Rational design of lanosterol 14α-demethylase for ergosterol biosynthesis in <i>Saccharomyces cerevisiae</i>.

ABSTRACT: Ergosterol is widely used in skin care products and drug preparation. Lanosterol 14α-demethylase (Erg11p, 14DM, CYP51) is the rate-limiting enzyme for the biosynthesis of various steroid compounds in Saccharomyces cerevisiae. Herein, Erg11p was engineered to extend the in vivo catalytic half-life and increase the turnover rate. Single mutations resulting in lower folding energy were selected, and mutant P201H had an ergosterol yield of 576.9 mg·L^-1. Through consensus design, single mutations resulting in higher sequence identity to homologs were tested and mutant K352L had an ergosterol yield of 677.9 mg·L^-1. The key residues for substrate binding were confirmed via alanine scanning mutagenesis and mutant F384A had an ergosterol yield of 657.8 mg·L^-1

SUBMITTER: Liang R 

PROVIDER: S-EPMC11564469 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature