Purification and Characterization of Sarcosine Oxidase of Bacillus Origin(Biological)

概要

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Sarcosine oxidase (EC 1.5.3.1) produced by Bacillus sp. B-0618 was purified by ion exchange chromatography on diethyl aminoethyl-cellulose and gel filtration on Sephadex G-100 and G-150. The molecular weight of the enzyme was estimated to be 42000 by gel filtration on Sephadex G-l 50 and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme exhibited an absorption spectrum characteristic of flavoprotein. The enzyme showed the maximum activity at pH 8.5-9 and was stable at pH 7-10. The pi value was 4.7 as determined by tsoelectric focusing. Although sarcosine is the preferred substrate, the enzyme also oxidized N-methyl-DL-alanine, Af-methyl-L-leucine and N-methyl-DL-valine to lesser extents. The apparent K_m values of sarcosine, N-methyl-DL-alanine, W-methyl-L-leucine and W-methyl-DL-valine were 12.2, 6.8, 106 and 173 mM, respec-tively. The enzyme was inactivated by N-bromosuccinimide, Zn^<2+>, Fe^<3+> and Hg^<2+>, but not by -ethylenediaminetetraacetate, p-chloromercuribenzoate, monoiodoacetate or p-toluenesulfonyl-chloride.
公益社団法人日本薬学会の論文
1987-02-25