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Citation
Tags
HERO ID
6546662
Reference Type
Journal Article
Title
Automated Design of Efficient and Functionally Diverse Enzyme Repertoires
Author(s)
Khersonsky, O; Lipsh, R; Avizemer, Ziv; Ashani, Y; Goldsmith, M; Leader, H; Dym, O; Rogotner, S; Trudeau, DL; Prilusky, J; Amengual-Rigo, Pep; Guallar, V; Tawfik, D; Fleishman, SJ
Year
2018
Is Peer Reviewed?
1
Journal
Molecular Cell
ISSN:
1097-2765
EISSN:
1097-4164
Volume
72
Issue
1
Page Numbers
178-+
Language
English
PMID
30270109
DOI
10.1016/j.molcel.2018.08.033
Web of Science Id
WOS:000446317300017
Abstract
Substantial improvements in enzyme activity demand multiple mutations at spatially proximal positions in the active site. Such mutations, however, often exhibit unpredictable epistatic (non-additive) effects on activity. Here we describe FuncLib, an automated method for designing multipoint mutations at enzyme active sites using phylogenetic analysis and Rosetta design calculations. We applied FuncLib to two unrelated enzymes, a phosphotriesterase and an acetyl-CoA synthetase. All designs were active, and most showed activity profiles that significantly differed from the wild-type and from one another. Several dozen designs with only 3-6 active-site mutations exhibited 10- to 4,000-fold higher efficiencies with a range of alternative substrates, including hydrolysis of the toxic organophosphate nerve agents soman and cyclosarin and synthesis of butyryl-CoA. FuncLib is implemented as a web server (http://FuncLib.weizmann.ac.il); it circumvents iterative, high-throughput experimental screens and opens the way to designing highly efficient and diverse catalytic repertoires.
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