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15 December 2013, Volume 34 Issue 23
Functional Properties of Pea Protein Isolate Subjected to Phenol Removal and Alkali Treatment
JIANG Jiang1,ZHU Bo1,LIU Yuan-fa1,*,XIONG You-ling L.2
2013, 34(23):  1-5.  doi:10.7506/spkx1002-6630-201323001
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Pea protein isolate (PPI) was prepared from defatted pea flour by sequential treatment with methanol followed by
acetone and then subjected to alkali treatment. The effects of phenol removal (with methanol/acetone extraction) and alkali
treatment at pH 12 on the particle size, solubility and emulsifying properties of PPI were investigated. The amino acid profile
and protein subunit changes were examined to elucidate the relationship between protein structure and functionality. Gentisic
acid, hydroxybenzoic acid, coumaric acid and ferulic acid were identified using HPLC with phenolic compound standards.
Decreased protein particle size and improved solubility and emulsifying properties were observed in PPI after both treatments.
These processes also selectively removed some polypetides rich in arginine and cysteine. However, alkali treatment
was more effective to improve protein functionality than phenol removal.

Spectroscopic Analysis of Interaction between Bovine Serum Albumin and Anthocyanin
YAO Hui-fang,JING Hao*
2013, 34(23):  6-10.  doi:10.7506/spkx1002-6630-201323002
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The interaction between anthocyanin and bovine serum albumin (BSA) was studied by UV-Vis, fluorescence and
Fourier transform infrared spectroscopy (FT-IR). The results suggested that anthocyanin had a strong ability to quench the
BSA fluorescence in a static mode. The binding constants (KA) and site numbers (n) obtained at different temperatures were
2.218 × 104, 1.084 (298 K); 1.770 × 104, 1.065 (306 K) and 1.706 × 104, 1.086 (310 K), respectively. According to the
thermodynamic parameters, electrostatic forces played a dominant role in the interaction between anthocyanin and BSA. The
distance between donor and acceptor in anthocyanin-BSA complex was calculated as 2.1 nm, based on the equations from
Förster nonradiative energy transfer theory. The synchronous fluorescence spectra revealed that anthocyanin interacted with
tryptophan residues in BSA, and the vicinity of tryptophan residues was more hydrophobic. The FT-IR spectra revealed that
conformational changes of BSA were caused by anthocyanin, thus leading to structural changes of α-helix.

Rheological and Low-field NMR Characteristics of Chicken Liver Emulsion Composite Gel
WANG Peng,WANG Hua-wei,SHAO Jun-jie,SUN Jian,ZHAO Ying-ying,XIA Tian-lan,XU Xing-lian*,ZHOU Guang-hong
2013, 34(23):  11-14.  doi:10.7506/spkx1002-6630-201323003
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With the purpose of exploring the theoretical feasibility of new chicken liver products, chicken liver-chicken
breast meat-vegetable oil emulsion composite gels were prepared by shear emulsification and investigated for dynamic
rheological and low-field NMR characteristics. The results showed that the emulsion composite gels exhibited an obvious
weak gel property after heating treatment and possessed simultaneously a higher viscoelastic modulus and a better thermal
stability, which were the advantages of chicken meat emulsion and chicken liver emulsion, respectively. NMR studies
indicated four forms of water, bound (T2b), emulsified (T2b-1), immobile (T21) and free water state (T22) in chicken meat and
liver emulsions. The proportion of immobile water in mixed emulsions was as high as 78.87% showing good shelf-life
stability. The first three principal components could explain 91.96% of the total variance. The score plots for the first and
second principal components provided an overall and visual interpretation of the quality characteristics of the emulsion
composite gels. As apparent from the above results, chicken liver-chicken breast-vegetable oil composite gels have excellent
physical properties and stability.

The stability of lutein in organic solvents
PANG Hui-li1,LI Da-jing1,2,*,LIU Chun-quan2,3
2013, 34(23):  15-19.  doi:10.7506/spkx1002-6630-201323004
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The HPLC analysis method of lutein was carried out to monitor variations of lutein in original solvents to know the effects of the solvents, oxygen and temperature on the stabilities of lutein, which can provide some instruction to the extraction of lutein and the production of lutein-based formulations. The variations of lutein in ethanol, ethyl acetate, tetrahydrofuran and toluene at 30, 40, 50℃ was analysised by C30-HPLC-DAD, and then the rate constants were calculated respectively. The stabilities of lutein at the same temperature normally was ethanol > ethyl acetate> tetrahydrofuran > toluene. The rentention of all-(E)-lutein decreased during the heating treatment and the rentention of its cis isomers increased firstly and then decreased slowly. Higher stabilities were found in nitrogen-bubbled condition, especially for tetrahydrofuran. All-(E)-lutein was less stable as a function of temperature and it was more easy to its isomerization reaction. The thermal degradation of all-(E)-lutein followed a first-order kinetic model.

Effects of Heat Treatment on the Quality of Waxy Dough and Bread
SHANG Shan1,HUANG Wei-ning1,*,JIA Chun-li1,WANG Feng2,RAYAS-DUARTE Patricia3
2013, 34(23):  20-24.  doi:10.7506/spkx1002-6630-201323005
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The pasting and baking properties of waxy flour under different heat treatments at 100 or 120 ℃ for 60 , 90 min
and 120 min were analyzed by RVA, DSC and TA, respectively. The results showed that (1) compared with the control group
without heat treatment, the water-holding capacity and pasting properties of waxy flour were enhanced after heat treatments.
The pasting parameters (peak viscosity, breakdown viscosity and final viscosity) were higher than those of the control. But
when waxy flour was thermally treated under specific conditions (100 ℃), dough setback showed a downward trend. (2)
DSC results showed that both To and Tp increased with longer treatment time for all samples, and △H gradually decreased
along with heat treatment time. The gelatinization temperature range was narrowed after heat treatments. (3) after addition
of thermally processed waxy flour, the specific volume of waxy bread was increased. For 100 ℃/120 min heat treatment
group, the specific volume of waxy bread was increased from 4.26 mL/g to 4.87 mL/g by 14.32% when compared with the
control group. Heat treatment could be beneficial for reducing the hardness, gumminess and chewiness of waxy bread. The
lightness for bread made from heat treated waxy flour decreased with the extension of heat treatment time and increase of
heat treatment temperature. Bread with 30% substitution of waxy flour treated at 100 ℃ for 60 min showed the best sensory
quality.

Granular Structure of Buckwheat Starch and Resistant Starch
ZHOU Yi-ming1,2,LI Bao-guo1,CUI Lin-lin3,ZHOU Xiao-li2,*,CHEN Yue2
2013, 34(23):  25-27.  doi:10.7506/spkx1002-6630-201323006
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In the present study, some physical characteristics of buckwheat starch and resistant buckwheat starch such as
particle size and crystal properties were explored. The results showed that the buckwheat starch granules were irregular
polyhedrons or spheres in shape with diameters ranging from 7 to 8 μm. Its crystal type was typically type A, and the
crystallinity was 34.95% for tatary buckwheat starch and 26.92% for common buckwheat starch. The resistance buckwheat
starch granules showed undefined shapes with diameter of 150 μm and were amorphous.

Effect of High Pressure Processing on Thermal Characteristics of Connective Tissue Collagen of Beef
CHANG Hai-jun1,TANG Cui2,NIU Xiao-ying2,TANG Chun-hong3
2013, 34(23):  28-31.  doi:10.7506/spkx1002-6630-201323007
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The main objective of this study was to investigate the characteristic changes of intramuscular connective tissue
collagen of beef using high pressure processing (HPP) from 200 to 600 MPa for 10 min and 20 min, respectively. The
changes in mechanical strength, contents and thermal characteristics of connective tissue collagen from beef muscle were
analyzed during these treatments. The results indicated that with increasing pressure up to 300 MPa, the mechanical strength
of beef connective tissue collagen showed an increasing trend. Meanwhile, the content of extractable collagen increased initially
and then showed a downward when the pressure was increased up to 600 MPa. Higher thermal stability was observed for beef
connective tissue collagen treated with high pressure. Compared with the control group, the thermal denaturation temperature of
beef connective tissue collagen subjected to high-pressure treatment for 10 min and 20 min was also increased. A significantly
positive correlation between beef connective tissue mechanical strength and collagen content was observed, as well as between
onset denaturation temperature and peak temperature, and the correlation coefficients were 0.487 and 0.686, respectively.

Structural Identification of Components I and II from Pu-er Tea Theabrownins
DONG Wen-ming,TAN Chao,GONG Jia-shun*
2013, 34(23):  32-35.  doi:10.7506/spkx1002-6630-201323008
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In the present study, two components of the theabrownins extracted from Pu-er tea were obtained through
gradient methanol-water extraction, thin-layer chromatography and dialysis, and both components were identified by infrared
spectroscopy, high performance liquid chromatography, nuclear magnetic resonance and mass spectroscopy. The results
showed that compound I was a polyphenol polymer with carboxyl groups and a large number of polysaccharide residues
(mainly galactose residues) and compound II may be C3H8Na2O9P2.

Effect of Water Quality on the Main Components and Quality of Baked Green Tea Infusion
LIU Ping1,2,YIN Jun-feng1,*,XU Yong-quan1,WANG Fang1,LIU Pan-pan1,2
2013, 34(23):  36-40.  doi:10.7506/spkx1002-6630-201323009
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In order to explore the effect of water quality on green tea infusion quality, green tea infusions were
prepared with four kinds of commercial drinking water (pure, natural, artificial mineral and natural mineral water). The
pH, conductivity and metal ion content in water samples as well as green tea infusions were determined comparatively
as well as sensory and physiological characteristics of fresh and aged tea infusions. The results indicated that different
kinds of drinking water had no statistically significant influence (P < 0.05) on the contents of caffeine, amino acid,
P or Zn, but exerted a statistically significant impact on the contents of polyphenols, catechins, Ca or Mg. Pure water
and artificial mineral water K2 were better than other water samples. pH and conductivity can be the key standards for
selecting appropriate water.

Isolation and Characterization of Collagens from the Skin of Sturgeon
YANG Ling1,ZHAO Yan2,LU Liang2,WANG Zhong-wen2,SONG Bao-lei3,WANG Zhan-fang3,WANG Hai-bo1,*
2013, 34(23):  41-46.  doi:10.7506/spkx1002-6630-201323010
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In this study, acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) were extracted from sturgeon skin by the
methods of acid and acid-pepsin. The molecular structures and physicochemical properties of ASC and PSC were determined and
compared with those of bovine Achilles tendon collagen. The results indicated that the ASC and PSC were similar in their amino
acid composition and secondary structures, but they were different from those of bovine Achilles tendon collagen. The denaturation
temperatures for these collagens were determined to be 42.81 ℃ for bovine Achilles tendon collagen, 28.04 ℃ for sturgeon skin
ASC and 28.13 ℃ for sturgeon skin PSC, suggesting an obvious difference in thermostability between sturgeon skin collagen and
bovine Achilles tendon collagen. In the solution state, bovine Achilles tendon collagen could more easily form collagen polymers
and was more resistant to enzymatic hydrolysis than sturgeon skin collagen.

Analysis of Glycated Peptides by Neutral-loss-triggered CID MS3 Spectrometry
WANG Hui1,TU Zong-cai1,2,*,LIU Guang-xian3,SHA Xiao-mei1,XIAO Hui4
2013, 34(23):  47-50.  doi:10.7506/spkx1002-6630-201323011
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Identification of the glycated sites in proteins plays an important role in the field of protein glycation. In the
present study, we have established a CID neutral-loss-triggered MS3 method to analyze the glycated sites in proteins. ESICID-
MS was used to stimulate multi-site cleavage in glycated Amadori peptides. MS3 scans triggered by neutral losses of
3H2O and 3H2O + HCHO produced similar results in terms of glycated peptide identification. However, neutral loss of 3H2O
resulted in more accurate glycated peptide identification in multi-stage activation experiments. The results showed that the
K227 was more easily glycated by D-glucose when compared with the K229 in ovalbumin under dry heating conditions.
Overall, the multi-stage activation approach could identify the glycation site in peptides with one glycosylation site.

