期刊
ACS CATALYSIS
卷 7, 期 12, 页码 8524-8532出版社
AMER CHEMICAL SOC
DOI: 10.1021/acscatal.7b02954
关键词
biocatalysis; computational enzyme design; molecular dynamics; shortest path map; (retro)aldolases; directed evolution
资金
- Generalitat de Catalunya [2015-FI-B-00165]
- Ramon Areces Foundation
- Spanish MINECO [CTQ2014-52525-P, CTQ2014-59212-P]
- Ramon y Cajal contract [RYC-2014-16846]
- European Community [PCIG14-GA-2013-630978]
- European Research Council (ERC) under European Union [ERC-2015-StG-679001]
Enzymes exist as ensembles of conformations that are important for function. Tuning these populations of conformational states through mutation enables evolution toward additional activities. Here we computationally evaluate the population shifts induced by distal and active site mutations in a family of computationally designed and experimentally optimized retro-aldolases. The conformational landscape of these enzymes was significantly altered during evolution, as preexisting catalytically active conformational substates became major states in the most evolved variants. We further demonstrate that key residues responsible for these substate conversions can be predicted computationally. Significantly, the identified residues coincide with those positions mutated in the laboratory evolution experiments. This study establishes that distal mutations that affect enzyme catalytic activity can be predicted computationally and thus provides the enzyme (re)design field with a rational strategy to determine promising sites for enhancing activity through mutation.
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