Eur Food Res Technol (2012) 234:863872

DOI 10.1007/s00217-012-1700-4

ORIGINAL PAPER

Functional properties of gelation-like protein hydrolysates

from scallop (Patinopecten yessoensis) male gonad
Wen-Gang Jin Hai-Tao Wu Bei-Wei Zhu
Xu-Qin Ran

Received: 17 November 2011 / Revised: 10 February 2012 / Accepted: 17 February 2012 / Published online: 2 March 2012
Springer-Verlag 2012

Abstract Gelation-like protein hydrolysates from scallop

(Patinopecten yessoensis) male gonad (SMG) were
obtained by enzymatic hydrolysis using neutrase. Functional properties of SMG hydrolysates (SMGHs) with different degree of hydrolysis (DH: 4.94, 6.84, 7.53 and
11.86%, respectively) were evaluated with the objective to
investigate the relations between hydrolysis characteristics
and functionalities. The results showed that hydrolysis with
neutrase improved the gelation property, solubility, waterholding capacity (WHC), oil-holding capacity (OHC), and
surface hydrophobicity (SH), but not foaming capacity
(FC) of SMG. The SMGHs at high DH (11.86%) showed
better gelation property and solubility than that at low DH
(4.947.53%). However, the maximum values of WHC,
OHC, and SH of SMGHs were found at DH of 4.94%,
significantly higher than (p \ 0.05) or equivalent to
(p [ 0.05) that of soy protein isolate (SPI) for WHC and
OHC. Emulsifying capacity of SMGHs is independent of
W.-G. Jin  B.-W. Zhu
College of Food Science and Engineering,
Northwest A&F University, Yangling Shaanxi
712100, Peoples Republic of China
H.-T. Wu  B.-W. Zhu (&)  X.-Q. Ran
School of Food Science and Technology,
Dalian Polytechnic University, Dalian
116034, Peoples Republic of China
e-mail: zhubeiwei@163.com
H.-T. Wu  B.-W. Zhu
Engineering Research Center of Seafood,
Ministry of Education, Dalian 116034,
Peoples Republic of China
H.-T. Wu  B.-W. Zhu
National R&D Branch Center for Shellfish Processing (Dalian),
Dalian 116034, Peoples Republic of China

DH, but restricted by pH environment. The emulsifying

activity index of all SMGHs was significantly higher than
that of SPI in pH 5 (p \ 0.05) and slightly higher than or
equivalent to that of SPI in pH 7. Meanwhile, SMG and
SMGHs were abundant in glycine, lysine, alanine, glutamic
acid, and aspartic acid, containing all the essential amino
acids (41.6342.90% of the total amino acids). These
results imply that SMGHs might be utilized as multifunctional and nutritive ingredients in food industry.
Keywords Scallop (Patinopecten yessoensis) gonad 
Neutrase  Gelation  Protein hydrolysate  Controlled
hydrolysis  Functionality
Abbreviations
SMG
Scallop (Patinopecten yessoensis) male gonad
SMGHs Scallop (Patinopecten yessoensis) male gonad
hydrolysates
DH
Degree of hydrolysis
SPI
Soy protein isolate
WHC
Water-holding capacity
OHC
Oil-holding capacity
EAI
Emulsifying activity index
ANS
1,8-Anilinonaphthalenesulphonate

Introduction
Scallop (Patinopecten yessoensis) is an economically
important edible shellfish, widely cultured in East Asia.
The main edible part of scallop body is adductor muscle,
which is traditionally regarded as delicacy and has high
nutritive value. During processing, a large quantity of

