International Journal of Biological Macromolecules 178 (2021) 136–142
Contents lists available at ScienceDirect
International Journal of Biological Macromolecules
journal homepage: http://www.elsevier.com/locate/ijbiomac
Impact of ice structuring protein on myofibrillar protein aggregation
behaviour and structural property of quick-frozen patty during
frozen storage
Fangfei Li a, Xin Du b, Yanming Ren c, Baohua Kong b, Bo Wang b, Xiufang Xia b,⁎, Yihong Bao a,⁎
a
b
c
College of Forestry, Northeast Forestry University, Harbin, Heilongjiang 150040, China
College of Food Science, Northeast Agricultural University, Harbin, Heilongjiang 150030, China
Heilongjiang Province Agricultural Products and Veterinary Drug Feed Technical Identification Station, Harbin, Heilongjiang 150090, China
a r t i c l e
i n f o
Article history:
Received 5 December 2020
Received in revised form 24 January 2021
Accepted 21 February 2021
Available online 23 February 2021
Keywords:
Ice structuring protein
Quick-frozen pork patties
Myofibrillar protein
Average particle size
Structure unfolding
a b s t r a c t
The goal of this study was to explore the cryoprotective effect of ice structuring protein (ISP) on the aggregation
behaviour and structural changes of myofibrillar protein (MP) from quick-frozen pork patties during frozen storage. Frozen storage causes the formation of large protein aggregates and weakens MP structures. After adding ISP
into patties, MP had a more stable aggregation system, which was manifested by a uniform particle size distribution and significantly higher absolute zeta potential (11.71 mV) than the control (9.56 mV) (P < 0.05). Atomic
force microscopy results showed that the surface roughness of MP aggregation decreased by 9.78% with ISP
after freezing for 180 d. Additionally, compared to patties without ISP, the MP carbonyl content from the ISPtreated patty decreased by 32%, and the free amino content increased by 14.99% during frozen storage. Results
from circular dichroism spectroscopy and fluorescence spectroscopy showed that MP secondary and tertiary
structure stability in patties improved with ISP. Overall, ISP has the potential to improve MP aggregation and
structural stability during frozen storage.
© 2021 Elsevier B.V. All rights reserved.
1. Introduction
As a type of convenient food, quick-frozen pork patties should be
quickly frozen at −30 °C for 30 min and then placed at −18 °C for processing, conservation and sale circulation [1]. These patties are popular
with consumers due to their convenient characteristics, satisfactory
quality and sensory properties [2]. Frozen storage is one of the most useful methods for food preservation and can effectively prolong the shelf
life of meat and meat products by inhibiting microbial spoilage and reducing enzyme activity and biochemical reaction rates [3,4]. However,
irreversible changes and quality deteriorations of meat products induced by the formation of ice crystals, recrystallization, oxidative reactions [5] and protein denaturation unavoidably occur during frozen
storage [6,7].
Myofibrillar protein (MP), which accounts for approximately
55–65% of meat muscle total protein, is responsible for many physicochemical properties of meat products. Improper MP aggregation and
structural degradation are critical factors that reduce protein functionalities, leading to the quality diversification of meat and meat products
[8]. For example, the formation of protein aggregates has been known
to yield decreasing gel qualities, such as water holding capacity, gel
⁎ Corresponding authors.
E-mail addresses: xiaxiufang@neau.edu.cn (X. Xia), baoyihong@163.com (Y. Bao).
https://doi.org/10.1016/j.ijbiomac.2021.02.158
0141-8130/© 2021 Elsevier B.V. All rights reserved.
strength and microstructure, during frozen storage [9]. Unfolding of
the MP structure induced by a high denaturation temperature may
cause MP gel to soften and rot, which degrades the final juiciness and
organization status of meat products [10]. Also, the destruction of protein structural integrity results in the shrinkage of interfilaments [11],
which lead to water moisture and loss of patty after freezing and
thawing [4]. Zhang et al. [12] showed that MP oxidative denaturation
may lead to a reduction in shrimp texture properties and flavour.
Thus, MP properties are critical to improving meat product quality during processing.
