Abstract
In order to avoid the interference of natural GAD expressed by the own genome of Escherichia coli, the recombinant cellulose-binding domain glutamate decarboxylase (CBD-GAD) was firstly immobilized by regenerated amorphous cellulose (RAC), and then the RAC-CBD-GAD immobilized enzyme was isolated and used as an evaluation index to optimize the culture conditions for highly efficient expression of CBD-GAD in E. coli GDMCC60445 by one-factor-at-a-time and Taguchi methods. The results indicated that the optimal medium for E. coli GDMCC60445 to express CBD-GAD was modified Luria-Bertani (LB) medium, which was consisted of 8 g/L tryptone, 6 g/L yeast extract, 10 g/L sodium chloride, and pH 5.5. The suitable culture conditions were 37 ℃, 120 r/min and 24 h. Under the optimal conditions, the activity of RAC-CBD-GAD prepared from E. coli GDMCC60445 was (419.29±10.37) U/g, which was consistent with the predicted value of Taguchi and improved by (30.28±3.22)% as compared to the initial one.
Publication Date
7-28-2019
First Page
31
Last Page
38
DOI
10.13652/j.issn.1003-5788.2019.07.006
Recommended Citation
Shengyuan, YANG; Qian, LIN; Xiaoning, ZHANG; and Jiayi, ZHEN
(2019)
"Optimization of conditions for efficient expression of recombinant cellulose-binding domain-glutamate decarboxylase of Enterococcus faecium in Escherichia coli,"
Food and Machinery: Vol. 35:
Iss.
7, Article 6.
DOI: 10.13652/j.issn.1003-5788.2019.07.006
Available at:
https://www.ifoodmm.cn/journal/vol35/iss7/6
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