Purification of Escherichia coli Chloramphenicol Acetyltransferase by Affinity Chromatography

Abstract

Chloramphenicol acetyltransferase (CATase) was purified by affinity chromatography from Escherichia coli W677/HJR66, an R-factor-bearing mutant. The chloramphenicol aryl nitro group had to be reduced to an amino group prior to its coupling to the Sepharose 4B matrix. The single-step isolation procedure resulted in a 237-fold purification of CATase with over 65% recovery of the enzyme.