短乳杆菌DM9218肌苷水解酶基因的异源表达及活性检测

目的探讨短乳杆菌DM9218肌苷水解酶基因A0008的异源表达及其对肌苷的分解活性检测。方法克隆来源于短乳杆菌DM9218基因组的肌苷水解酶基因A0008,构建原核表达载体,转入大肠埃希菌BL21诱导重组蛋白表达并纯化,进行体外酶活检测。结果成功构建了肌苷水解酶A0008-pET28a原核表达载体,表达并纯化出重组蛋白,酶活结果显示该重组蛋白具有水解肌苷的能力。结论短乳杆菌DM9218基因A0008可能编码肌苷水解酶并参与DM9218对肌苷的分解。

Heterologous expression of an inosine hydrolase gene derived from Lactobacillus brevis DM9218 and detection of its inosine-degrading activity

Abstract: Objective To explore the heterologous expression and activity of the inosine hydrolase gene A0008 derived from Lactobacillus brevis DM9218. Methods The inosine hydrolase gene A0008 derived from the genome of Lactobacillus brevis DM9218 was PCR-amplified and ligated to a prokaryotic expression vector. This constructed vector was transformed into Escherichia coli BL21 to induce the expression of recombinant protein. The inosine-degrading activity of the purified protein was evaluated in vitro. Results The prokaryotic expression vector of inosine hydrolase A0008-pET28a was successfully constructed, and the recombinant protein was expressed and purified. The result of the detection of enzyme activity showed that the recombinant protein had the ability to hydrolyze inosine. ConclusionThe gene A0008 from Lactobacillus brevis DM9218 may encode inosine hydrolase and participate in the decomposition of inosine.