Emerging techniques, including high-intensity ultrasound [13], electric field-assisted freezing and high-pressure-assisted freezing [14],
have been used to inhibit meat quality deterioration by acting on the
MP structure. Cryoprotectants, such as alginate oligosaccharides [15],
xylooligosaccharides [16], ice structuring protein [17] and plant extract
[18], have also been reported to increase functional quality properties
when added to food. As a new type of antifreeze agent, ice structuring
protein (ISP) is a type of stress-tolerant protein [19] and has been discovered in different organisms, including bony fish [20], insects [21],
plants [22], fungi [23] and diatoms [24]. The protective function of
ISPs are based on their unique properties, which include ice plane affinity [25], thermal hysteresis [26], ice recrystallization inhibition ( [27]
and transient binding of an organism to ice [28]. ISP in frozen food can
control the growth morphology and aggregation of ice crystals [29,30],
F. Li, X. Du, Y. Ren et al.
International Journal of Biological Macromolecules 178 (2021) 136–142
2.6. Surface morphology
which can enhance cell integrity and reduce tissue damage [31]. In frozen meat systems, the inhibitory effect of ISP on meat quality deterioration [32] and peroxidation [30] during freezing processes has been
demonstrated. Previous studies have also shown that ISP can effectively
protect the quality of meat and meat products [33]. To date, information
concerning the impact of ISP on MP aggregation behaviour and structural properties in patties during frozen storage has not been reported.
The objective of this study was to identify the cryoprotective effect of
ISP on the aggregation behaviour and structural changes in MP and to
show the mechanism by which ISP maintains the MP characteristics of
quick-frozen pork patties during frozen storage.
The micromorphology of MP aggregation was measured using
atomic force microscopy (AFM). MP (40 mg/mL) was deposited onto
glass slides heated at 75 °C for 20 min and then immediately blowdried. Results were acquired using an AFM instrument (Veeco Instruments Inc., USA) in tapping mode.
2.7. MP primary structure
2.7.1. Carbonyl content measurement
The carbonyl group was measured using the method described by Li
et al. [4] and calculated using the absorption coefficient
(22,000 M−1 cm−1).
2. Materials and methods
2.1. Chemical
2.7.2. Free amino content determination
MP (0.25 mL, 1 mg/mL) was dissolved in a phosphate buffer
(pH 8.2), mixed with 2 mL of TNBS for 60 min, and kept away from
light at 50 °C. Absorbance was noted at 340 nm by a spectrophotometer,
and the contents of free amino acids were estimated using the L-leucine
standard curve.
The ice structuring proteins were procured by Nanjing Anfei Bio at a
purity above 95%. Piperazine-N,N′-bis-2-ethanesulfonic acid, sodium
chloride (NaCl), ortho-phthaldialdehyde, trichloroacetic acid,
hydrochloric acid, ethanol, ethyl acetate, guanidine, borax, 2,4dinitrophenylhydrazine, sodium dodecyl sulfate, guanidine hydrochloride, β-mercaptoethanol and L-leucine were purchased from Solabio
Corporation (Beijing, China). All chemicals were of analytical grade.
2.8. MP secondary structure
The extracted MP (0.2 mg/mL) was characterized on a circular dichroism (CD) instrument (Jasco J-815, Tokyo, Japan) with a scan rate
of 100 nm/min and a spectral range from 200 nm to 260 nm.
2.2. Patty preparation
Post-rigor pork shoulder and neck were purchased from Jiajiale
commercial abattoir (Harbin, Heilongjiang, China). Trimmed of visible
connectivity, the meat, ISPs (0% and 0.20%) and 12% ice water were
mixed for 5 min. The mixture was made into patties of approximately
100 g each (9 cm diameter and 2 cm thickness) and then kept at
−18 °C. A total of 30 patients were divided into 2 groups: the control
group (15 samples with 0% ISP) and the ISP group (15 samples with
0.20% ISP). Each group was randomly divided into 5 treatments and frozen for 0, 30, 60, 90 and 180 d. Samples were thawed at 4 °C overnight
before analysis.
2.9. MP tertiary structure
The MP tertiary structure was measured on a fluorescence spectrophotometer (Hitachi Co., Tokyo, Japan) under an excitation wavelength
of 283 nm, a slit width of 5 nm, a scanning wavelength of 300–400 nm
and a scan speed of 240 nm/min.
2.10. Statistical analyses
Data were analysed using by analysis of variance (ANOVA) with
Statistix 8.1 (Analytical Software, St. Paul, MN). The means ± standard
deviations were considered significant when P < 0.05. All graphs were
generated using SigmaPlot 12.5.
