Abstract:The breeding of fermentation strains affect the efficiency of sugar utilization on lignocellulosic ethanol,especially for increasing the xylose utilization efficiency to improve ethanol yield.This review summarized the development of fermentation strains,processes optimization and ethanol dehydration on lignocellulosic ethanol,and their advantages were discussed.Moreover,the future on the development of fermentation and ethanol dehydration are proposed.

Abstract:Three different enzymes are involved in lignin degration produced by the white rot fungi,they are lignin peroxidases,manganese peroxidases and laccase.Laccase,a member of the blue multi-copper oxidase family,catalyzes the oxidation of a variety of organic substrates coupled to the reduction of molecular oxygen to water.The abilities to degrade lignin by different laccases vary widely,which depend on the structure of laccases,especially the structure of the trinuclear copper site.The structure characteristics,active site and three-dimension structure of fungal laccase weresummarized in this review.

Abstract:The present study aimed at exploring the effect of Taihe Black-bone silky fowl active peptides and their 13 separated fractions on the proliferation of bone marrow nucleated cells from normal mice.MTT method was used to investigate the effect of active peptides on the proliferation of bone marrow nucleated cells from normal mice.HPLC precolumn derivation with 2,4-dinitrofluorobenzene method was used to determine the amino acid compositions of four active peptide fractions.When mice bone marrow nucleated cells were cultivated for 24 h,it was found that the fractions 7,8,9 and 10 separated from Taihe Black-bone silky fowl active peptides exhibited significantly strong activity(P<0.01) in stimulating mice bone marrow nucleated cells,and the proliferation rate could be up to 130%.Especially,at lower concentration(0.1 μg/mL),fraction 7 was able to significantly stimulate mice bone marrow nucleated cells(P<0.01) and revealed better reproducibility.Fraction 7 was capable of stimulating mice bone marrow nucleated cells proliferation by 298%.Amino acid composition analysis demonstrated that the main amino acids of fraction 7,8,9 and 10 consisted of tyrosine,glycine,glutamic acid,aspartate and alanine.It could be presumed that tyrosine,glycine,glutamic acid,aspartate and alanine may be the material base of active component in stimulating mice bone marrow nucleated cells.

Abstract:The objective of this study is to choose a novel antibactierial substance from Chinese herbs to control bacteriosis of fishes,based on the result of the agar well diffusion method and minimal inhibitory concentration(MIC).The result showed that the extracts from Tea polyphenol,Galla Chinensis,Rhubarb,Scutellaria and Terminalia chebula exhibited excellent antibacterial activity.And the optimum formation of Traditional Chinese herbs is Tea polyphenol,Galla Chinensis and Rhubarb at ratio 5:2:1(w/w/w),the minimum inhibitory concentration(MIC) is 78 μg/ml.The contents of epigallocatechin gallate(EGCG),epicatechin(EC),epigallocatechin(EGC),epicatechin gallate(ECG),catechin、gallic acid were 12.6%,7.57%,4.26%,4.19%,1.03%,1.18% in the compound,respectively.

Abstract:In this manuscript,10% and 16% Ascorbic acid(AA) was encapsulated in glassy isomalto-oligosaccharides by extrusion.The measuring feeder was set to obtain a flow rate of 1 kg/h of premixed material and screw speed was maintained at 60 r/min to study effect of barrel temperature,torque,and die head pressure on the AA yield,loading capacity and moisture content of the products.It was found that high barrel temperature could make motor torque and die head pressure decrease.At middle and low barrel temperature,torque decreased with increased AA.AA could be encapsulated successfully.The glass temperature was decreased with increased AA.The result of X-ray diffraction indicated that the formulation is a glassy solid solution.

Abstract:In this study,different levels of taurine(0,100,500 mg/kg) were added to the diets and the effects of taurine on production,immune responses and antioxidative ability of quails were investigated.A total of 300 quails were allocated to 3 dietary treatments.Each treatment comprised 4 replicates of 25 quails.The experimental period lasted 6 weeks.The results showed that dietary 500 mg/kg taurine increased body weight of 3 weeks and body weight gain(0~3 week),decreased feed conversion ratio significantly compared with the control(P<0.05).Dietary taurine increased the ability of stimulation B-lymphocyte proliferation significantly and 500 mg/kg taurine increased the ability of stimulation T-lymphocyte proliferation significantly(P<0.05).Dietary taurine increased serum IgG content significantly compared with the control(P<0.05).Dietary taurine of 100 mg/kg enhanced the activity of total antioxidant capacity(T-AOC) in serum,decreased the malondialdehyde(MDA) level in serum(P<0.05).The above results demonstrated that taurine can increase 1) performance of quails;2) antioxidant ability;3)immune function.

Abstract:This manuscript study the effect of 4-α-GTase on the digestibility of rice starch,then the corresponding mechanisms was elucidated by differential scanning calorimetry(DSC),X-ray diffraction(X-ray),and scanning electron microscope(SEM).It was found that: 1) The content of the slowly digestible starch(SDS) in starch increased to the maximum value when modified for 3～4 h by 4-α-GTase(50 U/g dry starch),and then decreased with increasing of the modification time;2)During of 2～6 h modification,the rate of retrogradation was accelerated,then gradually decreased after 6 h;3) The crystal structure of starch was changed from A-type to B-type after treated by 4-α-GTase;4) The surface of starch became rough and irregular with more and smaller fraction stacked.From the above results,it is can be concluded that the changes of the digestibility could be attributed to the changes of structure and property.

Abstract:This manuscript investigated effects of temperature and vacuum pre-cooling on the special enzyme activity of three kinds of edible fungi,such as Boletus aereus,Lentinus edodes and Agaricus bisporus.For Boletus aereus and Lentinus edodes,the temperature were set at 4 ℃,10 ℃ and 20 ℃,while Agaricus bisporus were controlled by vacuum pre-cooling.The changes of edible fungi specific enzymes activities such as peroxidase,polyphenol oxidase,catalase,as well as carboxymethyl cellulose enzyme,superoxide dismutase,poly-galacturonic acid with the storage time were demonstrated in this study.

Abstract:In vitro antioxidant activities of L-ascorbyl fatty acid esters were studied by the items of hydroxyl radical system,superoxide anion free radical system,system of DPPH·,and reducing power.Its antioxidant effects was further evaluated by adding into the lard and bead oil.The results indicated that L-ascorbyl fatty acid esters within the given concentration range in dose-effect relationship showed good antioxidant activities on both the clearance of hydroxyl radical,superoxide anion free radical and DPPH· and the reducing power.Furthermore,the antioxidant activities of L-ascorbyl fatty acid esters were comparable with that of L-ascorbyl palmitate.When 0.2%(w/w) of L-ascorbyl fatty acid esters were added in the lard for a 20 h forced oxidation at 100 ℃,the POV of lard was 42.8 meq/kg,which illustrated L-ascorbyl fatty acid esters had significant antioxidant activities.When 0.2%(w/w) of L-ascorbyl fatty acid esters were added in the soybean oil for a 14 h forced oxidation at 100 ℃,the POV of soybean oil was 11.9 meq/kg.The antioxidant ability of L-ascorbyl fatty acid esters for soybean oil was higher than that of TBHQ,but was close with that of L-ascorbyl palmitate.Thus,L-ascorbyl fatty acid esters were demonstrated to be potential food antioxidants.

Abstract:The cryoprotective role of emulsifiers combined with two molar soybean lecithin and one molar sucrose ester on baker’s yeast was examined in this study.The results showed that the survival ratio of yeast cultivated for 48 hours after frozen at-30 ℃ for 5 days in a culture medium with 4 g/dL combined emulsifers increased by 60% compared with that of the control group,and the adaptive phase shorten by 10 hours.The dough made from yeast dealed with emulsifiers after frozen at-30 ℃ for 60 days had a relatively stronger ability to leaven within a short time.The phenomenon of yeast congregated with enulsifiers,which was imaged by scanning electron microscope(SEM),indicated that there might be some interaction effects between the two substances.

Abstract:In this manuscript,five types of macroporous resins were selected for adsorption and separation of flavonoids from Bamboo Shell.It was found that resin HPD600 was an excellent adsorbent with higher adsorption and desorption capacity.The effects of concentration,pH and flow rate of the solution on the dynamic adsorption of resin HPD600 were also studied.The results showed that the optimum adsorption conditions are listed as following: 7 BV flavonoids concentration of extract solution at 3.89 mg/mL,pH 3.0,flow rate at 3 BV/h using 6 BV40% ethanol as the best desorption solvent,the total recovery of chlorogenic acid reached at 82.33%,and the content of flavonoids reached at 35.12% in the crude product.

Abstract:This manuscript developt a rapid detection method Elispot assay for Gentamicin sulfate,for this,Anti-GM polyclonal antibody through GM immunogen was obtained;Establishing milk GM-Elispot assay method by spotting coating antigen on PVDF membrane,and the coating antigen and GM in samples competitive binding the site of anti-GM polyclonal,then the HRP catalyzed chromogenic substrate,according to presence or absence of color and depth for interpretation of results.Results: The detection thre-shold of this method was 10 ng/mL,detection time was within 40 minutes and it can achieve semiquantitative detection.The strip of which was stored at 4 ℃ for 90 days could be used and no cross-reaction was found with several structural analogues.The results of samples analysis obtained from the Elispot assay were in a good agreement with that of ELISA method.

Abstract:This manuscript study the process parameters for juice made from full lotus root powder.The optimum composite levels were achieved:carrageenan 0.02%,β-cyclodextrine 0.08%,sodium alginate 0.05%.The juice flavor of full lotus root powder and fresh lotus root were both conducted with the aid of the electronic nose,it was found that both of them has the same flavor material type,but the flavor threshold value full lotus root powder juice was inferior to the fresh lotus root juice.

Abstract:The melamine deaminase gene MDA with optimized codon usage was obtained from vector pUC57-MDA,and subcloned into the expression vector 6HisT-pRSET generating recombinant expression vector 6HisT-pRSET-MDA.The recombinant strain BL-MDA was generated after 6HisT-pRSET-MDA was transformed into E.coli BL21(DE3).Expression of BL-MDA was analyzed by SDS-PAGE and the activity of recombinant enzyme was determined by indophenol blue spectrophotometric method.

Abstract:In this study,112 lactic acid bacteria strains were isolated from four kinds of traditional fermented meats.Through the study of fermentation properties and strain tolerance,the strain T1-1 was selected which grown well in MRS broth with 8 g/dL NaCl and 150 mg/kg NaNO2,and showed a high ability to survive at 80 ℃ water bath after 10 min.Then the strain was identified as Lactobacillus plantarum by using physiological-biochemical characteristics and 16S rRNA sequence.

Abstract:To study the native promoter of xylanase II of Trichoderma asperellum,cloning and sequence analysis of xylanase II structural gene and its 5’ flanking region was performed.On the basis of the genomic conserved sequence of Trichoderma spp.,degenerate PCR was designed to amplify the xylanase II structural gene and its 5′flanking region of Trichoderma asperellum.The product was cloned into T vector and confirmed by EcoR I digestion.Sequence analysis showed that cloned fragment was 1 875 bp long,comprising the 795 bp xylanase II structure gene and its 1 080 bp 5’flanking region.The structure gene encoded a polypeptide of 223 amino acids,where the Glyco-hydro-11 superfamily domains were detected.In the 5’ flanking region,core promoter region,transcriptional start site,CAAT-Box and TATA-Box were detected.Several typical transcriptional factor binding domain,such as CreI,AreA,XlnR,Ace1,AlcR were also found.The 1080 bp 5’ flanking region was a typical promoter.The isolation of this native promoter can benefit the development of an efficient Trichoderma asperellum host system for gene expression.

Abstract:To clone the RmlA gene in Lactobacillus rhamnosus JAAS8,degenerate primers were designed from the conserved domain of the exopolysaccharide biosynthetic genes of lactic acid bacteria(LAB).The distinctive PCR products with the expected size were amplified from L.rhamnosus JAAS8.The gene sequence(4.1 kb) of RmlACB and Orf1 were cloned by genomic walking method.Nucleotide sequence similarity analyses with GenBank sequences were performed using the BLAST and ClustalX software.GenBank database searching shows that the predicted protein products of the RmlACB genes from L.rhamnosus JAAS8 have highest similarity with the EPS biosynthetic genes of Lactobacillus.The genes RmlA、RmlC and RmlB are closely related to the biosynthesis of dTDP-L-rhamnose precursor.

Abstract:Expression of staphylococcus aureus enterotoxin A in Escherichia coli BL21(DE3) using lactose as inducer instead of IPTG was investigated.Effect of inducing conditions,including cell age of recombinant,concentration of lactose,inducing time and adding model of the lactose on the expression level were detaited studied and analyzed.The optimal inducing conditions listed as followed: 0.5 mmol/L(final concentration) lactose by one-time method at the mid-phase of cell growth(OD600=0.6) and then cultured bacteria at 37 ℃,180 r/min for 6 h.The yield of recombinant staphylococcal enterotoxin A induced by lactose was about 36.9% of the total cell solution protein.It was just a little below of that induced by IPTG(38.1%).The price of lactose is only 1% of that of IPTG.All of these suggested that the lactose is available to be effective inducer with lower cost in the process of large-scale production of recombinant protein.

