A method for assay of chloramphenicol acetyltransferase from crude cell extract. 1985

K Datta, and M K Majumdar

This is a method for detection and assay of CAT from Chloramphenicol resistant bacterial isolates. The procedure involved quantitative separation of both chloramphenicol acetate (CmA) and Chloramphenicol (Cm) from assay mixtures by adsorption chromatography on a neutral alumina column. The solvent system--2.5% methanol in benzene--was used for elution of CmA and methanol for Cm. This was followed by either colorimetric estimation of CmA by coupling with N--(1-Naphthyl)--ethylene diamine HCL after zinc hydrocholoric acid reduction or by microbiological assay. In the latter method CmA was deacetylated by enzyme from chicken liver acetone powder. As low as 1 microgram of CmA could be determined by the described microbiological turbidimetric assay method.

UI MeSH Term Description Entries
D008722 Methods A series of steps taken in order to conduct research. Techniques,Methodological Studies,Methodological Study,Procedures,Studies, Methodological,Study, Methodological,Method,Procedure,Technique
D000123 Acetyltransferases Enzymes catalyzing the transfer of an acetyl group, usually from acetyl coenzyme A, to another compound. EC 2.3.1. Acetyltransferase
D001419 Bacteria One of the three domains of life (the others being Eukarya and ARCHAEA), also called Eubacteria. They are unicellular prokaryotic microorganisms which generally possess rigid cell walls, multiply by cell division, and exhibit three principal forms: round or coccal, rodlike or bacillary, and spiral or spirochetal. Bacteria can be classified by their response to OXYGEN: aerobic, anaerobic, or facultatively anaerobic; by the mode by which they obtain their energy: chemotrophy (via chemical reaction) or PHOTOTROPHY (via light reaction); for chemotrophs by their source of chemical energy: CHEMOLITHOTROPHY (from inorganic compounds) or chemoorganotrophy (from organic compounds); and by their source for CARBON; NITROGEN; etc.; HETEROTROPHY (from organic sources) or AUTOTROPHY (from CARBON DIOXIDE). They can also be classified by whether or not they stain (based on the structure of their CELL WALLS) with CRYSTAL VIOLET dye: gram-negative or gram-positive. Eubacteria
D012988 Soil Microbiology The presence of bacteria, viruses, and fungi in the soil. This term is not restricted to pathogenic organisms. Microbiology, Soil
D015500 Chloramphenicol O-Acetyltransferase An enzyme that catalyzes the acetylation of chloramphenicol to yield chloramphenicol 3-acetate. Since chloramphenicol 3-acetate does not bind to bacterial ribosomes and is not an inhibitor of peptidyltransferase, the enzyme is responsible for the naturally occurring chloramphenicol resistance in bacteria. The enzyme, for which variants are known, is found in both gram-negative and gram-positive bacteria. EC 2.3.1.28. CAT Enzyme,Chloramphenicol Acetyltransferase,Chloramphenicol Transacetylase,Acetyltransferase, Chloramphenicol,Chloramphenicol O Acetyltransferase,Enzyme, CAT,O-Acetyltransferase, Chloramphenicol,Transacetylase, Chloramphenicol

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