Composition, Physicochemical and Functional

Properties of Reference Whey Protein Concentrates

C. V. MORR

ABSTRACT 50°C and spray dried in an Anhydro spray drier using 205°C inlet and
92°C outlet air temperature conditions.
Four reference whey protein concentrates (WPCs) were prepared from Cheddar cheese WPCs were prepared from commercial Cheddar
pasteurized and nonpasteurized acid casein whey and Cheddar cheese cheese whey. The whey was prepared from 72°C - 15 set pasteurized
whey using ultrafiltration/diafiltration and spray drying processes. The milk by conventional cheese making procedures using S. cremoris
WPCs exhibited comparable composition, protein solubility and PAGE starter culture and rennet coagulant. The curd was cooked at 50°C and
and reverse phase HPLC properties. Major differences were observed separated from the whey at 50°C. One part of the whey was pasteur-
in the viscosity and foaming properties of the reference WPCs as a ized at 72°C for I5 set and the other part was not pasteurized. Both
function of pH. The WPCs generally produced higher foam expansion lots of whey were concentrated 20: I by ultrafiltration and diafiltered
but lower viscosity and stability values than for liquid egg white pro- as above, except that the whey was not clarified prior to ultrafiltration
tein. The findings were discussed in terms of the need for additional, as with the acid whey. The pH of the retentate fractions was 6.28-
fundamental knowledge of the physicochemical structure and reactiv- 6.32. The retentatcs were stored for 5 I day at 5”C, warmed to 50°C
ity of whey proteins to understand the reasons for their poor functional and spray dried as above.
performance. Reference S-lactoglobulin was purchased from ICN Pharmaceuti-
cals, Inc. (Plainview, NY) and a-lactalbumin and bovine serum al-
bumin were obtained from Sigma Chemical Co. (St. Louis). Control
INTRODUCTION WPC was prepared at Clemson University by acidifying raw skim
THE CHEMICAL and physicochemical properties of milk pro- milk with O.lN HCI to pH 4.6, filtering, concentrating the whey
fraction 3.5:l by ultrafiltration with a DC2 hollow fiber dialyzer/
teins have been reviewed (Eigel et al., 1984; Swaisgood, 1982)
concentrator (Amicon Corp., Lexington, MA) equipped with a HlPlO,
and their functional properties have also been reviewed (Work- 10,000 M W cutoff membrane and freeze-drying the retentate fraction.
shop II, 1979; ADSA Symposium, 1984; Morr, 1982, 1983, Three commercial WPCs were obtained for the study: LACPRODAN
1984; Kinsella, 1984). Even so, the relationship between the 60 and 80 from Danmark Proteins (Dairy Specialties, Inc., Worthing-
basic physicochemical and functional properties of whey pro- ton, OH) and ENRPRO 50 (Stauffer Chemical Company, Dobbs Ferry,
teins as influenced by compositional and processing factors is NY).
still poorly understood. It is generally agreed that the rather
poor functionality of whey protein products in food product Analytical
applications is largely responsible for their limited utilization. Moisture was determined by drying samples 3 hr in a 10&105”C
Additional research is needed to determine the relationships atmospheric oven (AOAC, 1980). Ash was by ignition of oven-dried
between processing conditions and functionality of whey pro- samples in an electric muffle furnace at 550°C (AOAC, 1980). Milkfat
tein concentrates (WPCs) in model and commercial food sys- was determined by a modified Roese-Gotlieb ether extraction proce-
tems. This study was conducted to investigate the effect of dure for dried milk (16.156, AOAC, 1980). One-gram samples were
whey source, whey processing, whey protein fractionation and accurately weighed into Mojonnier extraction flasks and carefully dis-
other processing parameters upon the composition, physico- persed into IO mL of distilled water to minimize.lumping. The dis-
persion was dissolved by adding I .5 mL of NH,OH and heating in a
chemical and functional properties of WPC. Acid casein whey
60°C water bath with occasional shaking for 5-10 min. The test was
and Cheddar cheese whey were used as source materials and completed according to the above procedure, except that an extra IO
the proteins were fractionated and concentrated by a combi- mL of ethyl alcohol were added just prior to making the second ether
nation ultrafiltration/diafiltration process and spray dried. extraction to prevent gel formation. Protein was determined by micro-
Functionality was limited to solubility and foaming perform- Kjeldahl (AOAC, 1980) using a nitrogen conversion factor of 6.38.
ance testing.
Gel electrophoresis
MATERIALS & METHODS Discontinuous polyacrylamide gel electrophoresis (PAGE) was con-
ducted by the method of Laemmli (1970) using a model 220 vertical
Whey protein concentrates electrophoresis cell (Bio-Rad Laboratories, Richmond, CA). The
stacking gel contained 3% acrylamide in pH 6.8 buffer and the sep-
Reference WPCs were prepared from sulfuric acid casein whey and arating gel contained 9% acrylamide in pH 8.8 buffer. Ten milligrams
Cheddar cheese whey by the New Zealand Dairy Research Institute, of WPC were dispersed in 1 mL distilled water, filtered through a
Palmerston North, New Zealand. Sulfuric acid casein WPCs were 0.45 pm metricel membrane filter (Millipore Corp., Bedford, MA)
prepared by pasteurizing milk at 72°C for IS set, separating it at 72°C and diluted I:5 (v/v) with sample buffer. Fifty p,L of protein solution
and acidifying the skim milk fraction to pH 4.6 with dilute H2S04. were added to each well and electrophoresed 2.5 hr at 30 mA constant
The curd was cooked at 50°C and the whey was separated at 47°C. current. Gels were stained with Coomassie Brilliant Blue R 250 (Bio-
One part of the whey was pasteurized at 72°C for 15 set and the other Rad Laboratories, Richmond, CA) in a solution containing 454 mL
part was not pasteurized. Both lots of whey were clarified at 48-5O”C, of 50% methanol in distilled water (v/v) and 46 mL of glacial acetic
concentrated 20: I (v/v) by ultrafiltration and diafiltered with a 2.37: I acid. Gels were then destained in a solution containing 50 mL of
(v/v) water to retentate ratio using a DDS Type 35 ultrafiltration unit methanol and 75 mL of glacial acetic acid, diluted to IL with distilled
fitted with a 9 m2 GR 60 PP membrane at 47-50°C to provide a flux water.
rate of 52-57 L mm2 hr ‘. Retentate fractions containing about 16% SDS-PAGE was also conducted by the procedure of Laemmli’( 1970),
total solids and at pH 4.7-4.8 were held 1-2 days at 5°C. warmed to except that the concentration of SDS was increased to 0.2% (v/v) in
all solutions and O.OlM dithioerythritol (DTE) was substituted for
mercaptoethanol (Brooks and Morr, 1984). About 10 mg of WPC
were dispersed in I .O mL of sample buffer and 10 pL of the dispersion
Author Morr is affiliated with the Dept. of Food Science, Clem- were added to each well. The I .5 m m thick gels were electrophoresed
son Univ., Clemson, SC 29631. at 35 mA constant current, fixed with trichloroacetic acid and sulfo-

