Abstract:To explore the effects of tyrosine oxidation products（TOP） on the spatial learning and memory of rats and hippocampus. 40 SD rats are divided into four groups：Control（normal diet），Tyr group（add 0.44%Tyr），TOP1group（add 0.22% tyrosine oxidation products） and TOP2 group（add 0.44% tyrosine oxidation products）. After 24-week feeding，Morris water maze is used to test the spatial learning and memory of rats. Protein and lipid oxidation products in hippocampus are measured. Expressions of antioxidation- and inflammation- related genes are detected using RT-PCR. The results show that，after 24-week feeding，compared with Control and Tyr group mice，TOP1 and TOP2 group rats showed impaired spatial learning and memory in Morris water maze；hippocampal MDA，4-HNE，Dityr，3-NT and Aβ40 are significantly increased（p<0.05）；expressions of Nrf2、NF-κB and iNOS were significantly up-regulated（p<0.05）. Thus，tyrosine oxidation products induce oxidative damage to rats hippocampus，accumulation of lipid and protein oxidation products in hippocampus，and impair the spatial learning and memory of rats.

Abstract:This study aims to inactivate Bacillus subtilis in Polyethylene Terephthalate（PET） preforms by hydrogen peroxide vapour. Based on single factor experiment combined with Box-Behnken central combination design，sterilization processes，such as H2O2 flowrate，treatment and temperature，are optimized by response surface methodology. The optimum process parameters are：H2O2 flowrate= 14 mL/min，treatment time= 9 s，and temperature= 120 ℃.Under the optimized conditions，the reduction of Bacillus subtilis could be up to 5.40 log. It provides theoretical basis for the industrial application of hydrogen peroxide vapour sterilization.

Abstract:The crude trehalose synthase is immobilized on polyacrylonitrile membrane by adsorbtion. The optimal conditions of immobilization is research and the properties of immobilized enzyme by single factor method is analyzied. The results show that the optimum immobilization conditions are：pH 7.6，30 ℃ for 3 h with 0.1 mg trehalose synthase per square centimeter PAN membrane. The optimal reaction conditions of the immobilized trehalose synthase are 7.4，40 ℃ and initial substrate concentration 200 g/L. Compared with the free trehalose synthase，the thermal stability increases by 10 ℃，pH stability expande from pH 6.6 ~ 7.4 to pH 5.4 ~ 8.0. The max trehalose content is 50.9% when the reaction reached equilibrium，the by-product of glucose content is 9.1%，which reduced by 6 percentage points compared with the free enzyme and is the lowest byproduct content of the reported immobilization trehalose synthase. Under the optimum reaction conditions of pH 7.4，40 ℃，substrate concentration 200 g/L，the immobilized enzyme remains 64.4% of the initial enzyme activity after repeating used for 72 h.

Abstract:In order to achieve the high production of 2-keto-D-gluconic acid（2-KGA） by Serratia sp. FMME043，feeding strategy and fermentation conditions are optimized in 30 L fermentor，and fed batch fermentation is established. The optimal fermentation conditions are as follows：the dissolved oxygen is 40%，the quality of 64，80 g，and 96 g ammonium sulfate is added at 16，22 h，and 28 h，respectively，and the quality of 801 g glucose is supplemented with five times at the residual sugar concentration between 15~25 g/L. Under these conditions，at 44 h，the production and the yield of 2-KGA reached 268.5 g/L and 1.04 g/g，which are 58.4% and 9.9% higher than that not optimized，respectively.

Abstract:Effect of the N-terminus of a family 11 xylanase EvXyn11TS on its thermostability is systematically analyzed. Thirty xylanases that had the sequence homologies more than 55% aligned with EvXyn11TS and the known temperature characteristics are sought out from the GenBank database using the BLAST server. Then，the multiple alignment of 31 xylanase sequences is performed using the ClustalW2 program，and the phylogenetic tree is constructed using a MEGA 5.1 software. Eight representative thermophilic and mesophilic xylanases（including EvXyn11TS） are selected from the phylogenetic tree. Finally，the differences and similarities at the N-termini between EvXyn11TS and the other xylanases are analyzed using biology softwares. The analytical results indicate that the contents of amino acids favorable to thermostability，the β-strand A1 and the interactions between amino acids at the N-terminus of EvXyn11TS have certain contributions to its thermostability.

