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

Title
Directed evolution of the 3-hydroxypropionic acid production pathway by engineering aldehyde dehydrogenase using a synthetic selection device
Authors
Seok, Joo YeonYang, JinaChoi, Sang JinLim, Hyun GyuChoi, Un JongKim, Kyung-JinPark, SunghoonYoo, Tae HyeonJung, Gyoo Yeol
POSTECH Authors
Jung, Gyoo Yeol
Date Issued
May-2018
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
Abstract
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.
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.
Keywords
Aldehydes; Binding sites; Enzymes; Glycerol; Metabolism; Screening; Substrates; Transcription; Transcription factors; 3-Hydroxypropionic acid; Aldehyde dehydrogenase; Biological production; Catalytic efficiencies; Directed evolution; Evolutionary approach; Semialdehyde dehydrogenase; Synthetic biology; Metabolic engineering; aldehyde dehydrogenase; aldehyde derivative; hydracrylic acid; Article; binding site; catalysis; enrichment culture; enzyme engineering; enzyme inactivation; enzyme substrate; molecular evolution; molecular library; nonhuman; priority journal
URI
http://oasis.postech.ac.kr/handle/2014.oak/94522
DOI
10.1016/j.ymben.2018.03.009
ISSN
1096-7176
Article Type
Article
Citation
METABOLIC ENGINEERING, vol. 47, page. 113 - 120, 2018-05
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정규열JUNG, GYOO YEOL
Dept. of Chemical Enginrg
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