Purification, Physicochemical Properties and Immunomodulatory Activity of Polysaccharides from Porphyra yezoensis
QIU Wei-fen1,YANG Wen-jian1,XUE Mei1,YANG Fang-mei2,ZHAO Qing-qiu2,FANG Yong1,MA Ning1, AN Xin-xin2,ZHAO Li-yan2,HU Qiu-hui2,*
2013, 34(23):  51-56.  doi:10.7506/spkx1002-6630-201323012
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Polysaccharides from Prophyra yezoensis (PYP) were extracted with papain and purified using DEAE
cellulose-52 and Sephadex G-100 chromatography. Four fractions named as PYP-1, PYP-2, PYP-3 and PYP-4 were
obtained, and their physicochemical properties including total sugar content, molecular weight, sulfate and uronic acid
contents and monosaccharide composition were investigated. In addition, RAW264.7 macrophage cell model was employed
to evaluate the immunomodulatory activity in vitro. Results showed that total sugar contents of PYP-2, PYP-3 and PYP-4
were 91.83%, 96.93% and 97.66%, respectively. Uronic acid contents were 19.39%, 23.77% and 18.23%, respectively, and
sulfate group contents were 18.84%, 17.63% and 18.41%, respectively. All these three polysaccharides were composed of
galactose, glucose and xylose with molecular weight of 478.6, 314.4 kD, and 417.3 kD, respectively. Pyranose ring structure
and β-type glycosidic linkages were also found in them. PYP-2, PYP-3, and PYP-4 when used at a concentration ranging
from 25 to 250 μg/mL could significantly promote the proliferation and neutral red phagocytosis of RAW264.7 macrophage
cells in vitro, suggesting the potential immunomodulatory activities of PYP-2, PYP-3, and PYP-4.

Circular Dichroism Analysis of MRJP2 Purified from Royal Jelly
HU Yi-fan1,2,CHEN Zhong-zhou2,TIAN Wen-li1,2,ZHAO Ya-zhou1,PENG Wen-jun1,*
2013, 34(23):  57-61.  doi:10.7506/spkx1002-6630-201323013
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MRJP2, a member of the major royal jelly protein (MRJP) family, was extracted from fresh royal jelly, purified
by affinity chromatography, molecular sieve chromatography and ion exchange chromatography and identified by SDSPAGE
and mass spectrum. The MRJP protein was analyzed by synchrotron radiation circular dichroism (SRCD) to clarify
its secondary structure as a function of temperature, pH and ion concentrations. The results of this study showed that heat treatment
at elevated temperatures from 10 to 90 ℃ resulted in increased proportion of α-helix and decreased proportion of β-strand
and random coil, while the proportion of β-turn kept stable. As the pH varied from 4.0 to 9.0, the proportion of α-helix revealed
an initial decrease followed by an increase, the highest proportions of β-strand and random coil were found at pH 5.0 and 8.0,
respectively, and the proportion of β-turn remained basically unchanged. After different metal ions were added in the protein
solution, the proportion of α-helix was increased by Ca2+ but decreased by Mg2+ and Zn2+.

Comparative Studies on Beef Sensory and Processing Characteristics of Nanyang and Yanbian Yellow Cattles
WANG Fu-long1,LU Gui-song1,WANG-Na2,PENG Zeng-qi1,*,QI Xing-lei3,LI Yu-lin2, YAO Yao1,LI Jun-ke1,ZHU Yi1,WAN Ke-hui1,ZHANY Ya-wei1,HUI Teng1
2013, 34(23):  62-66.  doi:10.7506/spkx1002-6630-201323014
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This study was focused on a comparative evaluation of the sensory quality properties and processing
characteristics of striploin and knuckle meats from Yanbian and Nanyang yellow cattles. Results showed that the a* value
of Yanbian beef was significantly higher than that of Nanyang beef (P < 0.05), but the L* and b* values were significantly
lower than those of Nanyang beef (P < 0.05). The juiciness of striploin from Yanbian cattle was better than that of Nanyang
beef, while the juiciness of knuckle was worse than that of Nanyang beef. The cooking loss showed that Yanbian beef had a
higher yield during processing than Nanyang beef. The striploin of Yanbian cattle had better tenderness with shear force of
3.19 kg/cm2, which was suitable for frying and roasting. The gel property of Yanbian beef was better than that of Nanyang
beef, so Yanbian beef was suitable for preparing gel-type products. Nanyang beef had a better emulsifying property, and
thuswas suitable for preparing emulsification-type products.

Effects of Processing Conditions on Gel Performance and Microstructure of Fish Scale Gelatin
MAN Ze-zhou1,TU Zong-cai1,2,*,WANG Hui1,SHA Xiao-mei1,HUANG Xiao-qin2,HUANG Tao1,LI De-jun1
2013, 34(23):  67-71.  doi:10.7506/spkx1002-6630-201323015
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In this study, the effects of pH, inorganic salts, heating temperature and heating time on the gel strength and
melting temperature of fish scale gelatin were examined. The changes in its microstructure were also monitored. The results
indicated that the gel strength and melting temperature were increased at first and then decreased with increasing pH. When
pH value was 9, the gelatin had a smooth network structure and acquired the best gel performance. Different salts had
different influences on its gel properties; NaCl and MgCl2 reduced its gel strength and melting temperature while the addition
of (NH4)2SO4 led to the highest melting temperature and NaH2PO4 induced the highest gel strength. Heating temperature and
time also had some influences on the gel performance. The gel strength was the highest when heating temperature was 60 ℃,
and the gel performance was decreased along with increasing the heating time.

Selection of Factors for Evaluating Fruit Quality of Mid-early Ripening Apple Cultivars
MENG Xian-jun1,GONG Li-yan1,BI Jin-feng1,2,BAI Sha-sha3,LIU Xuan2,WANG Xuan2
2013, 34(23):  72-76.  doi:10.7506/spkx1002-6630-201323016
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In order to provide a theoretical basis for the selection of indicators for evaluating the quality of apple fruits,
22 quality parameters of 52 mid-early ripening apple cultivars were determined and analyzed by principal component
analysis and hierarchical cluster analysis. Results revealed either relative independence or close correspondence among the
characteristic indicators of fruit quality. The cumulative variance contribution ratio of the first seven principal components
of PCA was up to 72.8%. The 52 apple cultivars were classified as 8 major groups by HCA on the basis of the measured
parameters. Eight factors were suitable for a comprehensive system for the evaluation of apple quality, including the content
of total soluble solids, sugar/acid ratio, browning degree, the content of magnesium, fruit firmness, fruit shape index, single
fruit weight and pulp cell size.

Solubility of Lac Resin in Aqueous Ammonia and Physico-chemical Properties of Its Ammonium Salt
CHEN Qi1,2,ZHANG Hong1,ZHENG Hua1,*,SUN Yan-lin2
2013, 34(23):  77-82.  doi:10.7506/spkx1002-6630-201323017
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To improve the hydrophilicity of lac resin and broaden its application in the food industry, a chemical method
was used for preparing carboxyl-modified lac resin. Lac resin ammonium salt was prepared to improve the solubility. The
relationship between the quantity of dissolved lac resin and either total dissolved solids (TDS) or pH at 40 ℃ was elucidated
as well as the reaction mechanism between lac resin and aqueous ammonia. The physico-chemical properties of lac resin
and its ammonium salt were compared. The results indicated that the reaction between them was essentially an acid-base
neutralization reaction between RCOOH and NH3·H2O, leading to generation of water-soluble strong electrolytes. At
the ammonia concentration of 0.1 mol/L, 62.90% of the lac resin was achieved after reaction at 40 ℃. The swelling ratio
of lac resin ammonium salt film was as high as 275.03% at pH 5.0, and dissolving ratio was up to 67.04% at pH 7.0. In
addition, it was more soluble in acidic water than lac resin. The puncture intensity was 4.82-fold higher than that of lac resin,
accompanied with a 64.50% increase in toughness. The softening point of lac resin ammonium salt was elevated by over 10 ℃ and
it had significantly better heat resistance when compared to lac resin.

Isolation and Purification of Antioxidative Polysaccharides from Spirulina platensis
CAI Bing-na1,CHEN Chun-xin2,CHEN Xin2,SUN Hui-li1,3,CHEN Yan-yun2,PAN Jian-yu1,*
2013, 34(23):  83-87.  doi:10.7506/spkx1002-6630-201323018
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Crude polysaccharides with antioxidant activity were obtained from S. platensis through extraction with a hot
alkali solution followed by precipitation with 95% ethanol. The scavenging activities against hydroxyl and superoxide
anion free radicals of the crude polysaccharides were evaluated. Further studies were carried out to separate and purify
the crude polysaccharide extract by chromatography on a DEAE-52 cellulose column and measure the molecular masses
and IR spectroscopic features of fractions. The crude polysaccharides extracted from S. platensis had a potent free radical
scavenging activity, and the scavenging capacity was (37.01 ± 1.58) U/mL and (47.84 ± 2.46) U/mL for hydroxyl and
superoxide anion radicals when used at concentrations of 0.6 mg/mL and 0.4 mg/mL, respectively. Four water-soluble acidic
fractions designated as SP1-2, SP2-1, SP3-1 and SP4-1 were obtained with molecular weight of 4.18 × 104, 5.11 × 104, 2.80 ×
104 D and 3.76 × 104 D. The infrared (IR) spectrum of SP3-1 showed the characteristic absorption of glycoprotein. These
results indicate that S. platensis polysaccharides may be used as an effective antioxidant in functional foods.

Application of Grey Correlation Analysis for Evaluating the Overall Quality of Fuji Apples from Different Growing Areas
WANG Xuan,BI Jin-feng*,LIU Xuan,Lü Jian,YANG Ai-jin
2013, 34(23):  88-91.  doi:10.7506/spkx1002-6630-201323019
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This study aims to apply grey correlation analysis for mathematical modeling of the overall quality of Fuji
apples from 9 different growing areas in China as a function of 8 quality parameters. Our results in this study suggested that
weighted relevance values were more accurate than equal-weighted relevance values in the assessment of applied quality and
that developing the reference variety and providing different quality traits with weighting values were the critical steps in
the grey correlation analysis. On the basis of our results, Fuji apples from Xinjiang, Gansu and Liaoning revealed excellent
quality, followed by those from Shaanxi, Shandong and Henan, and those from Hebei, Shanxi and Jiangsu were poor.

Effects of Pyrus pyrifolia Nakai Polyphenol Oxidase Treatment on the Quality of Black Tea in Summer and Autumn
YE Fei1,GAO Shi-wei1,2,GONG Zi-ming1,2,*
2013, 34(23):  92-95.  doi:10.7506/spkx1002-6630-201323020
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Fermentation is a critical procedure in black tea manufacture. In order to improve the quality of black tea, we
attempted to utilize the polyphenol oxidase from Pyrus pyrifolia Nakai to improve the fermentation process and thus investigated
the sensory quality, color and main taste and aroma components of the resulting black tea in the present study. Results indicated
that this new treatment significantly improved the sensory quality of black tea. As compared with those of the control, the
brightness of tea infusion was increased significantly (P < 0.01) and the ratio of theaflavins to thearubigins and the content of
soluble sugars in black tea treated with a ratio of crude polyphenol oxidase solution to rolled tea were increased by 86.05% and
34.34%, respectively, whereas the ratio of polyphenol to amino and the content of flavonoids were decreased by 3.96% and 7.35%,
respectively. The aroma components β-ionone, trans-geraniol and α-copaene were increased.