Abstract:In this study，codon usage optimization and co-expression of molecular chaperone are used to improve the extracellular production of alkaline polygalacturonate lyase（PGL） in Pichia pastoris. According to the codon bias of P. pastoris，the gene of the stabilized mutant（K314M） of Bacillus sp. WSHB04-02 PGL is optimized and cloned into the EcoR I-Not I sites of pPIC9K-PGL，yielding the expression plasmid pPIC9K-PGL. pPIC9K-PGL is transformed into the P. pastoris GS115（his-），yielding the strain GS115/PGL 14#. GS115/PGL 14# produced 301.32 U/mL of extracellular PGL which is 25.1% higher than that of the strain with native PGL gene. On this basis，two molecular chaperones the protein folding factors endoplasmic reticulum oxidoreduction 1（ERO1） and ubiquitin-conjugating enzyme（UBC1） are co-expressed with the optimized K314M gene individually and simultaneously. It is showed that co-expression of ERO1 and UBC1 at the same time（GS115/PGLl-ERO1-UBC1 2#） increased the extracellular PGL by 49.3%，which is up to 450.12 U/mL. By using an exponential fed-batch strategy in 3L-fermenter，GS115/PGLl-ERO1- UBC1 2# produces 1 362.31 U/mL after the 96 h induction by methanol. The results here may benefit the industrial production and application of the PGL.

Abstract:To understand the characteristics of fungal community structure in the lesion tissues of Kuerle fragrant pear "Blackhead Disease", the fungal community structure of 12 samples from 4 different fresh-keeping storage in Kuerle area is studied using PCR-DGGE technique. Biodiversity is analyzed by DGGE fingerprint. The results indicat that there is some differencs in the community structure of all samples，which may be related to the planting environment. The 9th stripe in different samples has the highest dominance in the whole the lesion process. The representative strains can be considered as the main pathogenic bacteria in Korla pear "blackhead disease". According to the results of sequence analysis of DGGE dominant bands，it is phylogentically close to Alternaria. It preliminary reveals the variation regular of microbial communities in Korla fragrant pear diseased tissues，which provides a theoretical basis for studies of "Blackhead Disease" in Korla fragrant pear.

Abstract:The poly（solketal methacrylate）-block-poly（glycidyl methacrylate）（PSA-b-PGMA） copolymer is synthesized by atom transfer radical polymerization（ATRP） and functionalized with mannose by the "click" reaction to yield PSA-b-PGMA-Mannose. These copolymers are used to prepare polymeric micelles，and an anticancer drug，doxorubicin（DOX），is encapsulated into the micelles by oil-water method. Structure of PSA-b-PGMA-Mannose is confirmed by 1H NMR and FT-IR. The size and morphologie of PSA-b-PGMA-Mannose with different block ratios were characterized by transmission electron microscopy and dynamic light scattering. The in vitro cytotoxicity and cellular uptake against MDA-MB-231 cancer cells were evaluated by confocal laser scanning microscopy and MTT assay. The results show a decrease in the average size of micelles upon an increase in the hydrophilic/hydrophobic block ratio. The DOX-loaded micelles are efficiently trapped by MDA-MB-231 cancer cells via receptor-mediated endocytosis and releass loaded DOX into the cells.

Abstract:Silk fibroin contains 10% of tyrosine residues，however，the reactivity of fibroin is limited owing to the restricted accessibility. In this work，grafting of a tyrosine-containing polypeptide is carried out using a natural crosslinking agent of Genipin(GP)，aiming at promoting the tyrosinase-catalyzed reaction between fibroin and anime compounds. The changes in molecular weight，structure and performance are investigated by means of SDS-PAGE，FTIR and TG，are the efficiency of enzymatic grafting of ε-Poly-L-Lysine （ε-PL） is also concerned. The results indicate that the molecular weight for the fibroin proteins increas after GP treatment；the thermo stability and mechanical property for the fibroin-based membranes are also improved compared to the control. Based on methyl orange colorimetry，it is also verified that grafting of polypeptide could promote the tyrosinase-catalyzed coupling of ε-PL. It provides an efficient way to improve reactivity of fibroin via grafting of tyrosine-containing using GP.

