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How enzymes adapt: lessons from directed evolution
Author(s) -
Arnold Frances Hamilton
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.415.1
Subject(s) - directed evolution , natural selection , directed molecular evolution , molecular evolution , biology , function (biology) , computational biology , genetics , selection (genetic algorithm) , gene , phylogenetics , computer science , mutant , artificial intelligence
When making his case for the key role of natural selection in evolution, Darwin pointed to the enormous phenotypic variation that could be achieved in relatively few generations of artificial selection. Today, artificial selection (or ‘directed evolution’) applied to proteins allows us to observe how readily the functional molecules of life adapt in the face of defined selection pressures. Circumventing our profound ignorance of how sequence encodes function, directed evolution is a powerful approach to generating useful new biological molecules. I will describe our efforts to engineer a cytochrome P450 (CYP102A1 from Bacillus megaterium ) by directed evolution. Properties such as catalytic activity or stability can frequently be enhanced by single amino acid substitutions, and accumulating relatively few beneficial mutations (as little as 1–2% of the sequence) can make very significant changes to enzyme function. I will show how a P450 fatty acid hydroxylase has been converted into a whole family of catalysts for oxidation of small alkanes to carbohydrate synthesis and drug lead diversification. Where natural evolution has gone (e.g. impressive diversification of function in the P450 enzyme superfamily), directed evolution can follow. Even more interesting are the catalysts nature may not care about, but chemists dream of. While yielding useful biocatalysts for chemical synthesis, these studies provide new insights into the mechanisms underlying evolution of natural enzymes.