Fractional Purification and Structural Analysis of Polysaccharides Extracted from Camellia oleifera Seed Cake
LIU Xiao-ru,ZHANG Li-mei,HU Jiang-ning,LIU Rong,LI Hong-yan,FAN Ya-wei,LI Jing,DENG Ze-yuan*
2013, 34(23):  96-102.  doi:10.7506/spkx1002-6630-201323021
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In this study, we conducted fractional purification and structure analysis of polysaccharides (CSPs) extracted
from Camellia oleifera (C. oleifera) seed cake with the aim to provide experimental data for subsequent studies on the
structure-function relationship of CSPs. Methods: After removal of proteins, the crude CSPs were dialyzed, and then
fractionated and purified by DEAE-Sephadex A-25 anion exchange resin column chromatography. UV spectroscopy,
Sephacryl S-200 column chromatography and HPLC were used to detect the purity of the purified polysaccharides. A precolumn
derivatization HPLC method was used to analyze the monosaccharide composition. Infrared spectroscopy and a
nuclear magnetic resonance scanner were then used to study the configuration and structures of the purified CSPs. The
fractional purification led to three polysaccharides, namely CSP1, CSP2 and CSP3, as the main polysaccharides of CSP.
Both CSP1 and CSP2 were found to be neutral heteropolysaccharides whereas CSP3 a homogeneous, acidic one. CSP1was
composed of glucose, mannose and rhamnose with a molar ratio of 6.29:1.00:0.15; CSP2 was composed of monosaccharides
glucose, mannose and galactose with amolar ratio of 3.13:1.00:0.03; and CSP3 was composed of monosaccharides glucose,
mannose, galactose, arabinose, galactose, rhamnose, galacturonic acid and a small amount of rhamnose with a molar ratio of
0.42:1.00:1.09:0.96:0.43:0.19.

Effect of High Pressure Cooking on Bone Composition and Strength of Chicken Skeleton
CHAI Xiang-hua1,HE Wen-long1,WU Ke-gang1,*,LUO Ji1,WANG Sheng-li2,PAN Xian-zong2
2013, 34(23):  103-107.  doi:10.7506/spkx1002-6630-201323022
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High-pressure-cooked chicken skeletons were investigated for bone protein, fat and calcium contents and
firmness in comparison to raw chicken skeletons. The results showed that the total content of protein and fat as major
chicken bone components and calcium content were both lower in high-pressure-cooked chicken skeletons, especially when
high-pressure cooking in the presence of water was used. Moreover, bone strength became weak and the cellular structure
and small holes in the bone surface disappeared after high-pressure cooking under SEM observation. This finding explains
why the decrease in bone strength may be caused mainly by the losses of bone components and the structural collapse of
chicken bones as observed under SEM after high-pressure cooking of chicken skeletons. Orthogonal array design was used
to establish the optimal high-pressure cooking conditions for chicken skeletons: cooking with a bone/water ratio of 1:4 at
0.12 MPa for 45 min. The firmness of cooked chicken skeletons under the optimized conditions was 134.41 N.

Effect of Ultrafiltration on the Flavor of Silkworm Pupae Protein Hydrolysates
MU Li-xia,LU Zhen,LIAO Sen-tai*,ZOU Yu-xiao
2013, 34(23):  108-112.  doi:10.7506/spkx1002-6630-201323023
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Color and flavor characteristics of ultrafiltration fractions of silkworm pupa protein hydrolystaes and their
Maillard reaction products were evaluated to choose the most suitable raw material for flavoring bases. Five ultrafiltration
fractions were obtained including 30 kD MWCO retenate (PMHs-Ⅰ), 10 kD MWCO retenate (PMHs-Ⅱ), 5 kD MWCO
retenate (PMHs-Ⅲ), 1 kD MWCO retenate (PMHs-Ⅳ) and 1 kD MWCO filtrate (PMHs-Ⅴ). The results showed that PMHs-
Ⅳ had the best sensory characteristics and contained the highest amount of 1k-5k D peptides and the highest proportion of
amino acids responsible for umami and sweet tastes. At the same time, its Maillard reaction products had a strong and pure
meaty aroma and consisted of 32 flavor compounds as identified by GC-MS.

Effects of Different Drying Methods on the Content and Antioxidant Activity of Total Polyphenol from Banana Slices
WANG Yu-ting1,CHEN Yi1,*,LI Yu-bo2
2013, 34(23):  113-117.  doi:10.7506/spkx1002-6630-201323024
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Objective: To compare the effects of hot-air drying, vacuum drying and vacuum freeze drying on total polyphenol
content in banana slices and its antioxidant activity. Methods: Banana slices were dried by hot-air drying, vacuum drying and
vacuum freeze drying, respectively. The content of total polyphenol was determined by Folin-Ciocalteu method. The DPPH,
hydroxyl, superoxide anion radical scavenging capacities and total reducing power of total polyphenols in dried banana
slices were determined and compared. Results: The contents of total polyphenol in banana slices dried by freeze drying,
vacuum drying and hot-air drying were 2.33, 7.16 mg/g and 2.68 mg/g, respectively. The DPPH and hydroxyl scavenging
capacities of total polyphenols from banana slices dried by different methods followed the decreasing order: vacuum freeze
drying > vacuum drying > hot air drying. The superoxide anion radical scavenging capacity of total polyphenol from hot
air dried banana slices was the lowest, those from freeze dried banana slices had the highest capacity at lower concentration
and those from vacuum dried banana slices had the highest capacity at higher concentration. The reducing power of total
polyphenol from vacuum freeze dried banana slices was also superior to that of those obtained from the other two methods,
and no significant difference between vacuum drying and hot air drying was observed. Conclusion: Compared with hot-air
drying and vacuum drying, freeze drying has less influence on the content of total polyphenol and the antioxidant activity of
the extracted total polyphenol from banana slices.

Separation, Purification and Structural Analysis of Angiotensin-converting Enzyme (ACE) Inhibitory Peptides from Hydrolysate of Silkworm Pupae Protein
WU Qiong-ying1,DU Jin-juan1,XU Jin-ling1,JIA Jun-qiang2,YAN Hui1,GUI Zhong-zheng2
2013, 34(23):  118-122.  doi:10.7506/spkx1002-6630-201323025
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Objective: In order to investigate the structural characteristics of angiotensin-converting enzyme (ACE)
inhibitory peptides from silkworm pupae protein. Methods: The ACE-inhibitory peptides from hydrolysate of silkworm
pupae protein were separated and purified by ultrafiltration, DEAE-52 ion exchange chromatography and Sephadex G-50 gel
filtration chromatography. Then, the structural characteristics of purified ACE-inhibitory peptides were analyzed by liquid
chromatography-mass spectrometry (LC-MS). Results: Fraction 2 (IC50 0.072 mg/mL) with higher ACE-inhibitory activity
was obtained from silkworm pupae protein hydrolysate. This fraction was the major component of ACE-inhibitory peptides,
and its ACE-inhibitory activity was increased by 2.95 times over that of silkworm pupae protein hydrolysate. Conclusion: In
fraction 2, ACE-inhibitory peptides were mainly composed of the peptide fragments from dipeptide to octapeptide, and their
relative molecular weights were 226.34 to 983.61 u.

Effect of High Pressure Homogenization on Properties and Microstructure of Yuba
JIANG Mei,DONG Ming-sheng,CHEN Xiao-hong,LI Wei,RUI Xin
2013, 34(23):  123-127.  doi:10.7506/spkx1002-6630-201323026
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The effect of different homogenization pressures (20-140 MPa) on Yuba properties and microstructure was
studied. By increasing the homogenization pressure, the color of Yuba turned from light golden yellow to golden yellow. At
a homogenization pressure larger than 100 MPa, the formation of Yuba was speeded up, with enhanced cooking tolerance
and stable quality. TPA analysis showed that high-pressure homogenization improved the tensile strength and elongation at
break of Yuba. Scanning electron microscopy showed that the internal structure of Yuba was protein-fat complex forming
superimposed layers of mesh-like structure. Free fat globules appeared in the mesh-like structure of Yuba homogenized
at 20 MPa or 60 MPa rather than in the structure of degreased yuba. But the free fat globules were degraded when using
a homogenization pressure larger than 100 MPa, resulting in more homogenous and compact structure. In addition, Yuba
treated at 140 MPa had no significant change in microstructure before and after defatting treatments, and the protein-fat
complex was more stable. This structure reduced the “oil-exuding” phenomenon and prevented its oxidative rancidity,
thereby extending the shelf life.

Preparation and Isolation of Bioactive Sulfated Derivatives from Water-insoluble Glucan in Ganoderma arum
ZHANG Hui,WANG Jun-qiao,NIE Shao-ping,ZHANG Shen-shen,WANG Yuan-xing,XIE Ming-yong*
2013, 34(23):  128-132.  doi:10.7506/spkx1002-6630-201323027
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Objective: To improve the water solubility of a water-insoluble glucan from Ganoderma atrum by sulfated
modification technique and to investigate the in vitro antitumor activity and properties of sulfated derivatives. Methods: The
water-insoluble glucan was modified using chlorosulfonic acid-pyridine (CSA-Pyr) method to obtain sulfated products with
different degrees of sulfation (DS). The sulfated polysaccharides were characterized by gel filtration chromatography and
FT-IR spectroscopy. The cytotoxicity on S-180 sarcoma cells was measured to screen a fraction with significant antitumor
activity. The separated fraction with high antitumor activity was purified and its solution was subjected to molecular
characterization. Results: Nine sulfated products with different DS were prepared. AGAPS5 (DS 1.14) had significant
antitumor activity. The sugar content of AGAPS5 was 66.3%, and the inhibitory rate of AGAPS5 on the growth of S-180
sarcoma cells was 67.74%. The purified AGAPS5-1 showed a random coil chain in 0.1 mol/L NaNO3 solution. The weight-average
molecular weight (Mw) of AGAPS5-1 was 5.19 × 104 u and the number-average molecular weight (Mn) was 2.63 × 104 u. Conclusion:
Sulfated modification can improve the water-solubility and solution properties, and enhance the in vitro antitumor activity of
glucan from Ganoderma atrum.

Effectiveness of Catalytic Infrared Radiation in the Sterilization of Dehydrated Spinach
ZHANG Xin,QU Wen-juan,MA Hai-le*,PAN Zhong-li,WU Ben-gang
2013, 34(23):  133-137.  doi:10.7506/spkx1002-6630-201323028
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Dehydrated spinach was treated by catalytic infrared radiation to improve the microbial safety and meet the national food
hygiene standards. The effectiveness of infrared radiation and subsequent heat insulation to reduce total bacterial populations in dehydrate
spinach was evaluated in comparison to high-temperature steam sterilization. In addition, the changes in color and chlorophyll content
in dehydrated spinach before and after these treatments were analyzed. The results showed that after 180 s of high-temperature steam
sterilization and then 30 min of hot-air drying at 70 ℃, the logarithmic amount of residual bacteria was -1.96 ± 0.06, color difference
(ΔE) 21.61 ± 0.13, chlorophyll retention 64.98% and the observed results were -1.98 ± 0.05, 5.17 ± 0.83 and 84.62%, respectively,
after 180 s of infrared radiation and then holding at 70 ℃ for 45 min. These data demonstrate that the effectiveness of infrared
radiation followed by heat insulation for sterilizing dehydrated spinach is similar to that of high-temperature steam sterilization and
is more helpful to maintain its quality.