1406-JOURNAL OF FOOD SCIENCE-Volume 50 (1985)

salicylic acid, stained with Coomassie Brilliant Blue R-250, and de- RESULTS
stained by the method recommended for analytical electrofocusing
gels (Anon., 1984).
WPC composition
Gel filtration chromatography The four spray-dried reference WPCs contained comparable
WPC samples were dispersed in pH 7, 0. 1M phosphate buffer and concentrations of moisture (3.8-4.4%), protein (71-73.6%)
chromatographed on a 160 mL bedvolume Sephadex G-l.50 column and milkfat (4.6&6.2%) as shown in Table 1. However, the
with dimensions of 2.6 cm x 30 cm (Pharmacia Fine Chemicals, two acid casein WPCs contained lower mineral ash concentra-
Piscataway, NJ) using down-flow rates of 35-46 mL/hr. Protein elu- tions (I .54-l .68%) than the cheese WPCs (2.64-2.73%).
tion was monitored at 280 qrn using a model U.4-5 Absorbance/Flu-
orescence Monitor (Instrumentation Specialties Co., Lincoln, NB). WPC solubility
HPLC analysis Reference WPC solubility values ranged from 92-100% at
pH 3 and from 89-100% at pH 7 (Table 2). These data clearly
The reverse phase HPLC procedure of Pearce (I 983) was followed demonstrate that it is possible to produce highly soluble WPCs
using a 4.6 mm x 75 mm Ultrapore RPSD Protein Separation column by ultrafiltration/diafiltration and spray-drying processes.These
(Beckman Instruments, Inc., Berkeley, CA). A model 332 gradient
liquid chromatograph with Hitachi model IO&400 variable wave-
WPCs were considerably more soluble than those commonly
length detector (Altex Instruments, Berkeley, CA) was used to per- produced by commercial processing (Morr et al., 1973) or by
form the analyses. A model C-RIA integrator (Altex Instruments, the Spherosil-S adsorption process (Nichols and Morr, 1985).
Berkeley, CA) was used in its area normalization mode to quantitate
individual protein peaks. The total solvent flow rate was I .O mL/min Sephadex gel filtration
and a discontinuous, linear solvent gradient program was used to change
SephadexG- 150 gel filtration elution patterns for freeze dried
the proportion of Solvent B from 0 to 36% at 0 to 3 min (12% change/
min), from 36 to 48% at 3 to 27 min (0.5% changeimin) and from
control WPC and the four spray-dried reference WPCs are in
48 to 0% at 27 to 30 min (16% changeimin). Solvent A consisted of Fig. I. Control WPC proteins were eluted in two incompletely
0.15M NaCI/HCl at pH 2.1 and Solvent B was acetonitrile (Fisher resolved peaks that were completely accessible to the internal
Scientific, Atlanta, GA). volume of the gel matrix and thus have molecular weights of
WPC samples were dispersed in Solvent A at a concentration of 1 5 150,000 (Anon., 1979). In contrast, the proteins from the
mg/mL, dialyzed overnight against excess Solvent A, centrifuged to four spray-dried reference WPCs were only partially accessible
remove insoluble materials and filtered through a Metricel 0.2 pm to the internal volume of the gel matrix. Their proteins exhib-
membrane filter (Gelman Sciences, Inc., Ann Arbor, MI) prior to ited generally similar gel filtration elution patterns, which in-
injection onto the column using a 20 FL sample injection loop.
dicated that a major portion of their proteins had molecular
Protein solubility weights of 2 150,000. The fraction of reference WPCs pro-
teins that were accessible to the gel matrix eluted at V,/V,
The method of Morr et al. (1985) was used to determine protein ratios of I .&2.86. Gel filtration elution patterns for two com-
solubility. About 500 mg of WPC was accurately weighed into a 150 mercially prepared WPCs (Fig. 2) revealed that a relatively
mL beaker and carefully dispersed by adding 40 mL of O.IM NaCl
greater portion of their proteins were eluted in the aggregated
solution in small increments with intermittant stirring to avoid lump
formation. Immediately after obtaining complete sample dispersion
state (molecular weights 2 150,000) than for the above ref-
the pH was adjusted to 3.0 or 7.0 with O.lN HCI or NaOH and it erence WPCs.
was stirred for 1 hr on a magnetic stirrer at a rate that just failed to
form a vortex. The dispersion was quantitatively transferred to a vol- HPLC analytical data
umetric flask and made to 50.0 mL with 0. IM NaCl. The dispersion Data in Tables 3 and 4 demonstrate that the reverse phase
was then centrifuged 30 min at 20,000 X g and the protein content HPLC procedure of Pearce ( 1983) fractionated reference whey
of the supernatant was determined by micro-Kjeldahl (AOAC, 1980).
Percent protein solubility was computed as the portion of total protein
proteins and nonpasteurizedCheddar cheeseWPC proteins with
recovered in the supernatant fraction. good peak area and retention time reproducibility. Typical HPLC
patterns for reference WPCs and reference whey proteins are
Foaming properties in Fig. 3. The percentage distribution of the respective peak
areas in Tables 4 and 5 indicate that a-lactalbumin and p-
The WPC samples were dispersed in distilled water to provide a
6% protein concentration (w/v) and adjusted to pH 4.5 or 9.0 with
lactoglobulin peaks (peaks 6-8) accounted for upwards of 80%
0. IN HCI or NaOH. Viscosity was determined with a Brookfield of the total proteins in the four reference WPCs. This technique
viscometer (Stoughton, MA) equipped with a HA-I T-bar spindle and
a helipath stand. Dispersions were whipped at room temperature with
a Hobart K5-A mixer (Troy, OH) equipped with a wire whip beater Table l-Composition of reference whey protein concentrates, %a
at speed setting 6 for up to 15 min to determine maximum foam Whey protein concentrate Moisture Ash Protein Milkfat
expansion. Whipping was interrupted after each 3 min interval to
determine foam expansion and viscosity. Foam expansion was deter- Nonpasteurized acid
casein WPC 4.39 1.54 70.97 5.25
mined by level-filling a 100 mL plastic weighing boat with foam and
Pasteurized acid
weighing to t O.Olg. Foam expansion was computed using the casein WPCb 4.18 1.68 71.66 4.66
expression: Nonpasteurized Cheddar
cheese WPC 4.28 2.64 73.63 6.18
Unwhipped Pasteurized Cheddar
Foam dispersion wt (g) - Foam wt (g) x ,oo cheese WPCb 3.80 2.73 72.62 5.62
=
expansion, % &whipped dispersion wt (g) a Mean of duplicate determinations.
b Whey was pasteurized at 72”C-15 set prior to ultrafiltration and diafiltration
After the viscosity was determined on the foam in the weighing
boat it was returned to the bowl and whipping was resumed for an
additional 3 min period. Foam stability was determined by transferring
100 mL of maximum expansion foam into a Pyrex filter funnel with Table Z-Protein solubilitv for whev orotein concentrates. %a
dimensions of 7.5 cm inner top diameter, 0.4 cm inner stem diameter Whey protein concentrate PH 3 PH 7
and 7.0 cm stem length. A small plug of glass wool was placed in Nonpasteurized acid casein WPC 92.3 100
the top of the funnel stem to retain the foam but allow drainage of Pasteurized acid casein WPCb 94.4 93.7
the liquid. The time required for the first drop of liquid to drain from Nonpasteurized Cheddar cheese WPC 94.6 88.9
the funnel was determined as an index of foam stability. Also, the Pasteurized Cheddar cheese WPC? 99.9 98.1
time (min) for drainage of the entire foam was determined for low a Mean of duplicate determinations.
stability foams. b Whey was pasteurized at 72°C.15 set prior to ultrafiltration and diafiltration.

