Purification and Characterization of Sarcosine Oxidase of Streptomyces Origin(Biological,Chemical)

概要

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Sarcosine oxidase (EC 1.5.3.1) produced by Streptomyces sp. KB210-8SY was purified by ion exchange chromatography on DEAE-sepharose CL-6B, affinity chromatography on sarcosyl-AH-sepharose 4B and gel filtration on sephadex G-150 to electrophoretic homogeneity. The molecular weight of the enzyme was estimated to be 44000 by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and gel filtration on Sephadex G-150. The enzyme showed the maximum activity at pH 8 and was stable at pH 7-9. In addition to sarcosine, the enzyme oxidized N-methyl-L-leucine, N-methyl-DL-alanine and N-methyl-DL-valine to lesser extents. The apparent K_m values for sarcosine, N-methyl-L-leucine, N-methyl-DL-alanine and Nmethyl-DL-valine were 0.91, 0.58, 1.6 and 6.7 mM, respectively. The enzyme was inactivated by N-bromosuccinimide, hydroxylamine hydrochloride, SDS, Zn^<2+>, Ni^<2+> and Hg^<2+> but not by ethylenediaminetetraacetate, p-chloromercuribenzoate or p-toluenesulfonyl chloride. The taxonomic characteristics of strain K13210-8SY are closely related to those of Strepiomyces flavovirens and S. misakiensis.
公益社団法人日本薬学会の論文
1987-10-25