1. 毕业设计(论文)的内容和要求
毕业论文内容葡萄糖异构酶(glucose isomerase,EC 5.3.1.5),又称木糖异构酶(xylose isomerase),是一种胞内酶,参与对进入体内的木糖的利用,其最适的天然底物为D-木糖 ,被广泛用于高果糖浆(HFCS)的工业生产中。
本课题组通过基因挖掘的方法从NCBI数据库中筛选得到一株嗜热的葡萄糖异构酶(GI)产生菌株Bactillus thermoantarcticus,化学合成该酶的基因并实现其在大肠杆菌E. coli BL21(DE3)中的可溶性表达。
利用亲和层析实现了重组葡萄糖异构酶TPGI的分离纯化,并从最适反应温度、最适pH、最适金属离子及酶促反应动力学等方面研究了该酶的酶学性质。
2. 参考文献
[1] 邓辉, 陈存武, 韦传宝. 葡萄糖异构酶的结构特征及其工业(高产)用酶发掘[J]. 中国生物化学与分子生物学报, 2016, 32(5):510-517.[2] JIA Dong-xu,WANG Teng,SUN Jia-cheng,et al. Research on the immobilization conditions of recombinant Escherichia coli strain harboring refractory glucose isomerase[J].Food and Fermentation Industries,2018,44(6): 8-15.[3] 贾东旭, 周霖, 王腾,等. 耐高温葡萄糖异构酶重组菌发酵与转化条件研究[J]. 食品与发酵工业, 2017,43(7):13-19.[4] DAI Chen-xia, CAO Hui-fang, LUO Hui-ying, et al. Cloning, Expression and Conversion Application of Glucose Isomerase from Alicyclobacillus sp.A4[J].Biotechnology Bulletin, 2018,34(11):144-151.[5] JIN Li-qun, GUO Dong-jing, LIAO Cheng-jun, et al. Progress in immobilized glucose isomerase[J]. Bulletin of Fermentation Science and Technology, 2015, 44(3):47-51.[6] LIAO Cheng-jun, ZHENG Wei, HUANG Jian-feng, et al. Genome mining, expression and characterization of a hyperthermophilic glucose isomerase from Thermotoga petrophila RKU-1[J]. Bulletin of Fermentation Science and Technology, 2015, 44(4):19-23.[7] ZHAO Guo-qun, ZHOU Xue-yan, HU Tao, et al. Influence of Activation Conditions on the Enzyme Activity of Immobilized Glucose Isomerase[J]. Food Research and Development, 2012,33(10):156-159.[8] DENG Hui, CHEN Sheng, WU Jing, et al. Immobilization of Cells Producing Glucose Isomerases[J]. Food Science, 2013, 34(09):164-169.[9] DENG Hui, WANG Guang-lin, CHEN Nai-dong, et al. Construction of Fusion Protein to Improve the Catalytic Performance of Recombinant Glucose Isomerase[J]. Modern Food Science and Technology, 2016, 32(10):52-59.[10]邓辉, 陈存武, 孙传伯, 等. Thermobifida fusca 葡萄糖异构酶在枯草杆菌中的表达[J]. 食品与发酵工业, 2015,41(12):31-35.[11]JIANG Qi, SU Ling-qia, WU Jing, et al. Co-Expression of Phospholipase C and Glucose Isomerase Which Promoting the Extracellular Expression of Glucose Isomerase in E.coli[J]. Journal of Food Science and Biotechnology, 2017, 36(3):236-242.[12]张甫生, 胡国洲, 田美玲, 等. 固定化葡萄糖异构酶的微波活化及性质研究[J]. 食品与发酵工业, 2015,41(8):36-41.
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