2.3. Myofibrillar protein preparation
MP was extracted via the following procedure and stored in a refrigerator (4 °C). Briefly, the minced patty was homogenized 3 times with 4
vol cold-extracted solution (pH 7.0) and then centrifuged at 6500 ×g for
15 min at 4 °C. The obtained pellet was washed 3 times with 4 vol
extracting buffer (0.1 M NaCl) and centrifuged at 6500 ×g for 15 min
at 4 °C again. The obtained MP extractions were kept at 4 °C.
3. Results and discussion
3.1. Protein aggregation behaviour
3.1.1. Particle size
The effects of ISP and frozen storage on myofibrillar protein aggregation was assessed with dynamic light scattering and are shown in
Fig. 1A and Table 1. At the start of frozen storage, a relatively uniform
and wide particle distribution was observed in the control and ISP
groups, which showed that the distribution of fresh samples was uniform and stable. During frozen storage, the first clear trend was an increase in the average particle size of MP from all samples. For the
control, d43 and d32 increased by 41.82% and 41.55%, respectively, after
freezing for 180 d. The changes in the mean particle diameters were primarily caused by myofibrillar protein unfolding and an increase in protein surface area [9]. Additionally, the disulphide bridges, hydrogen
bonds, and hydrophobic interactions between proteins were also
strengthened, which promoted the aggregation of MP and increased
the average particle size [35].
Compared to the control, the particle size gradually decreased with
the addition of ISP. After freezing for 180d, d43 and d32 of the samples
with ISP significantly decreased by 4.57% and 4.26% compared to those
of the control (P < 0.05). Similarly, d0.1, d0.5 and d0.9 of the samples
with ISP significantly decreased by 4.91%, 4.73% and 5.31%, respectively
2.4. Particle size distribution
MP particle size was measured with 1.5 mL of MP-dissolving liquid
(1 mg/mL) in a Mastersizer 2000 instrument (Malvern, UK). The aggregate degree of the MP was determined by the following statistical parameters: d43-volume-mean diameter; d32-volume-surface-mean
diameter; and dV,0.1, dV,0.5, and dV,0.9 represented the size of the particle
for which 10%, 50% and 90% of the sample was below this size,
respectively.
2.5. Zeta potential
The MP zeta potential (0.1 mg/mL) was evaluated using a zeta potential analyser (Zeta Plus, Brookharen, Holtsville, NY, USA), and certain
modifications were made based on the method described by Beliciu and
Moraru [34]. After the MP concentration was adjusted by double
steaming wate, 1 mL of MP was transferred to the measuring tank,
and the average measurement was 6.
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International Journal of Biological Macromolecules 178 (2021) 136–142
Frozen storage (d)
0
6
0
30
60
90
180
5
-5
Zeta potential
4
3
2
1
0
1
Par
(A)
0
m)
90
-15
180
1000
Aa
Bb
Cb
(d)
180
ge
a
90
r
60
ol sto
30
-20
ntr
0
Co ozen
r
(B)
F
10
t icl
e si 100
ze (
μ
60
30
-10
Ca
Da
Eb Da
Aa
Ba
Db
Control
ISP
P
IS
Fig. 1. Effect of ISP on the particle size and Zeta potential of myofibrillar protein of pork patties during frozen storage. a–b indicate significant differences (P < 0.05) with the same frozen
storage of different treatments. A–E indicate significant differences (P < 0.05) with the same treatment after different frozen storage.
(Fig. 2D) by AFM [41]. As shown in Fig. 2, the bright and dark regions
(Fig. 2A and C) occur in convex and concave areas of the MP aggregation
surface, respectively. Additionally, the red arrows (Fig. 2B) indicate the
highest and the lowest locations on the cross-section of each image
[9,42].
The surface morphology of MP aggregation was smooth and homogeneous before frozen storage. The MP aggregation surface had a
brighter and concave area, and a larger distance between the highest
and lowest points after freezing for 180 d. A significant increase from
23.8 nm to 64.4 nm in the average roughness after 180 d was also observed in the control group (P < 0.05). These observations indicated
that the MP in patties underwent substantial molecular changes and
gradually formed protein aggregates during frozen storage [43]. The
consistent production of these aggregates was due to protein oxidation
and denaturation, which were caused by ice crystals growing after
freezing [44].
Compared to the control, samples from the ISP group had a
smoother and more homogeneous gel morphology and lower Rg after
the same storage time. Results showed that the behaviour of protein aggregation was inhibited by adding ISP. Kong et al. [31] found that ISP
added to patties could decrease the degree of muscle cell damage by
inhibiting recrystallization. Therefore, the adjunction of ISP could maintain protein structure stability to some extent by preventing protein oxidation during frozen storage [27].
