Cell-free protein engineering of Old Yellow Enzyme 1 from Saccharomyces pastorianus

[摘要] In protein engineering, cell-free transcription/translation of linear mutagenic DNA templates can tremendously accelerate and simplify the screening of enzyme variants. Using the RApid Parallel Protein EvaluatoR (RAPPER) protocol, we have evaluated the impact of amino acid substitutions and loop truncations on substrate specificity and stereoselectivity of Old Yellow Enzyme 1 from Saccharomyces pastorianus. Our study demonstrates the benefit of systematically assessing amino add variations including substrate profiling to explore sequence-function space. (C) 2016 Elsevier Ltd. All rights reserved.

[发布日期] 2016-11-17

[效力级别]  Proceedings Paper

[关键词] Old Yellow Enzyme;Flavoenzyme;In vitro transcription/translation;RAPPER;Mutagenesis