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by-products, such as shells, mid-gut gland, mantle lobe,

gonad (ovary and testis), accounting for more than 60% of
the scallop body, are usually discarded as industrial wastes
[1]. Gonads are also edible part of scallop body somewhere, for instance, 95% of scallops are processed as an
adductor muscle and gonad roe-on product in UK [2],
whereas adductor muscle product is the main processing,
consuming, and exporting commodity in China. Scallop
gonads, together with mid-gut gland and mantle lobe, are
deemed as low-valued by-products, which contain many
bioactive materials such as polysaccharide [3], enzyme [4]
and protein/peptide [5]. To reduce the burden on the
environment, effective utilization and disposal methods of
these wastes are required.
So far, efforts have been made for transforming scallop
wastes to value-added products. Mukhin et al. [6] studied
a protein hydrolysate enzymatically produced from scallop wastes as nutrient for microorganism cultivation. Lee
et al. [7] purified a lysozyme from the viscera of scallop
by ion exchange, gel permeation, and affinity chromatographies. Oyamada et al. [1] found three mycosporinelike amino acids from scallop ovaries, which might be
used in cosmetics and toiletries as a UV protectors and
activators of cell proliferation. Some biologically active
substrates such as carotenoid [8, 9], lipid [10] and polysaccharide [3] were also extracted from scallop viscera.
However, there are few studies focused on protein
hydrolysates obtained from scallop by-products for food
use and health benefit.
The use of enzyme technologies for protein recovery
and modification from food wastes and marine by-products
is widespread, which has led to the production of a broad
spectrum of novel food ingredients with improved functionalities [11, 12] and various biological activities such as
antioxidant activity [12, 13], anticancer activity [14],
antihypertensive activity [15], and antimicrobial activity
[16]. In this field, our previous studies have successfully
obtained protein hydrolysates from abalone (Haliotis discus hannai Ino) viscera and sea urchin (Strongylocentrotus
nudus) gonads, which exhibited good antioxidant activities
[17, 18].
In the previous attempts for preparing scallop (Patinopecten yessoensis) gonad hydrolysates, we found that
neutrase-treated male gonad showed a unique characteristic
of gelation [19]. This phenomenon urged us to study
functional properties of the resulting hydrolysate. Several
researchers reported that hydrolysis degree to a certain
extent affected the functionality of protein hydrolysates
[13, 20, 21]. Therefore, the present work was performed to
investigate the effect of hydrolysis degree on functional
properties of scallop (Patinopecten yessoensis) male gonad
hydrolysates (SMGHs).

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Eur Food Res Technol (2012) 234:863872

Materials and methods

Materials and chemicals
Scallops (Patinopecten yessoensis) were supplied by Hailin
Aquatic Products Foodstuff Co., Ltd (Dalian, China) in
March 2010. After dissection, the male gonads with milkwhite color were collected, vacuum freeze-dried, crushed,
and the gonad powder was stored at -20 C before use.
Neutrase (Bacillus subtilis var. amyloliquefaciens strain
1398) was purchased from Pangbo Biological Engineering
Co., Ltd (Nanning, China) and stored at 4 C until it was
used for the hydrolysis experiments. A commercial soy
protein isolate (SPI) was purchased from Sanwei Soy
protein Co., Ltd (Linyi, China). All other chemicals were
of analytical grade.
Preparation of protein hydrolysates
Freeze-dried SMG powder was mixed with deionized water
at a ratio of 1:20 (w/v). The mixture was heated at 95 C
for 10 min to denature native proteins. After cooling, the
mixture was adjusted to a required pH using 0.5 M NaOH
and pre-incubated at 50 C for 10 min. The enzymatic
hydrolysis was initiated by adding neutrase at a dose of
3,000 U/g protein with continuously stirring at 50 C. The
pH was maintained at 7.0 by adding 0.5 M NaOH during
the reaction. After 10, 20, 30, and 180 min of hydrolysis,
the enzyme was inactivated by heating at 95 C for 10 min.
When they were cooled at room temperature, the hydrolysates were obtained and lyophilized using VirTis
2KBTES-55 freeze dryer (Gardiner, NY, USA) and stored
at -20 C until use. Crude protein of SMG powder was
determined according to AOAC methods 981.10 [22].
Evaluation of degree of hydrolysis
Degree of hydrolysis (DH) was defined as the percent ratio
of the number of peptide bonds cleaved to the total number
of peptide bonds in the substrate studied. It was assayed
using pHstat method, and the calculation formula was
expressed as below [23]:
DH%

