Journal
METABOLIC ENGINEERING
Volume 47, Issue -, Pages 113-120Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2018.03.009
Keywords
3-hydroxypropionic acid; Directed evolution; Transcription factor; Screening; Synthetic biology; Metabolic engineering
Categories
Funding
- Advanced Biomass R&D Center (ABC) of Global Frontier Project [ABC-2015M3A6A2066119]
- National Research Foundation of Korea (NRF) - Ministry of Science and ICT, Korea [NRF-2015R1A2A1A10056126]
- Korea Institute of Energy Technology Evaluation and Planning (KETEP)
- Ministry of Trade, Industry & Energy (MOTIE) of the Republic of Korea [20174030201600]
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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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