Food Chemistry 228 (2017) 243248

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Effect of a multiple freeze-thaw process on structural and foaming

properties of individual egg white proteins
Xiang Duan a,1, Junyi Li a,1, Qinjun Zhang a, Tong Zhao a, Mei Li a, Xueming Xu b, Xuebo Liu a,
a
College of Food Science and Engineering, Northwest A&F University, Yangling 712100, PR China
b
State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, PR China

a r t i c l e i n f o a b s t r a c t

Article history: In this study, major albumen proteins (ovalbumin, ovomucoid, ovotransferrin, lysozyme and ovomucin)
Received 9 November 2016 were singly subjected to a multiple freeze-thaw process, and the resulting changes in structural charac-
Received in revised form 31 January 2017 teristics and foamability were investigated. Structural changes of proteins occurred during the process,
Accepted 1 February 2017
regarding by sulfhydryl-disulfide interchange and exposure of hydrophobic groups. The differential scan-
Available online 3 February 2017
ning calorimetry and scanning electron microscopy showed that these albumen proteins underwent
denaturation, dissociation and possibly aggregation. Correspondingly, the foaming ability of albumen
Keywords:
proteins improved after the freeze-thaw treatment, except for ovalbumin. The foaming ability of whole
Albumen proteins
Freeze-thaw
egg white was higher than that of each albumen protein, and improved after the multiple freeze-thaw
Structural characteristics process. This study extended knowledge of the relative contribution of each albumen protein to foaming
Foamability properties of whole egg white during a freeze-thaw process, and suggested that a multiple freeze-thaw
process is a promising technique for improving foaming properties of egg white proteins.
2017 Published by Elsevier Ltd.

1. Introduction proteins underwent a structural change during frozen storage

because of denaturation (Mori, 1971; Wootton, Hong, & Thi,
Based on its excellent foaming ability, egg white is widely used 1981). Therefore, we are interested in learning whether a multiple
in a variety of food products, including cakes, dessert shells and freeze-thaw (F-T) treatment can be implemented to enhance foam-
pies. For these food products, foams provide unique and desirable ing properties of white proteins via modifying their structures.
textures and largely affect the final quality. Thus, technologies that Actually, Zhao, Dong, Li, Kong, and Liu (2015) recently reported
can improve foaming properties of egg white are greatly desired that soy proteins treated with multiple F-T cycles exhibited an
for food industry. improved functional properties due to partial structural unfolding.
Foam comprised of millions of bubbles each encapsulated by a The effect of a F-T process on the functionalities of whole egg
protein film and separated by a thin water filled lamella. Egg white white has been extensively studied, though results were conflict-
foam is created as liquid egg whites are whipped. During this pro- ing (Herald & Smith, 1989; Vaclavik & Christian, 2014; Wootton
cess, air comes into solution to form bubbles, white proteins et al., 1981; Xu, Shimoyamada, & Watanabe, 1997). Egg white con-
adsorb rapidly at the air-water interface and undergo rapid confor- sists of a mixture of proteins, including ovalbumin, ovotransferrin,
mational changes to form cohesive viscoelastic films around bub- ovomucoid, lysozyme, ovomucin and others (Mine, 2008). The
bles. The spreading ability of a protein at the liquid surface structure of egg white allows it to perform well in foams because
depends on the protein conformation. For albumen proteins, a cer- each albumen protein carries out a different function (Stadelman,
tain degree of denaturation is benefit to their foam-forming capac- Newkirk, & Newby, 1995). Although the foaming properties of each
ity (Campbell, Raikos, & Euston, 2003; Johnson & Zabik, 1981a). albumen protein has been adequately investigated, further
Eggs are usually marketed as shell eggs. Due to the increase of research on the functional behaviors of individual albumen pro-
food industrys demand for eggs and egg products, egg white or teins during a multiple F-T process are little reported so far. Thus,
yolk in the form of liquid, frozen or dried are available for ease investigating the functional behaviors of individual albumen pro-
transport and storage. Previous studies reported that egg white teins subjected to a multiple F-T process is essential to evidence
the relative contribution of each albumen protein to functionalities
Corresponding author. of whole egg white. In this study, these individual albumen
E-mail address: [email protected] (X. Liu).
1
These authors contributed equally.

