Protein engineering of esterase E2 for synthesis of (R)-o-chloromandelic acid

• Published in: Biotechnology and bioprocess engineering Vol. 30; no. 2; pp. 354 - 362
• Main Authors: Gu, Jiali, Xiao, Li, Wang, Yajun
• Format: Journal Article
• Language: English
• Published: Seoul The Korean Society for Biotechnology and Bioengineering 01.04.2025; Springer Nature B.V; 한국생물공학회
• Subjects: Acids; Benzene; Biosynthesis; Biotechnology; Chemistry; Chemistry and Materials Science; Chromatography; Conjugation; Enantiomers; Engineering; Enzyme kinetics; Esterase; esterases; hydrolysis; Industrial and Production Engineering; Industrial applications; Molecular dynamics; Mutants; Optimization; Protein engineering; Proteins; Research Paper; Simulation; Solvents; Temperature; 생물공학; chloromandelic acid; Enantioselectivity; (; ); Molecular dynamics simulations; Semi-rational design; Asymmetric hydrolysis
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-024-00172-9

( R )- o -chloromandelic acid is one of the most preferred chiral building blocks. For efficient biosynthesis, esterase E2 was redesigned using semi-rational design strategy. After three rounds, mutant A150F/I228A with significant increase in R -enantioselectivity ( E  = 75.5) was obtained. The molecular dynamics simulation results demonstrated that π-π conjugation interaction existed between the benzene ring of ( R )-enantiomer and residue F150 in the mutant A150F/I228A. The optimum temperature and pH for mutant A150F/I228A were 30–35 °C and 8.0, respectively. Under the optimal conditions, the asymmetric hydrolysis of methyl ( R,S )- o -chloromandelate could be completed in 5 h, and the enantiomeric excess value of product ( ee p ) was 99%, which was potential in industrial application.