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

Murciano-Calles, Javier and Romney, David K. and Brinkmann-Chen, Sabine and Buller, Andrew R. and Arnold, Frances H. (2016) A Panel of TrpB Biocatalysts Derived from Tryptophan Synthase through the Transfer of Mutations that Mimic Allosteric Activation. Angewandte Chemie International Edition, 55 (38). pp. 11577-11581. ISSN 1433-7851. PMCID PMC5014574. doi:10.1002/anie.201606242.

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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.

Item Type:Article
Related URLs:
URLURL TypeDescription CentralArticle
Murciano-Calles, Javier0000-0002-8667-1651
Romney, David K.0000-0003-0498-7597
Brinkmann-Chen, Sabine0000-0002-5419-4192
Buller, Andrew R.0000-0002-9635-4844
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. Version of record online: 11 August 2016; Manuscript Received: 27 June 2016. The authors thank Dr. Jackson Cahn for the data on frequency of amino acids in each position of TrpB and Dr. Jennifer Kan for helpful discussions and comments on the manuscript. J.M.-C. gratefully acknowledges support from the Alfonso Martín Escudero Foundation. This work was funded through the Jacobs Institute for Molecular Engineering for Medicine and Ruth Kirschstein NIH Postdoctoral Fellowships F32GM117635 (to D.K.R) and F32G110851 (to A.R.B.).
Group:Jacobs Institute for Molecular Engineering for Medicine
Funding AgencyGrant Number
Alfonso Martín Escudero FoundationUNSPECIFIED
Jacobs Institute for Molecular Engineering for MedicineUNSPECIFIED
NIH Postdoctoral FellowshipF32GM117635
NIH Postdoctoral FellowshipF32G110851
Subject Keywords:allostery; enzymes; protein engineering; tryptophan synthase
Issue or Number:38
PubMed Central ID:PMC5014574
Record Number:CaltechAUTHORS:20160812-092420412
Persistent URL:
Official Citation:J. Murciano-Calles, D. K. Romney, S. Brinkmann-Chen, A. R. Buller, F. H. Arnold, Angew. Chem. Int. Ed. 2016, 55, 11577
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:69586
Deposited By: Tony Diaz
Deposited On:12 Aug 2016 16:35
Last Modified:25 Apr 2022 22:46

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