Abstract:Cider Beer is a kind of special beer fermented with apple juice. Through fermentation experiment and detection of fermentation degree，alcohol content，the sample 7 is selected as the raw material of cider beer. By comparing the growth situation of different yeasts under the conditions of poor nitrogen and the fermentation experiment results，the yeast C is selected as the best suitable yeast of cider beer. Using sensory quality of product as evaluation index，based on the orthogonal test，the optimal brewing technology conditions are：fermentation temperature 11 ℃，fermentation time 7 d and yeast inoculation quantity 8×106/mL. The product has fruit aroma and a unique taste style and its total sugar content is 13.28 g/L，the total acid content is 6.5 g/L，the alcohol content is 2.85%，the free amino acid content is 3.60 g/L，the polyphenol content is 1.04 g/L.

Abstract:With the purpose of elucidating the introduction of cytoplasmic reductive pathway in Saccharomyces cerevisiae on the production of L-malic acid， the reductive pathway for L-malic acid biosynthesis is constructed in S. cerevisiae by overexpressing genes Afpyc，Afmdh and Afmae from Aspergillus flavus. When gene Afpyc is kept in low level，the pyruvate titers of W003 are decreased 42% compared with control strain W001，respectively. To further introduced gene Afmdh in strain W003，the concentration of L-malic acid in W005 is up to 1.93 g/L，the DCW is increased by 350% and the pyruvate titers is decreased by 65.9% than that of the strain W004. Moreover，the malate efflux system is enhanced by introducting the tansporter gene Afmae and the L-malic acid titers of strain W006 is increased by 21.2%（2.34 g/L）. The concentrations of L-malic acid is increased from 2.34 g/L to 3.28 g/L with the initial OD600=2. It could be summarized that introducting the reductive pathway of overproducing L-malic acid strain A. flavus in S. cerevisiae is indeed producted L-malic acid，and this successful method provides a strategy to the metabolic engineering.

Abstract:The β-1，4-endoglucanase from Aspergillu sniger（AnCel5A） is a cellulase which is classified to glycoside hydrolases subfamily （GH） 5. It can hydrolyze the amorphous region of cellulose long chains by endo-action，which makes it one of the key enzymes in cellulose hydrolysis.This research aims at determination the crystal structure of AnCel5A by screening and optimization its protein crystals. The signal peptide truncated AnCel5A is successfully expressed in P. pastoris X33. Purified proteins are obtained by using ion-exchange and hydrophobic chromatography. The crystal screen of AnCel5A is completed by using sitting-drop vapor diffusion method and screen kits bought from the Hampton Research company. Initial crystals are found in the reservoir solution of cryoI kit （A6）. After further optimization，better crystals are obtained. The crystals diffracted to 1.8 ? and dataset is collected at beam line BL15A1 of the National Synchrotron Radiation Research Center （NSRRC，Hsinchu，Taiwan）.

Abstract:Botulinum toxin（BoNT） is a neurotoxin produced by Clostridium botulinum，which is one of the most powerful toxin. To study the activity and function of BoNT/A in yeast，BoNT/A LC in yeast is expressed，observing the yeast growth and BoNT/A protein expressing level by western blot. Expression of LC of BoNT cerotype A caused a growth defect in yeast cells，and the toxicity is due to the hydrolytic activity. It is because catalytically inactive BoNT/A LC does not cause a growth defect in yeast cells. Overexpression of a yeast SNAP-25 homologue gene，SEC9，could rescue the growth defect in BoNT/A LC expressing cells. The results demonstrate that，in BoNT/A LC expressing yeast cells，the hydrolytic activity can be monitored through their growth. Thus，such cells can be used to analyze BoNT/A.

Abstract:The purpose of this study is to screen high protease production strain from the squid，with generation protease hydrolysis circle and protease activity determination method. The typical strains are identified by 16S rRNA sequencing，and subject to phylogenetic analysis. Meanwhile，the properties of crude protease enzyme are researched. The results show that after screening 10 strains had a high yield protease activity，identified belong to Bacillus sp.，Providencia sp.，Pseudomonas sp.，Among them SW5 strains produced highest enzyme activity up to 257.67±2.44 U/mL，identified as Bacillus methylotrophicus. Investigation on the crude enzymology characteristics further indicate that the enzyme exhibites the highest activity at pH 8.0 and the optimum temperature is 40 ℃． The protease activity is greatly inhibited by Mn2+、Ba2+ and Ca2+，but is not inhibited by Fe2+ and Zn2+ at the final ion concentration 1 mmol/L.