Abstract:Microbial community and its changes in pickled mustard tuber were investigated at the low salinity by using single-strand conformation polymorphism(SSCP) method.Effects of gel concentration,glycerol addition,foretreatment of electrophoresis and silver staining methods on SSCP pattern were also examined in this manuscript.The results demonstrated that an ideal SSCP method was obtained as the optimum conditions,gel concentration of 22 g/dL at the crosslinking degree of 29∶1(a ratio of acrylamide and bisacrylamide),glycerol concentration of 6%,voltage of 250 V,electrophoresis buffer of 1×TBE,electrophoresis at 4 ℃ for 18 hours,and using silver staining method of strong alkaline solution and formaldehyde as reducing agent.Based on the optimized SSCP method for pickled leaching liquid measurement,it was found that the Leuconostoc mesenteroides is the dominant microorganism in the initial stage of pickling,then Lactobacillus plantarum and Lactobacillus brevis grown up with the alteration of environmental conditions.At the later stage,the dominant microorganism are Lactobacillus plantarum and Lactobacillus versmoldensis,respectively.The contribution of lactic acid bacteria was confirmed by microbial community structure analysis in pickled mustard tuber with low-salinity,which is carried out by combining the sensory evaluation method.

Abstract:Malate dehydrogenase(MDH) gene was amplified via PCR from the chromosome of Escherichia coli in this manuscript.The PCR product was cloned into the expression vector pET-28a(+).The resulted recombinant plasmid was transformed into E.coli BL21(DE3).Induced by 0.5 mmol/L IPTG,MDH,a 36KDa protein,was successfully expressed in E.coli BL21(DE3).An active MDH was purified by Ni-NTA column affinity Chromatography,with the specific activity of 112.5 U/mg,the purification multiple of 2.62,and the recovery rate of 59%.In a preliminary study,the enzymatic properties of the purified His-tagged enzyme were characterized.It was found to have pH and temperature optima of 37 ℃ and 6.0,respectively.The enzyme was stable when pH and temperature kept in the range of 2.0 to 6.0 and blow 42 ℃,respectively.Its activity was activated by K+ dramatically,inhibited by Cu2+,seriously inhibited by Zn2+ and Hg2+.Although alcohols have little effect on this enzyme,glycerol could dramatically improve the thermal stability of MDH.When oxaloacetic acid was used as substrate,the enzyme kinetic constants of Km and Vmax was 0.235 mmol/L and 0.47 μmol/(L·min),respectively.

Abstract:The objective of this study was mainly to screen the bacteria that could remove aflatoxin B1 in the peanut soil and moldy meal.After the first screening by the nutrient-specific method,the effect of cells and the supernatant fluid of the seven strains on aflatoxin B1(200 μg/L) was investigated.The result demonstrated that among the seven strains,the strain TRS-3 can remove aflatoxin B1 effectively,with 48.26% and 53.56% of the removal ratio of AFB1 by viable and dead cells,respectively.The degradation ratio of the supernatant fluid reached 78.55%,which was significantly higher than that of others.The strain TRS-3 was finally identified as Bacillus megaterium.

Abstract:Vibrio parahaemolyticus(Vp) has been considered as one of the most threatening foodborne bacterial pathogens.The aim of this study is to uncover the situation of contamination of Vibrio parahaemolyticus which isolated from the shellfish in the retail market of Shanghai,and provide some guidelines for Vp prevention.For this,we carried out the international method GB/T 4789.7-2008 to isolate bacteria and analyze its physiological and biochemical characteristics.Additionally,some primers for tlh specific genes and virulence genes(tdh,trh) recommended by FDA were used for the PCR analyze.The result shows that 45 strains of Vibrio parahaemolyticus were isolated from 161 samples which come from the shellfish market of Shanghai The average detection rate is 27.95% Based on the PCR detection of tdh and trh gene,there were only 7 tdh gene found among the 45 strains,but none of them contain the trh gene.45 strains of Vp consisted of 7 serum groups:O1 2.2%(1/45),O3 24.4%(11/45),O4 15.6%(7/45),O5 20.0%(9/45),O9 4.4%(2/45),O10 8.9%(4/45),O11 26.7%(12/45).