Composition Analysis and Antioxidant Activity of Auricularia auricula Melanin
ZOU Yu1,YIN Dong-mei2,GANG Jie1,DU Xiong-wei1,LIU Cheng-hui1,GU Zhen-xin3,*
2013, 34(23):  138-141.  doi:10.7506/spkx1002-6630-201323029
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By Sephadex G-100 column separation, components F1 and F2 of A. auricula melanin were obtained with
molecular weights of 404.97 kD and 20.69 kD, respectively. The UV-visible spectra of both melanin components were
very similar to that of the typical melanin. The IR spectra indicated that the structural composition of F1 and F2 included
OH, NH2, C=O, C=C, CH, CH2 and aromatic groups. Elemental composition analysis showed that F1 and F2 belonged to
phaeomelanin due to S element besides C, H, N and O elements. F1 was darker than F2, but both of them possessed the same
solubility and redox properties. They exhibited a strong antioxidant activity. When used at a concentration of 1.0 mg/mL, the
superoxide radical-scavenging activity exceeded 80% and the hydroxyl radical scavenging activity was close to 40%.

Separation and Identification of A-type and B-type Dimers of Procyanidins from Peanut Skin by RP-HPLC-ESI-MS/MS
CHEN Yang,WANG Bing,YU Jia-nan,LI Bao-li,WEN Yun,LIU Rui*,TAN Jun*
2013, 34(23):  142-146.  doi:10.7506/spkx1002-6630-201323030
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A-Type and B-type oligomeric proanthocyanidins were prepared from the 70% ethanol extract of peanut skin
by purification on AB-8 macroporous adsorption resin and two fractions PSP-3 and PSP-5 were separated and identified
by reversed phase high-performance liquid chromatography-tandem mass spectrometry (RP-HPLC-MS/MS). Peanut skin
procyanidins (PSP) with a purity of over 98.0% and a recovery of 9.8% were obtained after the purification. PSP-3 was
composed of 2 B-type procyanidolic dimers and PSP-5 was composed of 4 A-type procyanidolic dimers.

Correlational Analysis between Appearance and Eating Quality of Indica Rice Harvested at Different Times
XU Xing-feng,ZHONG Ye-jun*,XIA Wen,LIU Cheng-mei,LIU Wei,ZUO Yan-na,AI Yi-min
2013, 34(23):  147-150.  doi:10.7506/spkx1002-6630-201323031
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In the present study, the effects of harvesting time on color parameters and sensory characteristics of indica
rice (early indica rice “Zaoxian 167” and late indica rice “Gan Wanxian 923”) were evaluted, and the relationships
between appearance quality and eating quality were established by regression analysis. The results showed that
when the picking time was increased from the thirtieth day to the forty-second day after heading, the length, width
and whiteness of the grain revealed a gradual increase, while the yellow degree was reduced. The eating quality as
evaluated by hardness, stickiness, sweetness, deliciousness and overall judgement analysis of both rice cultivars
showed that the early-harvested rice was better than the ordinarily-harvested rice, especially on “sweetness” and
“deliciousness”. According to correlational analysis, the width of the grain did not have significant correlation with
the eating quality, which had negative correlation with the length and whiteness, but had positive correlation with the
yellow degree. As for the appearance indexes, the width of the grain had no significant correlation with the length,
yellow and white degree; the length had positive correlation with the white degree and negative correlation with the
yellow degree and there were negative correlations between yellow degree and white degree.

Effect of Coarse Grain Powder on Gelation Characteristics of Eggs
HU Rui,LI Chun,ZHOU Wen-qian,ZHAO Zhi-feng,LU Xiao-li*
2013, 34(23):  151-154.  doi:10.7506/spkx1002-6630-201323032
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The effects of coarse powders of corn, sweet potato and soybean on gelation characteristics of eggs were investigated
through cooking loos (CL), water-holding capacity (WHC) and texture profile analysis (hardness, springiness, cohesiveness
and chewiness). Results showed that with increasing amount of added coarse grain powders, for egg gel with corn
powder, the CL decreased sharply initially and then gradually increased, while WHC increased until reaching a plateau. For
egg gel with sweet potato, there was a peak level at 6% for CL, and the lowest point appeared at 4% for WHC. For egg gel
with soybean, its CL exhibited an initial decrease followed by an increase, while WHC exhibited an opposite changing trend.
The correlational analysis showed that different coarse grain powders were positively correlated with the springiness of egg
gel. The amount of coarse grain powders was positively correlated with the hardness, WHC and chewiness of egg gel but
negatively correlated with its springiness. Addition of appropriate amounts of coarse grain powders may improve the quality
and nutrition of egg gel.

Effects of Sulfhydryl Content and Hydrophobicity on Gel and Emulsifying Properties of Pork Proteins
SHAO Jun-hua1,2,WU Ju-qing2,ZHOU Guang-hong2,*,WEI Chao-gui2,XU Xing-lian2,LIU Deng-yong1,SONG Li1,JIA Na1
2013, 34(23):  155-159.  doi:10.7506/spkx1002-6630-201323033
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The effect of changes in sulfhydryl content and surface hydrophobicity induced by chemical modification with
β-mercaptoethanol and Tween-80, respectively, on the emulsifying and gel properties of salt-soluble proteins of pork muscle
were examined and the roles of disulfide bonds and hydrophobic interactions in forming emulsions and gels were elaborated.
Free sulfhydryl content of salt-soluble protein solutions was increased with the addition of β-mercaptoethanol as a sulfhydryl
blocking agent and delayed gel formation and reduced overall viscoelasticity were observed, suggesting that blocking
the formation of disulfide bonds can deteriorate the heat-induced gelation characteristics of pork proteins. However, no
significant correlation between the free —SH content and emulsifying properties of pork proteins was observed. On the other
hand, higher surface hydrophobicity resulted in formation of less stable emulsions without significantly affecting the gel
properties. These data suggest that an appropriate degree of hydrophobicity may be of great significance for the formation of
stable emulsions and that disulfide bonds presumably contributes to good gel properties.

Isolation, Purification and Compositional Analysis of Agaropectin from Gracilaria lemaneiformis
LIU Qiu-feng,SU Yong-chang,WU Cheng-ye*
2013, 34(23):  160-164.  doi:10.7506/spkx1002-6630-201323034
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Three polysaccharides named as SPS-1, SPS-2 and SPS-3 were separated by gradient elution on a DEAEcellulose
column from the agaropectin extracted from Gracilaria lemaneiformis. Their chemical analysis including sulfate
base content, relative molecular mass, purity, characteristic bonds and monosaccharide composition were characterized byion
chromatography, high performance liquid chromatography, ultraviolet spectroscopy and infrared spectroscopy. The results indicated that the
yields of SPS-1, SPS-2 and SPS-3 were 2.7%, 47.2% and 2.2%, with sulfate base contents of 5.1%, 9.3% and 6.3%, respectively.
The agaropectin contained no protein, amino acid or nucleic acid. The content of SPS-2 consisting of polysaccharides of similar chain
length of was 92.14%. The mass-average and number-average relative molecular masses of SPS-2 were 367686 and 230434,
respectively. SPS-2 was a group of complex sulfated polysaccharide with 17.34% galactose, 2.72% raffinose, 8.82% saccharose
and 1.18% arabinose in the monosaccharide composition.

Rheological Characteristics of Kudzu (Pueraria lobata) Root Syrup
LI Chun,ZHOU Wen-qian,HU Rui,ZHAO Zhi-feng,LU Xiao-li*
2013, 34(23):  165-169.  doi:10.7506/spkx1002-6630-201323035
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The rheological behavior of various concentrations of kudzu (Pueraria lobata) root syrup with different types
and amounts of thickening agent, sugar content, salt (NaCl) content and pH was investigated using a rotary viscometer
(DNJ-1 type) at different temperatures and rotating speeds. The results showed that higher concentrations of kudzu root
powder could result in higher apparent viscosity of kudzu root syrup at the same temperatures and rotating speeds. The
changing trend was more obvious when the rotating speed was lower. Kudzu syrup with the addition of xanthan gum had
a higher apparent viscosity than that with the addition of agar, sodium alginate, carboxymethyl cellulose (CMC), or konjac
glucomannan. The apparent viscosity was higher when the amount of thickening agent was increased. The apparent viscosity
of kudzu syrup was increased by a certain concentration of sugar but reduced by salt. pH had minor influence on the apparent
viscosity of kudzu syrup, which was a non-New tonian and thixotropic fluid.

Screening of Bacteriocin-producing Bifidobacterium Strain and Culture Coditions for Bacteriocin Secretion
SHANG Nan,WANG Yang,REN Fa-zheng,LI Ping-lan*
2013, 34(23):  170-175.  doi:10.7506/spkx1002-6630-201323036
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Bifidobacterium L-SN capable of producing an antimicrobial substance that was effective against Lactobacillus
plantarum PL-2 was screened from 32 strains of Bifidobacterium isolated from the feces of centenarians in Bama county
of Guangxi region. By eliminating the effects of organic acid and H2O2, we confirmed the antimicrobial role because of
bacteriocin. Based on morphological, physiological and biochemical characteristics and 16S rRNA sequence analysis,
the strain L-SN was identified as Bifidobacterium animalis subsp. lactis. We investigated the secretion characteristics of
Bifidobacterium animalis L-SN and established the optimum incubation conditions for producing bacteriocin: 28 h of culture
at 30 ℃ and initial medium pH 6.5. The bacteriocin had a wide spectrum of antibacterial activities against Gram-positive and
Gram-negative bacteria, especially against Staphylococcus aureus, Escherichia coli and Listeria monocytogenes showing a
potential application prospect.

Mutation of Saccharomyces cerevisiae Protoplasts for Enhanced Production of GSH and Gene Expression of GSH Over-accumulating Mutants
SUN Jiang1,ZHU Yi-bo2,WANG Li-mei2,DENG Da-qing2,QI Bin2,*
2013, 34(23):  176-179.  doi:10.7506/spkx1002-6630-201323037
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In this study, Saccharomyces cerevisiae Y518 was improved for enhanced GSH production by protoplast
mutagenesis. The mortality of Saccharomyces cerevisiae protoplasts after being treated with 0.03 mol/L HNO2 for 3 min
was 85.96%. After 4 repeated treatments, a mutant was obtained which produced 14.85 mg of GSH per g dry weight, a 24%
increase over the original strain. The gene GSH1 encoding the rate-limiting enzyme in the synthesis route showed a 2.26-fold
increased expression, thus resulting in increased GSH production from the mutant strain.

Comparison of Methods for Protein Extraction from Traditional Fermented Yongchuan Douchi and Degradation Pattern
SUO Hua-yi1,2,QIAN Yu1,3,KAN Jian-quan1,2,*
2013, 34(23):  180-183.  doi:10.7506/spkx1002-6630-201323038
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This study compared the effectiveness of four different methods, TCA/acetone precipitation, phenol extraction,
Tris-HCl extraction and homogenization for extracting proteins from traditional fermented Yongchuan douchi (Mucor type),
a soybean product originated in China. The patterns of protein degradation at different fermentation stages of Yongchuan
douchi were investigated as well. SDS-PAGE showed that the TCA/acetone precipitation method was the best of the four
methods. Protein macromolecules in the Mucor-type douchi were rapidly degraded into small molecules by steaming and
proteases from Mucor, and the effect of steaming was more obvious.