Abstract:The "Nanshu 88" polysaccharide（N88P-1） is obtained from the dried stems and leaves of "Nanshu 88" by hot water extraction，ethanol precipitation，and then purified by DEAE-cellulose 52 column chromatography. The structure of "Nanshu 88" pure polysaccharide（N88P-1） is identification by Infrared spectroscopy（IR），High Performance Gel Penetration Chromatography（HPGPC），High Performance Liquid Phase Chromatographic（HPLC） and nuclear magnetic resonance spectra（1H NMR and 13C NMR）. At the same time，the antioxidant and immune regulation effects of N88P-1 are also carries out. The results show that N88P-1 has molecular weight of 14 328 D. The infrared spectra shows the characteristic peaks of O-H stretching vibration appears in 3 444.81 cm-1，and the characteristic peaks of C=O stretching vibration appears in 1 640.20 cm-1. 1H NMR spectra showed anomeric hydrogen signals at δ4.93~δ4.86. High performance liquid chromatography（HPLC） chromatography shows N88P-1 is composed of galactose and mannose with the ratio of 1∶1. The activity study shows the N88P-1 has ability to clear the ABTS + free radical in a certain range of concentration，and the IC50 value is 0.48 mg/mL. The DPPH - free radical scavenging activity of N88P-1 is positively correlated with concentration in the range of 0.25~3 mg/mL，the IC50 value is 0.96 mg/mL. When the concentration of N88P-1 is 3 mg/mL，the scavenging activity of ABTS + and DPPH - free radical both reach the highest value. The proliferation activity of the immune cells results show that the proliferation effect on B cells and T cells are very significant（P<0.01） in the concentration of 20~80 mg/mL；while the effect of macrophage proliferation was very significant（P<0.01） in the concentration of 10~40 mg/mL；It is worth noting that the proliferation effect of the three kinds of immune cells all reached the highest value in the concentration of 40 mg/mL.

Abstract:The present study aims at using error Back Propagation（BP） neural network and genetic algorithms to improve lycopene production from semisynthetic medium by Blakeslea trispora. The effects of different kinds of carbon source，nitrogen source and vegetable oil on lycopene concentration and biomass are analyzed to confirm the component of medium. The establishment of BP neural network model is based on 49 group of samples data. In this model，Corn flour，corn steep liquor，soybean oil，monopotassium phosphate，magnesium sulfate are set to inputs，and lycopene volumetric production as output. Then，the genetic algorithms is applied to search the optimal value using BP network model as fitness function. After optimization，the maximum predicted value of lycopene production is 1.27 g/L. And the error of predicted value and actual value is less than 5% by experimental verification. Lycopene production in optimized medium is 31.6% higher than that in initial media. The optimized medium containes 41.2 g/L corn starch，8.93 g/L corn steep liquor，26.5 g/L soybean oil，1.39 g/L KH2PO4，0.46 g/L MgSO4. The combination of BP neural network with genetic algorithms is a power tool to obtain optimized lycopene fermentation medium.

Abstract:The effects of Cyclocarya paliurus（Batal.） Iljinskaja Extracts on glucose consumption of insulin resistance HepG2 cell and activity of alpha-glucosidase is studied. HepG2 cell with high concentration insulin（10 μg/mL） is induced to establish insulin resistance cell model and divided model cell into Cyclocarya paliurus polysaccharide（CPP），Cyclocarya paliurus flavone（CPF） and dimethylbiguanide groups. After 24 h administration，glucose assay is used fit to measure the gross glucose consumption of cells（ΔGC）， glucose consumption of unite cell（ΔGC/OD） is detected by MTT assay. The inhibitory activity on alpha-glucosidase of CPP and CPF is evaluated，compared with acarbose，in vitro. Compared with model group，ΔGC and ?驻GC/OD of IR-HepG2 cell is increased by CPF and CPP significantly. CPP and CPF have inhibitory effect on the activity of alpha-glucosidase. Those results indicat that CPF and CPP have effects on increasing the glucose consumption of cell and decreasing the activity of alpha-glucosidase，thus alleviating hyperglycemia.

Abstract:Four bacterial strains with antibacteria activity are isolated from Jianmen harbour seafloor mud in Taizhou of Zhejiang Province，China. Thus，further studying on one of the stronger antibacteria activity strain is undertaken. This strain belongs to the genus Bacillus amyloliquefaciens，named as Bacillus amyloliquefaciens MBRC1 through molecular biology，（G+C） mol% content，colony and morphological identification. The separation position also has been identified by the analysis on phylogenetic tree with such softwares：BLAST Clustal W（V 1.83） and Mega（V 5.1）. Fermented Bacillus amyloliquefaciens MBRC1 with different media and then investigates the effect of antimicrobial in extra using the method of oxford cup. The results show that Bacillus amyloliquefaciens MBRC1 could generate better antimicrobial substances and has good antimicrobial effect on S. Aureus、B. Subtilis and E. coli.