B  Nb 1
1
 
 100
a htot
Mp

where B is the amount (mL) of NaOH consumed to keep

pH constant during enzymatic reaction; Nb is the normality
of NaOH; Mp is the mass of protein (g); htot is the total
number of peptide bonds in the substrate, which is assured
to be 7.5 meq/g according to Mahmoud et al. [24]; a is the
average degree of dissociation of the a-NH2 groups
released during hydrolysis and expressed as below:

Eur Food Res Technol (2012) 234:863872

10pHpK
1 10pHpK

where pH and pK are the values at which the proteolysis

was conducted.
Assay of functional properties
Gelation properties
Gelation properties were determined by the method of
Severin and Xia [11] with slight modifications. Sample
suspensions of 3070 mg/mL with increments of 5 mg/mL
were prepared in 3 mL of deionized water. The test tubes
containing the suspensions were heated for 1 h in a boiling
water bath followed by rapid cooling under cold, running
tap water. The test tubes were further cooled for 3 h at
4 C. The least gelation concentration was determined
when the sample from the inverted test tube did not fall or
slip.
Nitrogen solubility

865

measured, and OHC was calculated as the mL of oil

absorbed per gram of sample.
Emulsifying activity index
Emulsifying activity index (EAI) was determined according to the method of Zhao et al. [26] with slight modification. Briefly, 30 mg of various samples was dissolved in
30 mL of deionized water, and the solutions were adjusted
to pH 3.0, 5.0, 7.0, and 9.0 with 0.5 M HCI or 0.5 M
NaOH. 10 mL of soy oil was added to each sample and
homogenized in an Ultra-Turrax T25 (IKA-Labortechnik,
Staufen, Germany) for 1 min at 10,000 rpm. Fifty microliters of emulsion was taken from the bottom of the
homogenized emulsion, immediately after homogenization, and diluted (1:100, v/v) in 0.1% (w/v) SDS solution.
After shaking in a vortex mixer for 5 s, the absorbance of
the diluted emulsions was recorded at 500 nm using a UV
Vis spectrophotometer (Purkinje General Instrumental Co.,
Beijing, China). EAI was calculated using the following
equation:
EAIm2 =g

Solubility was determined over a range of pH from 2.0 to

10.0 as described by Balti et al. [21] with slight modification. Samples (0.1% w/v) were dispersed in deionized
water, and the pH was adjusted using 0.5 M HCl or 0.5 M
NaOH solution. The dispersions were stirred at 25 C for
30 min and centrifuged at 9,5009g for 20 min. The
nitrogen content in the supernatant was determined by the
biuret method. Sample nitrogen concentration was determined by Kjeldahl method (AOAC 980.10) [22]. The
solubility was expressed as the formula below:
Supernatant nitrogen concentration
Solubility%
 100
Sample nitrogen concentration
Water-/oil-holding capacity
Water-/oil-holding capacity was determined according to
the method described by Wasswa et al. [25] with minor
modifications. For water-holding capacity (WHC), briefly,
samples (0.5 g) were dissolved with 20 mL of deionized
water in centrifuge tubes and dispersed with a vortex
mixer for 30 s. The dispersion was allowed to stand at
room temperature for 1 h and centrifuged at 2,0009g for
20 min. Difference between the initial volume of deionized water added to the sample and the volume of the
supernatant was determined. The results were expressed
as mL of water absorbed per gram of sample. For oilholding capacity (OHC), the procedure was the same as
that of water-holding capacity except that the 20 mL of
deionized water was replaced by 10 mL of soybean oil.
The volume of oil separated from the sample was

