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15 September 2013, Volume 34 Issue 17
Purification and Molecular Mass Determination of Lily Polysaccharides
CHEN Zhi-gang,ZHU Quan,WANG Fen
2013, 34(17):  1-4.  doi:10.7506/spkx1002-6630-201317001
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Microwave-assisted hot-water extraction method was used to extract lily polysaccharides. Fractions LLPS-1,
LLPS-2 and LLPS-3 were isolated from the aqueous polysaccharide extract and purified through deproteinizaiton by Sevag’s
method, dialysis, DEAE-Sepharose Fast Flow chromatography and Sephadex G-100 column chromatography. Results from
coomassie blue staining, UV and high-performance gel permeation chromatography (HPGPC) showed that the purified
product contained neither protein or nucleic acid. The molecular weights of LLPS-1, LLPS-2 and LLPS-3 were 350.5,
403.3 kD and 146.2 kD, respectively.

Effects of Arabic Gum and Carboxymethylcellulose(CMC) on Rheological and Baking Properties of Buckwheat Dough
LIU Yan1,HUANG Wei-ning1,*,JIA Chun-li1,WANG Feng2,RAYAS-DUARTE Patricia3
2013, 34(17):  5-9.  doi:10.7506/spkx1002-6630-201317002
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Buckwheat bread was made with 30% flour substitution. The effects of arabic gum (AG) and sodium
carboxymethylcellulose (CMC) on rheological properties of dough and baking properties of breads were investigated, with
the addition of 0.5%, 1.0% and 1.5% of AG and CMC respectively. The two gums had different effects on rheological
properties. Both viscous modulus and elastic modulus of buckwheat dough were decreased after AG was added, while the
results showed a positive correlation with CMC. Compared with the control, the total volume of buckwheat dough did not
change much after AG and CMC were added, whereas maximum fermentation height (Hm), maximum gas production height
(H’m) and retention coefficient (R) were elevated and the start time for dough to lose CO2 (Tx) was delayed. Those results
were more significant when CMC was added. With the increase of AG and CMC, the specific volume of bread was elevated
from 4.20 mL/g for the control to 4.47 mL/g and 4.78 mL/g, respectively. As the substitution was up to 1.5%, the hardness
of bread was decreased by 27.9% and 51.5%, and the cohesiveness and resilience were also improved. Besides the area
fraction and cell density of buckwheat bread became higher when hydrocolloids were added.

Antioxidant Capacity and Isozymes of Polyphenol Oxidase from Toona sinensis during Different Growth Periods
WANG Cheng,ZHANG Jing-fang*,ZHANG Yan,YAN Ming-ming,WANG Li-mei
2013, 34(17):  10-14.  doi:10.7506/spkx1002-6630-201317003
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In order to understand the antioxidant capacity and the changes of polyphenol oxidase (PPO) from Toona sinensis
during different growth periods, the antioxidant activity of the extract from Toona sinensis was evaluated by three methods,
and the contents of total phenols and total flavonoids as well as PPO activity were analyzed. Furthermore, the isozymes
of PPO were also compared by polyacrylamide gel slab electrophoresis. Results indicated that Toona sinensis at the third
growth period had the highest antioxidant activity, followed by the second and the first growth periods. The antioxidant
activity and phenolic components revealed a dose-dependent manner. The decreasing order of PPO activity in Toona sinensis
during different growth periods was the first, third and second growth periods. The contents of total phenols showed a
gradually increasing tendency during the entire growth periods (P < 0.05). Toona sinensis at the first growth period had
the lowest content of total flavonoids, and no significant difference between the second growth period and the third growth
period was observed (P < 0.05). Totally 5 kinds of isozymes (A1—A5) were separated, and A3 and A4 were the particular
enzyme bands of Toona sinensis PPO. A1, A2 and A5 showed different enzymatic activities during the three growth periods.
The changes of PPO activity and antioxidant activity at the same growth period were correlated with isozyme bands.

Effect of Garlic Powder on Properties of Heat-induced Gel from Porcine Salt-soluble Protein
LI Jun-guang,LI Zeng,JIANG Ai-min*,ZHOU Quan,QU Zhi,WU Lan-fang
2013, 34(17):  15-18.  doi:10.7506/spkx1002-6630-201317004
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The effect of garlic powder on properties of heat-induced gel prepared from porcine longissimus dorsi (PLD)
salt-soluble protein was investigated. The textural properties, water holding capacity, color and rheological properties of
the gel were determined by texture analyzer, centrifugation method, colorimeter and dynamic rheometer, respectively.
Changes of chemical interactions in the gel heating process were studied by chemical methods. The results showed that the
hardness, chewiness and storage modulus G’ of the gel were the highest with the addition of 3% garlic powder, and it also
demonstrated excellent color and water holding capacity; and hydrophobic interactions were the main chemical interactions
that maintained the stable structure of pork salt-soluble gel. Addition of 3% garlic powder enhanced the hydrophobic
interactions and improved the gel strength.

Effect of Oxidation Degree on Heat-induced Gel Formation and Physico-chemical Properties of Myofibrillar Protein
HU Zhong-liang1,ZOU Yu-feng1,LIN Yu-hai2,PU Qi3,XU Xing-lian1,*
2013, 34(17):  19-22.  doi:10.7506/spkx1002-6630-201317005
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Chicken breast myofibrillar protein was oxidized to different extents after 24 h of incubation in hydroxyl radicalgeneration
systems (10 μmol/L FeCl3, 0.1 mmol/L ascorbic acid, and 0 to 5 mmol/L H2O2) and physico-chemical properties
of the different oxidation products and properties of heat-induced gels prepared from them were comparatively investigated.
The results showed that carbonyl content steadily increased and thiol group content declined with increasing H2O2
concentration, indicating continuous oxidation of the myofibrillar protein. Heat-induced gel hardness increased significantly
as the concentration of H2O2 increased from 0.00 to 0.10 mmol/L while protein solubility and surface hydrophobicity
declined. When H2O2 concentration increased up to 5 mmol/L, protein solubility as well as gel hardness and springiness
significantly decreased (P < 0.05), whereas the converse trend was observed for surface hydrophobicity. These results
suggest that mild oxidation could improve the heat-induced gel formation of myofibrillar protein.

Isolation and Characterization of Collagens from Skin of Grass Carp (Ctenopharyngodon idellus) and Snakehead (Channa argus)
WANG Zhong-wen1,WANG Hai-bo1,*,LIANG Yan-ping1,LI Yun-yan1,WANG Min1,WANG Hai-ying2
2013, 34(17):  23-27.  doi:10.7506/spkx1002-6630-201317006
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In this study, the molecular structure and physico-chemical properties of collagens from herbivorous fish (grass
carp) and carnivorous fish (snakehead) were compared. Acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC)
extracted from grass carp and snakehead skins were characterized comparatively for their compositional, structural and
physiochemical properties and thermal stability and the results obtained were compared with those of pig skin collagen. Results
indicated that all five extracted collagens were classified as type Ⅰ collagen which comprised two different α chains (α1 and
α2). Collagens from different sources had considerably varying amino acid composition and protein patterns as well as their
primary structure. The denaturation temperatures of different collagens were: PSC from pig skin (41.58 ℃), ASC from grass
carp skin (35.62 ℃), PSC from grass carp skin (35.81 ℃), ASC from snakehead skin (33.85 ℃) and PSC from snakehead
skin (34.06 ℃). The enzymatic hydrolyzability of different collagens decreased in the order of snakehead skin PSC, grass carp
skin PSC, snakehead skin ASC, grass carp skin PSC and pig skin PSC. Collagens extracted from grass carp and snakehead had
lower denaturation temperatures and higher enzymatic hydrolyzability than those from pig skin. Grass carp skin collagens were
more resistive to collagenase and showed higher denaturation temperature compared with snakehead skin collagens. In addition,
ASC was more resistive to collagenase compared to PSC.

Molecular Weight and Antioxidant Activity of Enzymatic Hydrolysates of Silver Carp
LI Xiang-hong,CHEN Zhi-jun,LIU Yong-le*,YU Jian,WANG Fa-xiang,WANG Jian-hui
2013, 34(17):  28-32.  doi:10.7506/spkx1002-6630-201317007
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The molecular weight (Mw) and antioxidant properties of enzymatic hydrolysates of silver carp with different
degree of hydrolysis (DH) were studied. The antioxidant properties evaluated included total antioxidant capacity (T-AOC),
radical scavenging activity against hydroxyl and superoxide anion free radicals and lipid oxidation inhibitory effect. The
hydrolysates were prepared by respective use of protamex and alcalase. The results showed that the Mw of the hydrolysates
produced by alcalase were generally lower than those of the hydrolysates obtained with Protamex, and at DH 10%, over
95% of peptides fractions had Mw lower than 1500 u. The hydrolysates prepared by each enzyme preparation had excellent
antioxidant activities. The hydrolysates with DH 10% prepared by alcalase possessed the highest T-AOC and hydroxyl
radical scavenging capacity, which were 1.025 U/mg pro and 21.31 U/mg pro, respectively. The hydrolysates with DH 10%
prepared by protamex displayed the strongest superoxide anion radical scavenging ability and lipid oxidation inhibitory
effect, which reached 24.92 U/mg pro and 27.88% at a concentration of 5.0 mg/mL, respectively. The antioxidant activities
of these hydrolysates might be related to their molecular weights despite being not significant.

Screening of Key Indicators to Evaluate the Eating Quality of Emulsified Mutton Sausage
WANG Lin-chen,WANG Zhen-yu,XIA An-qi,CHEN Li,ZHANG De-quan*
2013, 34(17):  33-37.  doi:10.7506/spkx1002-6630-201317008
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Sensory evaluation was carried out on 13 kinds of emulsified mutton sausage, according to the Chinese national
standard (GB/T 16861—1997 Sensory Evaluation) and the results obtained were analyzed by M value method, principal
component analysis and correlation analysis to identify critical indicators to evaluate the eating quality of emulsified mutton
sausage. Appearance, texture, flavor and color were selected as critical quality traits of emulsified mutton sausage. The criteria for
appearance evaluation were surface uniformity and color and cross-sectional compactness and smoothness, the texture properties
were chewiness, springiness, firmness, gumminess and juiciness, flavor evaluation was based on mutton odor and aftertaste, and
the criterion for color evaluation was light pink. Correlation analysis showed that surface uniformity was significantly correlated
with surface color, cross-sectional compactness and smoothness (r = 0.785, 0.791 and 0.828, respectively), chewiness exhibited
a significant correlation with springiness, firmness and gumminess (r = 0.911, 0.764 and 0.927, respectively), and there was a
significant correlation between mutton odor and aftertaste (r = 0.891). It was concluded that the eating quality of emulsified mutton
sausage could be evaluated by surface uniformity, chewiness, springiness, juiciness, mutton odor and light pink.

Determination of Fermentation End-point of Tempeh by Image Analysis
LIANG Jing-dong1,SUN Ji-xiang1,FAN Juan2,LI Wei2,DONG Ming-sheng2,*
2013, 34(17):  38-42.  doi:10.7506/spkx1002-6630-201317009
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Tempeh is a fermented food originated in Indonesia. Its nutritional quality depends on the extent of mycelial
growth on its surface. A method based on image analysis was proposed to determine the fermentation end-point
automatically, quickly, reliably and non-destructively. Tempeh's surface images were collected by using a digital camera
during its fermentation process and a new uniformity evaluation of tempeh’s surface image was formulated through analysis
of texture features and combination of gradient and luminance information. The indicator reached the minimum value at the
fermentation end-point (22 h, determined by sensory evaluation) and showed a significant change near the fermentation endpoint.
When the pixel of tempeh’s surface image was 2592 × 3314, the indicator's value was 0.054 at 21 h, 0.044 at 22 h
and 0.055 at 23 h. However, no significant change was observed in the traditional uniformity evaluation of tempeh’s surface
image (standard deviation of luminance) in which the values were 8.3 at 21 h, 8.2 at 22 h and 8.2 at 23 h. The experimental
results showed that the method could quickly and accurately determine tempeh’s fermentation end-point based on surface
images with different pixels, had a good stability and was consistent with artificial sensory evaluation.

