Overproduction of α-Lipoic Acid by Gene Manipulated Escherichia coli.
Overproduction of α-Lipoic Acid by Gene Manipulated Escherichia coli.
复制标题
基因操纵大肠杆菌过量生产α-硫辛酸
DOI：
10.1371/journal.pone.0169369
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Wang H
中科院分区：
文献类型：
--
作者：
Sun Y;Zhang W;Ma J;Pang H;Wang H
α-硫辛酸（LA）是生物体广泛使用的重要酶辅因子，也是用于治疗由低水平内源性抗氧化剂驱动的病理的天然抗氧化剂。为了建立一个更安全，更有效的生产LA的过程中，我们开发了一种新的生物方法的基础上，硫辛酸生物合成LA的新兴知识。我们首先克隆了lipD基因，该基因编码丙酮酸脱氢酶E2亚基的硫辛酰结构域，允许高水平的LipD生产。随后利用各种载体和启动子构建含有用于LA生物合成的基因的质粒以产生高水平的LA。将这些质粒转化到大肠杆菌菌株BL 21中。辛酸（OA）被用作LA合成的底物。一株携带lipD、lplA和利帕的菌株YS 61产生的LA水平比野生型菌株高200倍以上，表明LA可以在E.大肠杆菌中进行基因工程改造。
Alpha-lipoic acid (LA) is an important enzyme cofactor widely used by organisms and is also a natural antioxidant for the treatment of pathologies driven by low levels of endogenous antioxidants. In order to establish a safer and more efficient process for LA production, we developed a new biological method for LA synthesis based on the emerging knowledge of lipoic acid biosynthesis. We first cloned the lipD gene, which encodes the lipoyl domain of the E2 subunit of pyruvate dehydrogenase, allowing high levels of LipD production. Plasmids containing genes for the biosynthesis of LA were subsequently constructed utilizing various vectors and promotors to produce high levels of LA. These plasmids were transformed into the Escherichia coli strain BL21. Octanoic acid (OA) was used as the substrate for LA synthesis. One transformant, YS61, which carried lipD, lplA, and lipA, produced LA at levels over 200-fold greater than the wild-type strain, showing that LA could be produced efficiently in E. coli using genetic engineering methods.