Abstract:Blueberry anthocyanins are the major active ingredients of blueberry with a variety of biological activities and widely used in the fields of food，medicine，cosmetics，and so on. The deep investigation of their extraction，separation，and purification has been made in recent years and this article reviewed the advance. The research status of blueberry anthocyanins was summerized and prospected. This work would provide reference for further application and industrial production of blueberry anthocyanins.

Abstract:The ubiquitination mechanism of key regulator Stp1p was investigated in Saccharomyces cerevisiae. The ubiquitination detection vector was constructed based on the BiFC（Bimolecular fluorescence complementation） technology to assay the ubiquitination process of Stp1p. The site-directed mutagenesis on the potential ubiquitination sites were performed to verify the effect on its ubiquitination regulation and the nitrogen utilization. The fluorescence levels of mutant strains were down-regulated compared to the wild type strain. Furthermore，the triple mutant Stp1K49，129，113R achieved the lowest level among the all of the strains. The results indicated that the process of ubiquitination was significantly repressed by removing the potential ubiquitination residues. In addition，the amino acid utilization assay implied the ubiquitination sites played a vital role on its ubiquitination process. The results presented here demonstrated that the ubiquitination process was invovled in the regulation of Stp1p by site-directed mutagenesis of potential ubiquitination sites and thus impact the nitrogen utilization process.

Abstract:In order to achieve efficient production of the trehalose synthase from Thermus thermophilus，the optimization of fermentation conditions was investigated in E.coli BL21（DE3） harboring the plasmid pET24a（+）-TreS. The optimized fermentation conditions were as follows：the inducing temperature is 32 ℃，the adding rate of lactose is 0.2 g/（L·h） When OD600 reached 50. Under the condition，the enzyme activity reached 472 U/mL at cultivation of 35 h，was 2.3 times than before. In addition，the fermentation was performed in 30 L bioreactor under the condition of preceding narrative，the enzyme activity reached 356 U/mL. Because the thermal stability of trehalose synthase is good， E.coli cell was broken up through high temperature. The results showed that treated it 30 min under 80 ℃ the yield of trehalose synthase reached 72.3％. It have industrial application prospect.

Abstract:L-valine is one of the branched chain amino acids widely used in food，feed，and drug industries. In this study，the global regulator Lrp was found to improve L-valine production in C. glutamicum，but slightly inhibit the growth. RT-PCR analysis showed that expression of lrp increased the transcriptional levels of genes related to L-valine biosynthesis （ilvA，ilvBN，ilvC，ilvD） and export （brnFE），which might be the reason for the improved L-valine production. A single amino acid substitution （Arg39Trp） in Lrp reduced the growth inhibition and led to higher L-valine production，especially in L-valine production strain C. glutamicum VWB-1 which contains mutations in the region between lrp and brnF. By co-expressing lrp1 and brnFE in VWB-1，L-valine production of fed-batch fermentation reached 42.1 g/L which was 48.9% increase.

Abstract:In order to lay a foundation for the development of new targets for typing and tracing，we investigated the characteristics of the variations based on the analysis of 16S rRNA gene sequences of Vibrio parahaemolyticus.We sequenced 16S rRNA gene of 111 strains of Vibrio parahaemolyticus which were isolated from fresh aquatic products，and aligned with standard sequence in GenBank. We also established phylogenetic tree of sequences which had a great variation，and verified the variation regularity of the genes by ERIC-PCR.The variability of different Vibrio parahaemolyticus of 16S rRNA genes was discovered. The variation mainly centralized in variable region V1 and V2. There were 38 base sites which similarity were less than 30% in the region of 50~270 bp，and 103 base sites which similarity ranged from 30% to 100%. In addition，there was one base which showed difference both at V3 and V8. The variability of different Vibrio parahaemolyticus of 16S rRNA genes can lay a foundation for the development of new targets for typing and tracing of Vibrio parahaemolyticus and for the study of micro-evolution.

Abstract:A codon-optimized gene（named xyn11EM），which encodes a thermotolerant xylanase belonging to the glycoside hydrolase family 11，was synthesized，and cloned into the expression plasmid pET-28a（+）. The recombinant Escherichia coli（designated E. coli BL21/xyn11EM），expressing the thermostable xylanase，was constructed by transforming the recombinant plasmid，pET-28a-xyn11EM，into E. coli BL21（DE3）. The recombinant E. coli BL21/xyn11EM was induced with IPTG to express reXyn11EM. The reXyn11EM activity reached 47.5 U/mL. The apparent molecular weight of reXyn11EM was estimated to be 24 800 by SDS-PAGE analysis. Its optimal temperature and pH were 70 ℃ and 7.0，respectively. It was stable at a temperature of 70 ℃ or below，and at a pH range of 5.0~8.0. Its activity was not significantly affected by most of metal ions tested and EDTA. The Km and Vmax of reXyn11EM toward birchwood xylan were 7.2 mg/mL and 54.7 U/mg. The excellent thermostability of reXyn11EM make it has great potential in industrial application.