Analysis of Glycated Peptide by CID-ETD Mass Spectrometry
LIU Guang-xian1,GUAN Shan-hong1, WANG Hui2,*,FENG Jian-xiong1
2013, 34(17):  43-46.  doi:10.7506/spkx1002-6630-201317010
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Identification of glycation sites in glycation protein plays an important role in the fields of food science and
medicine. In this work, liquid chromatography (LC) coupled with linear ion trap quadrupole-electrospray ionization tandem
mass spectrometry (LTQ-ESI CID/ETD MS-MS) was used to analyze peptides from glycated ovalbumin digested by pepsin.
The results indicated that primary identification of glycation peptides was carried out through CID MS spectrometry and site
identification through CID MS-MS fragment ions. However, new ion peaks from CID MS-MS analysis were very weak and
complex with some various degrees of neutral losses. ETD MS-MS was a technique more suitable for studying gaycated
peptide with medium molecular weight and 2 charges. Due to larger molecular gaycated peptides, it was hard to determine
the ETD fragments, so CID-NL-MS3 technique should be developed for the identification of the glycation site.

Correlation between Changes in Soluble Sugars and Anthocyanins Contents during Fruit Coloration
LIU Yu-lian,CHE Fei,GUO Yan-ping,LIANG Jun,ZHAO Zheng-yang*
2013, 34(17):  47-52.  doi:10.7506/spkx1002-6630-201317011
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In this study, fruits from the red apple cultivars ‘Starking’ and ‘Pink Lady’, the green cultivars ‘Granny Smith’,
and the yellow cultivars ‘Golden Delicious’ were analyzed by HPLC for changes in the kinds and amounts of soluble sugars
and anthocyanins after bag removal during coloration and correlation between these two components. The results showed
that the kinds of anthocyanins in apples varied among the different cultivars. Three anthocyanins, cyanidin 3-galactoside,
cyaniding 3-glucoside chloride and cyaniding 3-arabinoside were identified in the skin of the red apple cultivars after the
completion of coloration and two anthocyanins, cyanidin 3-galactoside and cyaniding 3-arabinoside in both green and
yellow cultivars in different amounts. Various anthocyanins were identified in significantly different amounts in the fruit
and cyanidin 3-galactoside was the most abundant in the skin. All the different apple cultivars contained the same soluble
sugars in different amounts, with fructose being the most abundant followed by sucrose, glucose and the smallest amounts
of galactose and sorbitol. Our analysis found that during the process of coloration, the changes in anthocyanin content,
especially cy-3-gal had an extremely significantly positive correlation with sucrose content. This study speculates that
sucrose might be responsible for the regulation of cy-3-gal biosynthesis in apple skin.

Effect of High Pressure Brining on Eating Quality of Chicken Breast
LENG Xue-jiao,ZHANG Lin,HUANG Ming*,XU Xing-lian,ZHOU Guang-hong
2013, 34(17):  53-56.  doi:10.7506/spkx1002-6630-201317012
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The objective of this study was to investigate the effect of brining combined with different pressure treatments on
eating quality of chicken breast. Chicken breast was brined under different pressure levels (0.1, 50,100,150, 200 MPa and
300 MPa for 20 min in 40 g/L salt solution. The salt content, moisture content, meat color, pH, shear force and water holding
capacity were measured afterwards. The results showed that chicken breast treated by high pressure brining gave higher salt
content compare to the control and the maximum salt content was obtained at 150 MPa. Brining with high pressure treatment
could also increase L* and b* but cause a reduction in a*. Treatments at a pressure level lower than 150 MPa showed less
effect on color of the chicken breast. The treatment at 50 MPa significantly reduced the pH while no significant effect was
observed under other pressures on the pH of chicken breast. High-pressure brining was not able to improve the tenderness
of chicken breast but could improve its water holding capacity. In general, brining at 150 MPa was the best treatment, and
showed no negative impact on eating quality of chicken breast.

Correlation between Meat Quality and Serum Biochemical Indices of Yak
DENG Li-juan1,GUO Zhao-bin1,BAO Shan-ke2,HAN Ling1,*,YU Qun-li1
2013, 34(17):  57-60.  doi:10.7506/spkx1002-6630-201317013
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The study was designed to evaluate meat quality traits and determine serum biochemical indices of Qinghai
yak using a NSA-400 type automatic biochemical analyzer. The results show that a significant increase in meat color and
cooking yield was found for yak meat in comparison with the meat of the local yellow cattle (P < 0.05). Moreover, serum
GOT and LDH activities as well as urea nitrogen, K+ and Ca2+ contents of yak were also significantly higher than those of
yellow cattle (P < 0.05). The correlations between AKP activity and pH at 1 h postmortem (pH1), between LDH activity
and marble score, between HDL content and cooking yield, between serum GOT activity, urea nitrogen level or creatinine
level and pH at 24 h postmortem (pH24), and between Mg2+ content and water loss rate were significantly negative, whereas a
significantly positive correlation was observed between LDH activity and cooking yield.

Application of Fuzzy Comprehensive Evaluation in Development of Wheat Bran Tea
ZHOU Li-ming,LUO Song-ming,ZHANG Zhi-qing*
2013, 34(17):  61-64.  doi:10.7506/spkx1002-6630-201317014
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Uniform design in combination with analysis of variance and fuzzy mathematics analysis was used to optimize
the cellulase-catalyzed hydrolysis and subsequent roasting of wheat bran for the development of wheat bran tea. The optimal
process conditions were determined as 0.2%, 10:1, 180 ℃ and 15 min for enzyme dosage, water/wheat bran ratio, roasting
temperature and time, respectively. Infusion of the resulting product possessed a strong and mellow wheat aroma, a bright
yellow color and a sweet and refreshing taste. Its DPPH free radical scavenging rate was 71.3%, which indicates higher
antioxidant activity compared with commercial tea products.

Antioxidant Capacity and in vitro Starch Digestibility of Cooked Buckwheat Noodles
MA Yu-jie,LIU Hang,XU Yuan-yuan,GUO Xu-dan,WANG Min*
2013, 34(17):  65-69.  doi:10.7506/spkx1002-6630-201317015
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Purpose: To evaluate the antioxidant activity and in vitro starch digestibility of cooked noodles made from two
tartary buckwheat cultivars (Xinong 9940 and 9909) and three common buckwheat cultivars (Dianbian red flower, Yuqiao
4 and Wensha) by comparison with wheat noodles. Method: Total flavonoids content was measured by NaNO2-Al(NO3)3
assay, and DPPH free radical scavenging capacity assay and reducing power method were employed to evaluate the
antioxidant capacity of the noodle samples. The content of resistant starch was measured through enzymolysis. Additionally,
total starch hydrolysis was tested at different times, and data were plotted as starch hydrolysis degree vs. time curves. Estimated
glycemic index (EGI) was calculated according to the curves. Results: The antioxidant property of buckwheat noodles was
significantly superior to wheat noodles. Moreover, the content of total flavonoids ranged from 600.76 to 183.33 mg /100 g and
tartary buckwheat noodles had higher total flavonoids content and antioxidant capacity and the best cultivar was Xinong
9909. Resistant starch content ranged between 2.78% and 6.07%, depending on the cultivar. Furthermore, EGI of buckwheat
noodles were significantly lower than that of wheat noodles, and different among cultivars. EGI of Dingbian red flower,
Wensha, Xinong 9909 and Xinong 9940 was less than 75, thus belonging to middle-EGI food. Conclusion: The quality of
buckwheat noodles is generally better than that wheat noodle and differs among cultivars and among the tested cultivars,
Xinong 9940 processes the best overall quality. As a result, buckwheat noodles can be served as daily food with the
preventing function against chronic disease such as diabetes.

Determination and Antioxidant Activity of Flavonoids from Winter Jujube
YUAN Ya-na1,2,ZHANG Ping-ping2,*,HE Qing-feng2,WU Hai-qing2,FAN Xiu-hua2,ZHANG Li-jin2
2013, 34(17):  70-73.  doi:10.7506/spkx1002-6630-201317016
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This study was aimed to analyze the flavonoids from winter jujube and the antioxidant activity was also assessed.
Winter Jujube was extracted with 60% ethanol and separated by macroporous resin chromatography to obtain crude
flavonoids (FW-1). FW-1 was further purified on silica gel column into FW-2. The composition and chemical structure of
FW-1 and FW-2 was analyzed by LC-MS, and their radical scavenging activity against ·OH and O2
-· and DPPH radicals
was measured. Results showed that total flavonoid contents in FW-1 and FW-2 were 55% and 94%, respectively. The
HPLC chromatogram of FW-2 showed that the main peak was E and based on mass spectra, it was identified as bisflavonoid
with a relative molecular weight of 790. Flavonoids at low concentration showed higher radical scavenging
capacity against ·OH than VC but less effective in scavenging superoxide anion radical than VC. The DPPH radical
scavenging rate of flavonoids at 200 μg/mL was higher than 85%, and the effect was dose-dependent. In conclusion,
the main component of the flavonoids extracted from winter jujube was bis-flavonoid and it demonstrated higher
antioxidant activity.

Interactions between Casein and Maltodextrin in the Formation of Microcapsule
SHI Yan1,LIU Fan1,ZHENG Yi2,ZHENG Wei-wan1,3,GE Hui1
2013, 34(17):  74-77.  doi:10.7506/spkx1002-6630-201317017
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The interaction between casein and maltodextrin during the formation of microcapsule wall was investigated
using fourier transform infrared spectroscopy combined with other mathematical treatments such as deconvolution,
second derivative spectroscopy and curve fitting. A preliminary investigation was conducted to explore the mechanism of
interaction between these two materials. Results showed that following heating and spray drying treatments, changes of the
amideⅠbands and the amide Ⅲ bands of casein occurred. The interaction between casein and maltodextrin was present in
the form of cross-linking and a strong hydrogen bond was then formed. In this study, the emulsifying capacity and emulsion
stability of casein were also measured with or without the addition of maltodextrin. The increased content of α-helix and
random coil contributed to the higher emulsifying capacity and stability of casein in the presence of maltodextrin.

Effect of Mixed Liquid Carrier on the Crystalline Structure of Corn Starch Granules after High Pressure Jet Homogenization
LIU Bin,ZHANG Yuan,FENG Tao,WU Xue
2013, 34(17):  78-81.  doi:10.7506/spkx1002-6630-201317018
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The effects of pure water and mixed (0.24% ethanol in water) media on the efficacy of high pressure jet
homogenization to micronize corn starch granules were explored. Under the identical operating conditions, water was the
least effective for damaging the crystalline structure of corn starch. However, in the aqueous ethanol medium, the crystalline
structure of corn starch was seriously damaged due to the enhanced cavitation and thus the increased gas–liquid–solid
impact. Scanning electron microscopic (SEM) observations indicated that surface flaking of corn starch granules happened
with micro-debris adhering to the surface of the granules. Polarized light microscopic (PLM) analysis exhibited the damaged
crystalline structure of corn starch. Differential scanning calorimetry (DSC) demonstrated that both the native granular
structure and crystalline arrangement (compact) were altered.

Changes of the Textural Characteristics of Yoghurt Made from Cow Milk and Goat Milk during Fermentation
ZHANG Yue1,GE Wu-peng1,*,YUAN Ya-juan2,CHEN Ying3,QIN Li-hu4,YANG Jing5
2013, 34(17):  82-86.  doi:10.7506/spkx1002-6630-201317019
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The changes of the textural properties of yoghurt produced from cow or goat milk during fermentation were
investigated comparatively. A texture analyzer (TA. XT plus, with Texture Exponent 32 software) was utilized for comparative
analysis of texture properties of set yoghurt and stirred yoghurt samples produced from cow and goat milk. The right back
extrusion probe and testing procedure were selected to quantify firmness, consistency, cohesiveness and viscosity during
milk fermentation. The results showed that during the per-fermentation period, all these four texture properties exhibited a
higher level in set cow yoghurt than set goat yoghurt, and the same results were observed during the post-fermentation period;
moreover, significant differences in each texture parameter were observed between both samples (P < 0.05). The texture
parameters of stirred cow yoghurt were lower than those of stirred goat yoghurt, but these differences were not significant
(P > 0.05). In summary, the texture properties of set cow yogurt are higher than those of set goat yoghurt, while there is an
improvement in texture properties of stirred goat yoghurt compared to stirred cow yoghurt.

Predictive Modeling of Simultaneous Growth of Escherichia coli and Pseudomonas spp. in Inoculated Pork longissimus dorsi Muscle
JIN Xin,YU Ying-ying,WU Ju-qing,XU Xing-lian,ZHOU Guang-hong*
2013, 34(17):  87-91.  doi:10.7506/spkx1002-6630-201317020
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Pork longissimus dorsi muscle was inoculated with both Escherichia coli and Pseudomonas spp., and then stored
at 10, 15, 20 ℃ or 25 ℃. The results showed that growth curves of E. coli and Pseudomonas spp. at different temperatures
could be well described by a modified Gompertz model. The influence of temperature on growth rate (μmax) and lag
phase (Lag) was well described by Belehradek model. The model developed in this study was confirmed to be reliable by
validation experiments at 13 ℃ and 22 ℃.

Pigments from Black Glutinous Corn Eliminate Nitrite and Block Nitrosamine Synthesis
SHENG Wei1,2,3,GAO Xiang2,3,XUE Jian-ping2,3,XIE Bi-jun1,*
2013, 34(17):  92-95.  doi:10.7506/spkx1002-6630-201317021
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The influence of pigment concentration and exposure time on the effectiveness of pigments from
black glutinous corn (PBGC) to eliminate nitrite and block nitrosamine synthesis was investigated in two kinds
of simulated human gastric juices and the results obtained were compared those of VC. At higher concentrations,
PBGC was more effective in eliminating nitrite and blocking nitrosamine synthesis. Both activities varied depending
on reaction time and different simulated gastric juices. In the simulated gastric juice B, 81.37% of the nitrite was
eliminated after 1 h of expoure to 0.10 mg/mL PBGC and nitrosamine synthesis was blocked by up to 57.03% by
PBGC. Comparing it with VC, we concluded that PBGC have a strong capability of eliminating nitrite and blocking
nitrosamine synthesis.

A Theoretical Study on the Interaction between Melamine and Acrylamide Functional Monomer in Molecularly Imprinted Polymers
LIU Jun-bo1,SHI Yang1,SUN Jia-ni1,TANG Shan-shan1,HU Yao-hui2,JIN Rui-fa3
2013, 34(17):  96-101.  doi:10.7506/spkx1002-6630-201317022
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The quantum chemical method was applied for theoretical investigation into the interaction between melamine
(MEL) and acrylamide (AM) in molecularly imprinted polymers (MIPs). The pre-assembly system of MEL as the imprinting
molecule and AM as the functional monomer in the MIPs was simulated based on the density functional theory (DFT) at the
pbe1pbe/6-31G(d,p) level using Gaussian 09 software. Moreover, the formation of hydrogen bonds, natural bond orbital (NBO)
charge and binding energy of MEL/AM complexes in different proportions were explored, and the mechanism and extent of
interaction between MEL and AM were illustrated. Results indicate that MEL/AM complexes with ordered and complementary
structure could be formed via hydrogen bonding interactions. The lowest binding energy was observed at MEL:AM ratio = 1:6.
Furthermore, the trend of charge transfer was the biggest overall and there were 9 active sites in the formed complex. This study
may provide a theoretical foundation for improved understanding of the principle of MEL molecular imprinting.

Optimization of Aqueous Extraction of Trypsin Inhibitor from Gourd and Trypsin Inhibitor Activity of Different Parts of Gourd
SHAO Yue,ZHANG Qi,ZHANG Cheng,MA Yong*
2013, 34(17):  102-105.  doi:10.7506/spkx1002-6630-201317023
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The aqueous extraction of trypsin inhibitor was optimized using orthogonal array design. Trypsin inhibitor activity
was determined from the standard curve generated using DL-BAPNA as substrate. Our results indicate that the optimal
extraction conditions were 1:4 of solid-to-solvent ratio, 70 ℃, pH 8 and 1 h. The trypsin inhibitor activity and content were 29.8
TIU/g and 38.66 μg/mL in seeds, 28.3 TIU/g and 37.06 μg/mL in rind, and 19.1 TIU/g and 25.33 μg/mL in pulp.

in vitro Free Radical Scavenging and Protein Oxidation Inhibitory Effects of Herbacetin
QIAO Yan,LIU Xue-bo*
2013, 34(17):  106-110.  doi:10.7506/spkx1002-6630-201317024
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The in vitro antioxidant activity of herbacetin was investigated in terms of scavenging activities against DPPH
and hydroxyl free radicals and inhibitory effects against oxidative protein damage and carbonylation induced by Cu2+-
H2O2 or AAPH. At a concentration between 10 μmol/L and 100 μmol/L, herbacetin exerted a marked scavenging effect on
DPPH and hydroxyl free radicals, with respective IC50 values of 49.28 μmol/L and 219.2 μmol/L. Meanwhile, herbacetin could
significantly inhibit oxidative protein damage and carbonylation induced by Cu2+-H2O2 or AAPH in a dose-dependent manner.
This study concludes that herbacetin has a strong ability to scavenge free radical and inhibit oxidative protein damage.

Farinograph Properties and Baking Characteristics of Dietary Fiber-fortified Wheat Dough
KONG Xiao-xue,WANG Ai,DING Qi-juan,DING Guang-qin,LIU Yang,LIU Chen,ZHENG Tie-song*
2013, 34(17):  111-115.  doi:10.7506/spkx1002-6630-201317025
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Farinograph properties and baking characteristics of soybean fiber-fortified dough were investigated in this
study. Different samples, 100% gluten dough and dough fortified with 10% soybean dietary fiber singly or together with
0.5% or 1% glyceryl monolinolenate or glyceryl monolaurate were examined comparatively. Results showed that addition
of soybean dietary fiber could shorten the stability time, increase the degree of softening and adhesiveness, and reduce
the hardness of dough. Moreover, Bread specific volume and springiness decreased and hardness increased. As a result,
the sensory quality of bread deteriorated. Addition of 1% glyceryl monolinolenate or glyceryl monolaurate resulted in a
significant decrease in water loss rate during storage of dietary fiber-fortified bread and a notable improvement in sensory
quality and baking characteristics. Thus we conclude that the negative impact of soybean dietary fiber added in dough
on farinograph properties and baking characteristics can be improved by further addition of glyceryl monolinolenate or
glyceryl monolaurate.

Extraction and Antioxidant Activity of Red Pigments from Pyracantha fortuneana
LI Peng-xiao1,MAO Guang-hua2,ZHAO Ting2,ZOU Ye2,REN Yue-na3,BAI Shi-qi1,WU Xiang-yang3,YANG Liu-qing3,*
2013, 34(17):  116-119.  doi:10.7506/spkx1002-6630-201317026
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Red pigments were extracted with 70% acidified ethanol (pH 3, HCl) and purified by C18 Sep-Pak column
chromatography. The radical scavenging activity of the purified pigments was assessed against •OH, O2
-• and
DPPH radical. The contents of anthocyanins, total phenols and total sugar in the crude extract and the purified product
were determined by pH differential method, Folin-Ciocalteau method and anthrone colorimetric method, respectively.
Results showed that the contents of anthocyanin, total phenols and total sugar in the crude extract were 1.16, 2.18 mg/100 g and
1095.34 mg/100 g, respectively, while those in the purified product were 20.61, 48.62 mg/100 g and 0 mg/100 g,
respectively. The IC50 values of the purified product against •OH, O2
-• and DPPH radical were 1.43, 3.13 μg/mL and
3.43 μg/mL, respectively. These results indicate that the C18 Sep-Pak column chromatography could effectively remove
sugar compounds and concentrate anthocyanins and total phenols in the crude extract. The purified red pigments had a
relatively strong antioxidant capacity.

Physico-chemical Properties of Konjac Fine Flour from Amorphophallus bulbifer
DENG Li-ling1,XU Xiao-qing1,YANG Yan-xun1,WANG Qiao1,ZHONG Geng1,2,*
2013, 34(17):  120-125.  doi:10.7506/spkx1002-6630-201317027
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Konjac fine flours from Amorphophallus bulbifer, Amorphophallu rivieri and Amorphophallu albus were
comparatively investigated for konjac glucomannan (KGM) content, apparent viscosity, color parameters, whiteness,
solubility and gelation properties. Meanwhile, a comparative analysis of functional groups in their structures was also carried
out by UV spectroscopy, infrared spectroscopy and nuclear magnetic resonance. Konjac fine flour from Amorphophallus
bulbifer had similar physico-chemical properties to those from Amorphophallu rivieri and Amorphophallu albus, but was
superior in terms of KGM content, viscosity, stability in solution and thermo-reversible gel blended with carrageenan.

Inhibitory Effect of Eriodictyol on Free Radical-induced Damage to Biological Macromolecules and Its Cytotoxicity on HepG2 Cells
ZHANG Yi-fan,LIU Gong-guan,LIU Qian,LIU Xue-bo*
2013, 34(17):  126-130.  doi:10.7506/spkx1002-6630-201317028
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The in vitro free radical-scavenging activity of eriodictyol was evaluated against DPPH and hydroxyl free radicals.
The protective effect of eriodictyol on free radical-induced damage to protein, lipid and DNA were evaluated based on oxidation
models of BSA, rat liver tissue and plasmid DNA induced by AAPH and its cytotoxicity on HepG2 cells was assessed by MTT
assay. The results showed that eriodictyol in the concentration range of 25 to 200 μmol/L had strong scavenging activity against
DPPH free radical with a median inhibitory concentration (IC50) of 95.7 μmol/L. However, the scavenging activity on hydroxyl
free radical was weaker. The inhibitory effect of eriodictyol on oxidative damage to protein, lipid and DNA induced by free
radicals was in a dose-dependent manner. Similarly, eriodictyol could significantly reduce the viability of HepG2 cells in a
dose-dependent manner. Therefore, eriodictyol has the capability to scavenge DPPH free radicals and inhibit macromolecular
damage induced by free radicals. It also can significantly reduce the viability of HepG2 cells.

Effect of 11S Globulin on Flour Rheological Properties and Bread Quality
YANG Chun-hua,SHI Yan-guo*
2013, 34(17):  131-135.  doi:10.7506/spkx1002-6630-201317029
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The effect of globulin on flour rheological properties and bread quality were studied. 11S globulin was extracted
from low-temperature defatted soybean meal by isoelectronic point and cooling precipitation. The extract of 11S globulin
was added to flour at various ratios to explore its rheological characteristics. The mixture flours containing 11S globulin
were used to prepare bread. The bread quality was evaluated by texture analyzer and sensory analysis. The experimental
results indicated that the addition of 11S globulin would strengthen the rheological properties and the quality of wheat flour.
The stability and farinose evaluation value reveal an obvious increase, the softness of dough was greatly reduced, the tensile
strength was increased and the gelatinization time is shortened. The quality of bread was the best by adding 11S globulin at
the ratio of 3%. The hardness, elasticity and sensory score under the conditions was 344.8 g, 0.92 and 46.7, respectively.

