Literature DB >> 33026156

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

Shunsuke Kato1, Akira Onoda1,2, Naomasa Taniguchi1, Ulrich Schwaneberg3, Takashi Hayashi1.   

Abstract

Directed evolution of Cp*RhIII -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*RhIII -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.
© 2020 The Authors. ChemBioChem published by Wiley-VCH GmbH.

Entities:  

Keywords:  C−H bond functionalization; artificial metalloenzymes; biohybrid catalysts; directed evolution; rhodium

Year:  2020        PMID： 33026156      PMCID： PMC7894531          DOI： 10.1002/cbic.202000681

Source DB:  PubMed          Journal:  Chembiochem        ISSN： 1439-4227            Impact factor:   3.164


  40 in total

1.  Controlling the enantioselectivity of enzymes by directed evolution: practical and theoretical ramifications.

Authors:  Manfred T Reetz
Journal:  Proc Natl Acad Sci U S A       Date:  2004-04-12       Impact factor: 11.205

2.  Single-step purification and immobilization of MBP-phytase fusion on starch agar beads: application in dephytination of soy milk.

Authors:  Mrudula Vasudevan Ushasree; Paramasamy Gunasekaran; Ashok Pandey
Journal:  Appl Biochem Biotechnol       Date:  2012-02-29       Impact factor: 2.926

3.  Chemomimetic biocatalysis: exploiting the synthetic potential of cofactor-dependent enzymes to create new catalysts.

Authors:  Christopher K Prier; Frances H Arnold
Journal:  J Am Chem Soc       Date:  2015-11-03       Impact factor: 15.419

4.  Genetic Engineering of an Artificial Metalloenzyme for Transfer Hydrogenation of a Self-Immolative Substrate in Escherichia coli's Periplasm.

Authors:  Jingming Zhao; Johannes G Rebelein; Hendrik Mallin; Christian Trindler; Michela M Pellizzoni; Thomas R Ward
Journal:  J Am Chem Soc       Date:  2018-10-03       Impact factor: 15.419

Review 5.  Towards the Evolution of Artificial Metalloenzymes-A Protein Engineer's Perspective.

Authors:  Ulrich Markel; Daniel F Sauer; Johannes Schiffels; Jun Okuda; Ulrich Schwaneberg
Journal:  Angew Chem Int Ed Engl       Date:  2019-02-11       Impact factor: 15.336

6.  Rhodium(III)-catalyzed heterocycle synthesis using an internal oxidant: improved reactivity and mechanistic studies.

Authors:  Nicolas Guimond; Serge I Gorelsky; Keith Fagnou
Journal:  J Am Chem Soc       Date:  2011-03-31       Impact factor: 15.419

7.  A robust protein host for anchoring chelating ligands and organocatalysts.

Authors:  Manfred T Reetz; Martin Rentzsch; Andreas Pletsch; Andreas Taglieber; Frank Hollmann; Régis J G Mondière; Norbert Dickmann; Birte Höcker; Simona Cerrone; Michaela C Haeger; Reinhard Sterner
Journal:  Chembiochem       Date:  2008-03-03       Impact factor: 3.164

8.  Incorporation of a Cp*Rh(III)-dithiophosphate Cofactor with Latent Activity into a Protein Scaffold Generates a Biohybrid Catalyst Promoting C(sp2)-H Bond Functionalization.

Authors:  Shunsuke Kato; Akira Onoda; Alexander R Grimm; Kengo Tachikawa; Ulrich Schwaneberg; Takashi Hayashi
Journal:  Inorg Chem       Date:  2020-09-11       Impact factor: 5.165

9.  Neutralizing the detrimental effect of glutathione on precious metal catalysts.

Authors:  Yvonne M Wilson; Marc Dürrenberger; Elisa S Nogueira; Thomas R Ward
Journal:  J Am Chem Soc       Date:  2014-06-11       Impact factor: 15.419

10.  Expression of proteins in Escherichia coli as fusions with maltose-binding protein to rescue non-expressed targets in a high-throughput protein-expression and purification pipeline.

Authors:  Stephen N Hewitt; Ryan Choi; Angela Kelley; Gregory J Crowther; Alberto J Napuli; Wesley C Van Voorhis
Journal:  Acta Crystallogr Sect F Struct Biol Cryst Commun       Date:  2011-08-13
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  1 in total

1.  Directed Evolution of a Cp*RhIII -Linked Biohybrid Catalyst Based on a Screening Platform with Affinity Purification.

Authors:  Shunsuke Kato; Akira Onoda; Naomasa Taniguchi; Ulrich Schwaneberg; Takashi Hayashi
Journal:  Chembiochem       Date:  2020-11-09       Impact factor: 3.164
  1 in total

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