A Panel of TrpB Biocatalysts Derived from Tryptophan Synthase through the Transfer of Mutations that Mimic Allosteric Activation
| Autor: | Sabine Brinkmann-Chen, Javier Murciano-Calles, Frances H. Arnold, Andrew R. Buller, David K. Romney |
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| Rok vydání: | 2016 |
| Předmět: |
Protein subunit
Allosteric regulation Tryptophan synthase Plasma protein binding Protein Engineering 010402 general chemistry 01 natural sciences Article Catalysis Allosteric Regulation Tryptophan Synthase chemistry.chemical_classification biology 010405 organic chemistry General Chemistry Protein engineering General Medicine biology.organism_classification 0104 chemical sciences Pyrococcus furiosus Kinetics Protein Subunits Enzyme chemistry Biochemistry Biocatalysis Mutagenesis Site-Directed biology.protein Function (biology) Protein Binding |
| Zdroj: | Angewandte Chemie. 128:11749-11753 |
| ISSN: | 0044-8249 |
| DOI: | 10.1002/ange.201606242 |
| Popis: | Naturally occurring enzyme homologs 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 homolog. We demonstrate that a small set of mutations of the tryptophan synthase β-subunit (TrpB) from Pyrococcus furiosus, which mimic the activation afforded by binding of the α-subunit, has a similar activating effect in TrpB homologs 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 this collection of stand-alone enzymes, we identified a new catalyst that displays a remarkably broad activity profile in the synthesis of 5-substituted tryptophans, a biologically important class of compounds. This investigation demonstrates how allosteric activation can be recapitulated throughout a protein family to efficiently explore natural sequence diversity for desirable biocatalytic transformations. |
| Databáze: | OpenAIRE |
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