditions of heating): by
reducing the pH, adding CaCl2, or by acidification (to
pH 4,6) followed by re-neutralization to the original pH
of the milk
• The main reason for severely heating cheese milk:
– incorporation denatured whey proteins into the
cheese: yield increases
– flavour defects, too high moisture content, does not
melt
– not for ripened cheese
 Acid coagulability
• Milk acidification
– colloidal calcium phosphate (CCP)
progressively solubilizes
– aggregation of the casein occurs near the iso-
electric point (pH 4,6)
– acid casein production (syneresis)
– formation of a gel network with good water-
holding capacity
 Acid coagulation
• Milk heating prior to acidification:
– production of fresh cheeses, fermented milk products and
yogurt
– incorporation of whey proteins: influences the rheological
properties and mechanism of structure formation
– destruction of pathogenic organisms and phages
– reduction total number of microorganisms: increased shelf life
– improves the culture medium for the starter bacteria:
• formation of growth-promoting substances (such as formic
acid)
• lowering the redox potential
• inactivating antimicrobial substance
– improved nutritional status: better digestible denatured proteins
– possible destruction of some heat-labile vitamins
 Nutritional Quality
• Pasteurization:
– virtually no change in nutritional quality
– during storage: possible loss of light-sensitive vit A and
riboflavin
– nutritional quality of proteins and amino acids unaltered
• UHT treatment:
– possible loss of folacin, ascorbic acid, vitamin B12 and
thiamine
– during storage: folacin, vit B6, vit A and some vit B12 can be
lost
– nutritional quality of proteins and amino acids unaltered
• Sterilization:
– Maillard reactions (destruction of essential amino acids)
– considerable destruction of vitamins
 Sensorial quality
• Two major flavors characterize heat treated milk:
– cooked, caramelized flavors: during heat treatment
– stale and oxidized flavors: during storage
• Flavor of heat treated milk affected by:
– sterilized > indirect UHT > direct (injection) UHT
> direct (infusion) UHT ≈ pasteurized > raw milk
– milk quality
– packaging material
– storage time and temperature
– oxygen content during heat treatment and
subsequent storage
Milk and Milk Products

Fermented dairy products

 Fermented milk products
• Definition - description
• Mechanism of growth
• The manufacture of yogurt
– Ingredients
– Processing of yogurt
– Structural and chemical changes
• Other fermented milks
 Definition- description
• Fermented milk is the non-leaked product obtained by
coagulating skimmed milk, semi-skimmed milk or full fat milk
by inoculation with lactic acid bacteria, with or without the
association of yeasts
• Yogurt is a fermented milk, obtained by a simultaneous
operation of L. bulgaricus and S. thermophilus and in which the
two specific cultures remains active (= living) until consumption
o Dry matter on fat free product > 8.2%
o Yoghurt: min 3% fat
o Partially skimmed yoghurt: between 1 and 3% fat
o Skimmed yoghurt: <1% fat
- By fermentation a characteristic mild clean lactic flavour and
typical aroma is produced
 Definition-description
• Thermal treated fermented milk is fermented milk that is
submitted to heat treatment after fermentation to enhance
• shelf life, but by which the specific flora is destroyed
• Thermal treated yogurt doesn't exist
• Among fermented milk also churned (butter-) milk is sold
• Other examples of fermented milks
– bioyogurt
– acidophilus milk
– bifidus milk
– kefir
– koumiss
– sour cream
 Mechanism of growth
• The growth of yogurt bacteria shows some specific
aspects
• Both organisms can grow separately in milk, but
stimulate each others growth when brought together
(synergism)
• Symbiosis of yogurt bacteria
– S. thermophilus produces formic acid and CO2 that
stimulates L. bulgaricus
– L. bulgaricus produces low-molecular peptides and
amino acids that stimulate S. thermophilus
– The oxygen content is very critical, if it is higher than 4
mg/kg growth is delayed
– The formed lactic acid is important as inhibition factor;
the ratio bars to cocs is stabilized
 The manufacture of yogurt
• Ingredients
– Milk ingredients
– Yogurt sweeteners
– Yogurt fruit preparations
– Stabilizers and thickeners
– Flavours and colours
– Preservatives
– Vitamin and mineral fortifications
– Yogurt cultures
 Milk ingredients
• The main ingredient of yogurt is milk or ingredients derived
from milk
• By combining milk ingredients with different composition
with respect to milk solids, fat and protein and even protein
type and functionality, it is possible to tailor products with
specific desired properties and composition
• The most commonly used milk based ingredients are
– fresh whole milk
– fresh skimmed milk
– fresh cream (30-48% butterfat)
– skimmed milk concentrate (evaporated or UF)
– skimmed milk powder
– milk protein concentrates and isolates
 Yogurt sweetners
• The sweetness source is commonly sucrose,
either in powder or in liquid form
• Raw cane sugar or honey may be used as
well as de-colorized,de-flavorized, de-
acidified and deodorized fruit concentrates
• Low-calorific sweeteners are used in low-
calorie formulations and the favored one is
aspartame
 Yogurt fruit preparations
• Yogurt fruit is generally added to the cooled yogurt
after fermentation
• It is usually supplied as a pasteurized fruit
preparation to which, in addition to fruit, sugar,
stabilizers and thickeners, flavouring and colourings
have been added
• Traditionally a high brix 'jam', with a total sugar
content of around 60%, was used
• Nowadays a lower brix fruit preparation (30-50°) is
used
• Yogurt fruit preparation is typically added at a
dosage of 8-25% of the final recipe, with 10-15%
being most common
 Stabilizers & Thickners
• The most used stabilizers and thickeners are
– modified starches
– native starches
– gelatin
– pectin
– locust bean gum
• Sometimes used
– agar
– carrageenan
– guar gum
 Stabilizers & thickners
• Main functions
– they provide viscosity and body in the final product and
