[Isolation and purification of the chloramphenicol-acetyltransferase from Y. pestis EV cells with extrachromosomal resistance to the antibiotic by affinity chromatography].
Purification of chloramphenicol acetyltransferase isolated from the cells of Y. pestis EV-R with extrachromosomal resistance to chloramphenicol included ultrafiltration on the microporous membranes Diaflo-XM 300 and biospecific chromatography on columns with cepharose 4B covalently bound with the reduced chloramphenicol. Elution of chloramphenicol acetyltransferase was achieved by means of addition of native chloramphenicol to the buffer solution. Such native chloramphenicol had higher affinity to the enzyme than the reduced antibiotic. As a result the enzyme preparations with 250 times higher purity levels and 70 per cent of the initial activity were obtained. The disc electrophoresis of the sample in polyacrylamide gel produced a single protein zone capable of chloramphenicol acetylation in the presence of Ac approximately CoA.
D000123:Acetyltransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015500:Chloramphenicol O-Acetyltransferase; D002846:Chromatography, Affinity / Q000379:methods*; D004352:Drug Resistance, Microbial; D004591:Electrophoresis, Polyacrylamide Gel; D005111:Extrachromosomal Inheritance; D011815:R Factors; D015010:Yersinia pestis / Q000201:enzymology* / Q000235:genetics
D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase
Vydelenie i ochistka khloramfenikol-atsetiltransferazy iz kletok Y. pestis EV s vnekhromosomnoĭ rezistentnost'iu k antibiotiku metodom affinnoĭ khromatografii.