2  2:303  A  DF
C  /  h  10; 000

where DF is the dilution factor (100), C is the initial

concentration of protein sample (g/mL), is the optical
path (1 cm), h is the fraction of oil used to form the
emulsion (0.25), and A is the initial absorbance of the dilute
emulsions.
Foaming properties
Foaming capacity (FC) and stability were evaluated
according to the method described by Mohamed et al. [27].
Briefly, 30 mL of various sample dispersion (3%, w/v)
was mixed thoroughly using an Ultra-Turrax T25 (IKALabortechnik, Staufen, Germany) at 9,500 rpm for 2 min
in a 250-mL graduated cylinder. After 30 s, total volume of
the liquid was measured. Difference between the initial
volume and the total volume was expressed as foam volume. The cylinder was placed at room temperature for
10 min or 30 min, and foam stability was estimated by
measuring the residual foam volume.
Surface hydrophobicity
Surface hydrophobicity (SH) was determined using
1,8-anilinonaphthalenesulphonate (ANS), as a fluorescent
probe, according to the method of Mu et al. [28] with slight
modification. Briefly, sample solutions were prepared in
0.01 M phosphate buffer (pH 7.0) with various concentrations (0.2, 0.15, 0.12, 0.06, 0.03, 0.015%, w/v). Twenty
microliters of 8 mM ANS in 0.01 M phosphate buffer (pH

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7.0) was added into each of 4.0 mL of sample solutions,

and mixed well by vortexing for 10 s. Fluorescence
intensity of these solutions was measured at 390 nm of
excitation and 484 nm of emission using an F-2700 Fluorescence Spectrophotometer (Hitachi High Technology
Corporation, Tokyo, Japan). The initial slope of the fluorescence intensity versus sample concentration plot was
calculated by linear regression analysis and used as an
index of surface hydrophobicity.
Amino acid analysis
The amino acid profiles of SMG and SMGHs were determined according to the method of Sanger [29] with slight
modification. Amino acid composition was determined by
high-performance liquid chromatography equipped with a
ODS amino acid column (4.6 mm 9 250 mm), Elite-AAK
(Dalian Elite Analytical Instrumental Co. LTD). Total
amino acid residues were determined after hydrolysis at
110 C for 24 h with 6 M HCl prior to the derivatization
with 2,4-dinitrofluorobenzene. The concentrations of the
specific amino acids were determined from their respective
absorption intensities, which were calibrated to the known
concentrations of amino acid standards (Elite, Dalian,
China). Alkaline hydrolysis was also done for determination of tryptophane level.
Statistical analysis
All the experiments were performed in triplicate, and the
results are expressed as mean standard deviation
(n = 3), except for amino acid composition analysis. The
independent sample t test in SPSS 11.5 Software for
Windows was used to determine the level of significance
(p \ 0.05).

Results and Discussion

The dried SMG containing 81.66% of protein was hydrolyzed by neutrase at required conditions. SMGHs at
hydrolysis period of 10, 20, 30, and 180 min were prepared, and DH value of each protein hydrolysate was 4.94,
6.84, 7.53, and 11.86%, respectively (Fig. 1a). All the
protein hydrolysates of SMG indicated gelation-like profiles (Fig. 1b). Thus, gelation properties of SMGHs were
further investigated according to the inverted test tube
method as comparison with SMG and SPI. As shown in
Table 1, SMG and SPI did not form gel over a concentration
range from 30 to 70 mg/mL. However, all SMGHs (DH:
4.94, 6.84, 7.53, and 11.86%, respectively) formed good gel
when the sample concentration reached 65 mg/mL. Moreover, the minimum sample concentration to form gel

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Eur Food Res Technol (2012) 234:863872