Volume 50 (1985)--JOURNAL OF FOOD SCIENCE-1407

 WHEY PROTEIN FUNCTIONALITY. ..
Table &Reproducibility of HPLC analysis of nonpasteurized Cheddar
cheese WPCa
Protein Retention time Mean peak areab
Peak number component Imin) ‘(%)
Unknown 3.59 0.73 k 0.08
Unknown 5.79 12.46-cl.23
Unknown 6.89 7.71?0.74
Unknown 7.79 4.82 + 0.35
BSA a.89 6.45 ‘- 0.42
u-La 10.5 16.60 k 0.91
7 P-b A 12.3
46.40 + 5.45
8 B-La I3 12.9
9 bnl;nown 15.2 1.51 ? 2.24
10 Unknown 17.9 1.392 3.38
11 Unknown 19.7 1.94k1.55
a Sample solution contained 1 mg/mL nonpasteurized Cheddar cheese WPC.
b Percentage of total integrator area, _i standard deviation, n = 4.

E‘“TON VOLUME, rnL -

Fig. I-Sephadex G- 150 gel filtration patterns from control WPC

and reference WPCs in pH 7 phosphate buffer (0.7 Ml: (A) con-
trol WPC; (B) nonpasteurized acid casein WPC; (Cl pasteurized
acid casein WPC; (0) nonpasteurized Cheddar cheese WPC;
and (E) pasteurized Cheddar cheese WPCa. Flow rate was 46
mLlhr. a Whey was pasteurized at 72% 15 see prior to ultrafil-
tration and diafiltration.

6P
Y
ELUTION VOLUME, mL -
Fig. 2-Sephadex G- 150 gel filtration patterns from commercial
WPCs eluted in pH 7phosphate
60; and (B) LACPRODAN-80.
buffer (0.1 M): (A) LACPRODAN-
Flow rate was 35 mLlhr.
L
rl
b:L TIME,min --w

