Directed evolution of a cyclodipeptide synthase with new activities <i>via</i> label-free mass spectrometric screening
Songya Zhang, Jing Zhu, Shuai Fan, Wenhao Xie, Zhaoyong Yang, Tong Si
Abstract
) values of 190 possible DKP molecules within a mass window of 115-373 Da. An F186L CDPS mutant was isolated to produce cyclo(l-Phe-l-Val), which is undetectable in the product profile of the wild-type enzyme. This robotic, label-free MS screening approach may be generally applicable to engineering other enzymes with new activities in high throughput.
Topics & Concepts
WorkflowDirected evolutionHigh-throughput screeningChemistryComputational biologyEnzymeATP synthaseComputer scienceCombinatorial chemistryBiochemistryBiologyGeneDatabaseMutantMicrobial Natural Products and BiosynthesisChemical Synthesis and AnalysisBiochemical and Molecular Research