decreased from 65 to 50 mg/mL as DH increased from 4.94

to 11.86%.
It is generally thought that hydrolysis leads to a decrease
in gelation properties, as it reduces the molecular weight
and increases the hydrophilicity of the proteinaceous
material by introducing charged groups. On the other hand,
upon limited hydrolysis the effective hydrophobicity of
certain globular proteins could be increased through
exposure of buried apolar residues, resulting in aggregate
formation. Creusot and Gruppen [30] have pointed out that
the enhancing effect of hydrolysis on protein gelation is a
specific property of a hydrolysate, since it is only observed
within specific combinations of substrates, enzymes, and
degrees of hydrolysis. Gelation also depends on disulfide
bonds, hydrophobic interactions, and charged groups [11].
Depending on the fact whether hydrolysis is limited or
extensive and whether the substrate is native or not, it is
assured that different enhancing mechanisms are involved
[30]. We found that the denatured SMG suspensions
incubated at 50 C in pH 7.0 for 180 min did not form gel,
and we got the same results when SMG suspensions were
hydrolyzed with trypsin and pepsin (data not shown).
These results suggest that gelation property of SMGHs is
perhaps the concerted work of protein type and enzyme
specificity.
Like gelation properties of SMGHs, it has been reported
that enhanced aggregation and gelation after proteolysis of
whey, soy, and sunflower proteins can be obtained with
many proteases. For example, the hydrolysis of a whey
protein isolate by alcalase [31, 32] or Bacillus licheniformis
proteinase [33, 34] led to gelation. Zhong et al. [35] found
that papain and alcalase also induced the gelation of soy
protein isolate. In addition, the gelation properties of sunflower protein hydrolysate obtained from trypsin has been
reported by Sanchez and Burgos [36, 37]. The gelation
property of SMGHs may be of interest for food applications, whereas the influence factors and underlying mechanism of neutrase-induced gelation of SMGHs still
deserves further study.
Figure 2 presents the nitrogen solubility profile of SMG
and SMGHs with different DHs (4.94, 6.84, 7.53 and
11.86%) in the pH range of 2.010.0. All hydrolysates
showed relatively flat curve without obvious isoelectric
points, whereas the SMG and SPI showed U-shaped profiles across the pH range from 2.0 to 10.0. The SMGHs had
a slightly higher solubility in alkaline pH than in acidic pH,
which might be explained by the difference of amino acid
net charge after enzymatic hydrolysis as it would affect the
solubility of pH change [38]. Figure 2 also demonstrates
that the solubility of hydrolysates increased with the
increasing of DH, indicating a positive relationship, which
is in agreement with the previous reports [21, 27, 39]. It has
been suggested that an increase in the solubility of protein

Eur Food Res Technol (2012) 234:863872

867

12

DH: 7.53%

Degree of hydrolysis /%

10

DH: 11.86%
DH: 6.84%

DH: 4.94%
6

0
0

30

60

90

120

150

180

Time /min

SMG
(50 , pH7.0-180min)

SMG
(DH: 0%)

SMGH
(DH: 6.84%)

SMGH
(DH: 4.94%)

SMGH
(DH: 7.53%)

Fig. 1 Hydrolysis profile of SMG and gelation-like profiles of

SMGHs with different degree of hydrolysis. a SMGHs with different
degree of hydrolysis were obtained through the controlled hydrolysis
for 10, 20, 30, and 180 min using neutrase, and the arrows represented

SMGH
(DH: 11.86%)

the corresponding DH (4.94, 6.84, 7.53, and 11.86%). b SMGHs with

different DH were cooled and photographed. The denatured SMG
suspensions either incubated at 50 C in pH 7.0 for 180 min or not
(DH: 0%) were used as negative controls

Table 1 Gelation properties of SMG, SPI, and SMGHs with different degree of hydrolysis
Samples

Concentrations (mg/mL)
30

35

40

45

50

55

60

65

70

SMG (DH:0%)

DH: 4.94%

9/H

9/H

9/H

9/H

9/H

DH: 6.84%

9/H

DH: 7.53%

9/H

DH: 11.86%

9/H

SPI

9/H

9/H

9/H

9, the sample slipped from the inverted test tube

H, the sample did not fall from the inverted test tube
9/H, slight turbidity of the sample

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Eur Food Res Technol (2012) 234:863872

Nitrogen solubility /%

120
100

SPI
SMGHs (DH: 4.94%)

SMG
SMGHs (DH: 6.84%)

SMGHs (DH: 7.53%)

SMGHs (DH: 11.86%)

Table 2 WHC and OHC of SMG, SPI, and SMGHs with different
degree of hydrolysis
Samples

