Purification and characterization of chloramphenicol acetyltransferase from Flavobacterium CB60.
From the highly chloramphenicol-resistant cytophaga-like bacterium Flavobacterium CB60, which can both acetylate chloramphenicol and degrade it in co-metabolism, the chloramphenicol acetyltransferase (CAT) was purified to homogeneity and characterized. The purification included fractional precipitation with ammonium sulphate and two affinity chromatography steps, eluting CAT the first time with 5 mM-chloramphenicol and the second time with a linear gradient (0-10 mM) of chloramphenicol. The purification was 3979-fold. Properties of this CAT were investigated and compared with CATs from other bacteria. Although CAT from Flavobacterium CB60 shares some properties with the enzymes from Escherichia coli and other Gram-negative bacteria--especially with CATII and CATIII--it has distinct properties like extreme heat lability and the inability to produce diacetylchloramphenicol, so that it might be regarded as a new variant.
Nolte
G
G
Institut für Mikrobiologie, Universität Hohenheim, Stuttgart, FRG.
Süssmuth
R
R
D000123:Acetyltransferases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000493:pharmacokinetics; D015500:Chloramphenicol O-Acetyltransferase; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000201:enzymology; D005417:Flavobacterium / Q000201:enzymology*; D007525:Isoelectric Focusing; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D013439:Sulfhydryl Reagents; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase