Engineering bacterium with high L-aspartic acid alpha carboxylase activity, and its application in production of beta-alanine

一种高L-天冬氨酸α羧化酶活性的工程菌及其在生产β-丙氨酸中的应用

Abstract

本发明公开了一种工程菌及其在生产β-丙氨酸中的应用。本发明提供的工程菌,其构建方法依次包括如下步骤:(1)将目的蛋白的编码基因插入载体pET-30a(+)的多克隆位点,得到的重组质粒pET30-panDB;所述目的蛋白如序列表的序列6所示;(2)将步骤(1)得到的重组质粒pET30-panDB导入大肠杆菌BL21(DE3),得到所述工程菌。本发明还保护一种生产β-丙氨酸的方法,以L-天冬氨酸为底物,采用所述工程菌进行生物转化,得到β-丙氨酸。采用本发明提供的方法制备β-丙氨酸,无需添加诱导剂,整个过程无需高温、高压,同时具有条件温和、操作简单、对环境友好等优点,已达工业化应用水平。
The invention discloses an engineering bacterium and its application in the production of beta-alanine. A construction method of the engineering bacterium comprises the following steps: 1, inserting the coding gene of a target protein the multiple cloning site of a vector pET-30a(+) to obtain a recombinant plasmid pET30-panDB, wherein the target protein is represented by a sequence 6 in a sequence table; and 2, introducing the recombinant plasmid pET30-panDB obtained in step 1 to Escherichia coli BL21(DE3) in order to obtain the engineering bacterium. The invention also discloses a method for producing beta-alanine. The engineering bacterium is biologically converted by using L-aspartic acid as a substrate to obtain beta-alanine. The method for preparing beta-alanine has the advantages of no need of an inducer, no need of high temperature or high pressure in the whole process, mild conditions, simple operation and environmental protection, and reaches the industrial application level.

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