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

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Title: Directed Evolution of a Cp*Rh-III-Linked Biohybrid Catalyst Based on a Screening Platform with Affinity Purification
Authors: Kato, Shunsuke Browse this author
Onoda, Akira Browse this author →KAKEN DB
Taniguchi, Naomasa Browse this author
Schwaneberg, Ulrich Browse this author
Hayashi, Takashi Browse this author →KAKEN DB
Keywords: artificial metalloenzymes
biohybrid catalysts
C−H bond functionalization
directed evolution
rhodium
Issue Date: 15-Feb-2021
Publisher: Wiley-Blackwell
Journal Title: Chembiochem
Volume: 22
Issue: 4
Start Page: 679
End Page: 685
Publisher DOI: 10.1002/cbic.202000681
Abstract: Directed evolution of Cp*Rh-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 Cp*Rh-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.
Rights: https://creativecommons.org/licenses/by/4.0/
Type: article
URI: http://hdl.handle.net/2115/80549
Appears in Collections:環境科学院・地球環境科学研究院 (Graduate School of Environmental Science / Faculty of Environmental Earth Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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