重组L-谷氨酸氧化酶固定化细胞的制备  

Preparation of recombinant L-glutamate oxidase immobilized cells

作　　者：李力 王亚平[1] 饶犇[2] 马立新[1] LI Li;WANG Ya-ping;RAO Ben;MA Li-xin(Hubei Key Laboratory of Industrial Biotechnology,Hubei University,Wuhan 430062,China;National Biopesticide Engineering Research Center,Hubei Academy of Agricultural Sciences,Wuhan 430064,China)

机构地区：[1]湖北大学湖北省工业生物技术重点实验室,武汉430062 [2]湖北省农业科学院生物农药工程研究中心,武汉430064

出　　处：《湖北农业科学》2019年第17期132-135,共4页Hubei Agricultural Sciences

基　　金：国家生物农药工程技术研究中心及农业部农业微生物资源利用华中区域科学观测试验站开放基金资助项目(JF-NBCOES-1807)

摘　　要：为了提高表达L-谷氨酸氧化酶(LGOX)的重组大肠杆菌(pET-23a-lgox)的底物催化耐受性,尝试了对重组菌进行固定化的研究。结果表明,产LGOX的重组大肠杆菌工程菌种的最佳固定化条件为:海藻酸钠浓度2.5%,CaCl2溶液浓度13g/L,pH6.5,OD600nm为14的工程菌与海藻酸钠的最佳体积比1∶40,此时得到的固定化工程菌机械强度最好。通过酶学性质比较发现,固定化的细胞与游离细胞在底物催化过程中相比,同等条件下前者温度耐受性、pH耐受性、使用频次等均高于后者。In order to improve the substrate catalytic tolerance of recombinant E.coli(pET-23a-lgox) expressing L-glutamate oxidase(LGOX),this study attempted to study the immobilization of recombinant bacteria.The results showed that the best immobilization conditions were 2.5% sodium alginate,13 g/L CaCl2 solution,pH 6.5,and the optimal volume ratio of engineering bacteria(OD600 nm=14) to sodium alginate was 1∶40.At this time,the mechanical strength of the stationary engineering bacteria was the best.Through the comparison of enzymatic properties,it was found that the immobilized cells were more tolerant than free cells in the substrate catalysis process.And under the same conditions,their temperature tolerance,pH tolerance,and the use of frequency were higher than free cells.

关 键 词：固定化细胞 海藻酸钠 L-谷氨酸氧化酶 

分 类 号：Q814.2[生物学—生物工程]