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조회911

연세대학교, 현 건국대학교

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Nat. Commun., Jan 02 2024, 15 (1) 49 | https://doi.org/10.1038/s41467-023-44218-7 Reprogramming biocatalytic futile cycles through computational engineering of stereochemical promiscuity to create an amine racemase

Sang-Woo Han^1,2, Youngho Jang¹, Jihyun Kook¹, Jeesu Jang¹ & Jong-Shik Shin¹

¹Department of Biotechnology, Yonsei University, 50 Yonsei-Ro, Seodaemun-Gu, Seoul 03722, South Korea.

²Present address: Department of Biotechnology, Konkuk University, Chungju, South Korea.

Corresponding author : Correspondence to Jong-Shik Shin.

Abstract

Repurposing the intrinsic properties of natural enzymes can offer a viable solution to current synthetic challenges through the development of novel biocatalytic processes. Although amino acid racemases are ubiquitous in living organisms, an amine racemase (AR) has not yet been discovered despite its synthetic potential for producing chiral amines. Here, we report the creation of an AR based on the serendipitous discovery that amine transaminases (ATAs) can perform stereoinversion of 2-aminobutane. Kinetic modeling revealed that the unexpected off-pathway activity results from stereochemically promiscuous futile cycles due to incomplete stereoselectivity for 2-aminobutane. This finding motivated us to engineer an S-selective ATA through in silico alanine scanning and empirical combinatorial mutations, creating an AR with broad substrate specificity. The resulting AR, carrying double point mutations, enables the racemization of both enantiomers of diverse chiral amines in the presence of a cognate ketone. This strategy may be generally applicable to a wide range of transaminases, paving the way for the development of new-to-nature racemases.

논문정보

형식Research article
게재일2024년 01월 (BRIC 등록일 2024-01-17)
연구진국내 연구진
분야 생명과학 > 생물공학

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