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Revealing Additional Stereocomplementary Pairs of Old Yellow Enzymes by Rational Transfer of Engineered Residues
Author(s) -
Nett Nathalie,
Duewel Sabine,
Richter Alexandra Annelis,
Hoebenreich Sabrina
Publication year - 2017
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201600688
Subject(s) - protein engineering , enzyme , rational design , directed evolution , computational biology , protein design , sampling (signal processing) , biochemistry , directed molecular evolution , stereochemistry , biology , chemistry , combinatorial chemistry , genetics , computer science , protein structure , gene , filter (signal processing) , mutant , computer vision
Every year numerous protein engineering and directed evolution studies are published, increasing the knowledge that could be used by protein engineers. Here we test a protein engineering strategy that allows quick access to improved biocatalysts with very little screening effort. Conceptually it is assumed that engineered residues previously identified by rational and random methods induce similar improvements when transferred to family members. In an application to ene‐reductases from the Old Yellow Enzyme (OYE) family, the newly created variants were tested with three compounds, revealing more stereocomplementary OYE pairs with potent turnover frequencies (up to 660 h −1 ) and excellent stereoselectivities (up to >99 %). Although systematic prediction of absolute enantioselectivity of OYE variants remains a challenge, “scaffold sampling” was confirmed as a promising addition to protein engineers' collection of strategies.