Abstract:The antibiotic resistance and resistant genes of lactic acid bacteria （LABs） were investigated on the fresh hot pepper used to fermentation. In current study，218 isolates suggested that about 2.18×104 CFU/g LABs were on the fresh hot pepper used to fermentation. These isolates were assigned to Weissella cibaria（61.93%），Lactococcus lactis（16.06%），Leuconostoc mesenteroides （6.88%），Enterococcus mundtii（5.5%），Enterococcus faecalis（3.67%），Enterococcus hirae （3.67%） and Leuconostoc holzapfelii（2.29%）. Of the 218 isolates，no one displayed the resistance of erythromycin（ERY），but 30 isolates （13.76%） were found to be against streptomycin （STR） and tetracycline（TET） with one or two resistance，including 10 W. cibaria（4.59%），2 Lac. lactis （0.92%），1 E. mundtii（0.46%），2 Leu. mesenteroides（0.92%） and 1 Leu. holzapfelii（0.46%） with resistance to STR，And 9 W. cibaria（4.12%），2 Lac. lactis（0.92%），1 E. faecalis（0.46%）and 2 E. hirae（0.92%） with resistance of TET and STR. In the TET and STR resistant strain W. cibaria CT024，none of TET resistant genes was found. While，the other TET and STR resistant strains had harbored one or more corresponding resistant genes. In 5 STR resistant genes had the highest detection rate with 60.00%，but aad6 with only 16.67%. Among 11 TET resistant genes，tetB and tetC had the highest detection rate，85.71%，and tetZ was found with 7.14% lowest detection rate. For the current antibiotic resistant strains，the STR or TET resistant phenotype of different strains in the same species was not directly related to the types or quantities of the corresponding antibiotic resistant genes.

Abstract:Phenyllactic acid was one of the most important compounds with wide application in pharmaceuticals and biological preservatives. Five ketoacid reductases capable of asymmetric reduction of phenylpyruvic acid（PPA） into phenyllactic acid（PLA） were obtained from genome databases，and were heterogeneously over-expressed in E. coli BL21（DE3）. Among 5 enzymes，the recombinant ketoacid reductase from Lactobacillus casei（LcKAR） displayed the best biocatalytic performance，with the highest specific activity of 0.32 U/mg of purified enzyme and enantioselectivity of >99%. This LcKAR obeys Prelog rule in the asymmetric reduction of PPA into（R）-PLA. The fermentation medium composition and culture conditions of recombinant E.coli BL21（DE3）/pET28a-LcKAR were optimized to be seed age of 6 h，3% inoculation，induced with 0.6 mmol/L IPTG at 30 ℃ for 5 h. Under above optimal conditions，the LcKAR production could reach 2.17 kU/L in a 3 L bioreactor.

Abstract:A sensitive and rapid monoclonal antibody-based sandwich enzyme-linked immunosorbent assay（ELISA） was established to detect Pantoea stewartill subsp.Stewartii（Pss）. Three murine hybridomas（12B4/10B1/11C2） were obtained from the spleen cells fused produce. A pair of monoclonal antibodies（mAb） was selected for the sandwich ELISA method. The mAb 10B1 was conjugated with horseradish peroxidase（HRP） as the detection antibody and the mAb 11C2 was used as the capture antibody. The limit of detection of this method was 1.5×104 cfu/mL，and the linear range from 4.57×105 to 1.11×108 cfu/mL.

Abstract:This article aimed to optimize protoplast formation conditions of industrial citrate-producing Aspergillus niger to establish genetic transformation system. As protoplasts were formed by enzyme digestion，components of enzyme mixtures，digestion conditions and hyphae growth conditions were studied. The optimized enzyme mixture contained 5 mg/mL lysing enzyme，0.2 U/mL chitinase and 460 U/mL glucuronidase. The best conditions for protoplasting were using 0.7 M KCl as osmotic stabilizer，thickness of pellets hyphae for 50 ?滋m，cell weight of 0.003 g/mL. Furthermore，addition of Mn2+ in culture medium helped reduce the chitinase concentration necessary for protoplasting. In addition，glucuronidase in culture medium facilitated conidia separation during germination，further improved protoplast formation. Finally，two cassettes were inserted into genome simultaneously.