Abstract:D-tagatose 3-epimerase(DTE) is the most effective enzyme for the biological production of D-psicose,a novel functional factor.In this study,a novel gene encoding DTE from Clostridium cellulolyticum H10 was cloned.pET-22b(+) was for the plasmid,E.coli BL21(DE3) acted as host cells,then the recombinant strains were constructed.The target protein was over-expressed by IPTG induction;the recombinant DTE purified to electrophoretical homogeneity with affinity chromatography was analyzed by SDS-PAGE,showing approximately 31 000 characteristic protein.The result of activity detection revealed that the recombined enzyme had a higher transforming activity.

Abstract:Due to the importance of NAD+ and NADH levels in the yeast metabolism,the determination of intracellular NADH/NAD+ is a very interesting for ethanol production.In this study,Three NADH./NAD+ extracting methods,including heating,freeze-thaw cycles,shaking bead,for the disruption and extraction of the NAD+ and NADH in yeast cells were investigated and compared.Finally an efficient process for extraction and detection of NAD+ and NADH in yeast cells was established as follows.Yeast cells were disrupted by shaking bead while NAD+ and NADH were extracted by acid or alkali,then the extracts were detected under the favorable enzymatic cycling system Ⅱ.

Abstract:The activities of β-glucanase,xylanase,protease and cellulase produced by 40 strains of Geotrichum candidum were studied in this manuscript.The weighted mark method was used to analyze activities of the above enzymes,and four G.candidum strains including No.3,No.12,No.18 and No.39 were selected as the malting microbial strains.The malting trials indicated that four G.candidum strains could improve the modification of malt and No.12 was the best one.Compared with that of the control,β-glucan content in final malt produced by G.candidum No.12 was decreased by 17.8%,and filtration volume in sixty minutes,diastatic power and free amino nitrogen content increased by 35.4%,10.3% and 7.3%,respectively.Meanwhile,other characteristics of the final malt met the requirement of QB1686-2008.The results of fermentation experiment shown that the brewing characteristics of microbial malt was better than that of the control malt,and the beer brewed with microbial malts or control malts both met the requirement of GB4927-2008.

Abstract:In this study,rice was used as a raw material in the preparation of cordycepic rice by solid state fermentation,and the cordycepic acid contents of Cordyceps militaris rice were determined.Single factor test and orthogonal test results showed that the optimum conditions were as follows: rice as raw material,material/water ratio 1∶0.6(g/mL),the amount of inoculum 10%(with rice dry weight as basis),the temperature 22 ℃.Under the optimal conditions the cordycepic acid content was achieved at 6.04 mg/g.Cordycepic rice was bright with uniform golden yellow colour.

Abstract:The effect of Fe2 + concentration on the enzymes during methane generation from Taihu blue-green algae was studied in this manuscript.It was found that the maximum methane yield reached at 986.7 ml when Fe2 + concentration was 3 mg/L,and it was about 43 times more than that of the control group.When 0.5 mg/L Fe2+ added to the fermentation medium,the activity of dehydrogenase,and BAA-hydrolysing protease was 118 U and 3 126.9 U,respectively,and increased 76% and 83.4%,when compred with the corresponding value of the control.The activity of β-glucosidase enzyme reached 47 493 U,when Fe2 + concentration was 1 mg/L,and it was about 6.3 times more than that of the control group.The coenzyme F420 content was 0.2 mmol/g when Fe2 + concentration was 3 mg/L,which was about 4.3 times more than that of the control group.

Abstract:Bdellovibrio which could grown at high temperature conditions was screened and purified in Tianjin four main rivers.The biological characteristics were tested by double deck agar plate,including its host spectra,pH resistance,and cultivation temperature.The results showed that the Bdellovibrio with high temperature resistance growed well at pH 7.0~7.5,and it could prey on the dead bacteria earlier compared with the living bacteria,such as Escherichia coli,Aeromonas hydrophila,Pseudomonas fluorescens,mass of gram negative bacteria and some gram positive bacteria could be preied on by the Bdellovibrio,which is an obligate parasite.

Abstract:In this study,the improved two-step carbodiimide method was used to connecte doxycycline hapten with carrier protein BSA,and doxycycline with the carrier protein OVA to prepare the artificial antigen of doxycycline(BSA-DC) and coating antigen(OVA-DC),respectively.UV scanning analysis and animal immune tests confirm the success in preparation of artificial antigen,and the combination ratio of doxycycline and BSA is 3∶1 by calculating.At the same time,the results showed that high titers of 2.048×106 anti-DC serum were produced by BSA-DC immunized rabbit.It provided a necessary condition for ELISA detecting doxycycline residues.