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ALTERNATIVE WAYS TO PROCESSING WHEY
Alternative Ways to Processing Whey
Alok Chatterjee, S. K. Ka na wjia a nd Yogesh Khetra
Cheese and Fermented Foods Lab,
Dairy Technology Division, National dairy Research Institute
Karnal-132001, Haryana
Introduction
Yeste ryears have witnessed significant potential of
science in resurre cting whe y. Not too long back,
whe y; the greenish colore d liquid discharge from
manufacturing of various mainstream dairy products;
was a potential threat to the e cology and problem to
the industrial dispense be cause be ing high on
biological ox yge n demand. But with science and
te chnology inte rve ning, the bane turned itself into
boon
with
te chnofunctional and biofunctional
abilities. Give n the pe rsistent increase in demand of
the products; i.e . chee se in prime; resulting in rising
volum inous gene ration of whey, more growth in
utilization of it is not only expe cte d but ine vitable
too. Whe y processing and application today are
yielding a wealth of quality products that are
increasingly seen as ingre die nts in formulations that
have re cognized positive health benefits (Onwulata.
2008). Due to the presence of scarce and
indispensable am ino acids (lysine, tryptophane,
me thionine , threonine and cyste in), whe y staffs itse lf
as an ex tremely important by-product of dairy
produce s (Kondratovych et al., 2013). Qualitative
composition of amino acids present in whe y have
confe rred various health be nefits and are pouring in
reports and studies in scie ntific community.
Innovative processing te chniques are ousting olde r
me thods, yie lding products with be tte r applica bility.
Bypassing of cheese -making step is possible for
producing separate prote in fractions from skim m ilk
by use of corre ct pore sizes in microfilte rs .
High Hydrostatic Pressure Processing
According to Pascal’s law, pre ssure acts in stantly,
isostatically and homogenously, independe nt to the
size and shape of the mate rial. High pre ssure
processing of m ilk, on microflora, appeared in
th
lite rature in 19 ce ntury (Hite, 1899). The reafte r,
the medie val years did not witness much of high
pre ssure processing of m ilk . Afte r 1903, the reports
of high pressure processing started appearing at
freque nt inte rvals and it was only in the latte r half of
20th century whe n studies on this field started
be com ing freque nt. The main reason of the long
inte rm ission in the fie ld of high pressure investigation
was the lack of appropriate e quipment (Huszar,
2008). High-pre ssure processing has been shown to
have
marginal
influence
on
the
nutritional
characte ristics of milk , hydrolysis, or stability of
vitamins. Se ve ral publications support β-Lg be ing
more sensitive to pre ssure ove r α-La. Forfe iture in
solubility at pH 4.6 explains de naturation of whe y
proteins. W ith HP me thod α -La was de natured at
pre ssure s highe r than 400 MPa, and β -Lg at
pre ssure s highe r than 100 MPa. Given that the re are
four intra -mole cular disulphide bonds, the supe rior
barostability of α -La is ex plaine d ove r β -Lg tat has
only two. Also, β-Lg was found to de nature more by
HP than α-La in milk , but less so in whe y. Howe ve r,
removal of colloida l calcium phosphate from m ilk
ne gative ly impacted HP -induced de naturation of α -La
and β-Lg. Huppe rtz et al., (2005) explained
de naturation of β -Lg and α-La at pressures greate r
than 100 MPa, with increase in the ir association
within the se rum phase milk fa t globule membrane.
Re ve rsible e ffe cts have been see n on β-Lg for
pre ssure s up to 300 MPa and non -occurence of
Maillard browning for pressures up to 600 MPa.
Digestibility remains unaffe cted due to de naturation
of prote ins resulting from HP proce ssing (Mes sens et
al., 2003). Though, at highe r pressures (400-800
MPa), occurrence
of
relative ly
little
furthe r
de naturation has been given by Scollard (2000), the
ex tent of HP-induced denaturation of α -La and β-Lg
has bee n shown to increase with increase in holding
time , tempe rature , and pH of milk by se ve ral
researche rs (Huszar, 2008). Unfolding of β-Lg due to
HP processing exposes its free sulphydril group.
Felipe et al., (1997) suggested formation of small
aggregates of de natured β -Lg during HP treatment of
milk whe reas worke rs like Nee ds et al., (2000) and
Scollard et al., (2000) sugge ste d inte raction with
case in m ice lles. Adding to the suggestion by Fe lipe et
al., (1997), Dumay et al., (1994) and Van Camp et
al., (1997) suggeste d partial re ve rsibility of
aggregated β-Lg, on subseque nt storage . In HP
treating of whole m ilk may result in association of
some α-La and β-Lg with the m ilk fat globule
membrane (Ye et al., 2004). Discussed nex t is the
me chanism for high pre ssure induced denaturation of
α-La and β-Lg in m ilk as we ll as in whe y as given by
Huppe rtz, 2006 and produced by (Huszar, 2008): βLg unfolds unde r high pressure , which results in the
exposure of the free sulphydryl group in β-Lg. This
free sulphydryl-group can inte ract with othe r m ilk
proteins (κ -case in, α-La or β-Lg, and pe rhaps αs 2-
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ALTERNATIVE WAYS TO PROCESSING WHEY
case in), through sulphydryl-disulphide inte rchange
reactions. On release of pressure , unfolde d α -La and
β-Lg mole cules, that have not inte racte d with
anothe r protein, may re fold to a state closely re lated
to that of native fo rm of these prote ins. The close
structural sim ilarity of monome ric untreated, and
HHP treated β-Lg indicates that the sulphydryldisulphide inte rchange reactions occur during HHP
treatment, since the free sulphydryl-group of β-Lg is
not available for inte raction afte r high pre ssure
treatment. Diffe rent pressure stability of isoforms A
and B of β-Lg we re sugge sted by Botelho et al.,
(2000) with B form be ing more sensitive to pre ssure
than A. The possible explanation was rested on the
ex istence of of a core cavity in β-Lg B.
Ultrasound Processing
As give n by Mason et al., (2005), the major
me chanical effe cts of ultrasound are provided when
the powe r is sufficie ntly high to cause cavitation.
Sim ilar to any sound wave , ultrasound too
propagates via a compression and rare faction wave
se ries induced in the mole cule s of the passage
medium. Whe n the powe r is sufficie ntly high, the
attractive forces of the mole cules of the liquid are
ex ceeded by rare faction cycle, thus form ing
cavitation bubbles. These bubbles continue to grow
and this phenomenon; known as re ctifie d diffusion, is
explained by e nte ring of small amounts of gas or
vapor from the medium in the bubble during its
expansion phase and incomple te expulsion during
compre ssion. The bubbles that are distributed
throughout the liquid grow ove r the pe riod of a fe w
cycles to an equilibrium size for the particular
freque ncy applied. If the bubble s we re only subje ct
to that particular fre quency the y would remain as
oscillating bubble s, howe ve r, the acoustic field that
influences an individual bubble among the many
thousands ge ne rated in a cavitating fluid is not
uniform . Each bubble will slightly affe ct the localized
fie ld expe rience d by neighbouring bubbles. Unde r
such circumstance s the irregular fie ld will cause the
ca vitation bubble to be come unstable and collapse. It
is this collapse that gene rate s the ene rgy for
chemical and me chanical effe cts. For example , in
aqueous systems at an ultrasonic frequency of 20
kHz, each cavitation bubble collapse acts as a
localized ‘hotspot’ ge ne rating tempe ratures of about
4000 K and pressures in ex cess of 1000
atmosphe res. This bubble collapse , distributed
through the medium, has a varie ty of effe cts within
the system de pending upon the type of mate rial
involved. Sonication, has been applied to m ilk
products for varying applications. Ultrasound was
used to break bacte rial “clump” which was mask ing
total bacte rial count in m ilk. Huhtane n, (1966)
suggested sonication at ele vated tempe ratures
im proved the de sired iso lation of bacte ria in raw
milk . The treatment of milk with low fre quency
sonication increased the total bacte rial counts, but
the heat produced by ultrasonic treatment did not
account entire ly for its effe ct. In presence of heat,
the syne rgistic effe ct of processing on prote in
increases the denaturation of whe y proteins α-La and
β-Lg; but case ins remain unaffe cte d as the highest
tempe ratures reporte d are below 76ºC . Ele ctrical and
sonic forces have been shown to modify the filtration
pe rformance of membrane filtration of whe y.
Reduction
in
membrane
fouling
and
thus
enhancement in flux, have bee n demonstrated by
Muralidhara et al., (1986), Tarle ton et al., (1992)
and Tarle ton (1988) by both e le ctric and ultrasonic
fie lds. The y obse rved a syne rgistic e ffe ct when both
the fie lds we re applied simultaneously. Effe ctive use
of stand-alone ultrasound has been demonstrated by
various othe r re searche rs to enhance the pe rmeate
flux (Kobayashi et al., 2003; Lamminen et al., 2004).
Muthuk umaran et al., (2005a) showed restoration of
initial
pe rmeate
fluxes;
during
membrane
processing; with the assistance of ultrasound. The y
also concluded that ultrasound does not damage the
membrane surface or increase the pore size of the
membranes (Lamminen et al., 2004). Their ye t
anothe r work re veale d the e ffe ctiveness of ultrasonic
enhancement by use of space rs irre spe ctive of flow
current concluding the possibility of doubling of
pe rmeate flux with the combined e ffe ct of space rs
and ultrasound. Increase in the acoustic stream ing
and me chanical vibration, we re utilized by them for
resting the main mechanisms involve d in flux
enhancement. Howe ve r, the influe nce of acoustic
cavitation cannot be comple te ly ex cluded. The
ultrasonic irradiation acts to reduce the resistance of
both the initial prote in deposit and the growing cake,
reducing the compressibility of the se deposits. The
mass transfe r coefficie nt within the concentration
polarization laye r also increases. The low powe r
le vels for sonication also imply that damage to the
membrane surface itself can be minim ized and
indeed. (Kondratovych et al., 2013) showed whe y
ultrasonic treatment as an e ffe ctive method of
product disinfe ction, allowing the doubling of its
storage time . The y described the kine tics of
microorganism de struction in whe y is by first orde r
k ine tic equation and concluded that ultrasound
treatment of whe y destructs the polyme rs, increases
the numbe r of amino and carbox yl groups due to the
protein hydrolysis and does not causes e vident
ox idation of organic compounds. In their work, the
polypeptides mole cular mass de crease d from 17.10 3
to 5.10 3 g/mol unde r ultrasound treatment.
Membrane Processing
The membrane te chnology is a novel, non-the rmal,
environmental frie ndly te chnology with a minimum
adve rse effe ct on product. The “membrane filtration”
is a separation process whe re spe cific sem ipe rmeable membrane filte rs are used to concentrate
or fractionate a liquid (Winston and Sirkar, 1992) by
se le ctive pe rmeation of some compounds through
membrane and re taining the othe rs. The liquid that is
able to pass the membrane is known as “pe rmeates”
and the re taine d liquid is known as“re tentate ” or
“conce ntrate”. The hydrostatic pre ssure gradients or
the trans-membrane pressure across the membrane
and concentration gradient of the liquids de te rm ine
the e fficacy of membrane . O ccasionally, membrane
efficacy is also affe cted by e le ctric potential (Winston
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Dairy Year Book (2014-15)
ALTERNATIVE WAYS TO PROCESSING WHEY
and Sirkar, 1992). W ide ly used membrane separation
components, and ce llular re ce ptors through its highly
te chnologies are m icro -filtration, ultra -filtration
positive ly charged N-te rm inus (Nuijens et al., 2005).
(Balanne c et al., 2005), nano -filtration (Vourch e t al.,
Modulation in immune functions is brought about by
2005) and re ve rse osmosis (Balanne c et al., 2005;
imm unoglobulins, and the y act as antibacte rial
Vourch et al., 2005). MF membranes are ge ne rally
age nts. Inhibition of bacte rial growth by catalyzing
used to separate fine particle s of 0.1 to 10.0µm size
thiocynates and othe r halides and by deple tion of
whe reas, in UF membranes with 1 to 100 nm pore
hydrogen
pe roxides
is
carried
out
by
size , prote ins and othe r macromole cule s are re tained
lactope roxidases. GMP and bovine se rum albumin
and (Van and Zydne y, 2007) wate r and low
(BSA) are available am ino acids (Ha and Zeme l,
mole cular we ight
solutes pass
through
the
2003).
membrane . The mole cular cut-off of UF is 10,000 MW
Future Possibilities
and ope rating at 40 psig, and at 50 to 60ºC with
polysulfone membranes. Nano-filtration has a Whe y proteins and components of whe y along with
membrane of slightly smalle r pore size than UF. It
modified forms foste r use ful nutritional and othe r
re tains the divalent ions. Re ve rse osmosis membrane
supplements with health mainte nance and healing
be ing more pressing . Contemplation fore runs that
pore size allows only small amounts of ve ry low
mole cular we ight solute s to pass through the
advanced processes for purifying and modifying whe y
membranes and is used mainly for concentration.
products upon de ve lopment can potentially increase
the nume rals of products that can be made. The y can
Diafiltration (DF) is anothe r spe cialize d type of
ultrafiltration process in which de ionized wate r is
smoothly fit into ne w products such as be ve rages,
adde d in the re tentate continuously to reduce the
confe ctione ry items (e .g., candies), convenience
foods, desse rts, baked goods, sauce s, infant food
concentration of lactose and m ine ral in re te ntate with
an increase in the concentration of re tained
and formulae, ge riatric foods, animal feeds, and as
components. Membrane filtration has provide d a
drug constitue nts, and plastics.
wide
scope
for whe y processing such as
Referenc
es:
concentration, fractionation, purification e tc.
Balanne c, B., Vourch, M., Rabille r-Baudry, M. and
Modification of Whey Proteins for Health
Chaufe r, B. (2005). Comparative study of diffe rent
Benefits
nanofiltration and re ve rse osmosis membrane s for
dairy effluent treatment by dead -end filtration. Sep
Past two de cades have witnessed steady growth in the
Purif Te chnol., 42: 195–200.
studie s relate d to the physiological be nefits from
spe cifically de fragmente d am ino acid se quence s,
Bothe lo, M. M., Valente -Mesquita, V. L., Olive ira, K.
be tte r known as Biologically Active Peptide s. The
M. G., Polikarpov, I. and Fe rre ira, S. T. (2000).
te rm Peptidome have often been associated with
Pressure denaturation of β-Lg. Diffe rent stabilities of
such k ind of studies. The se seque nces or peptides
isoforms A and 104 B, and an investigation of the
remain encrypted within the parent protein chain and
Tanford transition. European Journal of Biochemistry,
are re leased by processes as enzymatic/ acid/ alkali
267: 2235-2241.
hydrolysis or m icrobial action. The re leased peptides
Dumay, E. M., Kaliche vsk y, M. T. and C hefte l, J. C.
have been confirmed to confe r various health
(1994). High pressure unfolding and aggregation of
be nefits as be ing hypotensive , immunomodulating,
be ta -lactoglobulin and the baroprote ctive e ffe cts of
antiox idative, anticarcinogenic, m ine ral binding, etc.
sucrose . Journal of Agricultural and Food Chemistry,
In vitro, in vivo and human trials as we ll have been
42: 1861–1868.
conducte d with positive results. Biologically active
Felipe , X., Cape llas, M. and Law, A. J. R . (1997)
fractions of
whey; β-Lg,
α-La,
lactofe rrins,
Comparison of the e ffe cts of high -pre ssure treatment
glycomacropeptides, and immunoglobulins; function
and heat pasteurization on the whe y prote ins in
as
antiox idants,
antihype rte nsive,
antitumor,
goat’s milk . Journal of Agricultural and Food
hypolidem ic, antiviral, antibacte rial, and che lating
Chem istry, 45: 627-631.
age nts. β-Lg carrie s small hydrophobic mole cules and
re tinoic acid
modulating
lymphatic
re sponse
(Marshall,
2004).
Depression
in
body
fat
accumulation, acce le ration of loss in we ight and fat
and satiation enhancement has been shown by whe y
proteins. Calcium mediates the me chanism of
accomplishment of adiposity function by whe y
protein. (Ha and Zemel, 2003). Korhonen, (2002)
reported success in de riving anti-hype rtensive
peptide s from whe y prote in hydrosylates. It is
thought also that the iron binding capacity of
lactofe rrins reduces oxidative damage cause d by
unbound iron in tissues (Marshall, 2004). Apart
biologically active peptides, distinctive constituents of
whe y proteins such as Ig, Lf, GMP & LP improve
shie ld against cellular ox idative stresses the reby
be nefiting
cardiovascular
system.
Lactofe rrins
sequeste r iron, inte ract with m icrobial cell wall
Ha, E., and Zeme l, M. B. (2003). Functional
prope rties of whe y, whe y components, and essential
amino acids: Me chanisms unde rlying health benefits
for active people (Re vie w). J. Nutr. Biochem. 14:
251–258.
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disaggregation of m ilk bacte ria. Journal of Dairy
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L. (2005). High pressure-induced changes in bovine
milk proteins: a re vie w. Biochim. Biophys. Acta
1764(3):593–598.
Huszar. K. P. (2008). Protein changes of various
types of milk as affe cted by high hydrostatic pre ssure
processing. Ph.D. thesis subm itted to Faculty of Food
158
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ALTERNATIVE WAYS TO PROCESSING WHEY
Science , Department of Re frige ration and Livestock
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Kobayashi, T., Kobayashi, T., Hosaka, Y. and Fujii, N.
(2003). Ultrasound enhanced membrane -cleaning
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41(3): 185–190.
Kondratovych, O ., Koval, I., Kysle nko, V., She vchuk,
L., Predzumirska, L. and Maksymiv, N. (2013). Whe y
disinfe ction and its prope rties changed unde r
ultrasonic
treatment.
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&
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Te chnology, 7(2):185-190.
Korhonen, H. (2002). Te chnology options for ne w
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Kumar, P., Sharma, N., Ranjan, R ., Kumar, S., Bhat,
Z. F. and Jeong, D. K. (2013). Perspe ctive of
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Lamminen, M. O., Walke r, H. W. and Weave rs, L. K.
(2004). Me chanisms and factors influencing the
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ce ramic
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P. M. and Kelly, A. L. (2000). Plasm in activity, βlactoglobulin denaturation and proteolysis in high
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425–432.
Van-camp, J., Messe ns, W ., Clement, J., and
Huyghe bae rt, A. (1997). Influence of pH and calcium
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Vourch, M., Balanne c, B., Chaufe r, B. and Dorange,
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Ashokk umar, M., Mawson, R., Ste vens, G. W. and
Griese r, F. (2005a). The optimisation of ultrasonic
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Consumers’ Acceptance For
Buffalo Milk
Muthuk umaran, S., Kentish, S. E., Ashokkumar, M.
and G. W . (2005b). Me chanisms for the ultrasonic
enhancement of dairy whe y ultrafiltration. Journal of
Membrane Scie nce, 258: 106–114.
Nee ds, E. C ., Cape llas, M., Bland, A. P., Manoj, P.,
Macdougal, D. and Paul, G. (2000). Comparison of
heat and pressure treatments of skimmed milk,
fortified with whe y protein concentrate, for se t
yoghurt preparation: effe cts on milk prote ins and gel
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Nuijens, J. H., van-Be rke l, P. H. C . and Schanbache r,
F. L. (2005). Structure and biological actions of
lactofe rrins. J. Mammary Gland Biol. Neoplasia. 1(3):
285–295.
The Indian population has a great liking
for buffalo milk, which forms a thick cream layer
(malai). This layer thickens further after boiling
and storage. The high viscosity of buffalo milk
exerts an additive influence on the consumer’s
preference.
It is known to impart a distinct
whitening effect to tea and coffee because of higher
quantity of whey proteins and casein. Boiling of
buffalo milk causes the release of high amounts of
sulphydryl compounds, which contribute to nutty,
cooked flavour leading to its high acceptance as a
drink. Full cream buffalo milk is sold at premium
price because of its flavour and its ability to
produce
good
quality
products
(Rajorhia,
2000).bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
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