Abstract:Glucose oxidase （β-D-glucose：oxygen 1-oxidoreductase；EC 1.1.3.4，GOD） has gained considerable commercial importance in the last few years due to its multitude of applications in the chemical，food，pharmaceutical，beverage，biotechnology and other industries. In this study，the fermentation medium of the recombinant Yarrowia lipolytica strain 1-28 that was constructed to secrete GOD was optimized through single-factor and orthogonal tests. The optimal fermentation medium was consisted of 20 g/L glycerol，2.64 g/L yeast extract，2.64 g/L NH4Cl，0.13 g/L MgSO4，0.32 g/L KH2PO4，3.34×10-4 g/L Vitamin B1 at pH 6.0. The yield of GOD corresponded to 11.0 U/mL when cultured in the optimized medium，which was 72% higher than that at initial conditions. When fermented in the 3 L tank at 28 ℃ for 190 h，the enzyme activity of GOD reached 81.6 U/mL at pH 5.0 and glycerol feeding at the concentration of 30 g/L.

Abstract:To identify the origin，succession and potential function of lactic acid bacteria（LAB） during the fermentation process of Luzhou-flavor liquor，we compared the LAB community structures among Daqu，fermented grains，and pit mudusing 16S rRNA gene high-throughput sequencing，revealed the succession of LAB community in fermented grains，and predicted the potential function of LAB during fermentation processvia PICRUSt technology. Results showed significant differences of LAB community structure among Daqu，fermented grains and pit mud. The structureof LAB community in fermented grains changed dramatically in the first week of fermentation. Furthermore，PICRUStanalysis showed that LAB mainly participated in glycolysis，and possessed the functional potential to generate butanoate and propanoate.

Abstract:4-α-glucanotransferase can catalyze intramolecular glucan transfer reactions of amylose to produce cycloamylose. In this study，a recombinant E. coli BL21（DE3）/pET-24a（+）-AaAM derived from Aquifex aeolicus was constructed for intracellular production of 4-α- glucanotransferase. The AaAM produced in culture medium activity was 0.99 U/mL after 27 h in shake flask. Furthermore，the condition for cycloamylose preparation with potato starch by this enzyme was optimized. The optimized conditions for the conversion as follows: substrate concentration was 1 g/dL debranched potato starch, the initial pH was 7.0, the reaction temperature was 75 ℃, 4-α-glucanotransferase concentration was 20 U/g starch. Under this conditions, the cycloamylose yield is 24.8% for 10 h, which paved the way for indnstrial production of cycloamylose.

Abstract:Cathepsin B is an enzymatic protein that was closely related with tumor cell invasion and metastasis. The network of Cathepsin B is very important for research and development of new anticancer drug，for example，new Cathepsin B inhibitor. Using yeast two hybrid technique，the proteins that interacting with Cathepsin B were identified. Results：After screening analysis，15 positive clones of putative interactions were obtained and they were identified by sequencing and utilizing of the data online（http：//www.ncbi.nlm.nih.gov）. There were 7 genes including BCL2-associated athanogene 6（BAG6），filamin A，alpha（FLNA），　ras homolog family member G（RHOG），　metallothionein 2A（MT2A），　ZXD family zinc finger C（ZXDC），　DEP domain containing 7（DEPDC7），A kinase（PRKA） interacting protein 1（AKIP1）.

Abstract:We screened and identified the cold-adapted bacteria from Mingyong glacier.The bacterial strains were isolated and purified by the soil suspension coating and spread plate methods. Based on physiological and biochemical properties，homology and phylogenetic analysis of 16S rRNA gene sequences，all isolates were classified. In total 22 bacterial strains were isolated，based on the 16S rRNA gene sequence analysis：MY0504 strain was most closely related to Flavobacterium pectinovorum with 97.6%，MY1413 and MY14015 strains were most closely related to Arthrobacter psychrophenolicus and A. parietes with 98% ，while MY14011 strain was most closely related to Chromobacterium fluviatile with 97%，and other 18 strains of bacteria were identified as Pseudomonas spp. The research is preliminary exploration on the microbiological resources in Mingyong glacier，and provides reference information for further studies in this area.

Abstract:To achieve this study，MTT assay is used to measure the anti-breast cancer activity in different extracts from Inonotus obliquus；Change of cellular morphology is observed as the apoptosis detection；Cell scratch test is used to determine the effect of drug on cell migration ；LC-MS/MS is used to analyze the active constituents in drug. MTT shows that the petroleum ether extract fraction has the strongest cell growth inhibitory effect on MDA-MB-231 cells in a dose and time dependent manner（P＜0.05）；the IC50 value for 24 hours is 76.17 μg/mL.Cell morphology observation shows that apoptosis effect of petroleum ether extract fraction is increased in MDA-MB-231 cells in a dose manner. Scratch test shows that the petroleum ether extract fraction has a significant inhibitory effect on cell migration. LC-MS/MS data preliminarily characterizes 14 chemical components，of which 10 components are firstly identified in Inonotus obliquus. Although the other 4 substances have not been identified，the molecular weight and molecular formula have been characterized. The petroleum ether extract fraction is considered as the main anti-breast cancer active fraction of Inonotus obliquus. It can inhibit the growth of MDA-MB-231 cells and induce apoptosis，which provided the scientific basis for further development and utilization of the food and medicinal resources of Inonotus obliquus.

