Improving the Enantioselectivity of CHMO Brevi1  for Asymmetric Synthesis of Podophyllotoxin Precursor

In this study, we use a strategy called "focused rational iterative site-specific mutagenesis" (FRISM) at residues ranging of 6 Å from substrate. The mutations by using a restricted set of rationally chosen amino acids allows the formation of a small mutant library. By generating and screening less than 60 variants, we achieved high ee of 96.8%. Coupled with the cofactor regeneration system, 9.3 mM substrate was converted completely in a 100-mL scale reaction. Therefore, our work reveals a promising synthetic method for (R)-β-piperonyl-γ-butyrolactone with the highest enantioselectivity, and provides a new opportunity for the chem-enzymatic synthesis of podophyllotoxin.PMID:37728423 | DOI:10.1002/cbic.202300582
Source: Chembiochem - Category: Biochemistry Authors: Source Type: research