Putrescine-producing Property of Escherichia coli YSY1 and Related Genes
XU Wen-juan1.2,LIU Fang2,WANG Dao-ying2,ZHU Yong-zhi2,ZHOU Tao1,*,XU Wei-min2
2013, 34(17):  136-139.  doi:10.7506/spkx1002-6630-201317030
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HPLC was used to detect the levels of putrescine and spermine produced by Escherichia coli YSY1, and PCR was
used to amplify decarboxylase gene. The results showed that the level of putrescine was increased during fermentation, but the
level of spermine revealed a decrease at the same time. The content of spermine between 1 h and 6 h of fermentation changed
from 198.04 μg/mL to 156.33 μg/mL, whereas the amount of putrescine varied from 14.61 μg/mL to 18.83 μg/mL. Ornithine decarboxylase
gene was successfully amplified from Escherichia coli genome, while arginine decarboxylase gene and other related genes were not detected.

Biodiversity and Quantitative Analysis of Lactic Acid Bacteria in Traditionally Fermented Milk Products from Seven Pasturing Areas of Tibet
CHEN Zhi-lan1,YANG Ji-xia2,LI Meng-han1,LI Xiao-wei1,NAN Zhi-qiang1,GONG Wen-feng1
2013, 34(17):  140-145.  doi:10.7506/spkx1002-6630-201317031
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To investigate the biodiversity of lactic acid bacteria from traditionally fermented milk products in Tibet, 56
samples of milk products were collected from different herdsman families in seven pasturing areas of Tibet. Colony counting
was conducted for lactic acid bacteria and yeasts separately by dilution and pour-plate method. Suspected strains of lactic acid
bacteria were selected according to morphology and simple biochemical tests, and further identified by traditional taxonomy
methods and 16S rRNA sequence analysis. Results showed that the average population of lactic acid bacteria in these fermented
milk samples was (6.38 ± 1.54) (lg (CFU/mL)) compared to (5.27 ± 1.63) (lg (CFU/mL)) for yeast. A total of 110 lactic acid
bacteria were found in all the tested samples, including 96 Lactobacillus strains, 10 Lactococcus strains, 2 Enterococcus strains
and 1 Pediococcus strain. The 96 Lactobacillus strains were assigned to 8 species, including Lactobacillus casei
(48), L. plantarum (18), L. parabuchneri (9), L. fermentum (7), L.delbrueckii subsp. bulgaricus (5), L. breris (4), L. acidophilus
(2) and L. diolivorans (3). The 14 coccus strains were assigned to Lactococcus (2), Enterococcus (1) and Pediococcus (1).
Lactobacillus casei was the predominant species, accounting for 43.64% of the total isolates. This study may provide sufficient
data to elucidate a huge biodiversity in lactic acid bacteria from Tibetan traditionally fermented milk products.

Identification of Sterile Endophytic Fungus Isolated from Salicornia bigelovii
WANG Xiao-min,WANG Hui,LIU Tian-xing,XIN Zhi-hong*
2013, 34(17):  146-149.  doi:10.7506/spkx1002-6630-201317032
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The sequences of 18S rDNA and ITS gene of a fungal strain named Sal54 isolated from Salicornia bigelovii
were cloned and sequenced, and a 18S rDNA gene sequence of 1715 bp in length and an ITS gene sequence of 544 bp in
length were obtained. The accession number of the ITS gene sequence in GenBank was JX292134. Homology analysis was
conducted. A phylogenetic tree was constructed by using these sequences and the neighbor-joining method. Phylogenetic
analysis revealed that the isolated strain was identified as Phoma betae.

Isolation and Identification of Lactic Acid Bacteria from Natural Yak Yogurt in Tibet Plateau Pastoral Areas of Tibet and Western Sichuan
WU Jun1,ZHAO Xiao-juan1,CHEN Jia-xin1,DU Mu-ying1,2,3,*,KAN Jian-quan1,2,3
2013, 34(17):  150-155.  doi:10.7506/spkx1002-6630-201317033
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Totally 56 strains of lactic acid bacteria were isolated from self-made yoghurt samples collected from Tibet
plateau pastoral areas of Tibet and western Sichuan. Ten strains with better fermentation performance LAB were obtained
through preliminary screening by genetic stability, coagulation time, titratable acidity and sensory evaluation. Finally,
7 trains of LAB with excellent fermentation performance were selected. Coagulation time and titratable acidity at the
coagulation time of these 7 strains were 6-8 h and 69.99-92.31 °T. During cold storage, the titratable acidity was 8.46-
20.94 °T, and the viable bacterial pollution was 107-1011 CFU/mL. The contents of produced acetaldehyde and diacetyl
were 14.21-30.76 μg/mL and 0.51-1.55 μg/mL. Based on 16S rDNA sequence analysis, TG1-1 was identified as
Lactobacillus casei or Lactobacillus paracasei; TG1-11 as Lactobacillus paracasei, TG3-2 and DLDQ2-1 as Lactobacillus
casei, TG2-4 and KDLL-2 as Enterococcus faecium; and TG3-4 as Enterococcus durans.

Purification and Characterization of X-Prolyl Dipeptidyl Aminopeptidase from Lactobacillus helveticus
GUO Yu-xing1,ZHU Qian1,ZHU Kun1,ZHOU Hui-min1,SUN Yang-ying1,2,ZENG Xiao-qun1,2,PAN Dao-dong1,2,*
2013, 34(17):  156-160.  doi:10.7506/spkx1002-6630-201317034
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X-prolyl dipeptidyl aminopeptidase (Pep X) from Lactobacillus helveticus was purified and characterized. The
Pep X was released from Lactobacillus helveticus cells by sonication. The cell-free extract was purified by ammonium
sulfate precipitation, chromatographed on Sephacryl S-300 HR and native-PAGE gel. The molecular mass of purified Pep X
was measured by SDS-PAGE. The specific activity of the enzyme was 62.24 U/mg. The purified Pep X had monomer
molecular mass of approximately 26 kD. Optimal activity was observed at pH 7.0 and 37 ℃. It was activated by Co2+
and inhibited by Zn2+, Ni2+, Mn2+ and Fe3+. It was a metallopeptidase and serine proteinase that was inhibited by EDTA
and PMSF.

in vitro Degradation of Resistant Starch by Bifidobacteria
FAN Yuan-yuan1,WANG Shu-xiang1,TAN Mei-qing1,YANG Gong-ming2
2013, 34(17):  161-164.  doi:10.7506/spkx1002-6630-201317035
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This study aimed to investigate the in vitro degradation of resistant starch Ⅱ (RS2) from banana by
bifidobacteria. The residual starch in the degraded product was measured. Response surface methodology (RSM) was
applied to study the effect of bacterial concentration, substrate concentration and fermentation time on the degradation
of RS. A mathematical regression model was established between residual rates of RS and the three factors. The optimal
degradation condition of banana RS was obtained as follows: fermentation time 14 h, bacterial concentration 1011 CFU/mL
and substrate concentration 4 g/mL.

Optimization of Culture Medium Formulation for γ-Aminobutyric Acid-producing Lactobacillus plantarum MJ0301
HUANG Gui-dong1,MAO Jian1,2,*,JI Zhong-wei1,FU Jian-wei2,ZOU Hui-jun2
2013, 34(17):  165-170.  doi:10.7506/spkx1002-6630-201317036
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The aim of the present study was to investigate the optimization of formulation of culture medium for γ-aminobutyric
acid (GABA)-producing lactic acid bacteria. The effects of carbon sources, nitrogen sources, K2HPO4, Tween-80 and monosodium
L-glutamate (L-MSG) on the growth of L. plantarum MJ0301 and the yield of GABA were explored by single factor test. The
results showed that glucose, nitrogen (tryptone and yeast extract) and L-MSG significantly influenced the growth of L. plantarum
MJ0301 and the yield of GABA (P < 0.05). The response surface methodology was designed based on glucose, nitrogen (tryptone
and yeast extract) and L-MSG, and the yield of GABA was considered as response value. The optimal formulation of the culture
medium consisted of glucose 16.0 g/L, mixed nitrogen sources 10 g/L, L-MSG 30 g/L, ammonium citrate 2 g/L, K2HPO4
2 g/L, sodium acetate 5 g/L, magnesium sulfate 0.1 g/L, manganese sulfate 0.05 g/L, and Tween-80 1 mL/L. The resulting yield of
GABA was 1.59 g/L, which was 11.12-fold higher than before the optimization (143 mg/L).

An Enrichment Broth for Simultaneous Growth of Salmonella, Shigellosis and Staphylococcus aureus
WANG Yong-zhi1,YU Yi-gang1,HUANG Xiu-li2,XIAO Xing-long1,*,WU Hui1
2013, 34(17):  171-175.  doi:10.7506/spkx1002-6630-201317037
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In order to develop an enrichment broth for the simultaneous growth of Salmonella, Shigellosis and
Staphylococcus, several candidate agents were added to pre-formulated broth to observe their growth speeds. The
optimal performance was obtained with the enrichment broth composed of 15 g of tryptone, 5 g of soya peptone, 2.5 g of
monopotassium phosphate, 2.5 g of glucose, 35 g of sodium chloride, 0.5 g of lithium chloride, 0.1 g of bile salt, 2 g of
mannitol and 0.3 g of potassium tellurite in 1000 mL of distilled water. Results showed that Salmonella, Shigellosis and
Staphylococcus aureus could concurrently enrich in this broth and the concentration of bacteria could increased from
103 CFU/mL to 107 CFU/mL during 18 h. The growth of non-target bacteria was suppressed in this broth. Therefore, it may
be a promising broth for simultaneous detection of the three pathogens in a single platform.

Community Structure of Prokaryotes in Pit Mud of Lu-Flavor Liquor from Luzhou Prefecture Based on Culture-independent Approach
YE Guang-bin1,2,LUO Hui-bo1,2,YANG Xiao-dong1,LI Dan-yu1,WANG Yi1,NI Bin3
2013, 34(17):  176-181.  doi:10.7506/spkx1002-6630-201317038
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The community compositions of bacteria and archaea in mature pit mud of Lu-flavor liquor were detected
through establishing 16S rRNA gene clone library of bacteria and archaea, sequencing and analyzing phylogenetic tree. Data
analysis showed that the biodiversity of bacteria in pit mud was quite rich, reaching 34 OTUs, which were mainly distributed
in 14 families and 2 unidentified families of Clostridia and Bacteroidia. And also, most of them were restricted anaerobes.
Among them, Clostridia strains were absolutely dominant in the bacterial community of pit mud, and its ratio of clone
number was 68.2%. In addition, there was appropriate distribution for Bacilii (21.6%), Bacteroidetes (8.0%), Tenericutes
(1.1%) and Synergistetes (1.1%). Archaeal community composition was quite simple, and only 4 OTUs were detected
and mainly distributed in Methanoculleus, Methanosarcina, Methanobacterium and Methanomassiliicoccus with clone
distribution ratios of 58.8%, 29.4%, 5.9% and 5.9%, respectively.

Fermentation of Low-salt Pickled Radish by Pure Strains of Lactobacillus plantarum
WANG Li-ping1,2,WANG Xin1,AI Lian-zhong2,WU Zheng-jun2,ZHAO Yong1
2013, 34(17):  182-186.  doi:10.7506/spkx1002-6630-201317039
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In order to find an appropriate starter for the fermentation of pickled radish, four pure strains of
Lactobacillus plantarum isolated from traditionally pickled radish were used for the fermentation of fresh radish and
compared with Lactobacillus planterum B110 and naturally fermented pickle. The results indicated that Lactobacillus
plantarum STW-4 was the best culture starter due to the shorter fermentation time and lower nitrite content. The
fermentation conditions were optimized and as a result, the fermentation cycle was reduced to 60 h from 7 days when
compared with naturally fermented pickle under the conditions of 2% inoculation, room temperature and 8 g/100 mL salt.
In order to reduce the salt concentration but not destroy the brittleness, Lactobacillus plantarum STW-4 was inoculated
in the presence of 4 g/100 mL salt, and the brittleness of pickled turnips was 22.98 N, which revealed an improvement
from 15.67 N. This improvement may be related to the increased water loss and the inhibited activities of pectinase and
cellulose under a high salt concentration.

Isolation, Purification and Characterization of Acid Phosphatase from Chinese Chives
SUN Fang,REN Mei-feng,HU Rui-bin,TANG Yun-ming*
2013, 34(17):  187-191.  doi:10.7506/spkx1002-6630-201317040
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Fresh chives were homogenized, extracted with buffer and precipitated by ammonium sulfate. The precipitant
was dissolved in HAc-NaAc and then purified by CM-Sepharose ion-exchange chromatography and superdex-200 gel
filtration chromatography. A new type of electrophoretically pure acid phosphatase (ACP) from fresh Chinese chives was
then obtained. The enzyme activity of the purified ACP reached 645.83 U/mg. The purification factor was 454.81 and the
activity recovery was 10.50%. Characterization studies showed that the molecular mass of this enzyme was approximately
58.97 kD and the optimum reaction temperature and pH value were 65 ℃ and 5.4, respectively. The Km value for this
enzyme was 0.896 × 10-3 mol/L when using nitrophenyl phosphate disodium (pNPP) as the substrate. High concentrations
of Mg2+ and Mn2+ showed stronger activation of the enzyme while the higher concentration of ascorbic acid, methanol,
ethanol, chloroform, isopropanol, Cu2+ and Ag+, the stronger the inhibition of the enzyme.

Optimization of Fermentation Medium for Producing Uridine Phosphorylase by Lactobacillus brevis
WANG Wei-jie1,LI Hong-mei1,*,DENG Long-hua1,GAO Lu-jiao2,HUANG Yan-qing2
2013, 34(17):  192-196.  doi:10.7506/spkx1002-6630-201317041
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The culture medium and fermentation conditions for producing uridine phosphorylase by Lactobacillus brevis was
optimized by response surface methodology. On the basis of glucose-yeast extract culture medium, four factors such as glucose
concentration (P = 0.002), yeast extract (P = 0.054), phosphate group (P = 0.059) and uridine (P = 0.001) were identified as
important factors that affect the production of uridine phosphorylase by single factor and Placket-Burman designs. These factors
were further examined through steepest ascent path method to find the maximum response region and optimized by response
surface Box-Behnken design. The optimal fermentation medium comprised NaCl 5 g/L, glucose 18.30 g/L, yeast extract 15.71 g/L,
peptone 15 g/L, phosphate group 3.99 g/L, carnine 20 mmol/L and uridine concentration 21.38 mmol/L at initial pH of 8.0.
The optimal culture conditions were shaking speed of 110 r/min, culture temperature of 32 ℃, fermentation time of 14 h and
inoculum amount of 1.0%. Under the optimized conditions, the production of uridine phosphorylase by Lactobacillus brevis
was 1.325 U/mg wet bacteria, which exhibited an increase by 50.0% compared with non-optimization.

Biological Characteristics of Lactobacillin XH1 Bacteriocin Produced by Lactobacillus acidophilus
YANG Tian-you1,DUAN Gai-li2,ZHAO Rui-xiang2,*,NIU Sheng-yang2,YUAN Zheng2
2013, 34(17):  197-200.  doi:10.7506/spkx1002-6630-201317042
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Without the presence of organic acids or hydrogen peroxide, some biological characteristics such as antibacterial
activity, thermal stability, optimal pH and protease sensitivity of the bacteriocin lactobacillin XH1 produced by Lactobacillus
acidophilus were explored. The results indicated lactobacillin XH1, a broad-spectrum antimicrobial substance, could inhibit
Escherichia coli, Staphylococcus aureus and Bacillus anthraci. The optimal activity was detected under the condition of
pH 1.0-5.0 and the optimal pH was 2.0-3.0. Subjected to treatment at 121 ℃ for 20 min, lactobacillin XH1 remained unchanged
and revealed certain thermal stability. Meanwhile, lactobacillin XH1 was sensitive to trypsin, pepsin and papain, but insensitive to
proteinase K and neutral protease. Its molecular weight was approximately 30 kD as identified by Tricine-SDS-PAGE.

Optimization of Natural Fermentation Conditions for Yellow Serofluid
ZHAO Gui-li,LUO Ai-ping*,LIAO Ya-fan,WU Hong-man,YANG Jie,SONG Zhi-min
2013, 34(17):  201-204.  doi:10.7506/spkx1002-6630-201317043
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Natural fermentation of soybean yellow serofluid for acid production was investigated. Fermentation
conditions including temperature, time, inoculum amount and glucose addition were optimized by uniform design
to obtain the optimum acid production and pH. Quadratic polynomial regression analysis showed that the optimum
fermentation conditions were 37 ℃, inoculum amount of 3%, 46 h, 2 g/100 mL glucose, initial pH 6.2. The maximum acid
production under these conditions was 0.7815 g/100 mL.

Process Optimization for Antioxidant Peptide Preparation from Eucommia Seed Meal Protein by Enzymatic Hydrolysis
HUANG Qun1,2,YANG Wan-gen2,YU Ji2,MA Cheng-jin1,2,WU Li-ya2,ZENG Li-ting2
2013, 34(17):  205-209.  doi:10.7506/spkx1002-6630-201317044
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Eucommia seed meal protein was hydrolyzed with protease to prepare antioxidant peptides. Based on degree
of hydrolysis and superoxide anion radical scavenging activity, a Box-Behnken factorial design combined with response
surface analysis (RSA) was applied to optimize enzymatic hydrolysis conditions for antioxidant peptide preparation. The
results showed that the optimum hydrolysis conditions were 7.40 g/100 mL, 12292 U/g, 2.6 h, 7.0 and 50 ℃ for substrate
concentration, enzyme dosage, hydrolysis duration, initial pH and temperature, respectively. The superoxide anion radical
scavenging rate of the antioxidant peptide obtained under the optimized conditions was 56.60%.

Isolation, Identification and Culture Characteristics of Cellulase-producing Exiguobacterium sp. YT9
MIAO Jing,LI Tan-tan,CHENG Shi-wei?,LIANG Hui-hui,CUI Hai-yang
2013, 34(17):  210-214.  doi:10.7506/spkx1002-6630-201317045
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Strain YT9 with high-yield cellulase was screened from coastal soil samples, and the morphological,
physiological and biochemical characteristics as well as 16S rDNA sequence homology of the selected strain were
studied. Finally, the strain was identified as Exiguobacterium sp. Meanwhile, medium composition and culture
conditions for cellulase production were optimized. The optimal medium for higher production of cellulase consisted
of 20 g/L glucose, 0.5 g/L CMC, 8 g/L tryptone, 2 g/L ferrous alum, 2 g/L K2HPO4, 2 g/L NaH2PO4, and 1 g/L MnSO4.
The maximum activity of CMCase was 226.27 U/mL at pH 6.5, 30 ℃ and 4% inoculum. At the same time, the activities of
cellobiohydrolase and β-glucosidase were lower than the endoglucanase activity.

Analysis of Bacterial Diversity in Chinese Traditional Fermented Sichuan Pickles Using 16S rRNA Genes
TIAN Wei1,2,ZHANG Qi1,2,DENG Zhen-zhen1,2,LIU Sen1,2,LI Ming-yuan1,2,CHE Zhen-ming1,MA Li1,XIANG Wen-liang1,2,*
2013, 34(17):  215-218.  doi:10.7506/spkx1002-6630-201317046
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The bacterial diversity in Sichuan pickle samples was investigated by using the constructed 16S rRNA gene
clone library. A total of 129 representative clones were recovered and all identified as lactic acid bacteria, which belonged
to different species of the two genera Lactobacillus and Pediococcus with the proportion of 88.4% and 10.1%, respectively.
The dominant species of bacteria in Sichuan pickles were L. pentosus (50.4%), L. plantarum (16.3%) and P. damnosus
(10.1%) while other bacterial species were also observed such as L. paralimentarius (7.8%), L. sunkii (4.7%), L. brevis (3.1%),
L. kisonensis (1.6%), L. acetotolerans (0.8%) and L. namurensis (0.8%). In addition, several species such as P. damnosus,
L. paralimentarius, L. sunkii, L. kisonensis and L. acetotolerans were found in pickle for the first time. These results
revealed the microbial diversity and the microbial community structure in Sichuan pickles, which contained quantities of
undiscovered bioinformation.

Genetic Analysis of Oleocellosis in Navel Orange ‘Banfield’
WEI Zhao-xin1,CHENG Chun-zhen2,CHENG Chang-feng1,WU Chun-qing1,ZHU Shi-ping2,QI Ju-rong1,HONG Lin1,*
2013, 34(17):  219-224.  doi:10.7506/spkx1002-6630-201317047
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Oleocellosis, a widely distributed citrus disease, has a significant impact on the appearance and
commercial value of citrus fruits. To clarify the molecular mechanism of citrus oleocellosis, both forward and
reverse cDNA suppression subtractive hybridization libraries were constructed using RNA extracted from navel
oranges ‘Banfield’ with oleocellosis and healthy ones. Results showed that the length of majority of the inserts
ranged from 100 to 700 bp. A total of 292 clones were successfully sequenced from 300 randomly picked
positive clones, among which 165 ESTs in 55 genes were identified to share high homology with known genes
with E-value < 1.0×10-3. GO and KEGG analysis showed that these differentially expressed genes (DEGs)
were mainly involved in redox stress, defense response, and biotic or abiotic stress. We therefore speculated that oxidative
stress might contribute to the formation of oleocellosis.

Optimization of Fermentation Conditions for Cellulase Production by Aspergillus niger Using Response Surface Methodology
LIU Song,LI Zhu*,ZHOU Li-hong,GE Yong-yi,LIU Wu-juan,ZHANG Chuan-ping
2013, 34(17):  225-229.  doi:10.7506/spkx1002-6630-201317048
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Response surface methodology was used to optimize the fermentation conditions for cellulase production by
Aspergillus niger to obtain the maximum carboxymethyl cellulase (CMCase) activity. By one-factor-at-a-time designs, we found
the optimum fermentation conditions to be inoculum age of 24 h and incoculum amount of 10%, and 30 mL of culture medium
composed of 2 g/100 mL glucose, 1 g/100 mL yeast extract and 0.1% PEG at initial pH 7.0 filled in a 250-mL triangular flask.
Inoculum amount and glucose concentration were identified by Plackett-Burman (PB) design as main factors that influence
CMCase activity, and their optimal regions were approached by the steepest ascent path. Finally, using central composite design
and response surface analysis, the optimal fermentation medium was determined as 37.5 mL of a modified PDA medium consisting
of 2.7 g/100 mL glucose, 1.25 g/100 mL yeast extract and 0.1 g/100 mL PEG in a 250-mL triangular flask, and the optimal
fermentation conditions were 8% (3.3 mL), 7.0, 28 ℃, 150 r/min and 96 h for inoculums size, initial pH, temperature, shaking
speed and time, respectively. The maximum CMCase activity under the optimized conditions was (63.5 ± 1.7) U/mL, indicating a
29.59% increase compared to (49.0 ± 1.7) U/mL obtained under the conditions of 30 mL of PDA with 2% glucose at initial pH of 7.0 in
a 250-mL triangular flask, 10% (3 mL) inoculum size with an age of 24 h, and culture at 28 ℃ for 96 h with a shaking speed of 150 r/min.