Abstract:Doxorubicin（DOX）-loaded hyaluronic acid（HA） modified Liposomes（DOX/LP/ HOA） were prepared to enhance receptor mediated endocytosis and improve anti-tumor effect. Octadecyl was chosen to conjugate HA backbone by amide bond to prepare hyaluronic acid-octodecane（HOA） polymers. DOX，as a model drug，was encapsulated into HOA modified liposomes by the film dispersion method and post-insertion method. The average particle size，zeta potential and encapsulation of DOX/LP/HOA were（50.17±0.60） nm，（-7.61±0.43） mV and（98.85±0.40）%，respectively. In vitro release and cytotoxicity results showed that DOX/LP/HOA could release DOX slowly and improve tumor targeting ability. DOX/LP/HOA with excellent anti-tumor activity can be used as the potential carrier for delivery of anti-tumor drugs.

Abstract:Transglutaminases（EC 2.3.2.13，TGase） is an industrial enzyme widely used in several domains that include food processing and textile industry. The amino acid sites that may reduce folding free energy were predicted by using PoPMuSiC-2.1 to improve the thermal stability of TGase，and the corresponding mutants were then constructed. As indicated by PoPMuSiC-2.1 analysis，mutation at P132 revealed the highest level of decreases in folding free energy of TGase.Based on this prediction，the mutants with decreased folding free energy was thus contracted，including P132I，P132G，P132M，and P132Q. The analysis of enzymatic property showed that the half-life of P132I can reach 5.0 min at 50 ℃ which was 31% higher than that of wild-type TGase，and the other mutants also exhibited enhanced half-life. Moreover，P132I and P132G showed 24% and 12.4% enhanced specific activities in contrast to wild-type enzyme，respectively；while the specific activities of the rest mutants did not change significantly. And the structure analysis indicated that P132I mutant formed additional two hydrogen bonds as compared with wild-type enzyme. These results suggested that site-directed mutagenesis based on analysis of folding free energy is an effective method to improve the thermal stability of TGase，and newly formed hydrogen bond may be one of the reasons for enhanced thermal stability of P132I.

Abstract:To study the distribution of yeast in raw milk in pastoral areas of Tacheng，Xinjiang. Seven samples of raw milk were collected from the areas. Using the traditional culture-dependent method，the isolation yeasts were identified according to the colonial morphology，biological properties and 5.8S rDNA sequence homology analysis. A total of 8 species belonging to 6 genera were identified from 33 strains isolated from the isolates，which were 20 strains of Pichia kudriavzevii（61%）were the dominant strains，2 strains Pichia fermentans（6%）；3 strains Metschnikowia pulcherrima（9%）；2 strains Kluyveromyces lactis（6%）；2 strain Kluyveromyces marxianus（6%）；2 strains Rhodotorula mucilaginosa（6%）；1 strain Hanseniaspora uvarum（3%）；1 strain Kazachstania unispora（3%）. The study showed that the yeast diversity of raw milk in this region was good，and has its unique yeast resources. This provides the basic datas for the study of the microbial resources in this area.

Abstract:In order to improve the tolerance of Actinobacillus succinogenes to succinate，a microfibrous bed bioreactor with immobilized Actinobacillus succinogenes is constructed in this thesis. Through adaptive evolution，a stain with enhanced salt-tolerance and productivity was obtained with succinic acid concentration of 38.4±3.14 g/L，which was a 23.8% increase compared to the original strain. Fermentative production of succinic acid with both the original and the evoluted strains were conducted in a 3 L fermentor. A fed-batch strategy was used and after 62 h of fermentation，the concentration，productivity and yield of succinic acid was 86.3 g/L，1.39 g/（L·h） and 78.7%，respectively for the evolved strain. Compared to the original strain，the succinic acid yield and productivity increased by 14.2% and 24.1% for the evolved strain.