http://dx.doi.org/10.1016/j.foodchem.2017.02.005
0308-8146/ 2017 Published by Elsevier Ltd.
244 X. Duan et al. / Food Chemistry 228 (2017) 243248

proteins were singly subjected to a multiple F-T treatment, and (Mine, 2008). Therefore, the five proteins were redissolved sepa-
their structural and foaming properties were determined. rately in 5.4% (wt/v), 1.1% (wt/v), 1.2% (wt/v), 0.35% (wt/v), and
The aim of present work was to elaborate on the effect of a mul- 0.34% (wt/v) in deionized water. These dispersions and fresh egg
tiple F-T treatment on foaming properties of individual albumen whites were separately stored at 20 C for 24 h, followed by
proteins. The relationships between changes in structural proper- thawing at 22 C for 12 h. The F-T process was repeated for 1, 3
ties and foaming properties of these proteins were elucidated by and 5 times, respectively. These samples were then lyophilized
determining free sulfhydryl groups (SH), surface hydrophobicity and stored at 20 C. The samples without the F-T treatment were
(Ho), thermal property, morphology and foaming properties as a considered as untreated samples.
function of F-T cycles.
2.4. Free SH group content
2. Material and methods
The free SH group content was measured according to the
method of Li, Zhu, Zhou, and Peng (2012). Ellmans reagent was
2.1. Materials
prepared by dissolving 40 mg of DTNB in 10 mL of Tris-Gly buffer
(10.4 g of Tris, 6.9 g of Gly, and 1.2 g of disodium ethylenedi-
Fresh hen eggs laid within 24 h were collected from a local farm
aminetetraacetic acid (EDTA) in 1000 mL of deionized water, pH
(Yangling, Shaanxi, China). Ovotransferrin (C0755), lysozyme
8.0). The lyophilized protein samples were dissolved with Tris-
(62971) and the testing chemicals including 1-
Gly-SDS buffer (45 mL Tris-Gly buffer containing 5 mL of 2.5%
anilinonaphthalene-8-sulfonic acid (ANS), 5,50 -dithiobis (2-
(wt/v) SDS aqueous solution) to a final protein concentration of
nitrobenzoic acid) (DTNB) were purchased from Sigma-Aldrich
10 mg/mL. Four milliliter of each protein sample was mixed with
Co. LLC. (Shanghai, China). All other reagents used were of analyt-
0.04 mL of Ellmans reagent solution. Subsequently, the mixture
ical grade (Shanghai Chemical Reagents Co., Shanghai, China).
was shaken and incubated in dark at room temperature for
10 min. The absorbance of supernatant was measured at 412 nm
2.2. Preparation of individual albumen proteins against the blank (without Ellmans reagent and sample). Absor-
bance values were converted to amounts of free SH groups using
The fresh eggs were broken manually and white and yolk were a calibration curve with reduced glutathione (Wang et al., 2014).
carefully separated. Ovalbumin was separated according to the
method of Geng et al. (2012). Briefly, the egg whites (270 g) were 2.5. Surface hydrophobicity (Ho)
stirred using a magnetic stirrer (IKA, IKA Works Inc., Wilmington,
NC, USA) at 4 C for 2 h with 3 volumes of 50 mM NaCl solution. Ho of albumen proteins was determined by following the
The pH of solution was adjusted to 6.0 with 2 M HCl, and PEG- method of Kato and Nakai (1980), with slight modifications. Each
8000 was added while stirring (final wt/wt: 15%). The dispersion sample was diluted to five concentrations between 0% and 0.1%
was allowed to stand for 2 h at 4 C, and then was centrifuged at protein (wt/wt) in 10 mM phosphate buffer (pH 7.0). Then, aliquots
15,000g at 4 C for 10 min. The supernatant (ovalbumin) was dia- (20 lL) of ANS (8.0 mM in the same buffer) were added to 4 mL of
lyzed against Milli-Q water at 4 C for 72 h with changes of water each protein solution. The solutions were rested for 3 min in dark
every 6 h, and then lyophilized. Ovomucoid was prepared with the and placed in the cell of a Hitachi F4500 fluorescence spectrometer
modified procedure of Lineweaver and Murray (1947). The egg (Tokyo, Japan), and the fluorescence intensity was measured at
whites were slowly diluted with 1 vol of freshly 10% (wt/v) tri- 470 nm, using excitation at 390 nm, slit width 5 nm. Appropriate
chloroacetic acid (pH 1.15) and stirred mildly. The pH was main- blanks corresponding to the buffer were subtracted to correct
tained at 3.5 by addition of 1 N NaOH or 1 N HCl. Following background fluorescence. Ho was determined according to the
standing for 4 h at 4 C, the dispersion was centrifuged at 3000g slope method of Alizadeh-Pasdar and Li-Chan (2000).
at 4 C for 30 min. The supernatant was filtered through cellulose
filter paper (pore aperture 3050 lm) and diluted with 3 volumes 2.6. Thermal analysis
of cold acetone and stored overnight at 4 C. The suspension was
centrifuged at 3000g at 4 C for 10 min and the precipitate (ovomu- Thermal properties of albumen proteins were analyzed by dif-
coid) was dissolved in water (1:10, wt/v). The solution was dia- ferential scanning calorimetry (DSC). DSC was carried out accord-
lyzed against Milli-Q water at 4 C for 72 h with changes of ing to Wang et al. (2014) using a DSC Q2000 instrument (TA
water every 6 h, and then lyophilized. Ovomucin was isolated Instruments, New Castle, DE, USA). The freeze-dried proteins (2
according to the method of Omana and Wu (2009). Briefly, the 3 mg) were weighed in aluminum pans, the pans were hermeti-
egg whites were homogenized using a magnetic stirrer for cally sealed. A peltier device was used to heat samples from 20
30 min at ambient temperature (22 C). Stirring speed was set at to 180 C at 10 C/min, while gas nitrogen (50 mL/min) was used
350 rpm to avoid foaming. The dispersion was diluted with 0.1 N to purge. An empty pan served as the reference. The enthalpy value
NaCl to triple volume. The pH of dispersion was then adjusted to was calculated from the thermogram using the DSC Q2000 V24.2
6.0 using 1 N or 0.1 N HCl, while stirring gently at ambient temper- Build 107.
ature (22 C) for 30 min. The dispersion was stored overnight at
4 C and separated by centrifugation at 15,000g for 10 min at 2.7. Scanning electron microscope (SEM)
4 C. The precipitate (ovomucin) was freeze-dried and stored at
20 C until analysis. Rather than native proteins in fresh eggs, The morphology of albumen protein samples were observed
these individual proteins extracted for analysis could be seen as with a SEM. The lyophilized samples were sputter-coated with
pre-denatured state. gold, and examined in a Hitachi S-3400N scanning electron micro-
scope (Hitachi, Tokyo, Japan) at 5.0 kV.
2.3. F-T treatment
2.8. Foaming properties
For hen eggs, protein portion constitutes about 10% of whole
egg white. Ovalbumin, ovomucoid, ovotransferrin, ovomucin and The foaming properties of each albumen protein was deter-
lysozyme occupied 54%, 11%, 12%, 3.5% and 3.4%, respectively mined by the method of Akintayo, Oshodi, and Esuoso (1999), with
 X. Duan et al. / Food Chemistry 228 (2017) 243248 245

slight modifications. Briefly, weighed amounts of freeze-dried pro-

teins (100 mg) were dispersed in 10 mL distilled water. The result-
ing solutions were homogenized at a speed of 10,000 rpm, using a
homogenizer (IKA Labortechnik, Selangor, Malaysia) to incorporate
air for 2 min at room temperature. The whipped samples were
immediately transferred into a 25 mL cylinder and total volume
was read after 30 s. The foaming capacity was calculated according
to the following equation:
AB
Foaming capacity %  100
B
where A was the volume after whipping (mL), B was the volume
before whipping (mL).
The whipped samples were allowed to stand at 20 C for 30 min
and the volume of whipped samples were then recorded. Foaming
stability was calculated as follows:
CB
Foaming stability %  100
B
where C was the volume after standing (mL), B was the volume
before whipping (mL).
Fig. 1. Free sulfhydryl (SH) group contents of untreated and treated albumen
proteins (1, 3 and 5 F-T cycles). Different letters (ac) in the same protein indicated
2.9. Statistical analysis significant differences (P < 0.05).

The experiments were run in triplicate. The results were

expressed as the mean standard deviation and were analyzed when the number of F-T cycle increased from 0 to 3, and then
using SPSS 16.0 software (SPSS, Inc., Chicago, IL, USA). One-way decreased to a value which was not significantly different from
analysis of variance (ANOVA) was used to analyze data, and Dun- the untreated sample. Ovomucin was considered as a biopolymer
cans multiple-range tests were conducted to determine differ- which was cross-linked by disulfide bonds (Hayakawa & Sato,
ences between the means. Results were considered statistically 1976). The result showed that few F-T cycles (1 and 3 F-T times)
significant at P < 0.05. could break these disulfide bonds and thus increased the SH con-
tent. Whereas, more F-T cycles (5 times) induced the new forma-
3. Results and discussion tion of disulfide bonds because of SH oxidation, which
decreased the free SH content in ovomucin. Similarly, the free
3.1. Free SH group content SH content in egg white increased significantly from 1 to 3 F-T
times, and then decreased after 5 F-T cycles to a value which
Disulfide bonds play a key role in maintaining structure of egg was lower than that of untreated sample. Howell and Taylor
proteins. Inter and/or intramolecular sulfhydryl-disulfide inter- (1995) proposed that the formation of disulfide bonds between
change reactions are involved in foam formation (Johnson & sulfhydryl groups enhanced foaming properties of globular pro-
Zabik, 1981a). The free SH contents in albumen proteins and teins. This meant that the foaming properties of egg white might
egg whites with different F-T cycles are presented in Fig. 1. For improve after a multiple F-T treatment.
ovalbumin and ovotransferrin, there was no significant difference
between the untreated and F-T treated groups. Among albumen 3.2. Surface hydrophobicity (Ho)
proteins, ovalbumin is the only protein that has free SH groups,
containing a single disulfide bond between Cys74 and Cys121. Ho is a structural characteristic to evaluate conformational
Ovotransferrin contains 15 disulfide bridges which maintain its change of proteins. The binding of fluorescent probe (ANS) to pro-
bilobal structure (Mine, 2008). Free SH contents of the two pro- teins occurs at hydrophobic cavities formed by grouping of nonpo-
teins were scarely influenced by a multiple F-T treatment, indicat- lar residues on protein surface, thus could give information on Ho
ing that disulfide bonds of the both proteins were stable during the of proteins (Wang et al., 2014). Fig. 2 shows Ho values of albumen
process. Meanwhile, the free SH content in ovomucoid signifi- proteins with different F-T cycles. For ovalbumin, a remarkable
cantly decreased from 0.92 lmol/g to 0.31, 0.50 and 0.29 lmol/g decrease was observed in Ho value after a F-T treatment. This indi-
after 1, 3 and 5 F-T cycles, respectively. The ovomucoid molecule cated that ovalbumin folded or aggregated and thus the hydropho-
is composed of three distinct domains crosslinked only by intrado- bic residues were buried in interior during a F-T process. The Ho
main disulfide bonds (Mine, 2008). The decrease in free SH con- value of ovomucoid increased after 3 F-T cycles, and then
tent was due to the alteration in SS/SH exchange reaction or SH decreased to a value after 5 F-T cycles, which was still higher than
oxidation and rearrangement (Chen et al., 2016). Contrarily, the that of untreated sample. Combined with the free SH content
free SH content in lysozyme significantly increased from result, the increase in Ho value of ovomucoid after multiple F-T
0.28 lmol/g to 0.61, 0.70 and 0.45 lmol/g after 1, 3 and 5 F-T cycles was originated from the molecular rearrangements, with
cycles, respectively. Native lysozyme contains four disulfide bonds an exposure of hydrophobic side-chain groups. For ovotransferrin,
(Johnson & Zabik, 1981a). Disulfide bonds are relatively weak cova- the Ho value fluctuated largely with increasing number of F-T
lent bonds, they could be broken during a F-T process because of cycles. Ovotransferrin contains 15 disulfide bridges, and
water redistribution and ice recrystallization (Wang, Jin, & Xu, Muralidhara and Hirose (2000) reported that the interdomain
2015; Zhao, Li, Liu, Liu, & Li, 2012). Thus, the increase in free SH disulfide bonds significantly decreased its conformational stability
content of lysozyme was attributed to the depolymerization effect in the C- lobe. Therefore, the less stability of structure might be a
of lysozyme through breakage of disulfide bonds during a F-T pro- reason that ovotransferrin underwent a large-scare conformational
cess. Furthermore, the free SH content in ovomucin increased transition during a multiple F-T process. Furthermore, the Ho value
246 X. Duan et al. / Food Chemistry 228 (2017) 243248

3.3. Thermal property

DSC analysis was carried out to investigate the effect of a F-T

treatment on thermal property of albumen proteins. The denatura-
tion temperature (Tp) and enthalpy (DH) are main parameters of
DSC profile. Egg protein denaturation is one major importance in
establishing cake structure, thus alteration of albumen proteins
denaturation behavior may affect final properties of egg products.
Table 1 showed Tp of individual albumen proteins with different F-
T cycles. Ovalbumin is the most abundant protein in egg but is rel-
atively heat-labile (Mine, 2008). The Tp of untreated and F-T trea-
ted ovalbumin was below 60 C. This was lower than the data
reported by Donovan, Mapes, Davis, and Garibaldi (1975) indicat-
ing that Tp of ovalbumin was 84 C. We guessed that the difference
was because that the protein samples in this work were in freeze-
dried powder rather than protein solution. For ovalbumin and ovo-
transferrin, no significant difference in Tp was seen between
untreated and F-T treated samples. This suggested that the both
proteins maintained thermal stability during a F-T treatment. This
result was in agreement with the SH result showing that disulfide
Fig. 2. Surface hydrophobicity (Ho) of untreated and treated albumen proteins (1, 3 bonds of the two proteins were stable during the process. For ovo-
and 5 F-T cycles). Different letters (ac) in the same protein indicated significant
mucoid, Tp increased from 112.06 C to around 120 C after a F-T
differences (P < 0.05).
treatment. Combined with above-mentioned SH and Ho results,
the increase in heat stability of ovomucoid was contributed to
of lysozyme increased from 205.35 to 234.56, 244.68 and 258.07 the enhanced intermolecular forces. Contrarily, Tp of lysozyme
after 1, 3 and 5 F-T cycles. This trend was in agreement with the decreased from 118.19 C to 112.42 C, 106.29 C and 102.11 C
free SH content result, suggesting that the increase in Ho value after 1, 3 and 5 F-T cycles, respectively. This might be due to
was originated from the unfolded structure and thus the fluores- breakup of disulfide bonds and less ordered structures. Moreover,
cent aromatic amino acids transferred from core to surface. In Tp of ovomucin fluctuated during a F-T process, and the Tp of 5 F-
addition, Ho value of ovomucin increased from 700.50 to 823.75 T treated sample (116.80 C) was significant lower than that of
after 1 F-T cycle, and then decreased to values which were lower untreated sample (123.63 C). This was attributed to its flexible
than that of untreated samples, reaching 402.83 and 107.57 after structure (Donovan et al., 1975), which was easily affected by a
3 and 5 F-T cycles, respectively. This suggested that ovomucin F-T process. Finally, it was notable that the Tp of F-T treated egg
molecules unfolded after 1 F-T cycle, and then began to aggregate white was lower than that of untreated sample (145.10 C), which
with increasing number of F-T cycles. Finally, it was notable that suggested that weaker protein-protein interactions and partly
the Ho value of egg white increased from 158.29 to 232.89, denaturation occurred during the process.
241.02 and 238.05 after 1, 3 and 5 F-T cycles. The spontaneous The DH values obtained from the thermograms of DSC generally
adsorption of proteins from solution to the air interface is of cen- reflect competition between endothermic unfolding and exother-
tral importance to their foaming properties (Wang, Tao, et al., mic aggregation (Kato, Ibrahim, Watanabe, Honma, & Kobayashi,
2014). Hydrophobic patches on a proteins surface initially drive 1990). The DH of albumen proteins with different F-T cycles are
this process, and surface hydrophobicity is correlated with shown in Table 2. There was no significant change in DH of ovalbu-
improved foaming properties (Townsend & Nakai, 1983). There- min before and after a F-T treatment. This trend was in agreement
fore, the increase in Ho after a F-T treatment enhanced amphipathy with the SH and Tp results. The DH values of ovomucoid
of egg white and thus improved its foaming properties. decreased significantly after 5 F-T cycles treatment. This suggested

Table 1
Denaturation temperature (Tp) of untreated and treated albumen proteins (1, 3 and 5 F-T cycles). Different letters (ac) in the same column indicated significant differences
(P < 0.05).

F-T cycles Denaturation temperature Tp (C)

Ovalbumin Ovomucoid Ovotransferrin Lysozyme Ovomucin Egg white
Untreated 58.91 0.85 112.06 1.02b 113.44 1.25 118.19 1.09a 123.63 1.01a 145.10 1.37a
1 59.51 0.78 119.40 1.32a 110.83 1.21 112.42 1.17b 120.67 0.94ab 120.93 0.55c
3 58.56 0.21 119.50 0.78a 114.68 1.30 106.29 0.63c 122.49 1.36a 130.49 1.20b
5 58.82 0.60 120.14 0.42a 115.24 1.14 102.11 1.45c 116.80 1.52b 128.05 1.26b

Table 2
Enthalpy of denaturation (DH) of untreated and treated albumen proteins (1, 3 and 5 F-T cycles). Different letters (ad) in the same column indicated significant differences
(P < 0.05).

F-T cycles Enthalpy of denaturation DH (J/g)

Ovalbumin Ovomucoid Ovotransferrin Lysozyme Ovomucin Egg white
Untreated 148.9 4.3 204.0 5.8a 181.1 4.2b 132.5 1.1a 127.2 3.1b 94.0 3.3d
1 154.6 2.7 204.0 3.9a 215.9 1.9a 131.5 3.0a 155.3 1.8a 198.6 2.8a
3 153.8 3.8 199.4 2.6a 158.4 4.0c 106.3 3.1b 52.7 1.6c 142.1 2.5c
5 152.2 2.4 185.7 2.2b 182.6 2.7b 69.1 1.6c 155.1 2.3a 161.0 3.0b
 X. Duan et al. / Food Chemistry 228 (2017) 243248 247

Fig. 3. SEM images (1.0 k  magnification) of untreated and treated albumen proteins (1, 3 and 5 F-T cycles).

that ovomucoid underwent a progressive denaturation after 5 F-T

treatment involving disruption of ordered structures (Wang, Xu,
et al., 2014). Similarly, the DH values of lysozyme decreased from
132.5 J/g to 106.3 and 69.1 J/g after 3 and 5 F-T cycles treatment,
which was in agreement with the Tp result showing that the heat
stability of lysozyme decreased with increasing F-T cycles. For ovo-
transferrin and ovomucin, the DH values fluctuated during the pro-
cess. Finally, the DH value of egg white increased from 94.0 J/g to
198.6, 142.1, and 161.0 after 1, 3 and 5 F-T cycles, suggesting that
egg white underwent an increasing level of structure folding or
denaturation during the process.