(P < 0.05). Results thus showed that ISP added to patties effectively decreased protein aggregation [36]. ISP inhibits the protein aggregation of
pork patties during frozen storage, which can be associated with less
damage to muscle tissue. ISP can also inhibit recrystallization and prevent irreversible mechanical damage to tissues from ice crystals. Additionally, the protein denaturation induced by freezing can be delayed
during frozen storage [27].
3.1.2. Zeta potential
The zeta potential is a critical stability indicator of myofibrillar proteins that can be used to evaluate the driving force for electrostatic interactions between charged biopolymers [37]. As shown in Fig. 1B,
control MPs exhibited a relatively low net negative charge
(−15.23 mV) because weakly acidic proteins, such as aspartic acid
and glutamic acid, form during frozen storage. As frozen storage lengthened, the zeta potential significantly increased to −9.56 mV (P < 0.05),
which could be due to partial protein denaturation and aggregate formation [38]. The results are similar to those in Fig. 1A, in which protein
aggregation occurred during frozen storage.
During storage, patties with ISP had significantly lower MP zeta potentials. Particularly after freezing for 180 d, the MP zeta potential with
ISP decreased by 22.48% compared to the control. Results showed that
the addition of ISP can inhibit MP aggregation after freezing because
the formation of ice crystals was controlled by ISP, thus decreasing protein freezing denaturation and aggregation [39]. The MP particle size
with ISP was more uniformly distributed and had a larger electrostatic
repulsive force, which could prevent protein aggregation [40].
3.2. Protein structural property
3.2.1. Carbonyl content
Carbonyl content is one of the most important characteristics to
evaluate changes in a protein's primary structure. The effect of ISP and
frozen storage on the carbonyl groups is shown in Fig. 3A. After being
3.1.3. Surface morphology
The surface state of MP aggregation is shown in 2D images (Fig. 2A),
cross-sections of images (Fig. 2B), 3D images (Fig. 2C), and Rq values
Table 1
Effect of ISP on the particle size of myofibrillar protein of pork patties during frozen storage.
Frozen storage (d)
Particle size (μm)
d32
d43
Control
ISP
0
30
60
90
180
0
30
60
90
180
Ea
86.41 ± 0.45
90.47 ± 0.77Da
96.20 ± 0.27Ca
109.28 ± 0.73Ba
122.54 ± 0.85Aa
86.64 ± 1.16Da
89.08 ± 0.66CDa
91.84 ± 1.92Cb
104.95 ± 1.42Bb
116.94 ± 2.03Ab
40.79
46.46
50.74
54.93
57.74
41.48
40.14
46.94
50.88
55.28
dV,0.1
±
±
±
±
±
±
±
±
±
±
Ea
0.89
0.44Da
1.32Ca
0.77Ba
0.19Aa
1.46Da
1.07Db
0.04Cb
1.48Bb
1.04Ab
20.21
22.73
24.34
26.13
27.31
19.91
21.19
23.19
24.26
25.97
dV,0.5
±
±
±
±
±
±
±
±
±
±
Da
0.41
0.54Ca
0.57Ba
0.71Aa
0.05Aa
0.82Ca
0.89Ca
0.60Ba
0.22ABb
0.76Ab
61.27
68.07
72.99
87.92
92.58
63.78
68.75
71.49
76.02
88.20
dV,0.9
±
±
±
±
±
±
±
±
±
±
Da
3.51
0.80Ca
1.00Ba
1.16Aa
0.84Aa
1.47Da
0.94Ca
1.12Ca
0.26Bb
1.28Ab
179.61
186.96
198.90
224.06
254.83
181.22
177.54
183.98
215.89
241.31
±
±
±
±
±
±
±
±
±
±
1.29Ea
0.13Da
0.96Ca
4.16Ba
3.43Aa
4.16Ca
2.04Cb
4.26Cb
1.17Bb
3.50Ab
indicate significant differences (P < 0.05) with the same frozen storage of different treatments. A–E indicate significant differences (P < 0.05) with the same treatment after different
frozen storage.
a–b
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International Journal of Biological Macromolecules 178 (2021) 136–142
Fig. 2. Effect of ISP on the surface roughness of myofibrillar protein (3.0 × 3.0 μm2 scan size) of pork patties during frozen storage. A: The scale displayed on the left ranges from 0 to 200 nm.
The straight line on the 2D image represent the incision location of cross-sections. B: The image cross-sections correspond to the section cut by the straight line on the 2D image. a–b indicate
significant differences (P < 0.05) with the same frozen storage of different treatments. A–C indicate significant differences (P < 0.05) with the same treatment after different frozen storage.
after 30 d of frozen storage. After freezing for 180 d, patties with ISP
had lower carbonyl contents than patties without ISP, with a significant
difference of 32% (P < 0.05). The large ice crystals formed and destroyed
the muscle, which caused MP freezing and oxidative denaturation [4].
With ISP bound, the mechanical damage of muscle was controlled by
inhibiting the growth of ice crystals [48]. Therefore, the degree of protein degeneration decreased.
frozen for 180 d, there was a significant increase in carbonyl contents in
all samples (P < 0.05). Protein oxidation occurs frequently and leads to
the formation of carbonyl compounds [45]. During long-term frozen
storage, the formation of ice crystals caused damage to the muscle cell
ultrastructure, which released free radicals. This process aggravates oxidative changes in proteins and the formation of carbonyl groups [6,46].
Additionally, the NH and NH2 of protein side chains easily undergo oxidative changes and form carbonyl compounds [16]. Cheng, Xu, Xiang,
Liu, and Zhu [47] also found that carbonyl accumulation in minced
pork slice proteins increased significantly during frozen storage.
Although no significant differences were detected at the beginning
of frozen storage in all samples (P > 0.05), the control sample showed
a sharp rise in carbonyl content compared to samples treated with ISP
3.2.2. Free amino content
Changes in the free amino contents of proteins can be used to evaluate alterations in MP primary structure [49]. As shown in Fig. 3B, all
samples had significantly decreasing free amino contents after 180 d
(P < 0.05). The free amino acid contents of the control group and the
139
Control
ISP
3
Aa
Ba
Ca
2
Ab
Ab
Bb
Da
1
Ea Ca
Cb
Free amino (nmol/mg protein)
International Journal of Biological Macromolecules 178 (2021) 136–142
Carbonyl contents (nmol/mg protein)
F. Li, X. Du, Y. Ren et al.
Control
ISP
Aa Aa
Bb Ba
Ca
Da
Cb
Da
Db
80
Eb
60
40
0
0
(A)
100
30
60
90
0
180
Frozen storage (d)
(B)
30
60
90
180
Frozen storage (d)
Fig. 3. Effect of ISP on the primary structure of myofibrillar protein of pork patties during frozen storage. a–b indicate significant differences (P < 0.05) with the same frozen storage of
different treatments. A–E indicate significant differences (P < 0.05) with the same treatment after different frozen storage.
recrystallization and changed ice crystal morphology. Thus, cellstructure integrity is maintained, and the degree of protein oxidation
decreased [17]. Considering the result regarding carbonyl content, ISP
can decrease oxidative damage to patties.
ISP group decreased 23.09% and 12.99%, respectively. Proteins are susceptible to attack by reactive oxygen species while frozen [50]. The
NH2 groups of amino acid residues could be transformed into carbonyl
groups through a deamination process during frozen storage. Additionally, derivatives then reacted with available NH2 groups, thus causing a
further decrease in free amino content [51,52]. Results thus responded
to changes in carbonyl contents.
Compared to the control, the ISP group showed a marginally lower
decrease in free amino contents during frozen storage. After the same
frozen-storage period, the samples with ISP had higher amino contents
than those without ISP. Particularly at 180 d, the difference in amino
contents was as high as 14.99%. Mechanical damage to muscle caused
by recrystallization leads to protein oxidative denaturation. The higher
amino contents in the ISP group indicated that ISP hindered
3.2.3. MP secondary structure
The stability of MP secondary structural characteristics can be described by circular dichroism (CD) far-ultraviolet spectra, which comprise α-helices, β-sheets, β-turns and random coils [57]. Fig. 4 shows
that the far-ultraviolet wavelengths of all samples exhibit typical negative peaks at 190–250 nm. With longer frozen storage, the secondary
structures of all samples showed a similar trend: the percentage of αhelixes and β-turns gradually decreased, while the fraction of β-sheets
and random coils significantly increased (P < 0.05). The protein
Fig. 4. Effect of ISP on the secondary structure of myofibrillar protein of pork patties during frozen storage. (Control: A and B; ISP: C and D). a–b indicate significant differences (P < 0.05)
with the same frozen storage of different treatments. A–C indicate significant differences (P < 0.05) with the same treatment after different frozen storage.
