Directed evolution of the 3-hydroxypropionic acid production pathway by engineering aldehyde dehydrogenase using a synthetic selection device

醛脱氢酶 代谢工程 生物化学 定向进化 合成生物学 脱氢酶 代谢途径 甘油 蛋白质工程 化学 工业微生物学 生物 计算生物学 催化作用 基因 突变体 发酵
作者
Joo Yeon Seok,Joseph Yang,Sang Jin Choi,Hyun Gyu Lim,Un Jong Choi,Kyungjin Kim,Sunghoon Park,Tae Hyeon Yoo,Gyoo Yeol Jung
出处
期刊:Metabolic Engineering [Elsevier]
卷期号:47: 113-120 被引量:52
标识
DOI:10.1016/j.ymben.2018.03.009
摘要

3-Hydroxypropionic acid (3-HP) is an important platform chemical, and biological production of 3-HP from glycerol as a carbon source using glycerol dehydratase (GDHt) and aldehyde dehydrogenase (ALDH) has been revealed to be effective because it involves a relatively simple metabolic pathway and exhibits higher yield and productivity than other biosynthetic pathways. Despite the successful attempts of 3-HP production from glycerol, the biological process suffers from problems arising from low activity and inactivation of the two enzymes. To apply the directed evolutionary approach to engineer the 3-HP production system, we constructed a synthetic selection device using a 3-HP-responsive transcription factor and developed a selection approach for screening 3-HP-producing microorganisms. The method was applied to an ALDH library, specifically aldehyde-binding site library of alpha-ketoglutaric semialdehyde dehydrogenase (KGSADH). Only two serial cultures resulted in enrichment of strains showing increased 3-HP production, and an isolated KGSADH variant enzyme exhibited a 2.79-fold higher catalytic efficiency toward its aldehyde substrate than the wild-type one. This approach will provide the simple and efficient tool to engineer the pathway enzymes in metabolic engineering.
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