influence processing viscosity;
– they influence structure and texture;
– they control the degree of gelation required in stirred yogurt;
– they help prevent serum release;
– they influence creaminess and mouthfeel;
– they enable calorific reduction without loss of quality;
– they replace milk solids and fat and enable formulations to be
more cost effective.
• Typical inclusion rates of 0,05-1% have dramatic effects
on the properties of the final product
 Flavors and colors
• Common natural colours are
– betamin from beetroot
– anthocyanin from grape skin
– carmine from cochineal
– turmeric from the turmeric plant rhizome
 Preservatives
• When used, the common preservative in
yogurt is sorbic acid added as its potassium
salt.
• It has a very selective action against yeasts
and moulds
• Another preservative used in fruit preparations
is sulphur dioxide
 Vitamins and mineral fortification
• These are generally added to milk before
fermentation but some could be incorporated
via the fruit preparation
 Yogurt culture
• The culture organisms used to manufacture yogurt
are usually controlled by national legislation, but are
most commonly mixed cultures of Lactobacillus
bulgaricus and Streptococcus thermophilus
• As with cheese cultures phage attack can be a
problem if cultures are not rotated
• The main functions of the yogurt culture are generally
– to produce lactic acid to sour the milk from pH 6,5-6,7
to below pH4,6
– to produce the characteristic flavour of yogurt
– to produce a firm yogurt curd that will provide stability
and viscosity in the final product
 Yogurt culture
• Between 0,9-1,2% lactic acid is produced by the
breakdown of lactose; sucrose added to sweeten the final
yogurt is not metabolized
• The characteristic flavour of yogurt is due largely to
acetaldehyde produced mainly by L. bulgaricus
• Slime-producing or filant cultures, e.g. Streptococcus filant:
produce exopolysacharide
– are commonly used to give extra body and glossy appearance
to yogurt, particularly low fat formulations
– do however also give the product a somewhat long or ropy
texture often considered undesirable.
– main advantage is cost saving, enabling a lower milk solids
level to be used without loss of viscosity
 Yogurt culture
• In recent years bio-active cultures have a an
increasing popularity and are being used as a
marketing element
• These cultures contain species of L. acidophilus or
Bifido-bacterium to provide organisms that can
survive and grow in human intestines and contribute
to general health and well-being
• Several products claim to have a positive influence on
human's health, although nothing scientifically has
been proven yet
 Processing of yogurt
• Phases important during yogurt manufacturing
– increasing dry matter: to increase waterbinding properties, 1,5 to
2,5% milk powder is added
– heat treatment: after homogenization, a heat treatment of 5-10
min.at 85°C is applied, aim is to better the structure of the
product and to destroy natural inhibition factors, sufficient de-
aeration is also needed, especially with continuous heating
– fermentation is carried out in tanks and afterwards filled for
stirred yogurt, or fermentation is carried out in the package for
set yogurt
• for set yogurt the yogurt milk is after pretreatment cooled to 40-
45°C, ented with 2-3% culture and incubated in the recipient,
herefore incubation rooms or tunnels are used, after
fermentation to pH 4,5 the products are cooled
• for stirred yogurt a short acidification of 3 h at 45°C or a long
acidification of --15h at 31°C is applied, to enhance slime
formation tw o cultures are usually used,after incubation to pH
4,5 there is stirred, cooled and filled, the mechanical treatments
have an important influence on the structure of the yogurt
 standardize
Flow diagram: Yogurt d milk

