Open AccessPurification and Characterization of Chloramphenicol Acetyltransferase from Flavobacterium CB60
Author(s) -
Gisela Nolte,
Roland Süßmuth
Publication year - 1987
Publication title -
microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.352
H-Index - 35
eISSN - 1465-2080
pISSN - 1350-0872
DOI - 10.1099/00221287-133-8-2115
Subject(s) - chloramphenicol , chloramphenicol acetyltransferase , flavobacterium , cytophaga , escherichia coli , bacteria , microbiology and biotechnology , biochemistry , enzyme , chemistry , biology , antibiotics , pseudomonas , gene expression , promoter , genetics , gene
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.