80

WHC (mL/g)
3.20 0.53A

SMG (DH: 0%)

SMGHs (DH: 4.94%)

60

10

pH

Fig. 2 Solubility profile of SMG and SMGHs with different degree

of hydrolysis as influenced by pH. Data are expressed as means SD
from triplicate determinations. A commercial soy protein isolate (SPI)
was used as a reference protein

hydrolysates over that of the intact protein is due to the

reduction of its secondary structure, the release of smaller
soluble polypeptides from native protein, and also the
balance of hydrophilic and hydrophobic forces of peptides
[27, 39, 40]. Thus, the controlled enzymatic hydrolysis
could be used as an alternative to improve the solubility of
scallop gonad protein. The excellent solubility of SMGHs
indicates that they may have potential application in formulated food systems.
Several reports have shown that protein hydrolysates of
seafood have excellent water-holding capacity (WHC) and
can increase the cooking yield when added to minced meat
[25, 41]. The water-holding capacity (WHC) and oilholding capacity (OHC) of SMGHs were evaluated for
possible use as a functional ingredient for meat products.
As shown in Table 2, the WHC of SMG was significantly
improved (p \ 0.05) after enzymatic hydrolysis. The
highest WHC was observed at DH of 4.94% and showed
significant difference with that of SPI (p \ 0.05). It has
been suggested that presence of polar groups such as
-COOH and -NH2 that increased during enzymatic hydrolysis had a substantial effect on the amount of absorbed
water [25, 41]. The OHC of SMGHs with DH of 4.94%
was significantly higher than that of SMG (p \ 0.05) and
equivalent to that of SPI (p [ 0.05) (Table 2). Wasswa
et al. [25] reported that WHC and OHC of hydrolyzed grass
carp fish skin was increased with the increasing of DH,
whereas in our case the higher WHC and OHC were
observed at lower DH (4.94%). Diniz and Martin [39]
found an inverse correlation between solubility and WHC
or OHC in shark protein hydrolysate. In the present study,
the solubility of SMGHs was increased with the increasing
of DH (Fig. 2). It seems that the extensive hydrolysis will
decrease the WHC and OHC of SMGHs due to growing
solubility. The SMGHs with lower DH may be promising

123

2.13 0.76ABC

3.80 0.72B

9.06 0.62

Data are expressed as means SD from triplicate determinations

Different capital letters in the same column indicate significant differences (p \ 0.05)

50

Surface hydrophobicity

1.47 0.12C

9.13 0.61

SPI
2

3.53 1.03AB

7.80 1.06

SMGHs (DH: 11.86%)

3.27 0.50B

9.20 0.20

SMGHs (DH: 7.53%)

20

2.00 0.53AC

11.26 0.90

SMGHs (DH: 6.84%)

40

OHC (mL/g)

40

c
30

d
a

20

e
10

0
DH:0%

DH:4.94%

DH:6.84%

DH:7.53%

DH:11.86%

Fig. 3 Surface hydrophobicity of SMG and SMGHs with different

degree of hydrolysis. Data are expressed as means SD from
triplicate determinations. Different small letters denote significant
differences (p \ 0.05)

ingredients for certain food products where water holding

and oil binding are important.
Protein surface hydrophobicity is known to be closely
related with technologically relevant properties of food
proteins. We measured the surface hydrophobicity of
SMGHs with different hydrolysis degree using ANS as a
fluorescent probe (Fig. 3). The SH values of the hydrolysates with DH (from 4.94 to 7.53%) were significantly
higher than that of SMG (p \ 0.05). The maximum SH
value was observed at DH of 4.94%, which can be attributed to release of some stable hydrophobic peptides from
SMG protein. The SH value of SMGHs decreased with the
increasing of DH, which is probably due to enzymatic
cleavage of hydrophobic clusters. Calderon et al. [42]
reported that proteolysis is accompanied by gain or loss in
hydrophobicity, depending mainly on the nature of the
hydrolyzed protein and molecular weight size of the
formed peptides, due to the shortening of peptide chains.
Our results propose that the SH change between SMG and
SMGHs is related to the hydrolysis extent.
The EAI is a function of oil volume fraction, protein
concentration, and the type of equipment used to produce