Abstract:To improve the ethyl caproate in daqu. A hyper ethyl caproate esterifying enzyme producer-SXG-Z-6，was isolated from Fen-Daqu. The analysis of morphological and molecular characteristics showed that this wild-type strain was closest to Aspergillus niger. The culture media and conditions were optimized using esterifying power as the reference index. As a result，the optimal culture conditions were summed up as follows：10 g wheat bran as culture substrate，1.5 g corn flour，2 g yeast extract，1 %（v/v）soybean oil，0.2 g（NH4）2SO4 and 10 mL H2O，at 36 ℃ for 44 h. The highest esterifying power could reach 615.8 mg/dL，which was improved by 35.7% than the control.

Abstract:Cell lysis is a process problem in the protein expression using bacterial host. Under the same fermentation condition，the differences in the expression of foreign protein and cell activity between engineered and wild-type Escherichia coli W3110 expressed by the humanized single antibody fragments （ScFv） were studied. The results showed that high expression and accumulation of heterologous protein in the engineered E. coli cells were found，most living cells entered viable but nonculturable（VBNC） state 6 h in advance and the occurrence of cell lysis compared with wild-type. Thought the analysis of transcription levels of both E. coli c，we found that the process of ScFv expression causing overshoot phenomenon of multiple stressed responses pathway genes included rpoH，dnaK，dnaJ，groES，groEL in heat stress response pathway，rpoS，gadE，gadX in acid stress response pathway，sodA，katE in oxygen stress response patheway. However，during cell lysis process in the fermentation tank，the expression of membrane protein gene ompA，ompC，ompW，ompX was significantly decreased，but rpoE did not appear to continue to be high expression. These findings provide us new ideas to improve the resistance of engineering bacteria cells and control cells lysis.

Abstract:N-acetylglucosamine（O-GlcNAc） modification on protein serines/threonines is a dynamic，inducible and abundant post-translational modification，which is found on numerous cytoplasm and nucleus proteins，regulating many cellular process. It has demonstrated that O-GlcNAc plays important roles in some human diseases，such as diabetes and neurodegenerative. O-GlcNAcylation cycle is regulated by O-GlcNAc transferase（OGT） and O-GlcNAcase（OGA） in vivo. There are three isoforms of OGT have been found in mammalian cell. They are nucleoplasmic OGT（ncOGT），mitochondrial OGT（mOGT） and shorter OGT（sOGT）. We have expressed mOGT in yeast cells with a final goal to study its biological function. A certain number of yeast proteins were clearly revealed in mOGT overexpressed cells，demonstrating the activity of mOGT in yeast cells. Furthermore，it was found that human mOGT protein inhibited the cell growth of yeast. This humanized yeast strain can be used for studying the biological function and the regulation mechanism of human mOGT.

Abstract:Pullulanase gene （GenBank Accession No：AX203843） was heterologous expression in BL21 （DE3） in shake flask and scale-down to the microtiter plate scale successfully. Based on microtiter plate，a high-throughput culture method and an efficient high-throughput detection method for pullulanase activity were established. Random mutagenesis on pullulanase gene was performed through error-prone PCR strategy. The error-prone PCR products were recombinated in expression vector pET-28a（+）-pelB and then introducted into BL21 （DE3） to construct mutant library. The High-throughput screening method for Pullulanase in study was used to screen the mutant library efficiently and optimum mutant 4A4 was screened，the pullulanase activity in supernatant improved 1.465 folds. This method not only is also applicable to high-throughput screening of amylase and cellulase，but also provides a new way for high-throughput screening of strain library and directed evolution of proteins.

Abstract:To estimate the influence of different heavy metals to the Potamogeton crispus L.，especially the growth and development of its winter buds，the stresses of Cd2+，Cu2+，Zn2+ and Pb2+ in different concentrations to the growth and development of winter buds of Potamogeton crispus L. were studied，through the quantitative determination of germinated number of winter buds，chlorophyll of the leaves，soluble protein，malondialdehyde（MDA），and antioxidant system. As a result，under the same concentrations，Cu2+ had the most obvious effect on physiological and biochemical indexes of the plant. The Cd2+ ranked the second and followed by Pb2+ and Zn2+. The plants cannot grow normally or even survive with concentration of Cu2+ or Cd2+ higher than 0.5 mg/L. At the same time，the plants can grow normally under 10 mg/L of Zn2+ or Pb2+ treatment，which provided effective basis for aquatic plant in the metal threshold for ecological repair.