湖北农业科学 ›› 2019, Vol. 58 ›› Issue (17): 132-135.doi: 10.14088/j.cnki.issn0439-8114.2019.17.035

• 生物工程 • 上一篇    下一篇

重组L-谷氨酸氧化酶固定化细胞的制备

李力1, 王亚平1, 饶犇2, 马立新1   

  1. 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
  • 收稿日期:2019-02-21 发布日期:2019-11-14
  • 作者简介:李 力(1994-),男,湖北安陆人,在读硕士研究生,研究方向为微生物学,(电话)18672762809(电子信箱)1042271228@qq.com;
  • 基金资助:
    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)

Preparation of recombinant L-glutamate oxidase immobilized cellsLI Li1,WANG Ya-ping1,RAO Ben2,MA Li-xin1

(1.Hubei Key Laboratory of Industrial Biotechnology, Hubei University, Wuhan430062, China;2.National Biopesticide Engineering Research Center, Hubei Academy of Agricultural Sciences, Wuhan430064, China)   

  1. Abstract: 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.
  • Received:2019-02-21 Published:2019-11-14

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13 g/L,pH 6.5,OD600 nm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。

关键词: 固定化细胞, 海藻酸钠, L-谷氨酸氧化酶

Key words: immobilized cells, sodium glginate, L-glutamate oxidase

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