Development of an improved phenylacetaldehyde reductase mutant by an efficient selection procedure

Applied Microbiology and Biotechnology
Yoshihide Makino, Nobuya Itoh

Abstract

Chiral alcohols are valuable as diverse chemicals and synthetic intermediate materials. Phenylacetaldehyde reductase (PAR) is an enzyme that converts a wide variety of ketones into chiral alcohols with high optical purity. When an alcohol such as 2-propanol is used as a hydrogen donor, PAR itself will also mediate the regeneration of the coenzyme NADH in situ. Perceiving a capacity for improvement, we sought to develop a PAR that is able to convert higher concentrations of substrates in the presence of high concentrations of 2-propanol. The selection procedure for mutants was re-examined and a procedure able to select an effective amino acid substitution was established. Two advantageous amino acid substitutions were successfully selected using the procedure. When high-concentration substrate conversion reaction was subjected with a mutant that integrated both the two amino acid substitutions, near-complete conversions of m-chlorophenacyl chloride (m-CPC) (2.1 mmol/ml) and ethyl 4-chloro-3-oxobutanoate (ECOB) (1.9 mmol/ml) were achieved.

References

Apr 6, 2004·Current Opinion in Chemical Biology·Wolfgang KroutilKurt Faber
Aug 9, 2005·Applied and Environmental Microbiology·Yoshihide MakinoNobuya Itoh
May 9, 2007·Applied Microbiology and Biotechnology·Katja GoldbergAndreas Liese
Oct 19, 2007·Accounts of Chemical Research·Stefaan M A De WildemanOliver May
Dec 7, 2007·Accounts of Chemical Research·Jeffrey C MoorePaul N Devine
Jan 17, 2008·Analytical Biochemistry·Wen-Chi TsengTsuei-Yun Fang
Nov 15, 2011·Biotechnology Advances·Yan Ni, Jian-He Xu

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Citations

Nov 10, 2017·Journal of Agricultural and Food Chemistry·Juhua LiuZhiqiang Jin

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