Isolation and properties of a puromycin acetyltransferase from puromycin-producing streptomyces alboniger

Abstract

Puromycin 2″-N-acetyltransferase was isolated from cell extracts of puromycin-producing Streptomyces alboniger Kcc S-0309 by ammonium sulfate fractionation, he t treatment to eliminate contaminant proteins and chromatography on Deaf-Toyope rl 6505. After Page (polyacrylamide gel electrophoresis) of the final fraction, a single protein band corresponding to puromycin 2″-N-acetyltransferase was detected. The molecular weight of the enzyme determined by Sds-Page and Sephadex G-150 chromatography was about 21,000 and 85,000, respectively, suggesting that the enzyme consisted of four subunits. The isoelectric point and the optimum pH for reaction were 6.2 and 7.7, respectively. The Km values for puromycin and acetyl coenzyme A were 40 μM and 67 μM, respectively. The enzyme was thermostable up to 70°C for 12 minutes. © 1985, JAPAN ANTIBIOTICS RESEARCH ASSOCIATION. All rights reserved.