Fluorescence-activated cell sorting and directed evolution of α-N-acetylgalactosaminidases using a quenched activity-based probe (qABP)

Chem Commun (Camb). 2013 Aug 21;49(65):7237-9. doi: 10.1039/c3cc42836b. Epub 2013 Jul 10.

Abstract

An α-N-acetylgalactosamine-containing quenched activity-based probe (qABP) was designed and successfully synthesized, and it was subsequently used in directed enzyme evolution experiments aided by fluorescence-activated cell sorting (FACS) for the discovery of an α-N-acetylgalactosaminidase variant with improved catalytic activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Directed Molecular Evolution*
  • Flavobacteriaceae / enzymology
  • Flow Cytometry
  • Fluorescent Dyes / chemistry*
  • Kinetics
  • Mutation
  • alpha-N-Acetylgalactosaminidase / genetics
  • alpha-N-Acetylgalactosaminidase / metabolism*

Substances

  • Fluorescent Dyes
  • alpha-N-Acetylgalactosaminidase