Fig. 3-Reverse phase HPLC patterns from: (A) reference whey

proteins; (B) control WPC; (C) nonpasteurized acid casein WPC;
Table 3-Reproducibility of HPLC retention time and peak area for ref- (0) pasteurized acid casein WPCa; (E) nonpasteurized Cheddar
erence whey protein9 cheese WPC; and (F) pasteurized Cheddar cheese WPCa. a Whey
Mean retention Mean peak area, was pasteurized at 72°C15 set prior to ultrafiltration and diafil-
Whey protein component time, min mV set x 10s tration.
Bovine serum albumin 8.9kO.18 67.2 k 4.4
a-lactalbumin 10.6?0.08 83.3~0.8
p-lactoglobulin A 12.1 to.21 58.1 k3.7
p-lactoglobulin 6 12.720.18 56.6i 1.8 that for the reference WPCs (Fig. 3). However, both LAC-
a Reference proteins were dissolved in Solvent A (0.15 M NaCliHCl at pH 2.11 at a PRODAN WPCs exhibited significantly altered patterns than
concentration of 1 mg/mL. 2 standard deviation, n = 3. the above. For example, u-lactalbumin was eluted at 11.6-
11.7 min and P-Iactoglobulin was eluted at > I4 min for both
LACPRODAN WPCs compared to respective values of 10.6
fractionated the WPC proteins into a total of 1 1 peaks, which and 12.1-12.7 min for reference WPCs and ENRPRO 50. p-
included the four reference proteins listed in Table 3 and Fig. Lactoglobulin was not resolved into its two components in any
3A, plus seven additional, unidentified peaks. The four ref- of the commercial WPC patterns.
erence WPCs exhibited similar HPLC patterns and peak area PAGE data
distributions, except that both Cheddar cheese WPCs contained
less P-lactoglobulin than the acid casein WPCs. The HPLC PAGE separation patterns in Fig. 5 reveal that control WPC
pattern (Fig. 4A) for ENRPRO-50 was generally similar to and all four reference WPCs contained generally similar pro-

1408--JOURNAL Of FOOD SCIENCE-Volume 50 (1985)

 Table 5-HPLC analysis of reference whey protein concentrate9
Peak numbe@
2 3 4 5 6 7&8
Retention time, min
5.8 6.8-6.9 7.8 8.8-8.9 10.3-10.5 12-12.9
Protein component
- - BSA a-La p-Lg A&B
Percentage total peak area (%)
Nonpasteurized acid casein WPC 2.2 7.5 6.5 3.1 21.2 58.4
Pasteurized acid casein WPCc 6.5 4.0 7.0 3.1 23.2 56.2
Nonpasteurized Cheddar cheese WPC 13.0 7.6 4.8 6.3 16.7 51.1
Pasteurized Cheddar cheese WPCC 11.4 8.2 9.6 5.1 20.0 41.5
8 Mean data from two determinations.
“Sam as in Table 4.
CWhey was pasteurized at 72°C.15 sac prior to ultrafiltration and diafiltration.