Determination of AChE Activity in Periplaneta americana Head and Its Susceptibility to Five Common Pesticides
HE Shao-zhi,LI Wei,WANG Yan,YANG Xiao,RAO Yu,YANG Wen-yu,CHEN Xiang-gui*
2013, 34(23):  184-188.  doi:10.7506/spkx1002-6630-201323039
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Objective: To explore the inhibitory effect of pesticides on the activity of acetyl cholinesterase (AChE) in the
head of Periplaneta americana, and to provide a new susceptible enzyme source for the quick detection of pesticides.
Methods: The activity of AChE was determined by Ellman’s colorimetric method. An orthogonal array design was used to
establish the optimum reaction conditions for determining AChE activity. The susceptibility of the AChE activity to organic
phosphorus pesticides and carbamate pesticides was measured using enzymatic inhibition by calculating IC50. Electric eel
AChE was used as a control under the same experimental conditions. Results: The IC50 values of trichlorfon, dichlorvos,
carbaryl, carbofuran and metolcarb to Periplaneta americana head AChE were 5.495, 5.943, 0.290, 0.160, 1.897 μmol/L,
respectively. The susceptibility of AChE from the head of Periplaneta americana to dichlorvos, carbaryl and metolcarb were
slightly higher than that from electric eel. The IC50 values of trichlorfon, dichlorvos, carbaryl, carbofuran, and metolcarb to
electric eel AChE were 3.990, 7.656, 1.002, 0.007, 7.568 μmol/L, respectively. Conclusion: Periplaneta americana head
AChE shows good susceptibility to five pesticides and thus can be used as a new susceptible enzyme.

Reactive Oxygen Species (ROS) Regulates Listeria monocytogenes Biofilm Formation
ZHANG Chao,CHEN Guo-wei,YANG Yu-ping,WU Shu-yan,LIU Qing*
2013, 34(23):  189-193.  doi:10.7506/spkx1002-6630-201323040
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In order to explore the role of ROS (reactive oxygen species) in Listeria monocytogenes (LM) biofilm formation,
we added either diphenyleneiodonium chloride (DPI) as a NADPH oxidase inhibitor or N-acetylcysteine (NAC) as an
antioxidant in combination with exogenous H2O2 to observe the changes of ROS and biofilm formation. The results showed
that both DPI and NAC were able to restrain the biofilm formation of LM by reducing the amount of ROS generation. To
some extent, the addition of H2O2 could promote biofilm formation. Our results suggest that NADPH oxidase exists in LM,
which is responsible for generating ROS. ROS plays a vital role in LM biofilm formation and may be an important signal
molecule.

Effect of Probiotics on Proteins in Fermented Milk Products
YAO Lu1,2,MIN Wei-hong1,JIANG Tie-min2,CHEN Li-jun2,*
2013, 34(23):  194-199.  doi:10.7506/spkx1002-6630-201323041
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In order to explore the effect of probiotics on proteins in fermented milk products, probiotics-fermented yogurt
was compared with traditional lactic acid bacteria-fermented yogurt in terms of the contents of protein, pH 4.6 soluble
nitrogen, amino nitrogen and free amino acids by SDS-PAGE electrophoresis, reversed-phase high performance liquid
chromatography and ultra performance liquid chromatography. In probiotics-fortified yogurt under the optimal fermentation
conditions established by response surface methodology, the protein brands of αs2-casein, αs1-casein and β-lactoglobulin were
relatively shallower, and those in Lactobacillus plantarum-fermented yogurt were the weakest; in addition, the contents
of pH 4.6 soluble nitrogen, amino nitrogen and free amino acid revealed an increase, especially a significant increase
in Lactobacillus casei-fermented yogurt. These results may be due to the more protein degraded by the protease from
probiotics. Moreover, different probiotics with different metabolic activities in the yoghurt fermentation process can lead to
the extension of protein degradation.

A Comparative Study of Recombinant Expression of Three Aminopeptidases in Escherichia coli
ZHANG Jie1,ZHANG Liang1,HUANG Wu-ning2,SHI Gui-yang1,*
2013, 34(23):  200-205.  doi:10.7506/spkx1002-6630-201323042
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The genes encoding aminopeptidase GSAP, BSAP and BLAP were respectively amplified from Geobacillus
stearothermophilus IFO12589, Bacillus subtilis 168 and Bacillus licheniformis 14580. These amplified DNA fragments
were individually inserted into the expression vector E. coli pET-28a to construct plasmids pET28a-BLAP, pET28a-BSAP
and pET28a-GSAP, respectively. The expression of these genes was confirmed by activity analysis, and then the expressed
enzymes were characterized. The activities of both recombinant BSAP and GSAP were 1500 U/L, which were higher than
that of BLAP. The optimum temperatures were 50, 75 ℃ and 60 ℃ for BSAP, GSAP and BLAP, respectively, and the
optimum pH values were all 9.0. They were stable at pH 8.5—9.0. BSAP was obviously activated by 0.l mmol/L Co2+,
reaching a maximum of 195.6% of its original activity, but inhibited to different extents by other divalent metal ions, with
Zn2+ being the strongest inhibitor. In addition, the recombinant plasmids pET28a-BSAP and pET28a-GSAP were more stable
than pET28a-BLAP in E. coli.

β-Glycosidase Activity of Oenococcus oeni
YANG Rui1,Lü Zhen1,WEN Yan1,LI Kai1,ZHU Cheng-long1,LIU Shu-wen1,2,*
2013, 34(23):  206-211.  doi:10.7506/spkx1002-6630-201323043
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In this study, 8 strains of Oenococcus oeni with the highest β-glycosidase activity were obtained by measuring
β-glycosidase activity in 26 wild Oenococcus oeni strains and the effects of temperature, pH, alcohol concentration,
glucose, fructose and wine-model conditions on β-glycosidase activity from the selected strains were investigated using
two commercial strains as controls. The results showed that strain CS-1b exhibited the best enzymatic properties among the
tested strains under the optimum conditions of pH 5 and 30 ℃ and it retained high β-glycosidase activity under wine-model
conditions of pH and temperature. The β-glycosidase activity was slightly activated by lower concentrations of glucose or
fructose but slightly inactivated at higher concentrations. Higher concentrations of alcohol strongly inhibited its activity.
Under wine-model conditions, the β-glycosidase activity of this strain was 3.755 μmol/(g·min), accounting for 34.55% of
the original activity.

Nanoparticle-based Immunomagnetic Separation of Listeria monocytogenes
ZHONG Zi-qing,XIONG Yong-hua,LAI Wei-hua*,XU Heng-yi,SHAN Shan,CHEN Qi,XIE Fang,YU Yang-fan
2013, 34(23):  212-215.  doi:10.7506/spkx1002-6630-201323044
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The influence of antibody-coupling methods such as direct binding and streptavidin-biotin system on the
capture efficiency of immunomagnetic beads of different sizes (180, 350 nm and 1150 nm) for Listeria monocytogenes was
compared. The results showed that the capture efficiency of immunomagnetic beads using streptavidin-biotin method was
approximately two-fold higher than that of direct binding method. Meanwhile, the size of magnetic beads had significantly
effect on the capture efficiency. When the dosage of immunomagnetic beads was 100 μg, the capture efficiencies of
beads 180, 350 nm and 1150 nm in diameter were 95%, 18% and 9%, respectively, during the separation of Listeria
monocytogenes. Therefore, the 180-nm-diameter immunomagnetic beads prepared with streptavidin-biotin system can
provide a low-cost, easy-to- use and efficient method for rapid enrichment and detection of Listeria monocytogenes.

Effects of Fermentation Conditions on content of Bioactive Ingredient in Fermented Mulberry-leaf tea
XIAO Hong1,HUANG Xian-zhi2,SHEN Yi-hong2,DING Xiao-wen1,*,ZHANG Ya-qiong1,LIANG Han-yu1,ZHANG Di1
2013, 34(23):  216-220.  doi:10.7506/spkx1002-6630-201323045
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Objective: To investigate the changes of 1-deoxynojirimycin (DNJ) and other bioactive compounds in fermented
mulberry leaf tea with different fermentation conditions. Methods: DNJ was determined by HPLC. Flavonoids, polysaccharides
and polyphenosl were determined by spectrophotometric methods. Results: Compared with those in mulberry leaves, the contents
of flavonoids, polysaccharides and polyphenol in mulberry leaf tea fermented at any of the fermentation temperatures tested were
decreased. DNJ content in mulberry leaf tea fermented at 30 ℃ was increased by 7.37%, whereas the content of flavonoids was
slightly decreased. DNJ content in mulberry leaf tea fermented for different times was increased, with the highest increase by
7.10% at 6 h, accompanied by a 6.45% increase in polysaccharide content. The content of polyphenols was increased by 20.00% in
mulberry leaf tea fermented for 4 h, while the amount of flavonoids became lower. The contents of DNJ and polysaccharides were
increased by 4.23% and 3.68%, respectively, but flavonoid and polyphenol contents were decreased in mulberry leaf tea fermented
with an inoculums amount of 4 × 107 CFU. Conclusion: Mulberry leaf tea fermented at 30 ℃, 6 h with 4 × 107 CFU of inoculum
has the most favorable functional properties.

Strain Screening of Pleurotus nebrodensis and Optimization of Submerged Culture Conditions for Biomass Prodution
JI Hong1,2, ZHAO Li-ming1
2013, 34(23):  221-227.  doi:10.7506/spkx1002-6630-201323046
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Pleurotus nebrodensis Tianshan No.2 from Wuan, Hebei province was selected as the fastest-growing of
19 strains. The culture medium for submerged fermentation of strain Tianshan No.2 were optimized for the maximum
biomass (dry cell weight, DCW) production using an orthogonal array design L9 (34), and three culture conditions including
temperature, shaking speed and time were optimized by response surface analysis based on a three-variable, three-level Box-
Behnken experimental design. The results showed that the optimal medium composition (per 100 mL) was 3.50 g of soluble
starch, 0.40 g of beef extract, 0.25 g of monopotassium phosphate and 0.25 g of magnesium sulfate. The mycelia biomass
of strain Tianshan No.2 was 1.6004 g/100 mL after 7.5 d (180 h) of shaking culture at 26 ℃ with a shaking speed of 160 r/min
using the optimized medium.

Screening and Mutation Breeding of Lactic Acid Bacteria with Enhanced Ability for γ-Aminobutyric Acid Production
LIANG Jin-zhong,LI Wen,WANG Feng-qing
2013, 34(23):  228-232.  doi:10.7506/spkx1002-6630-201323047
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Lactic acid bacteria were isolated from pickled Chinese cabbage using BCP medium. After two rounds of
screening, strain Lp-Lw-131 having a higher ability to produce γ-amino butyric acid (GABA) was obtained. The strain was
identified as Lactobacillus plantarum by morphological observation and physiological and biochemical tests. The yield
of GABA generated from the decarboxylation of L-monosodium glutamate catalyzed by glutamate decarboxylase (GAD)
from Lp-Lw-131 was determined by colorimetrically to be 0.917 g/L. This strain was irradiated with ultraviolet light and
then mutagenized with diethyl sulfate (DES) to improve its ability for GABA production. Based on its lethality, the optimal
conditions for mutagenesis in strain Lp-Lw-131 were UV irradiation for 90 s at a distance of 30 cm from the light source
followed by treatment with a 25% ethanol solution of DES for 20 min. A mutant strain named Lp-Lw-131-34 was obtained,
which maintained its genetic stability after 10 successive passages, providing an average yield of GABA of 1.815 g/L,
showing an 1.979-fold increase over the initial strain Lp-Lw-131.

Cross-linked Chitosan as a Carrier for Immobilization of Naringinase
LEI Sheng-jiao,YU Hai-li,HUANG Chao,ZHENG Xing-long
2013, 34(23):  233-238.  doi:10.7506/spkx1002-6630-201323048
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Naringinase (EC 3.2.1.40) was immobilized onto glutaraldehyde cross-linked chitosan beads by an adsorption
method. Using one-factor-at-a-time and orthogonal array designs, the optimal immobilization conditions were determined as
follows: gel microspheres prepared with 3.5 g/100 mL chitosan as the carrier, 1.0 g/100 mL NaOH as the coagulant, 7.0%
glutaraldehyde solution, 2.0 h of cross-linking, 2.0 mg/mL of naringinase concentration and 3.0 h of adsorption at 25 ℃.
The maximum specific activity of the immobilized naringinase under these conditions was 7.37 U/g (based on dry chitosan)
without showing obvious differences in the optimum pH and temperature compared to the free enzyme. After 7 cycles of
repeated use at 40, 50 ℃ and 60 ℃, the immobilized enzyme maintained above 70%, 60% and 50% of its original activity.