Abstract:ACE inhibitory peptides are prepared by enzymatic hydrolysis of protein of marine red algae Gracialriopsis lemaneiformis. The hydrolytic activities of four kinds of enzymes of flavourzyme，alkaline protease，trypsin and papain are investigated，and the inhibitory activity of their enzymatic hydrolysates on ACE，and optimization of the digestion process are studied in the paper. Trypsin is considered as the suitable enzyme to prepare ACE inhibitory peptides. Response surface experiments are used to optimize the enzymolysis technology of trypsin based on the results of single factor experiments. Results show that the optimum condition is pH at 8.0，enzyme dosage at 6%，reaction temperature at 35 ℃. Peptides （2.0 mg/mL） prepared under the optimal condition shows a ACE inhibitory activity of 72.37%.

Abstract:A novel external fibrous-bed bioreactor with rice bran as cell immobilized material for butyric acid production is established in this study. Using Clostridium tyrobutyricum as the fermentation strain，free-cell and immobilized-cell fermentation for butyric acid production are conducted. The results show that the productivity of immobilized-cell batch fermentation reaches 0.48 g/L/h，which is 55% higher than that of free cells. Furthermore，21.05 g/L butyrate is achieved by immobilized cells in the repeated batch fermentation and butyrate production is stable by using this system. Finally，the simulation experiments about adsorption of sodium butyrate by rice bran are conducted，and the ratio of equilibrium adsorption is about 0.084 g/g. The sodium butyrate in the fermentation broth could be adsorbed by the rice bran and the ratio of adsorption is 0.105 g/g. The rice bran in this study can be directly used as feed additive.

Abstract:BBS2 is a BBSome subunit and its loss causes Bardet-Biedl syndrome. Chlamydomonas reinhardtii is a model organism used for studying the Bardet-Biedl syndrome. To investigate the role of BBS2 in causing Bardet-Biedl syndrome，a prokaryotic expression vector，pET28a-bbs2 is constructed by inserting the 5' 399 bp fragment of the bbs2 gene into the pET28a plasmid. The recombinant vector is then transformed into Escherichia coli BL21（DE3） to induce the 6×His-BBS2 fusion protein expression. SDS- PAGE electrophoresis shows that this fusion protein is insoluble and expressed with a molecular weight of approximately 15 kDa as expected. In the presence of 8 mol/L urine，the fusion protein is purified by Ni-affinity purification and used to immunize New Zealand white rabbits. After the 5th immunization，the antisera are determined to have a titer of as high as 256 000. The antisera are then purified using protein A sepharose CL-4B. The polyclonal antibody prepared can specifically bind the C. reinhardtii BBS2 as shown in Western blotting assay and identified BBS2 in the basal bodies and in the flagellum as determined by immunofluorescent microscopy. The availability of this antibody will strengthen the research to investigate the role of BBS2 in causing Bardet-Biedl syndrome.

Abstract:The lactobacilli which has high cholesterol uptake and utilization rate are isolated after screening. Their cholesterol-reducing mechanism is detected in vitro，to provide a theoretical basis for cholesterol-reducing fermented food research and development. The probiotic strains with cholesterol-reducing activity are first obtained by screening. Then the saponification-colorimetric screening is performed to obtain lactobacillus strains with high cholesterol absorption efficiency. The lactobacilli has cholesterol-reducing rates ranged from（3.30±0.89）%~（75.64±1.12）%，with the average rate of 44.34%. Among them，14 strains have higher cholesterol-reducing rate than average. Lb. plantarum S11 is the strain with the cholesterol-reducing rate as high as（75.64±1.12）%. It is indicated that only 0.002 mg of cholesterol is adsorbed and precipitated by 4.20×109 cfu/mL of Lb. plantarum S11 cells by determine the content of cholesterol in the suspension of the disrupted cells（only 1.30%）. This results indicate that the cholesterol-reducing activity of Lb. plantarum S11 is achieved mainly by absorbing and assimilation，but not absorbing and incorporating in cell membrane.The screening and isolation methods in this study can be applied to select cholesterol-reducing probiotics from traditional fermented foods. And it is also helpful for follow-up development of new cholesterol-reducing lactobacilli fermented products.