Homogenize
55°C & 20 MPa

High Pasteurization
85°C for 5 min

Cool to 45°C Cool to

30-32°C

Innoculate Starter Starter Innoculate

2.5 % 0.025%

Packaging Incubate
16-20 Hours

Incubation Stirring
2.5 hours

Cool to 6 °C
Cool to 6°C Packaging

Cool to 6°C Packaging

set yogurt Stirred

yogurt
 Structural & chemical changes
• Yogurt shows a gel structure
• In the protein matrix the components are included, the
bacteria are bound to the protein by slime filaments
• During fermentation the following conversions take place
– Lactose is transformed for 20-30%; lactose is split into glucose and
galactose by the b-galactosidase of S. thermophilus and L.
bulgaricus
– Glucose is mainly homofermentatively transformed to pyruvic acid.
– Pyruvic acid is further transformed to lactic acid; L. bulgaricus
forms D-lactic acid and S. thermophilus forms L+lactic acid
– Pyruvic acid is further converted to small quantities of diacetyl and
formic acid, acetaldehyde is the main aroma component
– Proteins are coagulated and proteolysis is caused by L. bulgaricus.
Fats are not changed
– Vitamins are partly metabolized, partly synthesized
 Structural & chemical changes
• Yogurt is said to have an influence on intestinal flora
• It can inhibit tumors and gives a better resorption of
lactose
• Yogurt has a limited shelf life (storage temperature <
7°C) especially when infection with yeasts occurs;
acidification continue
• To improve shelf life, sometimes a heat treatment is
applied
– In function of the intensity apart from the yeasts and
moulds, also bacteria and enzymes are inactivated
 Other fermented milk
• Among the other fermented milks, churned
milk is the most important one
• Churned milk is obtained as a by-product
during churning of cream, during churning
of milk, during acidification of separated
milk with a butter culture
 Other fermented milk
• Fromage frais (fresh cheese) are cultured
products obtained by fermenting milk to pH 4.3 - 4.8 with
mixed mild cheese cultures, usually comprising
Streptococcus cremoris and Streptococcus lactis, and
sometimes Leuconostoc citrovorum and Streptococcus
diacetylactis
• This is followed by removal of whey and concentration of
the solids (by using a cheese cloth, centrifugation or
ultra filtration) to obtain a stiff, white to off-white,
spreadable paste that should be homogenous, short
textured and free from serum or graininess
• The products should have a clean lactic flavour and be
mildly acidic
Technology of Milk and Milk
Products
Fat rich dairy products
 Fat rich dairy products
• Cream
• Butter
• Butter oil and special products
 Cream
• Cream
= fraction that rises to the top of whole raw milk when left to stand
= o/w emulsion prepared out of milk by enrichment of the fat
content
• According to food legislation
– cream is the product with at least 20% fat
– whipping cream is the product with at least 40% fat
– thinned cream is the product with at least 4% at maximum 20%
fat
– Double cream contains at least 48% fat
– Clotted cream contains at least 55% fat
• An intermediate product for the manufacturing of butter
• Also for direct consumption after heat treatment
• (pasteurization, sterilization or UHT)
• Further distinction: coffee cream, double cream,
 Manufacturing cream
• Milk maintained < 5 °C until required for separation
• Milk heated for separation to 45-55 °C for reasons of
efficiency and product quality
– separation at temperatures < 40 °C will yield cream
with a high
viscosity, but may result in the development of off-
flavors due to lipolytic activity
– separation at temperatures > 55 °C will cause the
cream to thicken rapidly and excessively in storage
• Decreaming is very important for the profitability of a
factory
– fat losses in skimmed milk may maximally reach
0,05%
– decreaming is coupled to pasteurization
 Manufacturing of cream
• Usually a separate pasteurization is applied
– the milk is pre-warmed and centrifuged
– afterwards the two phases, skimmed milk and cream, are
separately pasteurized
– skimmed milk serves as pre-warming flow for the incoming
milk
– pasteurization of full milk followed by centrifugation is rarely
applied
• Desired fat content of the cream for further butter making
is 38 to 45%
• Cream is used for direct consumption after
– standardization
– thermal treatment
– mild homogenization (low pressure)
 Butter
• Definition
• Treatment of cream for butter making
– Pre-treatment of cream
– Cream ripening
• Butter production
– Churning process
– Structure of butter
– Production of butter
– Other processes
 Butter
• = a w/o emulsion mainly produced by churning of cream
• According to food legislation
– butter must contain at least 82% fat and at most 2% FFDM
– the salt content is limited to 1%
• Can be classified according to the type of processing
1. dairy butter out of unpasteurized cream farm butter out of
cream after spontaneous acidification
2. churned butter or sour butter (pH of serum < 5,5) continuous
butter or sweet butter (pH of serum > 5,5)
• Main consumer perceived benefits
– colour and appearance
– flavour
– texture and mouthfeel
– spreadability
– keeping quality
 Treatment of cream for butter
making

• Pre-treatment of cream
• Cream ripening
 Pre-treatment of cream
• Of great importance because the quality of the butter
depends almost entirely on the treatment of the cream
• The fat content aimed at depends on the further use
of the cream
• 25 - 35 % for churning in the butter churn
• 30 - 50 % for butter making by the continuous
process
• 36 - 40 % for ripened cream
• 38 - 42 % for sweet cream
• 82 % for butter making by the Alfa process
 Pretreatment ofcream
• Cream heat treatment
– high pasteurization: minimum claim is a negative phosphatase
test
– usually a higher heat treatment is applied: 95°C during 30-40
seconds or even 100-110°C during 15 seconds
formation of free SH-groups Antioxidative effect
inactivation of lipases and proteases
• Facultative treatments
– deaeration or degassing before pasteurization
to remove gasses and undesired flavours
the cream is brought at 75-85°C under vacuum
– desodorization
applied after pasteurization
carried out under vacuum at 95°C
 Cream ripening
• Covers a physical and a biological phase
• Physical cream ripening
• Biological cream ripening
 Physical cream ripening
• Aim = to obtain a directed crystallization
• Can as such influence the structure of the butter
• Is needed because fluid fat is difficult to churn and too many fat
losses can occur
• Soft milk fat (unsaturated - in summer) soft and greasy butter
• Hard milk fat (saturated – in winter) hard and stiff butter.
• Different temperature schemes can be followed
• Earlier the Swedish or the Alnaro procedure was followed
the temperature is adapted to the iodine value
T1 T2 T3
Sumer 19 16 8
Winter 8 19 16
• T1: crystallization temperature
• T2: temperature of primary ripening
• T3: temperature of second ripening
 Soft Winter butter
• When cooling quickly
– rapid formation of crystals.
– Triglycerides with low melting points are “trapped” in the same
crystals mixed crystals are formed.
– Low ratio of liquid to solid fat
– Hard butter
• This can be avoided
– The cream is heated carefully to a higher temperature
– The low-melting triglycerides are melted out of the crystals
– The melted fat is then re-crystallized at a slightly lower
temperature,
resulting in a higher proportion of “pure” crystals and a lower
proportion of mixed crystals.
A higher liquid-to-solids ratio and a softer fat.
 Biological cream ripening
• Was previously generally applied and is now limited
(sweet butter) or not applied at all
• Aim = an acidification and aroma formation
• Actual processes
– 3-5% starter culture is added
– during the warm period
• After the desired acidification, e.g. sour butter pH » 5,2,
cooling is applied to churning temperature
– T = 12°C during winter
– T = 9°C during summer
– or cooling is carried out to below 7°C to inhibit further
acidification
• The main aroma component diacetyl is only formed at pH
< 5,2 and so only occurs in aromatic butter
 Butter production
• There are four basic processes involved in the
manufacture of butter
1. concentration of the fat phase of milk
2. Crystallisation of the fat phase of milk
3. phase separation of the oil-in-water (o/w) emulsion
4. formation of a plastical water-in-oil (w/o) emulsion
• Churning process
• Structure of butter
• Production of butter
• Other processes
 Churning process
• Churning = intense mechanical movement + air inclusion
rapid motion of the fat globules in relation to each other
collisions with surfaces and high turbulence
fat crystals penetrate and disrupt the membranes
destruction of the 5 to 10 nm thick fat globule membrane
the emulsion is destabilized
liquid fat leaves the fat globules
coalescence to form fat agglomerates or butter granules
o/w emulsion of cream is transformed to w/o emulsion of butter
• Agglomeration is difficult
– at too low temperature when the proportion of liquid fat is too low
– at too high temperature when all the fat is in the liquid form
Milk and Milk Products

Concentrated and dried products

 Outlines
• Definitions
• Unsweetened condensed milk
• Sweetened condensed milk
• Other concentrated products
• Dried dairy products
• Dried dairy ingredients
 Definitions
• Concentrated and dried dairy products are milk products
with an extended shelf life
• Concentrated products: partial water removal
– no organisms survive due to
• Sterilization
• increased osmotic pressure
• Concentrated and dried milk products have several
advantages:
– Storage: requires small space under regular storage
conditions and retains high quality at the same time
– Economy: transport costs are reduced
– Use in emergencies: wars, epidemics, earthquakes
– Formulations: tailored food products, e.g. for sportsmen,
 Milk

Clarification
Sediments
Cooling 4°C
Unsweetened
condensed Storage 4°C
milk
First Standardization
Fat
Preheating (115-128°C for 1-6min)

Evaporation (45-70°C)
Water
Homogenization (P1: 15-25 MPa; P2: 5-
10 MPa
Stabilizers
Second standardization
Packages

