Directed Evolution of a Cp*Rh-III-Linked Biohybrid Catalyst Based on a Screening Platform with Affinity Purification

Kato, Shunsuke; Onoda, Akira; Taniguchi, Naomasa; Schwaneberg, Ulrich; Hayashi, Takashi

Directed Evolution of a Cp*Rh-III-Linked Biohybrid Catalyst Based on a Screening Platform with Affinity Purification

Kato, Shunsuke; Onoda, Akira; Taniguchi, Naomasa et al.

Chembiochem, 2021, 22(4), 679-685

Permalink : https://hdl.handle.net/2115/80549

このアイテムのアクセス数:417件（2026-08-12 06:02 集計）

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https://doi.org/10.1002/cbic.202000681			54

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アクセス権	open access
URI	https://hdl.handle.net/2115/80549
タイトル	Directed Evolution of a Cp*Rh-III-Linked Biohybrid Catalyst Based on a Screening Platform with Affinity Purification
著者	Kato, Shunsuke ; Onoda, Akira ; Taniguchi, Naomasa ; Schwaneberg, Ulrich ; Hayashi, Takashi
著者名 Kato, Shunsuke
e-Rad_Researcher 60366424 著者名 Onoda, Akira
著者名 Taniguchi, Naomasa
著者名 Schwaneberg, Ulrich
e-Rad_Researcher 20222226 著者名 Hayashi, Takashi
言語	英語
キーワード	artificial metalloenzymes
	biohybrid catalysts
	C−H bond functionalization
	directed evolution
	rhodium
発行日	2021-02-15
出版者	Wiley-Blackwell
収録物名	Chembiochem
巻(号)	22(4)
ページ	679-685
抄録	Directed evolution of CpRh-III-linked nitrobindin (NB), a biohybrid catalyst, was performed based on an in vitro screening approach. A key aspect of this effort was the establishment of a high-throughput screening (HTS) platform that involves an affinity purification step employing a starch-agarose resin for a maltose binding protein (MBP) tag. The HTS platform enables efficient preparation of the purified MBP-tagged biohybrid catalysts in a 96-well format and eliminates background influence of the host E. coli cells. Three rounds of directed evolution and screening of more than 4000 clones yielded a CpRh-III-linked NB(T98H/L100K/K127E) variant with a 4.9-fold enhanced activity for the cycloaddition of acetophenone oximes with alkynes. It is confirmed that this HTS platform for directed evolution provides an efficient strategy for generating highly active biohybrid catalysts incorporating a synthetic metal cofactor.
資源タイプ	学術雑誌論文
出版タイプ	Version of Record
権利情報	Creative Commons Attribution 4.0 International
PISSN	1439-4227
NCID	AA11497584
関連情報 (isIdenticalTo)	出版社版DOI https://doi.org/10.1002/cbic.202000681