Isolation, Purification and Characterization of Malate Dehydrogenase from Duck Heart
SUN Fang,HU Rui-bin,REN Mei-feng,TANG Yun-ming*
2013, 34(23):  239-244.  doi:10.7506/spkx1002-6630-201323049
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Two electrophoresis-pure isoenzymes (c-MDH and m-MDH) of malate dehydrogenase from duck heart were
obtained through the procedures including homogenization, ammonium sulfate precipitation, organic solvent precipitation,
DEAE-sepharose chromatography and Superdex-200 gel filtration. c-MDH present in the crude enzyme solution in a higher
amount was characterized. The results showed that the purity of c-MDH and m-MDH were increased by 69.07- and 130.40-
fold with activity recoveries of 25.54% and 10.98%, respectively. The molecular mass of the subunits in c-MDH and m-MDH
were 39.58 ku and 37.62 ku, respectively. Enzymatic characterization showed that the optimal reaction temperature and pH
for c-MDH were 55 ℃ and 7.2, respectively, and it was stable below 30 ℃ and at pH 6-9. Meanwhile, its Km using oxobutanedioic
acid as the substrate was 140.93 mmol/L. The enzyme activity could be strongly inhibited by oxalic acid, SDS,
Cu2+, Co2+ and Ag+, while it could be obviously activated by Mg2+, K+, and Ba2+.

Enzymatic Preparation of Crude Polypeptides from Rapeseed Meal
RAN Ren-sen1,CHEN Jin-ping1,*,MI Rui-fang1,YANG Xiao1,YANG Xiu-ling2,YANG Ya-juan1,CHEN Jie1
2013, 34(23):  245-249.  doi:10.7506/spkx1002-6630-201323050
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The current study aimed to develop an optimized procedure for preparing crude polypeptides by the alcalasecatalyzed
hydrolysis of rapeseed meal and subsequent dialysis. The results of optimization by orthogonal array design on
the basis of one-factor-at-a-time experiments showed that the optimal conditions for enzymatic hydrolysis of rapeseed meal
were found to be 40 min of reaction at 50 ℃ and initial pH 8.0 by the addition of 1200 U/g of alcalase. The hydrolysate was
dialyzed against a 6-fold volume of double distilled water at 5 ℃ for 48 h. After subsequent lyophilization, the lyophilisate
contained 96.78% polypeptides.

Eukaryotic Expression and Characterization of the Mutant H23A of Phosphotriesterase
ZHAN Dong-ling1,REN Yu-xue1,LI Ke-jian2,MIN Wei-hong1,LIU Yang3,ZHANG Ying-wen1,LIU Jing-sheng1,*
2013, 34(23):  250-255.  doi:10.7506/spkx1002-6630-201323051
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Through homologous sequence alignment and crystal structure analysis, it’s found that the site of histidine 23
(H23) from the phosphotriesterase (PTE)-encoding gene of Geobacillus kaustophilus HTA426 PTE was highly conservative
and was located near the metal ion binding sites. By using the Rosseta design program, the mutant H23A was overexpressed
in Pichia pastoris GS115. Through His-tagged ni-sepharose chromatography, non-denaturing electrophoresis and westernblotting,
the recombinant enzyme was identified as an intermediate form between a monomer and a dimmer. The primary
enzymatic properties indicated that the expressed recombinant enzyme was an allosteric enzyme with Vmax and hill
coefficient (h) of 92.45 U/mg and 1.98, respectively. The optimal temperature and pH were 70 ℃ and 10.0, respectively. The
recombinant PYE enzyme had a better heat tolerance with half-time of 5.1 h. Most divalent metal ions showed activating
effects on the enzyme activity.

Predictive Modeling of the Competitive Growth of Listeria monocytogenes and Pseudomonads spp. in Pork
CHEN Er-ning,BAI Yu,MA Kai,LIU Qing-jun,WU Hui-juan,GAO Li-juan*
2013, 34(23):  256-260.  doi:10.7506/spkx1002-6630-201323052
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To predict the shelf-life of pork, Baranyi model and Lotka-Volterra model were used to establish a model for the
competitive growth of Listeria monocytogenes and Pseudomonads spp. in pork. Interaction coefficients (α) were calculated
from the steady-state equation of a modified Lotka-Volterra model. The results indicated that at lower temperature,
L. monocytogenes could inhibit the growth of Pseudomonads spp. (α > 0), especially at 4 ℃, which represents the strongest
inhibitory effect, while Pseudomonads spp. showed no significant inhibitory effect on L. monocytogenes. To validate the
model, the L. monocytogenes growth data of in pork acquired from the ComBase database were used to calculate bias and
accuracy factors. At 4 ℃, the bias factor was 1.207, indicating the model to be fail-safe, and the accuracy factor was 1.362,
suggesting good prediction accuracy.

Optimization of Culture Conditions for Bifidobacterial Enrichment with Whole Powder of Jerusalem artichoke Tubers as Carbon Source by Response Surface Analysis
YU Ji-yang1,2,LI Xin-hua1,*,WANG Di3,ZHANG Ming2,4,QIN Ning1,ZHENG Feng-e2,WANG Lin1
2013, 34(23):  261-265.  doi:10.7506/spkx1002-6630-201323053
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The best carbon source for bifidobacterial growth was selected from inulin, glucose and whole powder of
Jerusalem artichoke Tubers, and the bifidobacterial enrichment culture conditions based on the selected carbon source
were optimized by experimental design using Design-Expert 8.0 and response surface analysis. Both inulin and glucose
were good bifidobacterial growth promoters when used as sole carbon sources;however, whole powder of Jerusalem
artichoke Tubers was more effective in promoting bifidobacterial growth, providing a 5.3-fold and 16.8-fold increase in
bifidobacterial populations after enrichment culture compared to inulin and glucose, respectively. When whole powder of
Jerusalem artichoke Tubers was used as the carbon source, the optimal culture conditions were 36 ℃, 21 g/L of carbon
source concentration and 10% of inoculum concentration.

Mechanism of Action of Bacillus atrophaeus in Improving Aspergillus flavus Resistance of Peanuts
LIU Ding,QIN Wen*
2013, 34(23):  266-270.  doi:10.7506/spkx1002-6630-201323054
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The relationship between the resistance of peanuts to Aspergillus flavus and the activities of defense enzymes
including superoxide dismutase (SOD), catalase (CAT), phenylalanine ammonium-lyase (PAL), peroxidase (POD) and
polyphenol oxidase (PPO) was studied. For this, peanuts were inoculated with fermentation broth of Bacillus atrophaeus +
spore suspension of Aspergillus flavus (inoculum 1), spore suspensions of both strains (inoculum 2), fermentation supernatant
of Bacillus atrophaeus + spore suspension of Aspergillus flavus (inoculum 3) or spore suspension of Aspergillus flavus alone
(inoculum 4) to observe the changes in the defense enzyme activities. The results showed that the maximum activity of SOD
in peanuts was obtained after 4 d of incubation with each inoculum and inoculum 2 resulted in the highest SOD activity of
5.3318 U/(g·min) among the four inoculums. On the second day after the inoculation, POD activity reached a peak and the
highest POD activity of 13.4710 U/(g·min) was observed in peanuts inoculated with inoculum 2. CAT activity, however,
showed a continuous decrease after the inoculation until reaching the minimum level of 5.5375 U/(g·min)on the fifth day.
The various inoculums had no obvious inductive effect on the activities of PAL or PPO. These results suggest that Bacillus
atrophaeus can enhance the Aspergillus flavus resistance of peanuts to a certain extent. Therefore, the activation of these
defense enzymes is involved in the action of Bacillus atrophaeus against Aspergillus flavus.

Effect of Ganoderma atrum Polysaccharide on cAMP/PKA Pathway in Mouse Colon Cancer Cells
ZHANG Shen-shen,NIE Shao-ping,XIE Ming-yong*
2013, 34(23):  271-275.  doi:10.7506/spkx1002-6630-201323055
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Ganoderma atrum polysaccharide-1 (PSG-1) has been reported to be natural chemopreventive in several types
of tumor cells. In our previous study, we found that PSG-1 could significantly inhibit the tumor growth. We hypothesized
that PSG-1 might exert its anticancer effect by activating cAMP/protein kinase A (PKA) pathway. The present study was
designed to analyze the effect of PSG-1 on the cAMP/PKA pathway in mouse colon cancer CT26 cells. The cells were
treated with 0, 50, 100 μg/mL, and 200 μg/mL PSG-1 or 20 μg/mL 5-fluorouracil, respectively, for 48 h. The ratio of
apoptotic CT26 cells was measured by flow cytometry analysis. The cellular cAMP content was tested using a cAMP assay
kit. The activity of PKA was measured using a PKA activity assay kit. Reverse transcript-polymerase chain reaction (RT-PCR)
was used to analyze the mRNA expressions of PKA and cAMP response element binding protein (CREB) of the cells. The
results showed that PSG-1 significantly induced the apoptosis of the CT26 cells and that cAMP concentration in the cells
treated with PSG-1 was dramatically increased by 20.18%, 38.67% and 56.25% (P < 0.05), respectively. The PKA activity
was increased by 15.42%, 18.76% and 38.26%, respectively. The mRNA levels of both PKA and CREB of the cells treated
with PSG-1 were increased. The current study suggests that PSG-1 activates the cAMP/PKA pathway in colon cancer cells
by inducing cAMP production and activating PKA protein.

Effect of L-malate on Gene Expression of Carrier Proteins and Antioxidant Enzymes in Rats
ZENG Xiao-cong1,2,WU Jun-lin2,XIAO Chun2,WU Qing-ping2,XU Ming-fang1,ZHANG Ju-mei2,*
2013, 34(23):  276-281.  doi:10.7506/spkx1002-6630-201323056
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To investigate the antioxidant mechanism of L-malate on mitochondria, real time RT-PCR was used to analyze
the gene expressions of two carrier proteins (AGC and OMC) in the inner membrane and two antioxidant enzymes (CAT
and GSH-Px) related to the malate-aspartate shuttle in rat liver and heart mitochondria. The mRNA expressions of these
proteins were examined by real time RT-PCR in hearts and livers of SD rats treated with L-malate. The results indicated that
the mRNA expressions of AGC, OMC, CAT and GSH-Px in the heart of rats treated with L-malate were 1.25, 1.39, 1.12, 1.01
times as much as those in the control group. The mRNA expression of AGC, OMC, CAT and GSH-Px in the liver of rats
treated with L-malate was 1.33, 1.02, 1.25, 0.94 times as much as those in the control group. These results imply that L-malate
might increase the antioxidant capacity of mitochondria by enhancing the mRNA expression of the proteins involved in the
malate-aspartate shuttle and the antioxidant enzymes.

