Abstract:Formate dehydrogenase（FDH） is an industrial enzyme widely used for NADH regeneration. However，the low activity and low catalytic efficiency are flaws of native FDH. In this study，amino acid sequences alignment of FDH from different sources was conducted. The result showed that the 93-site amino acid of Candida boidinii FDH（CbFDH） was valine（V），while that of others were isoleucine（I）. Moreover，according to the hydrophobicity，four mutant enzymes V93L，V93I，V93A and V93G were constructed. The enzymatic properties showed that the activity of mutants was improved as the enhancement of 93-site amino acids hydrophobicity，except mutant V93L. And the activity and catalytic efficiency（for NAD+） of V93I were increased by 22.6% and 133%，respectively. Mutant V93I and L-leucine dehydrogenase were used to synthesize L-norvaline by asymmetric reduction of α-ketovaleric acid. The results showed that the conversion efficiency of α-ketovaleric acid was improved with mutant V93I.

Abstract:Patulin（PAT） is one of toxic fungal secondary metabolites，mainly produced by Aspergillus sp. and Penicillium sp，it has been identified in various fruits and fruit products all over the world，and it has cytotoxicity，teratogenicity and genotoxicity，hence，it poses potential hazard to human and animals. In recent years，biological control has become a promising method to control the mycotoxin produced by fruits because of its advantages of safety，green and high efficiency. It has been reported that，some strains of yeast can directly inhibit the production of PAT，and even degrade PAT，however，the mechanism of controlling and degradation PAT underlying is not clear. Our previous study found that，Sporidiobolus pararoseus Y16 can significantly degrade PAT in vitro，but the possible mechanism is not clear，in this study，the mechanism of degradation of PAT by S. pararoseus using proteomic technology was investigated. This study can provide a new theoretical evidence for the biological degradation of mycotoxins.

Abstract:The objective of this study was to investigate the inhibition effet of sodium lactate（NaL） on biofilm formation by Listeria monocytogenes. First of all，the crystal violet staining assay was used to evaluate the inhibition effect of NaL at different concentrations（0%，2.5%，5%，10% and 20%） on the biofilm formation of L. monocytogenes. Furthermore，the effets of NaL on its biofilm structure，polysaccharides，extracelluar proteins，cell activity and membrane integrity of L. monocytogenes biofilm cells were also investigated. Finally，the detection of the expression level of the biofilm-associated genes motB，mogR，degU，flgE，dnaK，prfA and sigB was performed by using quantitative real-time PCR（qRT-PCR）. With the increase of the NaL concentration（2.5~20%），biofilm formation of L. monocytogenes was inhibited significantly（p<0.05） and the inhibition rate was 8.34 %，32.2 %，46.6 % and 55.2 %，respectively. Results of microscopy analysis showed that after 5 % NaL treatment，the structure of L. monocytogenes biofilm was forming instead of small clumps with reduction of thickness，and a significant decrease（p<0.05） was determined in total production of polysaccharides and extracelluar proteins of L. monocytogenes biofilm cells. In addition，5 % NaL inhibited the cell activity and membrane integrity of L. monocytogenes biofilm cells to suppress the development of new biofilms. qRT-PCR results revealed that NaL could downregulate biofilm-associated genes in L. monocytogenes and therefore to inhibit its biofilm formation. In conclusion，this study could provide a scientific basis for the effective inhibition of NaL on L. monocytogenes biofilm formation.

Abstract:Bacteria adhesion on surfaces often causes implanted infections，food spoilage and human illness. The control of the initial adhesion to materials is very important to avoid safety and health problems. In this study，materials with different surface wettability were fabricated via chemical grafting. Three model strains（Escherichia coli BL21，Staphylococcus aureus 6538 and Pseudomonas aeruginosa ATCC 9027） were imported to demonstrate the influence of surface wettability markedly. Besides，surfaces with negative charge were more resistant to bacterial adhesion. The results of this study provided new insights into the mechanism of bacteria adhesion，and could be effectively used in designing of anti-bacterial materials.

