Protein engineering of lipase A from Candida antarctica to improve esterification of tertiary alcohols

Karla Wagner; Anke Hummel; Jianing Yang; Satoshi Horino; Kyohei Kanomata; Shuji Akai; Harald Gröger

doi:10.1002/cbic.202400082

Protein engineering of lipase A from Candida antarctica to improve esterification of tertiary alcohols

Chembiochem. 2024 Apr 26:e202400082. doi: 10.1002/cbic.202400082. Online ahead of print.

Authors

Karla Wagner¹, Anke Hummel¹, Jianing Yang¹, Satoshi Horino², Kyohei Kanomata², Shuji Akai², Harald Gröger³

Affiliations

¹ Bielefeld University, Faculty of Chemistry, GERMANY.
² Osaka University, Graduate School of Pharmaceutical Sciences, JAPAN.
³ Bielefeld University: Universitat Bielefeld, Fakultät für Chemie Organische Chemie I, Universitätsstr. 25, 33615, Bielefeld, GERMANY.

PMID: 38670922
DOI: 10.1002/cbic.202400082

Abstract

Chiral tertiary alcohols are important organic compounds in science as well as in industry. However, their preparation in enantiomerically pure form is still a challenge due to their complex structure and steric hindrances compared with primary and secondary alcohols, so kinetic resolution could be an attractive approach. Lipase A from Candida antarctica (CAL-A) has been shown to catalyze the enantioselective esterification of various tertiary alcohols with excellent enantioselectivity but low activity. Here we report a mutagenesis study by rational design to improve CAL-A activity against tertiary alcohols. Single mutants of CAL-A were selected, expressed, immobilized and screened for esterification of the tertiary alcohol 1,2,3,4-tetrahydronaphthalene-1-ol. A double mutant V278S+S429G showed a 1.5-fold higher reaction rate than that of the wild type CAL-A, while maintaining excellent enantioselectivity.

Keywords: biocatalysis; kinetic resolution; lipase A from Candida antarctica; protein engineering; tertiary alcohols.