重组大肠杆菌产Bacillus subtilis 168来源γ-谷氨酰转肽酶的摇瓶发酵优化

Fermentation Optimization of Recombinant γ-Glutamyltranspeptidase from Bacillus subtilis 168 in E. coli

DOI：10.3969/j.issn.1673-1689.2015.08.003

中文关键词: 枯草芽孢杆菌（Bacillus subtilis） γ-谷氨酰转肽酶 重组大肠杆菌 发酵优化

英文关键词: Bacillus subtilis, γ-glutamyltranspeptidase, recombinant E.coli, fermentation Optimization

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中文摘要:

为实现来源于Bacillus subtilis的γ-谷氨酰转肽酶（GGT）高效胞外分泌表达，以基因工程菌E.coli BL21（DE3）/pET-20b（+）/ggt为出发菌株，对其发酵产酶的诱导条件和培养基进行优化。最终确定其最优发酵培养基为：10 g/L甘油，21 g/L酵母粉，7 g/L蛋白胨，130 mmol/L磷酸盐；发酵条件为：37 ℃、200 r/min培养4 h，加入0.2 mmol/L IPTG后转入25 ℃，诱导40~48 h，优化后酶活达到81.2 U/mL，是未优化前的1.9倍。本研究结果为目前国内报道大肠杆菌摇瓶发酵产γ-谷氨酰转肽酶的最高酶活，为该酶的工业化生产奠定了基础。

英文摘要:

In the present study，in order to improve the production of the recombinant Bacillus subtilis γ-glutamyltranspeptidase in E.coli BL21（DE3）/pET-20b（+）/ggt，the culture conditions and medium compositions were investigated and optimized in shake flasks. The optimal fermentation medium was as follows：10 g/L glycerol，21 g/L yeast extract，7 g/L peptone，and 130 mmol/L PO43+. In addition，the optimal culture conditions were firstly cultured at 37 ℃，200 r/min for 4 h，and then induced by 0.2 mmol/L IPTG at temperature 25 ℃ and 40~48 h. Under these conditions，the maximal enzyme activity reached 81.2 U/mL，which was 1.9 times as high as that not optimized.

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