Abstract:To prepare the CD44-targeted hyaluronic acid-deoxycholic acid（HAAD） amphiphile to form micelles for the encapsulation of model anticancer drug doxorubicin（DOX），and its properties and anticancer activity were characterized. The boronate derived deoxycholic acid was synthesized via amidation reaction. Afterwards，HAAD was prepared by conjugating hyaluronic acid and boronate derived deoxycholic acid via boronate ester bond. The DOX encapsulated HAAD micelles were prepared and their properties，such as encapsulation efficiency（EE），drug loading level（DL） of DOX，in vitro drug release behavior were characterized. The cytotoxicity and cellular internalization were evaluated against model tumor cells（MCF-7） and normal cells（COS7）. HAAD was successfully prepared and could self assemble into micelles with DOX loaded inside. The DL value of DOX was 9.15%. Moreover，the DOX-loaded HAAD micelles could show pH-responsive drug release behavior，the drug release rate was faster at acidic milieu. The flow cytometry analysis demonstrated the DOX-loaded HAAD micelles could be internalized by CD44-overexpressed MCF-7 cells more effectively. Additionally，the cytotoxicity assay showed DOX-loaded HAAD micelles had less toxicity against normal cells than that of tumor cells，showing selectivity.

Abstract:In order to study the characteristics of brewing properties of different rice varieties，four kinds of rice（round-glutinous，long-glutinous，japonica and indica rice） were used as the raw materials for Shaoxing Process brewing. By monitoring the basic physical and chemical indicators（alcohol content，reducing sugar，total acid，etc.） in fermentation process and the content of fermentation liquid alcohols，aldehydes，acids，esters of volatile flavor compounds were analyzed. The results showed that round-glutinous rice performed easier saccharification and fermentation with high alcohol yield；alcohol content reaches 18% at the end of fermentation，and the residual sugar content was relatively high. At the end of fermentation，the amino acids in rice wine fermentation liquor of different rice varieties were mainly sweet and bitter amino acids，and there were significant differences in the contents of sweet，bitter，fresh and astringent amino acids（p<0.01）. The fermentation liquid of amino acid content is the highest，followed by japonica rice. Japonica glutinous rice fermentation broth of volatile flavor compounds of the total content and alcohols content has obvious advantages，and indica rice in brewing wine esters content is the highest.

Abstract:Coenzyme NAD（H） is necessary for whole cell synthesis of L-phenylglycine，so intracellular NAD（H） level is critical for L-phenylglycine synthesis. In this study，the co-expression of the genes pncB and nadE encoding of nicotinic acid phosphoribosyltransferase and NAD+ synthase separately was conducted to increase intracellular NAD（H） level so as to accelerate the whole-cell transformation. The results showed that the NAD（H） level in whole cell was increased by 2.35-fold when pncB and nadE were co-expressed，and it was further increased by 2.42-fold with the addition of 20 mg/L nicotinic acid（NA）. Significantly，the whole cell catalytic activity and L- phenylglycine yield were increased by 94% and 36.6%，respectively，indicating that the transformation efficiency can be effectively improved by the increase of intracellular coenzyme levels in the coenzyme- dependent whole cell transformation systems.

Abstract:To investigate the effects of hexane extract fraction 3（Fr.3） from Antrodia camphorate on hepatic stellate cells（HSCs） activation induced by TGF-β1，as well as mechanisms of its anti-fibrotic action in vitro. We established a cell model of liver fibrosis by using CFSC-8B stimulated by TGF-β1. Cell proliferation，cell migration，extracellular matrix（ECM） production and protein expression in the TGF-β1 signaling pathways were detected by the MTT，WST-1 assay，cell migration assay，qRT-PCR and western blotting analyses. The IC50 value of Fr.3 was 54.56 μg/mL and we found that less than 20 μg/mL of Fr.3 might be safe for CSFC-8B cells. Fr.3（< 20 μg/mL） significantly inhibited CFSC-8B cell proliferation and migration stimulated by TGF-β1. Fr.3 also downregulated α-SMA，Col1，Col3 and Fn mRNA expression. Mechanistically，Fr.3 blocked TGF-β1 induced protein expression of α-SMA，Col1，p-Smad2，p-ERK，p-P38 and p-AKT. Fr.3 from A. camphorata exhibits anti-hepatic fibrosis activity through negatively modulating TGF-β1/Smad/PI3K/MAPK signaling pathways in CFSC-8B cells.

Abstract:The process of lipid biosynthesis in oleaginous microorganisms is basically clear：both the reducing power NADPH and the building block acetyl-CoA are required by fatty acid synthase（FAS） to synthesize fatty acids. In oleaginous microorganisms，the main enzymes involved in the provision of reducing power NADPH are malic enzyme（ME），glucose-6-phosphate dehydrogenase（G6PDH），6-phosphogluconate dehydrogenase（6PGDH），and isocitrate dehydrogenase（ICDH）. In this study，the genes coding for 6-phosphogluconate dehydrogenase（gnd） from the oxidative pentose phosphate pathway（oxPPP） were over-expressed in the oleaginous bacteria Rhodococcus opacus PD630 to analyze its effect on lipid accumulation. The results showed that over-expression of gnd increased the content of lipid（w/w） by 20%~30% compared with the control strain. Meanwhile，the activities and the mRNA levels of 6PGDH in the overexpressing strains were higher than that of the control. Therefore，it was concluded that 6PGDH enhanced the lipid accumulation through promoting the generation of NADPH in the oleaginous bacteria R. opacus PD630.

Abstract:Static adsorption and desorption of characteristics of different macroporous adsorption resin was studied，LX-60 has a good effect to adsorb and desorb. The static adsorption and desorption of quantity was 5.12 mg/g and 4.86 mg/g. And adsorption and desorption of condition was investigated for LX-60，products of rosemary acid was got in the end，purity was 15.6% and yield was 86.79%. The purity of rosemary acid was 42 times higher than the purity of 0.37%. This study indicated it was feasible to purify rosemary acid in the lavender，as well as this research provided theory and experimental basis for production.

Abstract:In this study，we employed a fast and efficient approach，Global Transcription Machinery Engineering（gTME），to improve the butanol tolerance of Escherichia coli. The results show that one mutant E. coli JM109/pHACMB8 harboring rpoD mutant B8 could tolerate 2%（v/v） butanol. Further studies showed that the mutant B8 also exhibited high tolerance to other organic solvents such as 3%（v/v） isopropanol，8%（v/v） ethanol，4%（v/v） cyclohexane and 0.3%（v/v） toluene. To further reveal the tolerance mechanism of the mutant，we investigated the physiological characteristics of both /i> E. coli JM109/pHACMWT and the mutant B8. The results show that the intracellular butanol concentration of mutant B8 was 55% lower and has improved acid tolerance compared with WT. In addition，Mg2+ was proved to have a beneficial effect on the butanol tolerance of B8 mutant. Our study provides experimental data and theoretical basis for constructing OST bacteria for industrial applications.

Abstract:L-citrulline isan importantintermediate metabolite inurea cycle. The enzymes encoded by gene argGH catalyze thereaction of L-citrullineto L-arginineresults in difficulty of L-citrulline accumulation via microbial metabolism. In this study，promoter P-argG wasreplaced by a weak promoter P- dapAB6 to down-regulate argGH expression. The recombinant strain Corynebacterium crenatum H-7-P dapAB6： argGH showed lower argGH expression level，with ASS（argininosuccinate synthase，encoding by argG） and ASL（argininosuccinase，encoding by argH） activities reducing by 91.80% and 55.35%，respectively，compared with the parent strain. Via fermentation in shake flasks，the L-citrulline titer，yield and productivity of Corynebacterium crenatum H-7-P dapAB6：argGH were 33.85 g/L，0.25 g/g and 0.34 g/（L·h），respectively，which were 4.91，5.00 and 4.86-fold of those obtained from the parentstrain. These results indicatedthat，by promoter engineeringapproach，the preliminary construction of L-citrulline biosynthesis pathway was achieved.

Abstract: Lycium barbarum L. plays roles in anti-aging，anti-fatigue，anti-fatty and immunity improvement. We used alloxan-induced diabetic mouse model to investigate hypoglycemic activity of Lycium Barbarum L. water extract（LBWE）. Metformin hydrochloride solution（Met） was served as positive drug. 28-day LBWE treatment significantly reduced mouse blood glucose compared to model group. LBWE strongly improved alloxan-induced weight loss，abnormal symptoms of food and water intake. The hypoglycemic activity of LBWE was further confirmed by the inhibition of serum glucose in glucose tolerance test. Moreover，LBEW-treated mice showed low MDA and high SOD and GSH-Px activity compared to model group，suggested that LBWE prevented oxidative stress associated with diabetes complications. In conclusion，LBWE has a hypoglycemic effect by preventing oxidative damage.

