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Angew Chem Int Ed Engl. 2016 Sep 12;55(38):11577-81. doi: 10.1002/anie.201606242. Epub 2016 Aug 11.

A Panel of TrpB Biocatalysts Derived from Tryptophan Synthase through the Transfer of Mutations that Mimic Allosteric Activation.

Author information

1
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, 91125, USA.
2
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, 91125, USA. frances@cheme.caltech.edu.

Abstract

Naturally occurring enzyme homologues often display highly divergent activity with non-natural substrates. Exploiting this diversity with enzymes engineered for new or altered function, however, is laborious because the engineering must be replicated for each homologue. A small set of mutations of the tryptophan synthase β-subunit (TrpB) from Pyrococcus furiosus, which mimics the activation afforded by binding of the α-subunit, was demonstrated to have a similar activating effect in different TrpB homologues with as little as 57 % sequence identity. Kinetic and spectroscopic analyses indicate that the mutations function through the same mechanism: mimicry of α-subunit binding. From these enzymes, we identified a new TrpB catalyst that displays a remarkably broad activity profile in the synthesis of 5-substituted tryptophans. This demonstrates that allosteric activation can be recapitulated throughout a protein family to explore natural sequence diversity for desirable biocatalytic transformations.

KEYWORDS:

allostery; enzymes; protein engineering; tryptophan synthase

PMID:
27510733
PMCID:
PMC5014574
DOI:
10.1002/anie.201606242
[Indexed for MEDLINE]
Free PMC Article

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