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International Journal of Biological Macromolecules 178 (2021) 136–142
0d
30 d
60 d
90 d
180 d
5000
6000
Fluorescence intensity
Fluorescence intensity
6000
4000
3000
2000
1000
5000
4000
3000
2000
1000
0
300
(A)
0d
30 d
60 d
90 d
180 d
320
340
360
380
0
400
300
320
(B)
Wavelength (nm)
340
360
380
400
420
Wavelength (nm)
Fig. 5. Effect of ISP on the tertiary structure of myofibrillar protein of pork patties during frozen storage. (Control: A; ISP: B).
protein aggregation, as shown by the MP particle size distribution,
zeta potential and AFM. Additionally, the lower carbonyl contents and
higher free amino contents of the ISP group showed that MP primary
structure degradation was inhibited by ISP. Concurrently, the secondary
and tertiary structures of MP from pork patties with ISP tended to stabilize after frozen storage. These results systematically showed the mechanism by which ISP inhibits the frozen denaturation of patties based on
the importance of the aggregation behaviour and structure of MP.
Therefore, this study provides useful information for the future study
of the cryoprotective effect of ISP on meat and meat products. Future research should focus on exploring the molecular interactions between
the structure of ISP and MP spatial conformation at the molecular level.
oxidation aggravation induced by freezing causes a decrease in the percentage of α-helical conformation [16]. Additionally, freezing destroys
hydrogen bonds and disorders the MP structure, which transforms the
helix structure to a random structure [40]. Jin et al. [53] showed that
the protein structure stability of duck breast muscle decreased due to
the hydrophobic interaction of protein during frozen storage.
When the storage time was the same, patties with ISP exhibited
more stable secondary protein structures, as shown in the significantly
fewer changes in the contents of α-helices, β-sheets, β-turns and random coils (P < 0.05). Results thus suggested that ISP decreased the degree of hydrophobic residue exposure and inhibited interaction among
amino acid side chains. ISP can modify ice crystal morphology and inhibit recrystallization, minimizing freezing and oxidation damage [31].
Nian et al. [27] also reported the same conclusion that antifreeze proteins could maintain the secondary structural stability of large-mouth
bass MP during F-T cycles.
Credit author statement
Fangfei Li: Methodology, Software, Validation, Formal analysis, Investigation, Writing-Original Draft, Funding acquisition.
Xin Du: Investigation, Software.
Yanming Ren: Formal analysis.
Baohua Kong: Conceptualization, Resources.
Bo Wang: Investigation, Formal analysis.
Xiufang Xia: Conceptualization, Writing - Original Draft, Supervision, Funding acquisition.
Yihong Bao: Conceptualization, Resources.
3.2.4. MP tertiary structure
Fluorescence intensity (FI) describes the changes in MP tertiary
structure via the reflection of tryptophan (Trp) residues towards the polarity of the microenvironment [54]. Changes in the fluorescence intensity of MP are shown in Fig. 5. All samples exhibited a broad spectrum at
336 nm, and as the frozen storage period increased up to 180 d, FI decreased significantly (P < 0.05). The exposed and denatured indole
side chains of Trp residues, which are located in the head and tail regions of myosin, cause the formation of protein aggregation and the
weakness of protein conformation affected by freezing [55]. Additionally, MP unfolding and partial changes in protein conformation can
lead to the exposure of Trp in amino acids. The decrease in FI of MP
was also due to increased steric hindrance, which was caused by the aggregation of MP and the rise in hydrophobic interactions. This result is
consistent with a study that showed that the reductions in FI could be
due to the exposure and denaturation of Trp residue indole side chains
towards the aqueous environment or polarity of the environment [56].
Compared to the control, ISP MP samples exhibited more stable FI
after frozen storage. After adding ISP, recrystallization in patties was
inhibited, which decreased the damage to protein structure. ISP
inhibited myofibrillar protein denaturation and strengthened the hydrogen interactions between protein molecules, which could control
the formation and growth of ice crystals during frozen storage [40]. Similar results have been described by Nian et al. [27], who showed that the
antifreeze protein could maintain a stable myofibrillar protein tertiary
structure after freezing and thawing.
Acknowledgements
This study was supported by Northeast Forestry University's research funding for talent introduced (grant no. 60201520109) and the
National Natural Science Foundation of China (grant no. 31571859
and 31771903).
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4. Conclusions
The goal of this study was to investigate the protective effect of ISP
on the MP aggregation state and structural changes in pork patties
during frozen storage. Results show that ISP decreased the degree of
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