Screening of High-yield DHA-producing Crypthecodium cohnii Mutant
SHE Jun1,2,TIAN Hua1,CHEN Tao3,HE Dong-ping1,2,*
2013, 34(17):  230-235.  doi:10.7506/spkx1002-6630-201317049
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The original strain C. cohnii ATCC 30772 was irradiated by 60Co-γ ray. Mutant 2.4k-2A2-5 was selected as a
desired positive mutant according to its growth and development, Sudan black B staining, biomass, oil content and DHA
content, following screening and rescreening. The average DHA production was 39.04%, which was 3.39% higher than the
original strain. The yield and content of oil produced by this mutant was 51.66% and 63.37%, respectively, which were also
higher than those by the original strain. The DHA yield (measured in 7.14 g/L fermented liquid) was increased by 56.92%
compared to that produced by the original strain.

Effect of Lactosucrose on Plasma Th1/Th2 Type Cytokines in Rats with Colonitis
ZHOU Yan1,2,RUAN Zheng1,2,*,HUANG Xiao-liu1,2,LI Ling1,2,LIU Wen-qun1,2,WEN Yan-mei1,2,MI Shu-mei1,2,WU Xin1,2,3,YIN Yu-long1,2,3
2013, 34(17):  236-240.  doi:10.7506/spkx1002-6630-201317050
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Objective: To explore effect of lactosucrose (LS) on blood immunity and Th1/Th2 type cytokines in rats with
colonitis. Methods: Experimental rat colitis was induced by trinitrobenzenesulfonic acid (TNBS) and randomly distributed
into three groups as follows: model control (MC) group, LS group and SASP group. The normal control (NC) group was
also included. All rats were killed on the 21st day. The activities of alkaline phosphatase (ALP), lactate dehydrogenase (LDH)
and inducible nitric oxide synthase (iNOS), and the contents of IFN-γ, IL-4 and IL-10, as well as IFN-γ/IL-4 ratio were
measured in experimental colitis rats. Results: Compared with NC group, the activity of ALP and the content of IFN-γ were
significantly increased (P < 0.01) and the content of IL-4 was significantly decreased (P < 0.01) in MC group. Moreover,
the content of IL-10 was significantly decreased (P < 0.05), the activities of LDH and iNOS and IFN-γ/IL-4 ratio were
significantly increased (P < 0.05). Compared with MC group, the activity of ALP and the contents of IL-4 and IL-10 in
LS group were significantly increased, and the activity of iNOS and the ratio of IFN-γ/IL-4 were significantly decreased.
Conclusion: The mechanism of LS in relieving inflammation in colitis may be achieved by increasing Th2 type cytokine
production and rebalancing the Th1/Th2 immune response.

Effect of Intragastric Dose of Zanthoxylum Essential Oil on Intestinal Health in Ovarietomized Rats
LIU Qing-qing,REN Wen-jin,Lü Jiao,LIU Xiong*
2013, 34(17):  241-245.  doi:10.7506/spkx1002-6630-201317051
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Objective: To explore the effect of different intragastric doses of Zanthoxylum essential oil on intestinal health in
ovariectomized rats. Methods: Totally 40 female SD rats were randomly divided into five groups, including a group of rats
with pseudo-resection of ovarian, and the rest 4 groups of rats subjected to ovariectomy. After a week of wound healing from
surgery, both of the pseudo ovarian resection group and ovariectomy group were fed on basic feed, and three ovariectomy groups
were orally administered with the essential oil at doses of 5, 10 mg/(kg·d) and 15 mg/(kg·d) body weight per day for four
consecutive weeks. All rats were sacrificed after feeding for 28 days to analyze the gain of body weight and cecal tissue weight,
the amount of free ammonia, short-chain fatty acids (SCFAs) and microbes in the cecum. Results: After intragastric administration
of Zanthoxylum essential oil, the wet weight of cecal wall of these rats was increased, pH was decreased, harmful bacteria and
free ammonia were reduced, and total SCFAs in medium-dose and high-dose groups were increased. Conclusion: Intragastric
administration of Zanthoxylum essential oil can promote rat intestinal fermentation, and thus improve the intestinal health.

Molecular Mechanism of DHA for Improving Learning and Memory in Rats
MA Lin,WANG Li-mei,FAN Yong-bo,WU Chen,TAO Xing-wu,LIU Zhi-guo,LIU Lie-ju*
2013, 34(17):  246-249.  doi:10.7506/spkx1002-6630-201317052
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This study was aimed to investigate the effect of docosahexaenoic acid on learning and memory abilities in rats
and to explore the relevant molecular mechanism. A total of 32 neonatal male SD rats were randomly divided into 4 groups:
negative control group, positive control group, low (370 mg/(kg·d)) and high dose DHA group (740 mg/(kg·d)). After
8 weeks of feeding, the enzyme activities of acetylcholinesterase (AChE) and nitric oxide synthase (NOS) in brain tissue
of rats were analyzed. The mRNA expressions of N-methyl-D-aspartate receptor subunit (NR1), cAMP responsive element
binding protein (CERB), immediate early gene (c-fos) in the hippocampus of rats were determined by RT-PCR. Results
showed that compared to the negative control group, DHA at both doses was able to significantly increase AChE and NOS
activities in brain tissue as well as the mRNA expression levels of NR1, CREB and c-fos while the low dose presented
higher effect. In summary, DHA could remarkably improve learning and memory abilities in rats in a dose-dependent
manner. The mechanism might be involved with the upregulation of NR1, CREB and c-fos mRNA expression by DHA in the
hippocampus of rats.

Effects of Glycoprotein and Glycopeptides from Jellyfish on Immune Function of Mice in vitro
REN Guo-yan,LIU Zhi-long,GUO Jin-ying,FAN Jin-ling,LIANG Wang-chun,SHAO Zheng,KANG Huai-bin
2013, 34(17):  250-253.  doi:10.7506/spkx1002-6630-201317053
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The immunoregulatory activity of glycoprotein and glycopeptides from jellyfish was explored through cell
proliferation and cell transformation of mouse spleen lymphocytes and comparatively evaluated under various cultivation
times and concentrations. Jellyfish glycoprotein and glycopeptides revealed significant increase in the proliferation and
transformation of spleen lymphocytes in a dose-dependent manner. The results demonstrated that glycopeptides had
better immunoregulatory activity in vitro when spleen lymphocytes were cultivated for 60 hours and the concentration of
glycopeptides was 50 μg/mL.

Dietary Exposure Assessment of Chlorpyrifos in Cowpea Before and After Cooking
WANG Xiang-wei1, QIU Hou-yuan1,CHEN Wen-xue1, ZHANG Zhi-heng2,YUAN Yu-wei2,WU Li-yu2,WANG Qiang 2,*
2013, 34(17):  254-258.  doi:10.7506/spkx1002-6630-201317054
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This study was aimed to investigate the application of processing factors to dietary exposure assessment of
chlorpyrifos in cowpea during cooking in different age groups of different genders. Subjects of different ages were divided
into 10 groups: 2-3 years old, 4-6 years old, 7-10 years old, 11-13 years old, 14-17 years old, 18-29 years old, 30-
44 years old, 45-59 years old, 60–69 years old and ≥ 70 years old. The results showed the processing factors of cowpea
were obtained in different cooking modes after cleaning such as stir frying, microwave cooking, blanching and marination.
The chronic exposure risk index (CRfD) range was 7.5741-23.5425, lower than 100 and thus acceptabl, whereas the CRfD
range was 57.3599-111.0397 without taking the processing factors into consideration and the values in groups of 2-3, 4-
6 and 7-10 of both genders ranged from 102.0552 to 111.0397, which were higher than 100 and highly risky for subjects
in these groups. Accordingly, CRfD values not based on processing factors were 5-10 times more than those based on
processing factors. The value of dietary exposure assessment was more precise by considering the influence of processing
factors on chlorpyrifos in cowpea. All the different cooking modes following cleaning were effective to remove chlorpyrifos
residues from cowpea and microwave cooking showed the best effect.

Comparative Analysis of Nutritional Compositions in Muscle of Female and Male Phoxinus lagowskii Dybowski
ZHANG Yong-quan,YIN Jia-sheng*,DU Jia,GENG Long-wu,BAI Qing-li,XU Wei
2013, 34(17):  259-262.  doi:10.7506/spkx1002-6630-201317055
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The nutritional composition of the muscle of male and female Phoxinus lagowskii Dybowski was analyzed. The
results showed that the contents of crude protein in the fresh muscle of female and male Phoxinus lagowskii Dybowski were
19.45% and 18.95%, crude fat were 3.69% and 2.97%, moisture were 75.73% and 76.93%, ash were 1.43% and 1.37%,
respectively. There were no significant differences between both sexes (P > 0.05). The total content of amino acids (TAA),
essential amino acids and delicious amino acids (DAA) of female Phoxinus lagowskii Dybowski was significantly higher
than that of male Phoxinus lagowskii Dybowski (P < 0.05). There was no significant difference in the contents of Cys, Phe,
Trp or His, while a significant difference in the content of 14 other amino acids was observed. The constitutional rate of the
essential amino acids accorded with the FAO/WHO standard. The contents of Lys and Leu were higher. According to nutrition
evaluation in amino acids score (AAS), the first limiting amino acid was Trp and the second limiting amino acid was Val,
whereas the first limiting amino acid was Trp and the second limiting amino acid was Met + Cys in chemical score (CS). The
essential amino acid (EAAI) indexes of female and male Phoxinus lagowskii Dybowski were 61.97 and 54.51, respectively. In
comparison with other economic fishes, Phoxinus lagowskii Dybowski could be regarded as a source of high-quality protein
and amino acids, and the nutritional value of female was better than that of male by comparative analysis.

Analysis of Nutritional Composition of Edible Parts of Cultellus attenuatus Dunker
XU Jia-tao,XU Guo-cheng,XU Xing-hong,YAN Bin-lun,ZHOU Hua-shan
2013, 34(17):  263-267.  doi:10.7506/spkx1002-6630-201317056
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The nutritional composition of edible parts of Cultellus attenuatus Dunker grown in Haizhou bay was
investigated and compared with other razor clams. The results showed that the yield of edible parts of Cultellus attenuatus
Dunker was 57.58 %, and the contents of moisture, crude protein, crude fat and crude ash in fresh edible parts of C.
attenuatus were 83.01%, 10.83%, 1.17% and 2.03%, respectively. A total of 17 types of amino acids were detected in the
edible parts of C. attenuatus. In dry samples, the total content of amino acids was 54.10%. Among them, the contents of
essential amino acids, half-essential amino acids, nonessential amino acids and delicious amino acids were 19.46%, 7.61%,
27.03% and 24.97%, respectively. The ratio of total essential amino acids to total amino acids was 35.97%, and the ratio of
total essential amino acids to total nonessential amino acids was 71.99%. This amino acid composition met the FAO/WHO
standards for high quality protein. According to nutritional evaluation based on amino acid score and chemical score, the
first limiting amino acid was Met + Cys and the second limiting amino acid was Val. The essential amino acid index was
66.83, which was much higher than that reported for Novaculina Chinensis (36.87). The ratio of branch-chain amino acids
to aromatic amino acids was 1.88, which was lower than that of other razor clams. A toal of 27 types of common fatty acids
were found in the edible parts of C. attenuatus, including 8 saturated fatty acids (SFAs), 9 mono-unsaturated fatty acids
(MUFAs) and 10 polyunsaturated fatty acids (PUFAs). The contents of SFAs, MUFAs and PUFAs were 32.72%, 30.80%
and 36.48%, respectively. In the PUFAs, the content of ω-3PUFAs was much higher than that of ω-6PUFA, and the content
of EPA and DHA was 71.22%. In conclusion, this investigation indicates that C. attenuatus has high nutritional value and
health benefits.

Mechanisms Underlying the Protective Effect of Saponins Extracted from Fermented Codonopsis lanceolata against Oxidative DNA Damage Induced by Diethylnitrosamine in Liver Cells of Mice
YU Xing1,ZHENG Chun-ji1,HAN Chun-ji1,2,*,JIANG Guo-zhe1,CUI Cheng-bi2
2013, 34(17):  268-272.  doi:10.7506/spkx1002-6630-201317057
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Objective: To explore the mechanisms by which saponins extracted from fermented Codonopsis lanceolata
(FCLS) protect against oxidative DNA damage induced by diethylnitrosamine (DEN) in mouse liver cells and consequently
provide useful references for chemical prevention of liver cancer. Methods: Fifty mice were randomly divided into five
groups including negative control group (intragastric administration of 10 mL of distilled water per kg body weight), DEN
group (intraperitoneal injection of DEN at a dose of 20 mg/kg every other day), and high, middle and low dose FCLS groups
(intragastric administration of FCLS at doses of 200, 100, 50 mg/(kg·d), and intraperitoneal injection of DEN at a dose
of 20 mg/kg every other day). The administration period for all the treatments was 6 weeks. The contents of 8-hydroxyl
deoxyguanosine (8-OHdG) and the activity of 8-hydroxy guanine-DNA glycosylase (OGG1) in mouse hepatic tissue were
determined by ELISA, and cytochrome P4502E1 (CYP2E1) protein expression was analyzed by immunohistochemical
method. Results: The 8-OHdG content and CYP2E1 protein expression in liver tissue from the DEN group were significantly
increased when compared with those from the negative control group (P < 0.01), and OGG1 activity in the DEN group was
significantly decreased when compared with the control group (P < 0.01). The 8-OHdG content of liver tissue in the high
and middle dose FCLS groups were significantly lower than that of DEN group (P < 0.01), and increasing the FCLS dose
resulted in a lower 8-OHdG content in the FCLS treatment groups. OGG1 content in liver tissue from the DEN group was
obviously lower than that from the negative control group (P < 0.05). OGG1 content in liver tissue from the three FCLS
groups were significantly higher than that from the DEN group (P < 0.05 or P < 0.01). The dose of FCLS revealed an obvious dependent relationship with the activity of OGG1. CYP2E1 protein expression in the FCLS middle and high dose groups
were significantly lower than that from the DEN group (P < 0.05 or P < 0.01). Conclusion: The protective effect of saponins
from fermented Codonopsis lanceolata against oxidative DNA damage induced by DEN may be related to the increased activity of
antioxidant enzymes and DNA damage repair enzymes, and reduced activity of metabolic enzymes for drugs.

Pectin Ameliorates the Impact of Capsaicin on Intestinal Fermentation Environment
Lü Jiao,SU Xin-feng,FANG Guo-shan,LIU Xiong*
2013, 34(17):  273-277.  doi:10.7506/spkx1002-6630-201317058
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The chili pepper has a wide range of consumers; however, the consumption of chili will cause a discomfort
reaction of intestinal environment with a potential hazard. As an intestinal prebiotic factor, pectin is beneficial to the
improvement of the intestinal environment. In this study, Ficus pumila Linn pectin was used as the raw material to explore
the improvement of pectin on capsaicin-induced intestinal discomfort reaction. Supernatants from cecal contents of
healthy rats were fermented on a thermostat shaker at 37 ℃ along with blank, 5% pectin, 5% pectin + 0.1% capsaicin and
0.1% capsaicin, respectively. Based on these experiments, we found that capsaicin significantly decreased the amount of
Lactobacillus and Bifidobacterium, increased Enterobacter, also reduced the concentration of intestinal short-chain fatty
acids, and increased free ammonia concentration and pH. The pectin resulted in a significant increase in the amount of
Lactobacillus and Bifidobacterium and an obvious decrease in Enterobacter; moreover the concentration of intestinal shortchain
fatty acids was increased and free ammonia concentration and pH were decreased. These results show that Ficus
pumila Linn pectin could ameliorate capsaicin-induced deterioration of the intestinal environment.

Effect of High-fat Diet on the Structure of Gut Microbiota in Human Flora-associated Mice
CHEN Xing-yun1,ZENG Ben-hua2,WEI Hong2,HU Xin-zhong1,3,*
2013, 34(17):  278-283.  doi:10.7506/spkx1002-6630-201317059
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In order to explore the relationship among diet, obesity and intestinal microbiota, high-fat diet was used to
establish obese human flora-associated (HFA) mouse model to investigate the effect of high fat diet on the structure of
gut microbiota in HFA mice. Totally 20 pathogen-free mice were inoculated with fecal suspension derived from a healthy
volunteer to obtain HFA mice. The HFA mice were fed a control or a high-fat diet for 8 weeks. Body weight, blood glucose
and blood fat were determined and the change in gut microbiota was analyzed by PCR-DGGE. Results showed that body
weight, liver weight, fat tissue weight and serum total triglyceride level revealed a significant increase (P < 0.01) in the
high-fat group when compared with the control group. Blood glucose level also increased significantly (P < 0.05) in highfat
diet-fed HFA mice. The gut bacterial diversity index of the high-fat group significantly increased (P < 0.05). The
abundance of the dominant bacteria at zero time reduced, but the abundance of bacteria that were not dominant at zero time
increased. DNA sequencing showed that high-fat diet may induce Staphylococcus lentus, Staphylococcus vitulinus and
Shigella flexneri to bloom in the gut of HFA mice. An obese HFA mouse model was successfully established by feeding a
high-fat diet and high-fat diet obviously altered the structure of gut microbiota in HFA mice. The composition and bacterial
abundance of intestinal flora in high-fat diet-feed HFA mice greatly changed, leading to intestinal dysbacteriosis. It suggests
that human-derived intestinal flora may be involved in the development of obesity caused by imbalanced dietary profile.

Acute Toxicity of Glutelin in Transgenic Rice Expressing Lysine-rich Fusion Protein Gene
ZHAO Xiang-xiang 1,LU Chang-li1,HU Xiao-lan1,JI Li-lian1,*,LIU Qiao-quan2,TANG Tang1,LIU Fu-xia1
2013, 34(17):  284-288.  doi:10.7506/spkx1002-6630-201317060
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Lysine-rich rice is a transgenic rice produced by inserting lysine-rich fusion protein gene into the germline of
rice seeds. According to the national standard of the People’s Republic of China, the food safety of glutelin extracted from
this transgenic rice by low alkali method was evaluated using acute toxicity test. In this study, glutelin was administered to
ICR mice by oral gavage at the dose of 5000 mg/kg. for continuous 14 d. Some indicators including body weight, routine
blood test, relative organ weights and histopathological examinations were conducted. The results showed that no toxic
symptom was found in tested mice in the acute toxicity test, and the LD50 for glutelin was higher than 5000 mg/kg. The
glutelin from transgenic GL gene rice, therefore, could be classified as actual nontoxic. No adverse effect on animal behavior
or mortality was observed during the experiment. There were no significant biological changes in body weight, routine blood
parameters or relative organ weights and no histopathological damage was detected. Our results suggested that the glutelin
from transgenic GL gene rice had no acute toxic effect on ICR mice.

Effect of Gender, Position and Feeding Time on Fatty Acid Composition of Rex Rabbit Meat
HUANG Ye-chuan1,2,LI Hong-jun1,*,HE Zhi-fei1
2013, 34(17):  289-294.  doi:10.7506/spkx1002-6630-201317061
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This study was aimed to investigate the effect of sampling position, gender and feeding time on fatty acid
composition of Rex rabbit meat. Six 90-day-old and six 150-day-old (half male and half female) French Rex rabbits were
selected randomly and the fatty acid composition and lipid content of leg muscle and dorsal muscle in these samples were
analyzed. The results revealed significant differences in the total intramuscular lipid content between both genders and
between two feeding times (P < 0.05), which might be mainly attributed to the content of triglyceride (P < 0.05). A
significant difference was observed for total fatty acid composition between both genders and between two feeding times
(P < 0.05) and this marked difference between two feeding times might be related to its intramuscular triglyceride content.
However, the difference between two genders might be caused by the differences in both triglycerides and phospholipids.
Therefore among these three factors studied in this paper, gender made the biggest contributions to fatty acid composition of
French Rex rabbits, followed by feeding time, and sampling position showed the least effect.

A Preliminary Investigation of Astaxanthin Metabolism and Accumulation in Artemia
BAI Yan,PEI Ling-peng*
2013, 34(17):  295-298.  doi:10.7506/spkx1002-6630-201317062
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This study was aimed to investigate the in vivo metabolism and accumulation of astaxanthin from Adonis
aestivalis in Artemia (brine shrimps). A total of 1400 48-h fasted adult brine shrimps were randomly divided into two groups
(700 brine shrimps per group): control and experimental groups. Shrimps were fed on dried powder of Adonis aestivalis
flower in experimental group and each group was extracted at 0.5, 1, 2, 3, 4 h and 5 h for C18-HPLC analysis. Results
showed that free astaxanthin started to accumulate in shrimps following feeding of Adonis aestivalis flower and reached the
maximum in 3 h. In addition, astaxanthin ethers in brine shrimps gradually disappeared. In conclusion, administration of
Adonis aestivalis flower powder to brine shrimps led to an increase in free astaxanthin. The astaxanthin eithers from Adonis
aestivalis flower were metabolically transformed into free astaxanthin and accumulated in vivo.

Effect of Cold-pressed Grape Seed Superfine Powder (CP-GSSP) on Antioxidative Functions in Aging Mouse Viscera
LIU Xia1,YANG Ji-hong1,2,3,WANG Hai-rui4,SUI Yin-qiang1,LI Hua1,2,3,WANG Hua1,2,3,*
2013, 34(17):  299-304.  doi:10.7506/spkx1002-6630-201317063
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Objective: To explore the effect of cold-pressed grape seed superfine powder (CP-GSSP) on visceral anti-oxidative
function in aging mice. Methods: Sixty mice were randomly divided into CP-GSSP low, medium and high dose groups (323,
646, 1938 mg/(kg•d)), VE group (25 mg/(kg•d)), aging model group and normal control group. The mice in the normal control
group were intraperitoneally injected with physiological saline, whereas, other groups were intraperitoneally injected with
D-galactose to establish aging mouse model. Along with intraperitoneal injection, the mice from the CP-GSSP groups, VE
group, aging model group and normal control group were intragastrically administered with CP-GSSP at various doses, VE,
edible oil and edible oil, respectively. Forty-five days later, the content of MDA, and the activities of SOD and
GSH-Px in heart, kidney and liver were detected. Results: CP-GSSP resisted the aging performance caused by D-galactose
without causing adverse effects in viscera, and significantly reduced the content of MDA, and improved the activities of
SOD and GSH-Px in heart, kidney and liver. Conclusion: CP-GSSP can inhibit lipid peroxidation and improve the activity of
endogenous antioxidases in aging mice. Therefore, it has a good antioxidant and anti-aging effect.

Immunoenhancing Activity of Mactra chinensis Meat Extract in Mice
SHI Qian1,WANG Xiao-jie1,*,ZHANG Zhi-wei1,LIU Xiao-chen1,LIANG Jian-guang1,WANG Shou-hai1,LIU Xin-sheng2,QU Yan-qing2
2013, 34(17):  305-308.  doi:10.7506/spkx1002-6630-201317064
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Objective: To observe the immunoregulatory effect of Mactra chinensis meat extract (EMC, containing 19.01 g
of polysaccharide, 1.25 g of taurine and 0.65 g of choline /100 g) in mice. Methods: Mice were orally administered with EMC
at doses of 0.5, 1.5, 3.0 g/(kg•d) for 10 d. A series of immune function indices such as the total number and classification of
white blood cells (WBC), the titer of antibody production in agglutination reaction and the quantity of plaque-forming cells
(PFC), macrophage phagocytosis test, and the weight of body and immune organs were analyzed. Results: Compared with
the control group, the relative number of lymphocytes, the activity of antibody production, the quantity of antibody-forming
cells, the phagocytic function of mouse peritoneal macrophages and the index of spleen in all the treated groups were improved
significantly (P <0.05). Conclusion: EMC enhances the immune activity of normal mice effectively.

Identification of Species and Nutritional Analysis of Alpine Thin-shelled Cipangopaludina from Qingyuan County, Zhejiang Province
LU Hong-fa1,ZHENG Shan-jian1,FANG Mei-juan1,PAN Li-xin1,CHEN Shu-fen2
2013, 34(17):  309-312.  doi:10.7506/spkx1002-6630-201317065
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By morphological and molecular identifications, alpine thin-shelled Cipangopaludina was speculated as a
variant or a novel species. Our determinations showed that the contents of crude protein and fat in a fresh sample were
(14.20 ± 0.26)% and (0.70 ± 0.03)%, respectively. A total of 17 types of common amino acids were found in thin-shelled
Cipangopaludina including 8 essential amino acids (EAA) and 2 semi-essential acids (SEAA). One gram of dried sample
contained 587.08 mg of amino acids, 280.08 mg of EAA (including SEAA), 27.37 mg of Val, 14.63 mg of Met and 26.02 mg
of Ile, which were significantly higher than the values reported for the dried samples of C. cathayessis and B. aeruginosa.
Phe and Lys were also higher than in C. cathayessis. The first limiting amino acid was Met + Cys, and the second limiting
amino acid was Val. Unsaturated fatty acid (UFA) accounted for 44.18% of the total fatty acids, of which the contents
of C18:1n-9, C18:2n-6,9, C18:3n-6,9,12 and C18:3n-3,6,9 were 6.15%, 13.11%, 9.42% and 5.54%, respectively. Moreover, thin-shelled
Cipangopaludina was rich in trace elements and minerals, especially Ca and Zn and their contents were up to (1762.667 ± 36.074)
and (5.017 ± 0.076) mg/100 g in a fresh sample, respectively. Thin-shelled Cipangopaludina, therefore, could be used as a
primary calcium and zinc supplement.

Xylanase from Pleurotus ostreatus SYJ-042 Degrades Hemicellulose and Ameliorates Intestinal Microeological Balance
LI Tian-ge1,YUE Xiao-yu2,3,*,LI Zi-gang2,CAI Qing-he3
2013, 34(17):  313-316.  doi:10.7506/spkx1002-6630-201317066
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Xylanase has a promising potential of applications in the feed industry as well as the food and pulp industry. These
applications of xylanase from Pleurotus ostreatus SYJ-042 were investigated. The results showed that the degradation of
hemicellulose in pear pericarp was gradually increased with reaction time. The degradation rate at 5 h reached 62.44%. Animal
experiments showed that the experimental group fed on xylanase-containing wheat diet had a higher food intake than the control
group and the difference was significant. The daily weight gain was also higher than that of the control group and a significant
difference was observed (P < 0.01). However, the dietary hemicellulose degradation rate in the experimental group compared
with the control group did not differ significantly. Administration of xylanase-containing diet also decreased the total bacterial
count in the jejunum and colon (P < 0.01) without resulting in a significant difference for the cecum (P < 0.01). In conclusion,
the xylanase was beneficial to remove the pericarp, promote the growth of rats, and maintain intestinal microecological balance.

Effect of Squid Ink Melanin-Fe on Hematopoietic Modulators in Rats with Iron Deficiency Anemia
LIU Dong,WANG Jing-feng,ZHANG Xiao-zhe,LONG Teng-teng,WANG Yu-ming,XUE Chang-hu*
2013, 34(17):  317-322.  doi:10.7506/spkx1002-6630-201317067
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Objective: To investigate the effect of squid ink melanin-Fe on hematopoietic modulators in rats. Methods: Iron
deficiency anemia (IDA) model rats were established by feeding with iron-deprived food and bleeding via tail vein. IDA rats
were administered with squid ink melanin-Fe at different dosages for 30 days. Selected indicators such as red blood cells
(RBC), hemoglobin (Hb), hematocrit (HCT), mean corpuscular volume (MCV) and serum erythropoietin contents were
monitored. Lung conditioned medium (LCM), spleen cell conditioned medium (SCM) and abdominal cavity macrophage
treated with squid ink melanin-Fe were prepared. The effects of the different conditional media on the proliferation of bone
marrow cells and the formation of hematopoietic progenitor cells colonies including CFU-GM, CFU-E and CFU-Meg
were evaluated. The mRNA expression of GM-CSF, EPOR and TPO were determined by semi-quantity RT-PCR. Results:
Compared with the model group, RBC, Hb, HCT, MCV and serum erythropoietin contents in squid ink melanin-Fe group
revealed a significant increase. The squid ink melanin-Fe could significantly promote the proliferation of bone marrow cells
and the colony formation of CFU-GM and CFU-E in normal rats, but each colony showed various features. The squid ink
melanin-Fe could increase the mRNA expression of GM-CSF and EPOR in bone marrow cells. Conclusion: The squid ink
melanin-Fe can promote the formation of hematopoietic progenitor cells and the differentiation of granulocyte-macrophage
and erythroid line by up-regulating hemopoietic growth factor in IDA rats.

Effects of Defatted Walnut Meal on Learning and Memory Ability and Antioxidant Capacity of Rats
FAN Yong-bo,TAO Xing-wu*,MA Lin,LIU Lie-ju,LIU Zhi-guo,WANG Li-mei
2013, 34(17):  323-326.  doi:10.7506/spkx1002-6630-201317068
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Objective: To explore the effect of defatted walnut meal on learning and memory ability and antioxidant capacity in
rats and its corresponding mechanisms. Methods: Totally 64 rats were randomly divided into eight groups: normal feed group,
defatted soybean meal (2.5 g/(kg•d)) group, walnut oil (2.5 mL/(kg•d)) group, walnut (3.0 g/(kg•d))) group and defatted walnut
meal groups at doses of 1.0, 2.0, 3.0 g/(kg•d) and 4.0 g/(kg•d). Following continuous feeding for 60 days, Step-down and
Morris water maze tests were used to evaluate the learning and memory ability of rats. The contents of superoxide dismutase
(SOD), malondialdehyde (MDA), acetylcholinesterase (AChE) and nitric oxide synthase (NOS) in brain tissues of rats were
determined. Results: Moderate consumption of defatted walnut meal could greatly improve the performance of Step-down and
Morris water maze tests, increase the activities of SOD and AChE in brain tissues of rats and reduce MDA level. Conclusion:
Defatted walnut meal can obviously improve the learning and memory abilities and antioxidant capacity of rats.

Synthesis and Antiproliferative Effect on Esophagus Cancer Cells and Interaction with DNA of Eu-Momordica charantia L. Polysaccharide Complex
ZHANG Cui1,2,SUN Xiao-bo1,SONG Ai-rong2,PAN Wei1,HAN Zhi-jian1,2,DING Lu-jun1,WANG Jin-ping1,2,*
2013, 34(17):  327-332.  doi:10.7506/spkx1002-6630-201317069
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Objective: Polysaccharides have antitumor activity due to the stimulation to immune functions of human
body, while rare earth elements have antitumor ability by killing tumor cells. In order to exploit food or healthcare
products with antitumor effect, Eu-Momordica charantia L. polysaccharide complex (Eu-MCPS) was synthesized
with the expectation of synergistic antitumor effects of the individuals. Methods: Eu-MCPS was synthesized with
homogeneous precipitation by using Momordica charantia L. polysaccharide as the template. The complex was
subjected to structural characterization employing FT-IR, scanning electron microscope and transmission electron
microscope and its antitumor activity was evaluated by MTT assay. In addition, the interaction between Eu-MCPS
and DNA was studied by UV-Vis spectroscopy and fluorescence spectroscopy. Results: Eu adhered to the twining
and helical chains of Momordica charantia L. polysaccharide in the form of Eu(OH)CO3 particles instead of forming
chemical bond with it. The particle size of Eu-MCPS varied from 340 nm to 540 nm. Eu-MCPS inhibited the
proliferation of esophagus cancer cells, and its inhibitory rate was 21.1% at the concentration of 100 μg/mL. The
interaction between Eu-MCPS and DNA may be a non-intercalation mode. These results can provide useful references
to develop special functional foods or healthcare products.

Impact of Climate Change on China’s Food Safety: Risk Source and Management
SUN Zhen-qing,JIA Xu
2013, 34(17):  333-337.  doi:10.7506/spkx1002-6630-201317070
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Global environmental changes have led to an increase in soil and water pollution, caused the fluctuations of
the number and types of pests, changed distribution and transmission of bacteria and posed a threat to food safety around
the whole industrial chain link. From the perspective of industry chain, this paper analyzed the food safety risks posed by
climate change. It was believed that the impact of climate change on food safety risk is growing. Necessary measures must
be taken to control the risk to a minimu

Advances in Research on Sprouting Inhibition of Potatoes
ZHAO Shuang,FU Mao-run*,LIU Xiu-he
2013, 34(17):  338-343.  doi:10.7506/spkx1002-6630-201317071
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Potato is one of the most important crops worldwide. Sprouting after the termination of dormancy brings huge
economic losses and food safety problems. Therefore, rising attention has been paid in sprouting inhibition technology in
recent years. Some of the techniques currently applied in sprouting inhibition, such as cool storage, chemical sprout inhibitor
and radiation, are summarized and compared briefly in this paper. In addition, development of some new sprouting inhibition
technologies is discussed in hope of providing references for further research and industrial development of potato sprouting
inhibition technology.

A Review of Studies on the Effect of Non-thermal Processing on Food Allergens
XIE Xiu-ling1,2,3,LI Xin1,3,GAO Jin-yan1,3,CHEN Hong-bing1,2,*
2013, 34(17):  344-349.  doi:10.7506/spkx1002-6630-201317072
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Non-thermal processing can modify the structures of food allergens so as to achieve the controlling of food
allergens via non-thermal treatment. Currently, common non-thermal treatments include physical methods, chemical
methods, biological methods and enzymatic modification. Physical methods mainly refer to ultra-high pressure, irradiation,
ultrasound and high-voltage pulse, which could influence the high-level structure and conformation of allergen epitopes.
Moreover, high irradiation dose is also able to modify the linear epitopes. Chemical methods, such as glycosylation,
methylation and phsophorylation, mainly destroy linear epitopes by modifying the primary structure of allergen proteins.
However, enzymatic modification is widely used to decrease the allergenicity by enzymatic cross-linking to aggregate or
degrade allergens. Biological methods are mainly realized by mutating allergenic epitopes of allergens or allergen protein
gene silencing via genetic engineering techniques, consequently leading to modification of epitopes and no expression of
allergens. This paper summarize the recent progress in research on non-thermal processing to control food allergens, in order
to provide a theoretical basis for development and production of desensitized food.

Advances in Toxicity, Detection and Detoxification of Trichothecenes
XUE Hua-li1,2,BI Yang1,*,WANG Yi1,GE Yong-hong1,LI Yong-cai1
2013, 34(17):  350-355.  doi:10.7506/spkx1002-6630-201317073
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Trichothecenes are a class of mycotoxins, widely present in grains and animal feed as well as human food
produced from contaminated grain. The mycotoxins pose a potential health threat to humans and livestock. Toxic effects,
analytical method and detoxification technique are reviewed in this paper and recent advance in techniques for their detection
and detoxification are also discussed in detail. This review is expected to provide helpful references for controlling the use of
trichothecenes in grain and produces.

Classification of Class Ⅱa Bacteriocins from Lactic Acid Bacteria
ZHAO Ai-zhen,XU Xing-ran
2013, 34(17):  356-360.  doi:10.7506/spkx1002-6630-201317074
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Class Ⅱa bacteriocins produced by lactic acid bacteria (LAB) belong to antimicrobial peptides and have
potential applications as food preservatives or therapeutic agents. In this article, 30 class Ⅱa bacteriocins are classified into
different groups according to native bacteria, production methods, antimicrobial functions, antibacterial spectrum, action
modes and structures.