Sarcosine Dehydrogenase from Pseudomonas putida: Purification and Some Properties

Abstract

A sarcosine dehydrogenase was purified to homogeneity from cell free extract of Pseudomonas putida aerobically grown in a medium containing creatinine or betaine as the carbon and nitrogen sources. The enzyme catalyzed dehydrogenation of N-methyl derivatives of some amino acids but was inert toward dimethylglycine, betaine and choline. Phenazine me-thosulfate, 2, 6-dichlorophenol indophenol, methylene blue, meldora blue, nile blue and potassium ferricyanide served as electron carriers. The maximal activity was observed at pH 8.0 ~ 9.0. The Km and values for sarcosine were 29 mM and 1.2 µmol/min/mg, respectively. The molecular weight was estimated to be about 170,000, presumably composed of four subunits. Spectrophotometric and fluorometric analyses indicated that the enzyme was a flavoprotein. © 1979, Japan Society for Bioscience, Biotechnology, and Agrochemistry. All rights reserved.