Academic Journal
A Panel of TrpB Biocatalysts Derived from Tryptophan Synthase through the Transfer of Mutations that Mimic Allosteric Activation
| العنوان: | A Panel of TrpB Biocatalysts Derived from Tryptophan Synthase through the Transfer of Mutations that Mimic Allosteric Activation |
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| المؤلفون: | Murciano-Calles, Javier, Romney, David K., Brinkmann-Chen, Sabine, Buller, Andrew R., Arnold, Frances H. |
| المصدر: | Angewandte Chemie International Edition, 55(38), 11577-11581, (2016-09-12) |
| بيانات النشر: | Wiley |
| سنة النشر: | 2016 |
| المجموعة: | Caltech Authors (California Institute of Technology) |
| مصطلحات موضوعية: | allostery, enzymes, protein engineering, tryptophan synthase |
| الوصف: | 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. ; © 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 MartÃn 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.). ; Accepted Version - nihms804501.pdf Supplemental Material - anie201606242-sup-0001-misc_information.pdf |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | unknown |
| Relation: | https://doi.org/10.1002/anie.201606242; https://www.ncbi.nlm.nih.gov/pmc/PMC5014574; eprintid:69586 |
| DOI: | 10.1002/anie.201606242 |
| الاتاحة: | https://doi.org/10.1002/anie.201606242 https://www.ncbi.nlm.nih.gov/pmc/PMC5014574 |
| Rights: | info:eu-repo/semantics/openAccess ; Other |
| رقم الانضمام: | edsbas.DF0FF690 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1002/anie.201606242 |
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