Abstract:Hot air drying（AD） of vegetables has many disadvantages including agglomeration，poor quality，high energy consumption and heavy pollutions. In this study，we used cabbage as the raw material，pulse spouted bed low frequency microwave drying（PSBLMD） as the drying method to build a pilot scale research platform. The quality，uniformity and energy consumption of the PSBLMD-dried cabbages were compared with those of the AD-dried ones. Results shown that the amount of ascorbic acid，chlorophyll and the rehydration ratio of PSBLMD-dried cabbages with 15% sugar addition（4.69 mg/hg，3.48 mg/hg and 26.22，respectively） was higher than those of the AD dried cabbages（1.46 mg/hg，3.14 mg/hg and 19.40，respectively）. The uniformity of the products regarding the moisture content，the color appeal and the shrinkage ratio of PSBLMD-dried cabbages were >90%. Compared to AD，PSBLMD not only effectively solved the agglomeration problem during the drying process，but also improved the quality of the dried products. It shortened the drying time by >50% and reduced the energy consumption by >40%.

Abstract:The effect of multi-frequency ultrasound assisted enzymatic（phospholipase A1） treatment on the degumming efficiency and quality of rapeseed oil was investigated in this study. The results showed that multi-frequency ultrasonic treatment can effectively improve the degumming efficiency of rapeseed oil and also reduce the ultimate phosphorus content in the rapeseed oil. In addition，the 20/50 kHz dual-frequency ultrasonic treatment had the best degumming efficiency of rapeseed oil. Compared with the rapeseed oil after degumming by phospholipase A1，the acid value and peroxide value of degummed rapeseed oil after ultrasonic treatment increased slightly，but they were all within the safe range stipulated by the national standard. Moreover，the color of degummed oil became lighter，and the change in the value of fatty acid composition was not obvious.

Abstract:To express active endo-beta-N-acetylglucosaminidase（ENGase）from methylotrophic yeast Ogataea minuta（Endo-Om） in Escherichia coli. The Endo-Om was overexpressed by using pET system. After optimizing host strainand culture condition，we purified the recombinant protein by using Ni2+ metal chelating column.Using the purified Endo-Om，ENGase activity against fluorescent-labeled oligosaccharide was measured. The recombinant protein was well expressed by IPTG induction，however，the majority remained in the insoluble fraction. The insolubility of the recombinant was improved by culturing the E. coli cells in Overnight ExpressTM Instant LB Medium（Merck Millipore） at 16 ℃. The purified protein showed hydrolysis activity to the fluorescent- labeled oligosaccharide. The functional Endo-Om was successfully obtained in this study，it could accelerate the study on its structural and biological function.

Abstract:Pathogenic bacteria inactivation efficiency irradiated by electron beam（EB） irradiation for refrigerated chicken was evaluated through survival rate，which was the irradiation dose to keep 10% microorganism alive（D10）. The sensory quality and changes of total numbers of bacterial colony in refrigerated chicken with EB irradiation of different doses during storage at 0~4 ℃ were investigated. The results showed that the D10 values of Salmonella typhimurium，Escherichia coli and Taphylococcus aureus in refrigerated chicken after EB irradiation were 0.483，0.932 and 0.715 kGy，respectively. The total numbers of bacterial colony in refrigerated chicken after 4 kGy and 6 kGy or more irradiation during storage at 0~4 ℃ for 14 d and 28 d were measured to below the limit（5×105 CFU/g） and their sensory quality were no significant effect.

Abstract:Electrophoresis-purity cellulase from Meretrix meretrix L was obtained through buffer solution extraction，ammonium sulfate precipitation，DEAE-Sepharose ion exchange chromato- graphy and Phenyl Sepharose 6 Fast Flow hydrophobic chromatography. The specific activity of purified cellulase was 40.33 U/mg with purification fold of 13.12. SDS-PAGE results revealed the molecular weights of the enzyme was 59 700. Its optimum temperature and pH were 45 ℃and 5.2，respectively. The cellulase was relatively stable in the range of 4~50 ℃ and pH 4~6. Furthermore，its Km value was 0.111 mmol/L under the optimal conditions. The study laid a foundation for studying the composition and structure of endogenous cellulase from animals.

Abstract:The effects of Metal packings on low-alcohol wine production by extraction of carbon dioxide were studied.Metal packings were metal rasching ring，metal pall rings，metal intalox tower packing，mental gauze corrugate packing and theta ring，respectively.The experiment results showed that extraction of low-alcohol wine by carbon dioxide was feasible and theta ring packing was suitable for low alcohol wine production ，by which the ability of metal packings to take off the alcohol and aroma components in low alcohol wine were observed.In the processing of low-alcohol wine production by theta ring，the contents of higher alcohols and carbon dioxide had largely variation and the other aroma components were not almost variation.The contents of higher alcohols fell from 355.83 mg/L to 200.97 mg/L ，which reduced 43.52% and the contents of carbon dioxide was up to 0.06 MPa. Alcohol content of wine was decreased from 11.0%（volume ratio） to 3.77%（volume ratio） when the optimal conditions were metal packing height 300 mm，temperature 20 ℃，pressure 4.0 MPa，proportion of air to liquid flow 20. The alcohol content dropped 65.73%.