3.4. Morphology

To have tangible evidence of the transformation of albumen

proteins microstructure, we employed SEM (Fig. 3). The untreated
samples developed spongy structure and had large pores, owing to
recrystallization. After a multiple F-T treatment, these albumen
proteins collapsed into irregular shapes and aggregated into larger
rigid structures, which was due to the growth of ice crystal and
redistribution of water. This suggested that partial rearrangement
and denaturation and aggregation occurred during a F-T process.
Combined with above-mentioned results, the forces involved in
rearrangement and denaturation of albumen proteins included
disulfide bonds, hydrophobic interactions and others. Foaming
properties of proteins are generally influenced by surface flexibility
and hydrophobic properties. Furthermore, proteins with random
coil flexible structure have better foaming properties than highly
ordered structures for its faster absorption rate into air interfaces
(Graham & Phillips, 1979). Therefore, a multiple F-T treatment
could lead to partial denaturation of these molecules, which poten-
tially contributes to the improvement of foaming properties.

3.5. Foaming properties

Fig. 4. Foaming ability (FA) and foaming stability (FS) of untreated and treated
albumen proteins (1, 3 and 5 F-T cycles). Different letters (ac) in the same protein
In this work, foaming properties of albumen proteins are deter-
indicated significant differences (P < 0.05). The foaming properties of ovomucin
mined and shown in Fig. 4, the foaming properties of ovomucin were not determined for its low solubility.
were not determined for its low solubility. For ovalbumin, it could
be seen that there was no significant difference in foaming ability
(FA) during a F-T process. This was in agreement with the SH and were lower than that of other proteins. This was in agreement with
DSC results indicating that ovalbumin maintained disulfide bonds previous study reporting that lysozyme exhibited low foaming
stability and thermal stability during a F-T process. Whereas the characteristics (Johnson & Zabik, 1981a). Meanwhile, a fluctuation
foaming stability (FS) of ovalbumin decreased after 1 F-T cycle, in FA and FS of lysozyme was seen during a multiple F-T treatment.
and then began to increase with increasing F-T cycles. For ovomu- Finally, it could be seen that the FS values of egg white were higher
coid and ovotransferrin, the FA gradually increased with increasing than that of other individual protein samples. This was because
F-T cycles. This was attributed to the unfolded and rearranged con- that egg white contained ovomucin, ovoglobulin and other albu-
formation due to a F-T treatment, enhancing the ability to absorb men proteins, which could complexed with each other via inter-
at air-water interfaces. Furthermore, the FA and FS of lysozyme molecular interactions. For example, ovomucin was reported to
248 X. Duan et al. / Food Chemistry 228 (2017) 243248

serve physical functions such as maintaining structure and viscos- Donovan, J. W., Mapes, C. J., Davis, J. G., & Garibaldi, J. A. (1975). A differential
scanning calorimetric study of the stability of egg white to heat denaturation.
ity of egg white albumen, which played a key role in improving FA
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1981a; Sunwoo & Gujral, 2015). Ovotransferrin also participates in of chicken egg white proteins using polyethylene glycol precipitation and
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7580.
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may not behave similarly in isolation and in mixture with other adsorption and surface denaturation. Journal of Colloid and Interface Science, 70
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24.1% to 28.0%, 31.7% and 32.6% after 1, 3 and 5 F-T cycles, respec- Herald, T., & Smith, D. (1989). Functional properties and composition of liquid
tively. The improvement in foaming properties was partially attrib- whole egg proteins as influenced by pasteurization and frozen storage. Poultry
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white contains a wide array of other components, including trace Vaclavik, V. A., & Christian, E. W. (2014). Eggs and Egg Products. In Essentials of Food
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and their potential interactions during a multiple F-T process is gluten-, glutenin-and gliadin-rich fractions. Food Hydrocolloids, 39, 187194.
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Acknowledgment 189198.
Wang, P., Tao, H., Wu, F., Yang, N., Chen, F., Jin, Z., & Xu, X. (2014). Effect of frozen
This work was supported by grants from the National Natural storage on the foaming properties of wheat gliadin. Food Chemistry, 164, 4449.
Wang, P., Xu, L., Nikoo, M., Ocen, D., Wu, F., Yang, N., ... Xu, X. (2014). Effect of frozen
Science Foundation of China (Nos. 31501426 and 31671859). storage on the conformational, thermal and microscopic properties of gluten:
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