tein compositions. Control WPC exhibited severalprotein bands Cheddar cheeseWPC dispersion had a high viscosity value of
near the origin which were absent from reference WPC pat- 17.6 cp at pH 9, the stable nonpasteurized Cheddar cheese
terns. WPC dispersion foam had the lowest viscosity of I .5 cp at pH
SDS-PAGE separation patterns in Fig. 6 reveal that all four 4.5.
reference WPCs contain similar molecular weight protein com-
ponentsin approximately equal concentrations.A major amount DISCUSSION
of the protein was in the 20-30 K molecular weight range
(Brooks and Morr, 1984). These patterns also revealed three AS INDICATED PREVIOUSLY (Morr, 1982; Kinsella, 1984;
additional bands with higher molecular weights which may Workshop II, 1979; ADSA Symposium, 1984), the generally
indicate immunoglobulins or aggregated proteins. The Spher- poor functional performance of WPCs is largely responsible
osil-S WPC pattern (Fig. 6E) exhibited severaladditional bands, for their limited use by the food industry. A better understand-
confirming previous observations that its proteins have under- ing of the relationship between the basic physicochemical .and
gone substantial denaturation/aggregation. Interestingly, these functional properties of whey proteins and the influence of
latter protein aggregateswere not dissociated by SDS and DTE composition and processing conditions used in their manufac-
in the gel, which normally dissociate protein complexes (Ni- ture is needed. It is generally believed that improved WPC
chols and Morr, 1985). functionality can be achieved by: increasing protein content;
decreasingresidual lactose, minerals and milkfat contents; and
Viscosity and foaming data by using fractionation and manufacturing processing condi-
tions that will minimize protein denaturation and interaction.
Foaming data for 6% reference WPC dispersions and liquid Certain basic information is also needed on the inherent phys-
egg white are in Fig. 7 and 8 and Table 6. Initial viscosity icochemicalpropertiesof the whey protein componentsof WPCs,
values for unwhipped WPC dispersions were higher at pH 4.5 e.g., their primary structure with respect to location of ionic
(Fig. 7) than at pH 9 (Fig. 8). Although viscosity of unwhipped and sulthydryl-disulfide groups, and secondary structure with
liquid egg white was only about I cp and independent of pH, respect to conformation state and the opportunity for forming
its viscosity increased at a much greater rate during whipping intra- and inter-molecular bonding through ionic and disulfide
than any of the reference WPC dispersions at both pH values. bond interchange mechanisms. Also, the influence of their hy-
The viscosity of reference WPC dispersions remained rela- drophobicity as influenced by processing conditions that pro-
tively constant or underwent a slight decreasewhen whipped mote unfolding of the respective whey protein molecules may
at pH 4.5, but increased from about I cp (unwhipped) to 8- be important.
16 cp when whipped for I5 min at pH 9 (Fig. 8). HPLC results obtained in this study were interpreted as in-
All reference WPC dispersions produced higher maximum dicating that the proteins in all four reference WPCs retained
foam expansion values at pH 4.5 than at pH 9 and higher foam their relative hydrophobicity values. For example, B-lacto-
expansion values than for liquid egg white at pH 4.5 (Table globulin, which eluted last from the hydrophobic column with
6). The order of decreasing maximum foam expansion values retention time of about I2 min is assumed to be the most
for reference WPC dispersions at both pH 4.5 and 9.0 was: hydrophobic whey protein. The similarity of reversephaseHPLC
nonpasteurizedacid casein WPC, pasteurized Cheddar cheese elution patterns for the four reference WPCs confirms that the
WPC, nonpasteurized Cheddar cheese WPC and pasteurized processing treatments used in their preparation did not cause
acid casein WPC. The maximum foam expansion values for substantial protein denaturation or protein-protein interaction
reference WPC dispersions tended to be inversely related to which would have resulted in alteration of their hydrophobic-
foam viscosity after I5 min whipping at pH 4.5, but this re- ity, as for two of the commercial WPCs. This approach, in
lationship was less clear-cut at pH 9 (Table 6). conjunction with PAGE data, may provide useful criteria for
Foam stability data in Table 6 were quite variable and lacked evaluating the extent of denaturation and aggregation of whey
an obvious relationship with pH, viscosity or maximum foam proteins in WPCs and thus provide an indication of their po-
expansion. For example, the least stable foams were produced tential functionality.
from nonpasteurized acid casein WPC at pH 4.5 and nonpas- Results from this study allow a comparison of the functional
teurized acid casein and Cheddar cheese WPC dispersions at performance of WPCs produced from pH 4.6 acid whey and
pH 9, which had intermediate foam viscosity and expansion from pH 6.2-6.3 Cheddar cheesewhey, with and without pas-
values. Nonpasteurized acid casein WPCs dispersion foams teurization. Although acid whey generally contains about 1.5
had a low stability of only 5 min at pH 4.5, even though it times as much minerals as Cheddar cheesewhey (Morr, 1984),
had the highest maximum foam expansion value of 1510%. Cheddar cheeseWPCs in this study contained about 1.6 times
Maximum foam stability was provided by the nonpasteurized more minerals than the acid casein WPCs. Apparently, the
Cheddar cheese WPC dispersion at pH 4.5 (60 min) and by minerals in acid whey are in a more diffusable state than those
pasteurized Cheddar cheeseWPC dispersion at pH 9 (68 min), in Cheddarcheesewhey. Although not determined in this study,
even though they exhibited only intermediate maximum foam Cheddar cheese WPCs would be expected to contain a signif-
expansion values. Although the highly stable pasteurized icant concentration of glycomacropeptide (GMP) fragment of

Volume 50 (1985)-JOURNAL OF FOOD SCIENCE-1409

WHEY PROTEIN FUNCTIONALITY. ..

BSA
CY-123

$-lg A/B

ABCDEF
Fig. B-PAGE patterns from: (A) control WPC; (B) reference whey
proteins; (C) nonpasteurized Cheddar cheese WPC; (D) pas-
teurized Cheddar cheese WPCa; (El nonpasteurized acid casein
WPC; and (F) pasteurized acid casein WPCa. a Whey was pas-
teurized at 7.X-15 set prior to ultrafiltration and diafiltration.

ABCDE
Fig. B--SDS-PAGE patterns from: (A) pasteurized acid casein
WPCa; (B) nonpasteurized acid casein WPC; (C) pasteurized
Cheddar cheese WPCa; (D) nonpasteurized Cheddar cheese WPC;
and (E) Spherosil-S WPC (Nichols and Morr, 1985). a Whey was
pasteurized 72”C- 15 set prior to ultrafiltration and diafiltration.

