Substrate-Specific Evolution of Amine Dehydrogenases for Accessing Structurally Diverse Enantiopure (<i>R</i>)-β-Amino Alcohols
Xinjian Yin, Wenzhong Gong, Yujing Zeng, Hulin Qiu, Lan Liu, Frank Hollmann, Bi‐Shuang Chen
Abstract
High Resolution Image Download MS PowerPoint Slide The biocatalytic oxidative deamination of β-amino alcohols holds significant practical potential in kinetic resolution and/or deracemization process to access ( R )-β-amino alcohols. This study exemplifies a notable instance of acquisition and utilization of this valuable oxidative deamination activity. Initially, the mutation N261M (M0) was identified to endow a native valine dehydrogenase with oxidative deamination activity toward a few ( S )-β-amino alcohols. Subsequently, a phylogenetic analysis-guided, double-code saturation mutagenesis strategy was proposed to engineer M0's side-chain binding site. This strategy facilitated the substrate-specific evolution of M0, resulting in the creation of a panel of mutants (M1–M4) with noteworthy oxidative deamination activity toward structurally diverse ( S )-β-amino alcohols. Using these engineered amine dehydrogenases, termed as β-amino alcohol dehydrogenases (β-AADHs), the complete kinetic resolution and even deracemization of a range of β-amino alcohols have been achieved. This work reports distinct biocatalysts and a synthetic strategy for the synthesis of enantiopure ( R )-β-amino alcohols and offers an innovative approach for substrate-specificity engineering of enzymes.