Abstract:In order to increase the biomass and productivity of Nannochloropsis oculata，the effects of three kinds of nitrogen sources (ammonium chloride，ammonium acetate and sodium nitrate) on physiology-biochemistry and production of N.oculata were compared in this research paper. The total cellular protein content of ammonium groups (ammonium chloride and ammonium acetate) were higher than that of sodium nitrate group，while cellular chlorophyll a content showed the opposite result； the total cellular oil content of ammonium acetate group was the highest，being 13.5%； the total cellular carbohydrate content of nitrate sodium group was the highest，being 16.5%. The polar lipid to neutral lipid ratio and the relative content of EPA of sodium nitrate group were the highest，being 5.9 and 26.6% respectively. The microalgal cell size of the ammonium groups were larger than that of sodium nitrate group； the number，relative volume and thylakoid density of chloroplast of sodium nitrate group were higher than those of ammonium groups； only the cells of ammonium acetate group contained oil bodies. The results showed that the biomass and productivity of ammonium acetate group were the highest，being 873 mg/L and 25.3 mg/(L·d) respectively.

Abstract:In order to rapidly predict the dynamic growth of spoilage microorganisms in fresh-cut lettuce and make quantitative evaluation of the impact of temperature on the shelf life of fresh-cut lettuce，thus providing an effective method for the rapid and effective estimation of shelf life. Pseudomonas fulva，which caused the spoilage of fresh-cut lettuce was chosen as the study object in this paper. We studied the effects of temperature on the growth of Pseudomonas fulva in fresh-cut lettuce，established and validated the corresponding microbial growth kinetics model and remaining shelf life prediction model of fresh-cut lettuce stored at different temperatures．Experiment results showed that the Gompertz equation fitting correlation coefficient was beyond 0.99，the bias factor and accuracy factor of secondary model were in the range of 0.9~1.05. The research results indicated that the primary model and secondary model were valuable for the growth of Pseudomonas fulva in fresh-cut lettuce. The remaining shelf life model was validated by the measured values of fresh-cut lettuce at 7 ℃ and 15 ℃，the relative errors between predicted and real shelf life were -12.53% and 9.49% respectively，which showed that the established model can predict the remaining shelf-life of fresh-cut lettuce quickly and reliably.

Abstract:To investigate the ameliorative effects of extract fraction 8（Fr.8） from Cephalosporium sinensis on insulin resistance of HepG-2 cells，in this study，a stable insulin resistance cell model in vitro was established to detect the glucose consumption，glucose uptake，relative mRNA expression and protein expression of Fr.8 on HepG-2/IR cells.The results showed that 5 μmol/L insulin was the optimal concentration to induce HepG-2 cells become HepG-2/IR cells. In the range of non-cytotoxic，Fr.8 could distinctly promote glucose consumption and uptake in HepG-2 / IR cells.Fr.8（>25 μg/mL） significantly upregulated IR，IRS1，IRS2，Glut2，AMPKα mRNA expression and obviously increased IR，IRS，Glut2，p-AMPKα and p-AKT protein expression.Therefore，we provide theoretical basis to demonstrate Fr.8 from C. sinensis ameliorated insulin resistance of HepG-2 cells，which in part，the mechanism may be associated with insulin receptor and substrate，glucose transporter protein 2，the activated protein of p-AMPKα and p-AKT.

Abstract:To develop a chemiluminescent immunoassay method for detecting serum ferritin with biotin-avidin amplified system and a sandwich way. A pair of antibodies were labeled with biotin and horseradish peroxidase respectively. The 96 well microplate was coated with streptavidin. The serum ferritin standard and international standard was evaluated. 300 clinical serum specimens wer detected with this method and a foreign serum ferritin kit（Roche），the correlation coefficient was 0.97. The linear measurement range of this method was 10~800 ng/mL. The intra-assay and inter-assay coefficients of variation were 3.4~5.5%， and 5.0~7.3%. The cross-reactivity rate with CEA（carcinoembryonic antigen），human albumin，human heart ferritin，and hemoglobin were all less than 1%. The developed method was stable 9 days at 37 ℃. This assay was easily operable，small amount of specimens needed and the reaction was quick，so it was suitable for clinical application.

Abstract:Levansucrase from Bacillus amyloliquefaciens was cloned and heterologously expressed in Escherichia coli BL21（DE3）. After purification，the enzymatic characterization of levansucrase was determined and analyzed. The optimum temperature of hydrolytic activity was 45 ℃ and the maximal transfructosylation activity was measured at 40 ℃. The optimum pH for hydrolytic and transfructosylation activities were 6.5 and 6.0，respectively. The Km and Vmax values for levansucrase were calculated to be 150.74 mmol/L sucrose and 21.23 μmol/（mL·min），respectively. Levansucrase-catalyzed reaction was investigated at pH 6.0 and 40 ℃. The result indicated that the content of levan was up to 34.05% in total products after 12 h.

