Exemplifying Natural (R)-β-Transamination Potential of Fold Type-IV Transaminase for Kinetic Resolution of rac-β-Amino Acids Synthesized from Aldehydes

被引：1

作者：

Pagar, Amol D. ^{[1
]}

Khobragade, Taresh P. ^{[1
]}

Giri, Pritam ^{[1
]}

Jeon, Hyunwoo ^{[1
]}

Cho, Sungchan ^{[1
]}

Jung, Seohee ^{[1
]}

Goh, Younghwan ^{[1
]}

Joo, Sangwoo ^{[1
]}

Sarak, Sharad ^{[1
]}

Munussami, Ganapathiraman ^{[2
]}

Lee, Sun-Gu ^{[2
]}

Yun, Hyungdon ^{[1
]}

机构：

[1] Konkuk Univ, Dept Syst Biotechnol, Seoul 05029, South Korea

[2] Pusan Natl Univ, Dept Chem & Biomol Engn, Busan 46241, South Korea

来源：

ACS SUSTAINABLE CHEMISTRY & ENGINEERING | 2024年 / 12卷 / 19期

基金：

新加坡国家研究基金会;

关键词：

Biomass; aldehydes; beta-amino acids; beta-transaminase; OMEGA-TRANSAMINASE; ASYMMETRIC-SYNTHESIS; SPHAEROBACTER-THERMOPHILUS; DIRECTED EVOLUTION; MALONIC-ACID; BIOCATALYSIS; AMINATION; GAMMA; AMINOTRANSFERASE; SUBSTRATE;

D O I：

10.1021/acssuschemeng.4c00794

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Sustainable synthesis of chiral beta-amino acids from renewable feedstocks is of significant importance. The Rodionov reaction is a popular method to synthesize rac-beta-amino acids from aldehydes, which are abundant byproducts of biomass. rac-beta-Amino acids are easily converted into the enantiopure (R)-form by kinetic resolution using (S)-beta-transaminases (TAs). However, the inaccessibility of natural (R)-beta-transaminases with a broader substrate scope limits the application of the Rodionov reaction to produce (S)-beta-amino acids. Here, we report the kinetic resolution of rac-beta-amino acids by employing fold type-IV transaminase from Thermobaculum terrenum (TATT) with natural (R)-beta-transaminase activity. Various rac-beta-amino acids were successfully resolved by TATT into (S)-form with excellent conversions (similar to 50%) and enantiomeric excess (>99%) using pyruvate as an amino acceptor. The directed evolution of TATT resulted in a variant M-58 (K112F/F114M) with similar to 2-fold higher activity and was able to perform large-scale kinetic resolution of 20 mM rac-3-amino-3-(3-fluorophenyl) propanoic acid (1h) with complete conversion and enantiomeric excess >99% within 24 h.

引用

页码：7226 / 7234

页数：9