Packaging

Sterilization (100-120°C for 15-20min or

140 °C for 3 Seconds)
Unsweetened concentrated milk

Storage @ 10°C
 Unsweetened condensed milk
• Based on evaporation
– partial removal of water from milk
– extends the shelf life by suppressing the
microorganisms present in the milk
• Milk quality for unsweetened condensed milk is
very important
– milk solids are concentrated
– product is planned for long storage
 Unsweetened condensed milk
• Preheating
– necessary for the stability of the condensed milk
during sterilization
– shortens the staying period of the milk in the
evaporator
– Produces a stabilizing effect caused by a decrease of
calcium and phosphorus during heating
• The preheating is carried out in continual heat
exchangers of plate or tubular type
– 93 to 100°C for 10 to 25 min.
– 115 to 128°C for 1 to 6 min.
 Unsweetened condensed milk
• Evaporation
• Evaporation is carried out under partial vacuum
– Reduction of the evaporation temperature
– Temperatures used are never below 45°C, in order to
eliminate the growth of staphylococci
– Tubular and plate evaporators
– Single-effect or multiple-effect of two or more units up
to eight
– The falling film tubular evaporator
• The leading evaporator in dairy industry
• Tubes are about 3 to 5 cm in diameter and 15 m long
• The tubes are heated with steam.
 Unsweetened condensed milk
• Homogenization
– Improvement of the stability of milk fat emulsion
– Decrease of the average diameter of milk fat globules
• Second standardization
– Adjustment of the ratio of milk fat to nonfat milk solids
(if the first standardization was not conducted)
– Standardization of the total dry matter is standardized
(if the first standardization has been carried out).
• Heat stability depends on its salt balance
– Addition of stabilizing salts
• calcium, potassium or sodium carbonates and
bicarbonates
• potassium or sodium citrates
• phosphates and other salts
 Unsweetened condensed milk
• Packaging
– Usually packaged in cans of various sizes
depending on use
• Sterilization
– Sterilized (100-120°C for 15-20 min.)
– Continuous flow sterilization of evaporated milk is
common (130- 140°C), followed by packaging under
aseptic conditions.
• Evaporated milk can successfully be stored up to a
year without any significant quality change at
temperatures of 6-8°C.
 Sweetened condensed milk
• Part of the water from fresh milk is evaporated
• Sugar is added to the concentrated milk in
order to extend shelf life
• By increasing the osmotic pressure, the
growth of microorganisms is prevented
 Sweetened condensed milk
• hydrolysis of lactose
– Milk is cooled to 5-10°C after pasteurization
– Lactose is hydrolyzed by β-galactosidase,
obtained from Saccharomyces fragilis.
– The sweetness of the final product is
approximately the same
– In order to prevent lactose crystallization, acid-
hydrolyzed sugar syrup may be added
 Heat treatment, evaporation, sugar addition
and standardization
• Main goal of heat treatment:
– Total inactivation of osmophilic microorganisms
– Inactivation of enzymes, particularly lipase and proteases
– Decrease of fat separation
– Inhibition of oxidative changes
– Influence on the viscosity of the final product
– The most frequently applied temperatures are 100 to 120°C.
• Age thickening
– A consequence of physicochemical changes in casein
• Sugar addition is the way to prolong shelf life of this product
– Sucrose, glucose, dextrose and others could be applied
– Addition of sucrose before heat treatment increases the thermo-
resistance of bacteria and their enzymes and significantly
intensifies age thickening during storage.
• During second standardization, total solids, sugar and fat
contents are controlled
 Cooling with crystallization
• During cooling of the product after evaporation and
sugar addition, lactose crystallization is induced
• This is caused by:
– Temperature decrease
– High lactose concentration
– Presence of high concentrations of added sugar
– Relatively small amount of water
• To avoid formation of crystals larger than 15 mm
(sandiness), inoculation with powdered lactose crystals
are used and the process is completed with rapid
cooling and simultaneous agitation
Milk and Milk Products

Dried dairy products

 Milk powder
• Milk powder
– =Dairy products from which the water has been
removed to the greatest extent possible which
prevents the growth of microorganisms
• Skimmed Milk Powder
– SMP produced by a 'low heat method' is simply
pasteurized
– SMP produced by a 'high heat method' requires
heating at 85 to
88°C for 15 to 30 min. in addition to pasteurization.
– Intensive heat treatment is applied to powder to be
used in the bakery industry, where a degree of milk
protein denaturation is desirable.
• Standardization is needed to adjust the ratio of milk fat
to total solids as required in the final product.
 Milk

Clarification
Sediments
Cooling (4°C)

Standardization

Fat

Heat treatment
88-90°C for 3-5min
or 130°C for several seconds

Evaporation until 30-35% TS;

40-50% TS
Water
Homogenization (5-15 MPa)

Drying
130-150°C; 180-240°C
Water

Milk powder
Packages
Packaging

Storage @ 20°C

Flow diagram of processing milk powder

 Heat treatment
– Carried out at temperatures higher than those required for
pasteurization
– Destroys all pathogenic and most of the non-pathogenic
microorganisms
– Inactivates enzymes
– Activates the reducing SH groups of β-lactoglobulin, increasing
the resistance of the powder to oxidative changes during
storage
• Evaporation
– For roller drying evaporation is performed to 33-35% total solids
– For spray drying is performed to 40-50% total solids
• Homogenization
– not obligatory
– usually applied in order to decrease the free fat content
– Fat globules depleted of protective membranes reduce milk
powder solubility and increase their susceptibility to oxidative
rancidity
 Drying

• Milk is most commonly dried by

roller drying or spray
drying in a stream of hot air:
• Roller drying
– Roller drying at atmospheric
pressure
– Vacuum roller drying
• Spray drying
– Centrifugal atomization
– Pressure atomization
– Foam spray drying
– Steam swept wheel atomization
– Venturi spraying
 Roller drying
• Roller drying
– Commonly used in the production of skim milk powder and
whole milk powder which find applications in other food
industries (confectionery, feed blends) because of low product
solubility
– Direct contact of a layer of concentrated milk with the hot
surface of rotating rollers adversely affects the milk
components and causes irreversible changes.
– Caramelization, Maillard's reaction and lactose degradation
occurs
• Vacuum roller drying
– Operating temperatures below 100°C
– Eliminates an oxygen effect
 Spray drying
• Spray drying
– Mainly used for drying milk and milk products
– Evaporated milk is atomized into fine droplets and
exposed to a hot air flow in a spray-drying chamber,
which may be in horizontal or vertical position
– Air is filtered and heated up to 150-300°C
– Air is usually indirect heated with steam or oil
• Relation of milk flow and air stream
– Concurrent flow
– Countercurrent flow
– Mixed flow
• Concurrent flow
• In spite of the inferior heat economy, is preferred in the
dairy industry, as it improves product quality
 Atomizers
• Basic function
– To provide a high surface-to-mass ratio, enabling quick heat
transfer with a high evaporation rate
– The two atomizing designs:
• centrifugal (rotary) atomizer
• pressure (nozzle) atomizer
• Powder particles gain spherical shapes during drying, with
trapped air, thus gaining a low bulk density
• Atomization parameters affect the properties of the
product:
– Bulk density
– Size and size distribution of powder particles
– Incorporated air content
– Moisture content
– Others
 Atomizers
A: Atomizing disc with curved channels B. Single
component nozzle C. Dual component nozzle
Standardization process
 Standardization
• 100 kg of milk with 4% fat are centrifuged
into 90.1 skimmed milk with 0.05 % fat and
9.9 kg cream with 40% fat. How much of
this cream will be added to the skimmed
milk to get the standardized milk with 3%
fat. How much standardized milk will be
obtained ? How much will the
excess/surplus cream be?
 Solution
• Mass balance:
• Skimmed milk + cream= Standardized milk
• Lets X and Y be cream and standardized
milk, respectively
• 90.1+ X=Y (1)
• Fat balance:
• 0.0005*90.1+ 0.4X= 0.03Y(2)
• Combining (1) and (2)
 Solution
• 0.0005*90.1+ 0.4X= 0.03 (90.1+ X)
• 0.04505+ 0.4X=2.703+0.03X
• 0.37X= 2.65795
• X= 7.18= 7.2 kg
• Standardized milk=
• 90.1 kg+ 7.2 kg= 97.3 kg with 3 % fat
• Cream in surplus= 9.9 kg - 7.2 kg= 2.7 kg
 Question 2
• How much skim milk containing 0.1 % fat
is needed to reduce the fat of 200 kg of
cream from 34% to 30% ? How much
standardized cream will be obtained ?