Eur Food Res Technol (2012) 234:863872

869

the emulsion. As shown in Table 3, the EAI of all SMGHs

was significantly higher than that of SMG in pH 5 and 7
(p \ 0.05), being higher than or equivalent to that of SPI. It
demonstrates that enzymatic hydrolysis by neutrase can
improve emulsifying capacity of SMG in mild acidic and
neutral conditions. At pH 3.0, the highest value of EAI
(11.79 0.16 m2/g) was found at DH of 11.86%, significantly higher than SMG and SPI (p \ 0.05). At pH 5.0 and
pH 7.0, the maximum EAI was both observed at DH of
7.53%. This observation suggests that high hydrolysis
degree is favorable to emulsifying property of SMGHs in
highly acidic condition, whereas further proteolysis led to a
loss of EAI in weakly acidic and neutral conditions. No
significant differences were observed for SMGHs and
SMG at pH 9.0, which is similar with Jamdar et al. [15],
who found that the emulsifying activity was independent of
DH at alkaline pH. For SMG (DH: 0%), the EAI at pH 3.0
and pH 5.0 was significantly lower than that at 7.0 and pH
9.0 (p \ 0.05), due to its nearness of protein isoelectric
point. While the EAI of SMGHs with DH of 4.94 and
6.84% increased with the increasing of pH, probably
because of improvement in solubility, as good solubility is
critical to emulsifying properties [21].

Proteins in dispersions cause a lowering of the surface

tension at the waterair interface, thus creating foam. The
foam capacity and stability of SMG, SPI, and SMGHs are
shown in Table 4. The foam capacity and stability of SPI
were far higher than that of SMG and SMGHs, and the
foam capacity and stability of SMG were significantly
higher than that of all SMGHs (p \ 0.05). This is in
accordance with the report by Jamdar et al. [15], who found
peanut protein isolate (PPI) showed higher foam capacity
than its hydrolysates at neutral pH, The foam capacity and
stability of SMGHs also showed a reduction trend as DH
increased from 4.94 to 11.86%, which is consistent with the
general viewpoint that the larger molecular size for a
protein the higher foam stability [12]. This is in line with
previous findings reporting that good film cohesiveness is
reached with high-molecular weight peptides or partially
hydrolyzed proteins [43]. Besides, it has been suggested
that the effect of enzymatic hydrolysis on foam capacity
was pH dependent [15] and substrate specific [44]. The
present results indicate that the hydrolysis of SMG is detrimental to foam capacity and stability.
The composition of total amino acids affects the nutritional value of food [45]. Table 5 outlines the total amino

Table 3 EAI of SMG, SPI, and SMGHs with different degree of hydrolysis as influenced by pH
Samples

EAI (m2/g)
pH 3.0

SMG (DH: 0%)

pH 5.0
Aa

2.33 0.19

Ba

pH 7.0

1.44 0.30

Ab
Bb

SMGH (DH: 4.94%)

3.22 0.09

11.91 0.67

SMGH (DH: 6.84%)

3.16 0.61ABCa

12.22 0.21Bb

SMGH (DH: 7.53%)

SMGH (DH: 11.86%)
SPI

Ca

4.11 0.51
11.79 0.16

Da

Ea

5.62 0.33

19.65 0.80

Cb

14.03 0.79

Dbc

8.51 1.36

pH 9.0
Ac

14.92 2.46Ad

BCb

12.19 1.78

15.23 0.37Ac

14.55 2.77BCbc

15.96 1.14Ac

7.43 0.68

Bc

14.46 1.81Ac

BCb

14.62 0.65Ac

Cc

41.45 3.66Bd

15.57 1.69
12.99 0.40

Eb

12.62 0.24

Data are expressed as means SD from triplicate determinations

Different capital letters in the same column indicate significant differences (p \ 0.05)
Different small letters in the same row indicate significant differences (p \ 0.05)