Maternal High-fat Diet Exposure Leads to Insulin Resistance and Impacts Skeletal Muscle and Adipose Tissue Gene Expression in Offspring-rats
LAI Peng,YANG Xiao,LIU Zhen-ping,TANG Yuan-mou,RAO Su,WEN Chun-juan,JIAO Shi-rong,LEI Ji,CHEN Xiang-gui*
2013, 34(23):  282-290.  doi:10.7506/spkx1002-6630-201323057
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To explore the molecular mechanisms of insulin resistance (IR) in offspring-rats under maternal high-fat diet
exposure, we investigated whether maternal high-fat diet consumption can result in offspring malprogramming and the onset
of metabolic diseases in adulthood, and can impact skeletal muscle and adipose tissue gene expression. Sprague-Dawley
female rats were randomly divided into two groups and fed either a normal rodent chow (NC) or high-fat diet (HF). The
body weights of the rats were monitored and plasma parameters, oral glucose tolerance test (OGTT), insulin sensitivity index
(ISI) were measured. After mating with normal adult male rats, the pups of HF and NC female rats were divided into four
groups according to dams and gender: male offspring of maternal HF (MOH), female offspring of maternal HF (FOH), male
offspring of maternal NC (MON) and female offspring of maternal NC (FON). Offspring-rats were fed NC until 18 weeks
after weaning and weighed weekly. Plasma parameters, OGTT and ISI of offspring-rats were obtained and the morphology
of nuclei and mitochondria from left quadriceps cells in offspring-rats was observed. During the embryonic and fetal period,
the expression of 10 marker genes related to adipogenesis and myogenesis was detected. The results showed that maternal
high-fat diet exposure revealed obvious IR in MOH and FOH. The expression of adipogenetic and myogenetic genes was
disrupted seriously in MOH and FOH. As a consequence, maternal high-fat diet exposure leads to IR in offspring rats
through impacting the expression of adipogenetic and myogenetic genes.

Effect of Cyclocarya paliurus (Batal.) Iljinskaja Polysaccharide on the Proliferation and Differentiation of 3T3-L1 Preadipocytes
LIU Yao,FU Ling-yun,YE Zhen-nan,LI Nan,WANG Wen-jun*
2013, 34(23):  291-295.  doi:10.7506/spkx1002-6630-201323058
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This study was conducted to investigate the effect of Cyclocarya paliurus (Batal.) Iljinskaja polysaccharide
on the proliferation and differentiation of 3T3-L1 preadipocytes. The crude polysaccharide extract of C. paliurus was
fractionated by using D301-R macroporous resin and DEAE-anion fiber resin, and a C. paliurus polysaccharide component
(CPC) was obtained. Gel permeation chromatography (GPC) and multi-angle laser light scattering (MALLS) were then used
to determine its molecular weight distribution. The proliferation and differentiation of 3T3-L1 preadipocytes treated with
different CPC concentrations were measured by MTT and oil red O staining, respectively. At the same time the glucose
consumption and extracellular glycerol content were determined. Results showed that CPC consisted of three different
components and their viscosity-average molecular weight (Mη) was determined to be 50300, 3951 g/mol and 1406 g/mol,
respectively. Within a certain concentration range, CPC affected the proliferation of preadipocytes in a concentrationdependent
manner. Compared with the control group, CPC could reduce the glucose consumption and extracellular glycerin
content of 3T3-L1 adipocytes with inhibition ratios of as high as 9.61% and 30.8% respectively. These observations reveal
that the C. paliurus polysaccharide is capable of promoting the proliferation of adipocytes in certain concentration range and
reducing glucose consumption and extracellular glycerin content.

Nutritional Assessment of Proteins in Highland Barleys Grown in High-incidence Areas of Kashin-beck Disease in Aba, Sichuan
ZHANG Lu,PU Biao*,CHEN An-jun,LIU Xing-yan
2013, 34(23):  296-299.  doi:10.7506/spkx1002-6630-201323059
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The nutritional value of proteins in highland barleys grown in several high-incidence areas of Kashin-Beck
disease (KBD) in Aba prefecture, Sichuan province was evaluated by internationally recognized methods in comparison to
those from medium-incidence and non-incidence areas in the same prefecture. The results showed that the ratio of essential
to total amino acids in highland barleys from medium-incidence areas was 33.69% compared to 33.99% for those from highincidence
areas and 36.05% for those from non-incidence areas. The amino acid score (AAS), essential amino acid index
(EAAI) and score of ratio coefficient of amino acid (SRCAA) of highland barleys from both high- and medium-incidence
areas were lower than those of highland barleys from non-incidence areas. The nutritional value of proteins in highland
barleys from non-incidence areas was higher, whereas those from high- and medium-incidence areas had an unbalanced am
ino acid composition, inferior protein quality and lower protein utilization. The first limiting amino acid in highland barleys
was Lys for the three areas, and the second limiting amino acids were Met and Cys for high- and medium-incidence areas
and Thr for non-incidence areas. The average amino acid scores of Lys in highland barleys showed a significant difference
between incidence and non-incidence areas (P < 0.05). In conclusion, proteins in highland barleys from high- and mediumincidence
areas of KBD in Aba, Sichuan have a low nutritional value and high dietary consumption of highland barleys may
lead to deficiency of sulphur-containing amino acids and Lys in the body.

Protective Effect of Polysaccharide from Ganoderma atrum on Fasting Blood Glucose, Serum Lipids and Arteriosclerotic Narrowing of Superior Mesenteric Arteries in Type Ⅱ Diabetic Rats
ZHU Ke-xue,NIE Shao-ping*,SONG Dan,LI Chuan,LIN Su-li,XIE Ming-yong
2013, 34(23):  300-304.  doi:10.7506/spkx1002-6630-201323060
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Objective: To explore the protective effect of polysaccharide from Ganoderma atrum on fasting blood glucose,
serum lipids and superior mesenteric arteries of type Ⅱ diabetic rats. Methods: An animal model of type-Ⅱ diabetes was developed
by feeding high fat diet/glucose to rats for a relative long period followed by an injection of low dose of STZ. The experimental rats
were randomly divided into six groups: non-diabetic control group, high fat diet/glucose treated diabetic group, non-treated diabetic
group, diabetic group treated with Ganoderma atrum polysaccharide (100 mg/(kg·d)), diabetic group treated with cyclosporine
A (CSA, 8 mg/(kg·d)) and diabetic group treated with N-acetyl-L-cysteine (NAC, 100 mg/(kg·d)). Fasting blood glucose levels
and food and water intake amounts were measured once a week. At the end of the 4-week experimental period, serum TC, TG and
HDL-C levels and the histology of superior mesenteric arteries were assayed. Results: Ganoderma atrum polysaccharide exerted
noticeable hypoglycemic and hypolipidemic effects (P < 0.05) in diabetic rats. Meanwhile, it could relieve symptoms including
polydipsia, polyphagia and protect against the injury of superior mesenteric arteries. All the above beneficial effects were better
than those of CSA and NAC. Conclusion: Ganoderma atrum polysaccharide may ameliorate arteriosclerotic narrowing of superior
mesenteric arteries via regulating the glucose and lipid metabolism in type Ⅱ diabetic rats.

Effects of A Glycoprotein from Jellyfish (Rhopilema esculentum) on Cytokine mRNA Expression in BALB/c Mice
REN Guo-yan,SHAO Zheng,CAO Li,CAO Li,ZHANG Fan,GUO Jin-ying,FAN Jin-ling,CHEN Xiu-jin, ZHAO Rui-hong
2013, 34(23):  305-308.  doi:10.7506/spkx1002-6630-201323061
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In a previous study, we isolated a glycoprotein named JGP-III from jellyfish (Rhopilema esculentum). The specific
aim of the present study was to investigate the roles of JGP-III in the immunoregulatory mechanism in BALB/c mice by
investigating its effects on the mRNA expression of the Th1- and Th2-cytokines in the spleen lymphocytes of these mice. A total of
54 healthy BALB/c mice (SPF, 18-22 g) were randomly divided into 6 groups with 9 mice in each group: normal control, model
control, positive control, low dose (10 mg/(kg·d)), moderate dose (50 mg/(kg·d)) and high dose (100 mg/(kg·d)) groups, respectively.
After 21 days of these treatments, lymphocytes were prepared from the spleen of these mice and total RNA was
isolated from their lymphocytes. Fluorescent quantitative real-time polymerase chain reaction (FQ-RT-PCR) technique was
used to analyze the relative mRNA expression of Th1-cytokines (IL-2, IL-12 and IFN-γ) and Th2-cytokines (IL-4, IL-6 and
IL-10). Our results showed that JGP-Ⅲ promoted the mRNA expression levels of Th1-cytokines but inhibited the mRNA
expression levels of Th2-cytokines. Moreover, our results suggest that JGP-Ⅲ can improve the body’s immune function
probably by regulating the mRNA expression levels of Th1-/Th2-cytokines in spleen lymphocytes.

Effects of Probiotic Bacteria on Serum Antioxidant Capacity and Intestinal Flora of Mice Gavaged with Alcohol
ZHOU Meng-jia,NI Xue-qin*,ZENG Dong,ZHANG Lu,TU Teng
2013, 34(23):  309-312.  doi:10.7506/spkx1002-6630-201323062
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Objective: To study the beneficial effects of the probiotic bacteria Lactobacillus casei K1 and Bacillus subtilis JS01 on
serum antioxidant capacity and intestinal flora of mice gavaged with alcohol. Methods: Sixty 5-week-old male Kunming mice were
randomly divided into five groups (n = 12). Group I was gavaged with alcohol alone as a positive control and groups II, III, IV and
V with alcohol plus normal saline, Lactobacillus casei K1 (1×108 CFU/mL) alone, Bacillus subtilis JS01 (1×108 CFU/mL) alone
and both probiotic strains, respectively. The mice from each group were gavaged for 8 consecutive weeks. Microbial populations
were enumerated in the intestinal flora of mice at the end of the experimental period and some serum antioxidant parameters were
examined as well. Results: Compared with group I, the mice from group III showed a significant reduction in MDA content and
a significant increase in SOD activity as well as an extremely significant difference in CAT activity (P < 0.01), and significant
changes in SOD and MDA levels were also observed for those from group. However, the mice from the group V exhibited no
significant difference in MDA level from those from the positive control group, though the differences in SOD and CAT activities
were significant (P < 0.05) and extremely significant (P < 0.01), respectively. In the mice from groups III and V, significantly
higher amounts of intestinal lactic acid bacteria, bifidobacteria and anaerobic bacteria were observed as compared with the positive
control mice (P < 0.05). Similarly, the mice from group IV presented a significant increase in the number of intestinal lactic acid
bacteria (P < 0.05), although there were no significant differences in bifidobacteria or anaerobic bacteria (P > 0.05). Conclusion:
Lactobacillus casei K1 and Bacillus subtilis can not only improve serum antioxidant capacity but also reduce hepatic damage and
intestinal flora imbalance in mice gavaged with alcohol.

Effect of Fumonisin B1 on the mRNA Expressions of Cell Cycle-related Genes in Human Normal Esophageal Epithelial Cells
WANG Shao-kang,WANG Ting-ting,HUANG Gui-ling,SHI Ruo-fu,YANG Li-gang,DONG Shu-nan,SUN Gui-ju*
2013, 34(23):  313-316.  doi:10.7506/spkx1002-6630-201323063
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Epidemiological studies have shown that fumonisin B1 (FB1), a fungal toxin, may be associated with esophageal
cancer, but the laboratory evidence is still limited. In this study, human normal esophageal epithelial cells were isolated and
identified in vitro. The influence of FB1 on the mRNA expression of cell cycle-related genes including Cyclin D1, Cyclin E, P16,
P21, and P27 was determined by reverse transcription-polymerase chain reaction (RT-PCR). The results showed that FB1
played an important role in regulating the expression of Cyclin D1, Cyclin E, P16, P21, and P27 genes in normal human
esophageal epithelial cells at the transcriptional level, leading to high mRNA expression of Cyclin D1 but low mRNA
expression of Cyclin E, P16, P21, and P27 genes. The abnormal expression of cell cycle-related genes is closely related with
cancer. This in vitro study has further confirmed that FB1 is related with the occurrence of human esophageal cancer.