Abstract:To explore basic information for ethanol production from sugarcane molasses in fed-batch fermentation，yeast cell vitality of the secondary fermentation was investigated with aim at determining optimal feeding time based on the cell vitality. By batch fermentation at different initial sugar，optimal initial sugar concentration（220 g/L） for fed-batch fermentation was obtained. The secondary fermentations were done by inoculating the same amount of cells from the primary fermentation at 0，6，12，18，24，30，36 h，with emphasis on analyzing cell activity，sugar utilization and ethanol production for determining time point of highest cell vitality in the primary fermentation. By ethanol fed-batch fermentation with feeding timing based on cell vitality，the results showed that the optimum fermentation parameters were observed with feeding at 24 h when the cell vitality was the strongest，namely，the highest ethanol concentration（136.62±1.10） g/L，volumetric productivity（2.53±0.02） g/（L·h） and total sugar fermentation efficiency（83.34%） were achieved，indicating that the cell vitality of yeast based on secondary fermentation can be used as a parameter of feeding time in the ethanol fed-batch fermentation.

Abstract:Armillaria mellea is a well-known edible and medicinal mushroom，Whereas little information is available in regard to A. mellea's anti-inflammatory activity. In this study we explored the anti-inflammatory activity of n-hexane，chloroform，ethyl acetate and n-butyl alcohol extracts from Armillaria mellea. Our work will provide a scientific basis for screening new natural anti-inflammatory substances and a new medical application of Armillaria mellea. MTT colorimetric assay was applied to measure the cytotoxic effects of different extracts. Their anti-inflammatory activities were evaluated via inhibition against production of lipopolysaccharide（LPS）-induced nitric oxide（NO），tumor necrosis factor alpha（TNF-α），interleukin-6（IL-6） and interleukin-1beta（IL-1β） in murine macrophage-like cell line RAW264.7 cells. Cytotoxic assay showed that only n-hexane and chloroform extracts in a high concentration（300 μg/mL） have slight cytotoxic effects（inhibited 10.7% & 17% of cell viability） in RAW264.7 cells. As to the anti-inflammatory activity，except to the n-butyl alcohol extract，the other three extracts all can dose-dependently inhibited LPS-induced NO，TNF-α，IL-6 and IL-1β production. Among them，the ethyl acetate extract own both safty and anti-inflammatory activity.

Abstract:Botryosphaeria spp. are important pathogens on a variety of fruit trees causing cankers and gummosis on almond trunks，and resulting in a significant decline in the health development of the planting industry. The strain H47（producing catechol-type siderophore） and the strain H114（producing hydroxamate-type siderophore） were isolated from the honey. The two strains identified as Bacillus amyloliquefaciens by 16S rRNA gene analysis and physiology biochemistry experiment. The strains were cultured with the MM9 medium，and the cell-free supernatants were extracted by organic solvent. The organic phases collected as partially purified siderophore were dried in a vacuum oven. The partially purified siderophore was inoculated along with the B. dothidea Ces. & De Not ACCC 38026 and the inhibition of the mycelial growth was test by vernier caliper. The results indicated that both catechol-type and hydroxamate-type siderophores had antifungal activity to pathogen，toxicity regression equations were y=1.432x+0.578 and y=1.744x+0.063，respectively. Thus these honey isolates had the potential as the bio-control agents，which were useful to inhibit the growth of B. dothidea.

Abstract:Fomitopsis pinicola is a medicinal mushroom and a kind of new resource of food to explore. Two water-soluble crude polysaccharides were extracted using hot water and 5% of NaoH solution.，and then purified FPS by DEAE-Sepharose FF and Sepharose Cl-6B chromatography. Four polysaccharide factions named as FPS1-1，FPS1-2，FPS2-1 and FPS2-2 were isolated and purified from FPS. The result of the MTT test about normal mice spleen lymphocyte proliferation promotion activity indicated that the four polysaccharide factions both have a immunological activity. The effect of FPS1-2 and FPS2-1 are in a dose-dependent manner，and the effect of FPS2-1 was better than others. The HPSEC analysis indicated that the two fractions were all homogeneous polysaccharides and their relative molecular mass were respectively 9.10×103 and 3.02×105. The analysis of monosaccharide composition，infrared spectrum and NMR spectroscopy indicated that FPS1-2 was a heteropolysaccharide，it have α-（1→6）glucosidic bond；and FPS2-1 was a heteropolysaccharide ，it have α-（1→4）glucosidic bond.