谷氨酰胺酶基因的克隆、表达和酶学性质研究

Cloning，Expression and Characterization of Bacillus Subtilis Glutaminase

DOI：10.3969/j.issn.1673-1689.2019.10.009

中文关键词: 枯草芽孢杆菌 谷氨酰胺酶 酶学性质

英文关键词: Bacillus subtilis,glutaminase,enzymatic properties

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中文摘要:

以枯草芽孢杆菌基因组为模板通过PCR扩增获得谷氨酰胺酶基因ylaM，构建重组质粒pMA5-ylaM，将其转化到枯草芽孢杆菌168中获得重组菌BS168/pMA5-ylaM，并在宿主菌成功得到表达，纯化后的谷氨酰胺酶比酶活大小为939.48 U/mg。谷氨酰胺酶的反应最适pH为7.5，反应最适温度为55 ℃，La³⁺、Zn²⁺、Fe³⁺和Al³⁺对谷氨酰胺酶活力具有一定的抑制作用，在NaCl浓度为15%~17.5%的条件下，该酶酶活力可以保留50%以上。在5 L罐中，通过分批补料发酵，其最高酶活力产量为215.06 U/mL。

英文摘要:

The glutaminase gene（ylaM）was amplified from Bacillus subtilis by PCR.Construction of pMA5-ylaM and expressed in168.The specific activity of purified glutaminase was 939.48 U/mg. The optimization activity of recombinant enzyme was at pH 7.5 and 55 ℃. La³⁺、Zn²⁺、Fe³⁺and A^l3+inhibited the glutaminase activity. More than 50% of the maximum activity was retained in the presence of 15%~17.5% NaCl.The highest production of glutaminase of fed-batch fermentation was 215.06 U/mL in the 5 L fermentation.

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