Effect of Hydroxysafflor Yellow A on the Oxidation of Intracellular Glutathione Induced by Hydrogen Peroxide in L02 Cells
WANG Xiao-na1,XU Xiao-min1,QIU Li-hong1,LI Gang2,XU Bo2,MA Cheng-jun2,WANG Zhen-hua1,2,*
2013, 34(23):  317-320.  doi:10.7506/spkx1002-6630-201323064
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The effect of hydroxysafflor A (HSYA) on the oxidation of intracellular reduced glutathione (GSH) induced
by hydrogen peroxide (H2O2) was investigated in L02 cells. 5,5’-Dithiobis(2-nitrobenzoic acid) (DTNB) was used as the
derivatizing agent and the reduced glutathione was measured by HPLC. The total glutathione was measured by the same
method after reduced by dithiothreitol (DTT). Oxidative glutathione can be calculated through reduced glutathione and total
glutathione. The L02 cells were pretreated with HSYA for 24 hours, and then exposed to H2O2 for 1 hour. The intracellular
GSH, GSSG contents and GSSG/GSH were analyzed by HPLC. The results showed that the standard curve was linear over
the range of 0.1-2 mmol/L. The minimum quantification limit was 0.1 mmol/L. The precision discrepancy of inter-batch
and intra-batch assays were both less than 10%. The recovery rates at low, medium and high spiked concentrations were
in the range of 95%-105%. The developed method proved to accurate, quick, precise and suitable for the measurement of
glutathione in cells. HSYA can significantly inhibit the increase of GSSG and GSSG/GSH, thus relieving the oxidative stress
induced by H2O2.

Effect of Oxidized Konjac Glucomannan on Inflammatory Cytokines in C57BL/6J Mice Fed a High-fat Diet
WU Ying-long,WANG Xin
2013, 34(23):  321-325.  doi:10.7506/spkx1002-6630-201323065
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Obesity is considered to be in a low-grade inflammatory state and the secretion of inflammation-related cytokines
rises as adipose tissue expands. Inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interlukin 6 (IL-6)
and monocyte chemoattractant protein 1(MCP-1), are modulated by adipose tissue and play an important role in metabolic
disease, which is one of the common characteristics of obesity-related disorders. In this study, the effects of oxidized konjac
glucomannan(OKGM) on inflammatory cytokines in C57/BL6J mice fed a high-fat (HF) diet were analyzed. Male C57BL/6J
mice were randomly divided into five groups with 12 mice in each group (n = 12): control (CL); a HF-diet and a HF-diet
containing 50, 25, 12.5 g/kg of OKGM, namely OKGN (H), OKGM (M) and OKGM (L) respectively, for 12 weeks. The
serum levels of FFA, TNF-α, IL-6 and MCP-1 of HF group were significantly higher than those of CL group. The size of
adipocytes and the serum levels of FFA, TNF-α, IL-6 and MCP-1 in HF-fed mice were significantly reduced by treatments
with increasing OKGM doses. These results suggest that OKGM can suppress the serum FFA and inflammatory cytokines
and also affect the size of epididymal adipocytes in C57BL/6J mice fed a high-fat diet, thus probably having a potentially
preventive effect on obesity-related metabolic diseases.

A Comparative Study of the Nutritional Effect of 12 Brands of Chinese and Foreign Infant Formula Milk Powder
XU Chong-hui,WEN Qiao-ling,HUANG Hua-jun,PAN Fang,YI Rong,LIU Hui-zhi,LI Hua-yan
2013, 34(23):  326-332.  doi:10.7506/spkx1002-6630-201323066
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Objective: Six domestic brands (3 made from domestic milk sources and 3 from imported milk sources) and 6
foreign brands (3 marketed in the Chinese mainland and 3 purchased from Hong Kong) of infant formula milk powder were
used to feed SD rats for a comparative study of their biological effects in vivo. Methods: After 30-day feeding, nutritional
indexes of newly weaned rats were detected. Results: The body weights and food utilization of rat pups fed the different
brands of infant formula were significantly higher than those of the corresponding blank controls (P < 0.05 or P < 0.01).
In addition, no significant differences in body weight gain, gain trend, food utilization, routine blood parameters or blood
biochemical indexes were observed among 6 infant formulas from different milk sources and among 6 ones marketed in
different regions. Conclusion: These 12 brands of infant formula have the same biological effects in weaned rat pups.

Research Progress of Yuba Stick Processing Technologies and Factors Affecting Its Quality
LI Yong-ji,ZENG Mao-mao,HE Zhi-yong,CHEN Jie*
2013, 34(23):  333-337.  doi:10.7506/spkx1002-6630-201323067
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Yuba stick is a traditional soybean product with a long history of consumption, which is popular among
Chinese consumers. The mechanism of Yuba formation and its internal structure are outlined in this paper, beginning with a
description of the main processing technologies currently available for Yuba sticks as well as the major commercial products
and their features. The emphasis is on discussing some major factors influencing the yield of Yuba sticks such as soybean
cultivar, soybean milk concentration, temperature and heating methods as well as those influencing the quality of Yuba sticks
as evaluated by color, mechanical properties, rehydration and cooking tolerance and corresponding control measures. In
addition, an overview is presented on recent applications and efficacy of food additives in Yuba. Finally, the current situation
of research on Yuba sticks is summarized and future trends in its development are prospected.

Research Progress in the Mechanism of the Biosynthesis of Conjugated Linoleic Acid Isomers in Mil
FU Jin-heng1,DING Yi-chun2,LI Hai-xing1,CHEN Yan1,LI Chao-bo1,CAO Yu-sheng1,LIU Xiao-hua1,*
2013, 34(23):  338-343.  doi:10.7506/spkx1002-6630-201323068
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Conjugated linoleic acid (CLA) is a collective term for geometric and positional isomers of linoleic acid with
conjugated double bonds. They have been found to be responsible for many biological properties that are related to human
health. c9,t11-CLA has anti-carcinogenic activity, and t10,c12-CLA seems to be specifically responsible for reducing body
fat and enhancing energy metabolism. CLA occurs naturally in milk fat in a relatively low level. Several CLA isomers such
as c9,t11-CLA and t10,c12-CLA are detected in milk. These CLA isomers are mainly biosynthesized by some bacteria in the
rumen. The structure and functions of CLA, analytical methods for CLA isomers, the composition and biosynthesis pathways
of CLA in milk, methods of enhancing CLA content in milk and microorganisms involved in the biosynthesis of CLA in the
rumen are reviewed in this paper.

Phytosterols in the Prevention and Treatment of Cardiovascular Diseases
ZHANG Lei,CHEN Qing-sen*,YAN Ya-li,PANG Guang-chang
2013, 34(23):  344-350.  doi:10.7506/spkx1002-6630-201323069
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Phytosterols, as a group of bioactive substances, have many important physiological characteristics and unique
nutritional properties and are widely used in the fields of functional food and medicine. They play a significant role in the
prevention and treatment of cardiovascular and cerebrovascular diseases. In this paper, some causes and pathological mechanisms
and cardiovascular and cerebrovascular diseases are outlined. Moreover, the structural classification, physiochemical properties
and mechanism of action of phytosterols as well as the current status of development of related products and new processing
technologies are systematically reviewed. This review is expected to provide guidelines for further studies.

Research Advances in Bio-control of Postharvest Diseases of Citrus with Antagonistic Microorganisms
GUO Juan-hua1,TU Qi-hong2,CHEN Chu-ying1,CHEN Jin-yin1,*
2013, 34(23):  351-356.  doi:10.7506/spkx1002-6630-201323070
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The major postharvest diseases of citrus are briefly introduced and the bio-control of postharvest diseases
of citrus with antagonistic microorganisms is summarized in this paper. The following aspects are covered in this
review:antagonistic microorganism species, methods for their application, bio-control mechanisms and the development of
bio-control products which can control infectious diseases of postharvest citrus. In addition, many methods to enhance the
efficacy of antagonistic microorganisms for controlling postharvest diseases of citrus discussed. This knowledge will provide
a reference for bio-control of postharvest diseases in citrus.

Development of Biosensors for the Detection of Containments in Foodstuffs
JIANG Xue-song1,2,XU Lin-yun1,LU Li-qun2,SHEN Fei3,ZHOU Hong-ping1
2013, 34(23):  357-362.  doi:10.7506/spkx1002-6630-201323071
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Biological and chemical pollution in foodstuffs is becoming an increasingly important issue in the field of food
safety. As a result, higher analytical requirements are needed for the detection of contaminants in foods. Biosensors are
designed based on specific biochemical reactions mediated by isolated enzymes, immune preparations, tissues and organelles
or whole cells to detect biological and chemical compounds usually by electrical, thermal or optical signals. Some recent
applications of biosensors in the detection of containments in foods, such as microorganisms, toxins, pesticides, heavy
metals and illegal additives are reviewed in this article. In addition, some key points and future trends in the application of
biosensors for food safety detection are also discussed.

Research Advances in Postharvest Physiology and Preservation of Fig Fruits
ZHANG Xue-dan1,AN Miao1,ZHANG Qian1,MENG Yan-ling2,XIN Li1
2013, 34(23):  363-369.  doi:10.7506/spkx1002-6630-201323072
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The fig fruits have high nutritional value and medicinal value and are perishable climacteric fruits due to the
difficulty in postharvest storage and preservation. Along with deep understanding of its nutritional value, the demand
for high-quality figs is growing. Postharvest physiology and storage of figs are critical for the fig- processing industry.
The current progress of storage physiology, pathophysiology and storage of postharvest figs is reviewed in this paper.
The major technologies currently available for postharvest storage and preservation of figs include 1-MCP treatment,
modified atmosphere packaging, SO2 or ClO2 treatment, and so on. This review will hopefully provide references for future
development and utilization of figs.

Bibliometric Analysis of SCI Papers on Plant Polyphenol Published during 2002—2012 and the Hot Topics in Biological Activity
LU Yu-miao1,MA Hui-ling2,*
2013, 34(23):  375-383.  doi:10.7506/spkx1002-6630-201323074
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Based on the SCIE database, bibliometric methodology was applied to analyze relevant records regarding
to plant polyphenols from 2002 through 2012, including years, journals, affiliations, authors, key words, discipline
distribution and research topics. Hot topics were extracted from the statistical result of key word frequency and
thematic brows. The results showed that research on plant polyphenols was active in the fields of food technology,
applied chemistry, nutrition, biochemistry, molecular biology and pharmacology. The hot topics on biological activities
of plant polyphenols focuses on: 1) in vitro and in vivo antioxidant activity of plant polyphenols; 2) anti-cancer, antiaging,
anti-cardiovascular disease activities of plant polyphenols and corresponding mechanisms; 3) absorption,
transformation and bioavailability of dietary polyphenols.

Bibliometric Analysis of SCI Papers on Plant Polyphenol Published during 2002—2012 and the Hot Topics in Biological Activity
LU Yu-miao1,MA Hui-ling2,*
2013, 34(23):  375-383.  doi:10.7506/spkx1002-6630-201323074
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Based on the SCIE database, bibliometric methodology was applied to analyze relevant records regarding
to plant polyphenols from 2002 through 2012, including years, journals, affiliations, authors, key words, discipline
distribution and research topics. Hot topics were extracted from the statistical result of key word frequency and
thematic brows. The results showed that research on plant polyphenols was active in the fields of food technology,
applied chemistry, nutrition, biochemistry, molecular biology and pharmacology. The hot topics on biological activities
of plant polyphenols focuses on: 1) in vitro and in vivo antioxidant activity of plant polyphenols; 2) anti-cancer, antiaging,
anti-cardiovascular disease activities of plant polyphenols and corresponding mechanisms; 3) absorption,
transformation and bioavailability of dietary polyphenols.