Table 4 Foam capacity of SMG, SPI, and SMGHs with different degree of hydrolysis
Samples

Foam capacity (%)

0 min

10 min

30 min

SMG (DH: 0%)

51.11 1.92

33.33 0.00

24.44 1.92Ac

SMGH (DH: 4.94%)

41.11 1.92Ba

25.56 3.85Bb

18.89 1.92Bb

Bb

15.56 3.85Bb

Bb

15.56 1.92Bc

Bb

4.44 1.92Cc

Cb

104.44 7.70Db

SMGH (DH: 6.84%)

SMGH (DH: 7.53%)
SMGH (DH: 11.86%)
SPI

Aa

BCa

36.67 3.33

23.33 3.33

Ca

35.56 1.92

20.00 0.00

Ca

34.44 1.92
128.89 3.85

Da

Ab

18.89 1.92
116.67 5.77

Data are expressed as means SD from triplicate determinations

Different capital letters in the same column indicate significant differences (p \ 0.05)
Different small letters in the same row indicate significant differences (p \ 0.05)

123

870
Table 5 Amino acid
composition of SMG and
SMGHs with different degree of
hydrolysis

Essential amino acid (EAA)

Eur Food Res Technol (2012) 234:863872

Amino
acids

Amino acid content (mol%)

SMG
(DH: 0%)

SMGHs
(DH: 4.94%)

SMGHs
(DH: 6.84%)

SMGHs
(DH: 7.53%)

SMGHs
(DH: 11.86%)

Asp

6.30

6.35

6.38

6.34

6.44

Glu

7.77

7.88

7.59

7.73

7.87

Hyp

0.20

0.13

0.13

0.13

0.13

Ser

4.70

4.83

4.80

4.77

4.71

Arg

5.96

5.98

5.90

5.83

3.96

Gly

26.11

25.35

25.67

25.91

26.39

Thra
Pro

4.39
3.45

4.43
3.48

4.44
3.49

4.44
3.49

4.46
3.58

Ala

8.41

8.46

8.51

8.51

8.65

Vala

5.58

5.66

5.63

5.57

5.82

Meta

1.65

1.62

1.58

1.63

1.66

Cys

0.31

0.35

0.34

0.35

0.35

Ile

3.71

3.86

3.87

3.80

3.89

Leua

5.32

5.42

5.44

5.40

5.51

Trpa

0.17

0.15

0.19

0.14

0.10

Phea

3.24

3.25

3.28

3.25

3.32

His

1.46

1.45

1.50

1.47

1.48

Lysa

9.32

9.30

9.27

9.24

9.59

Tyr

1.94

2.03

1.99

2.01

2.09

EAA

41.63

42.00

42.02

41.83

42.90

Total

100

100

100

100

100

acid compositions of SMG and SMGHs. It was clear that

the total amino acid composition of SMGHs changed
slightly and resembled to that of SMG, which was consistent with our previous findings [18] by using abalone
viscera as materials. SMG and SMGHs contained all the
essential amino acids (41.6342.90% of the total amino
acids) and were rich in glycine, lysine, alanine, glutamic
acid, and aspartic acid, suggesting SMG and SMGHs can
possibly be a dietary protein supplement to poorly balanced
proteins.

food ingredients for food manufacture. More detailed

studies on the influence factors and underlying mechanism
of neutrase-induced gelation of SMGHs will be reported
elsewhere.
Acknowledgments This work was financially supported by The
National High Technology Research and Development Program of China
(863 Program) (No. 2011AA100803), and Liaoning Key Laboratory of
Seafood Science and Technology, China (No.2008403003).

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Protein hydrolysates from scallop (Patinopecten yessoensis) male gonad, exhibiting better functionalities than the
raw materials, could be achieved by neutrase hydrolysis.
Some functional properties of SMGHs could be modified
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DH) is favorable to gelation property and solubility of
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DH, but restricted by pH environment. The scallop male
gonad hydrolysates with good functionalities and high
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