K-casein that would result from the hydrolytic action of rennet

upon casein micelles during cheese-making.Although this highly
acidic and glycosylated GMP fragment might affect the func-
tional properties of Cheddar cheese WPCs, it did not appear
to have been important in the present study. The other major
experimental variable used in this study was the effect of pas-
teurization of whey prior to ultrafiltration/diafltration and drying
of the WPCs. Results of this study demonstrated that whey
pasteurization had little effect upon the composition, solubility
and foaming properties of the resulting WPCs. However, both
pasteurized whey WPCs contained slightly greater mineral
TIME, min --w contents than the nonpasteurizedcounterparts. Solubility, PAGE
Fig. 4-Reverse phase HPLC patterns from: (A) ENRPRO 50; (B)
and HPLC data obtained in this study indicate that pasteuri-
LACPRODAN-60; and (Cl LACPRODAN-80. zation and spray drying treatments produced much less whey
protein denaturation than is generally observed with commer-
cial WPCs (Morr et al., 1973). These reference WPCs were,

141O-JOURNAL OF FOOD SCIENCE-Volume 50 (1985)

 35

8
30

l
/

25 /

\
l LIQUID EGG UHITE
/
0

LIQUIO EGG Y",TE

A-A

0 3 6 9 12 IS
YHIPPING TME, min

Fig. 7-Relative viscosity of reference WPC dispersions in dis- L I I 1 I I

tilled water (b% protein concentration and 25°C) and fresh, liquid 0 3 6 9 12 15
egg white at pH 4.5 as a function of whipping time. Data rep- WHIPPING TIUE, mln
resent averages of three replicate values. Sample identification:
NPACWPC, nonpasteurized acid casein WPC; PACWPC, pas- Fig. &Relative viscosity of reference WPC dispersions in dis-
teurized acid casein WPG; NPCCWPC, nonpasteurized Cheddar tilled water (b% protein concentration and25”Cl and fresh, liquid
cheese WPC; and PCCWPC, pasteurized Cheddar cheese WPCa. egg white at pH 9.0 as a function of whipping time. Data are
a Whey was pasteurized 72”C- 15 set prior to ultrafiltration and averages of three replicate values. Sample identification:
diafiltration. NPACWPC, nonpasteurized acid casein WPC; PACWPC, pas-
teurized acid casein WPCa; NPCCWPC, nonpasteurized Cheddar
cheese WPC; and PCCWPC, pasteurized Cheddar cheese WPCa.
= Whey was pasteurized 12%15 set prior to ultrafiltration and
dia filtration.
however, stored at refrigerator or freezer temperatures which
may inhibit protein-protein interactions that likely occur at el-
evated temperature storage of commercial WPCs to cause loss REFERENCES
of solubility and functionality.
It is generally believed that protein molecular complexes ADSA Symposium. 1984. I. Production and utilization of whey and whey
components; II. Assessing functionality of whey proteins. J. Dairy Sci.
must be solubilized, disaggregated and unfolded during the 67: 2e20
whipping process if they are to effectively promote foam ex- Anonymous. 1979. Gel filtration theory and practice. Pharmacia Fine
pansion and stability. The basic physicochemical properties Chemicals AB. Piscatawav, NJ.
Anonvmous. 1984. EIectrofo&inn Instruction 1804. LKB Instruments Inc..
mentioned above would obviously play a key role in deter- Rocicville, MD.
mining how well each WPC would function in the whipping AOAC. 1980. “Official Methods of Analysis.” Association of OfIicial Ana-
process. It is of interest to compare the function of the four lvtical Chemists. Washineton. DC.
Brboks, J.R. and Morr, C.Vr1984. Phosphorus and phytate content of soy-
reference WPCs with that of liquid egg white (Table 6 and bean protein components. J. Agric. Food Chem. 32: 672.
Fig. 7 and 8) in terms of viscosity and foaming properties. It Eigel, W.N., Butler, J.E., Emstrom, C.A., Farrell, H.M., Hat-walker, V.R.,
Jenness! R., and Whitney, R. McL. 1984. Nomenclature of proteins of
appears that liquid egg white protein possessesa low initial cow’s xmlk: fifth revision. J. Dairy Sci. 67: 1599.
viscosity that permits rapid air incorporation into the liquid, Kinsella, J.E. 1984. Milk proteins: physicochemical and functional prop-
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