title: Purification and properties of neuraminidase isozymes in Arthrobacter ureafaciens mutant.
abstract: An Arthrobacter ureafaciens mutant (M1057) capable of producing neuraminidase constitutively was isolated by NTG mutagenesis from A. ureafaciens KMS 3663. Four molecular species (L, M1, M2, and S) of neuraminidase isozymes were homogeneously purified from the mutant and parent strains by means of DEAE-cellulose, affinity chromatography, ammonium sulfate precipitation, chromatofocusing, and Ultrogel AcA44 gel filtration. The molecular weights of L, M1, M2, and S isozymes were shown to be approximately 88,000, 66,000, 66,000, and 52,000, respectively. The optimal pHs and Km values of these isozymes for N-acetylneuraminosyl-alpha,(2-6)-lactose were 4.5-5.5 and 0.6-0.8 mM. Neuraminidase L, M1, M2, and S were able to hydrolyze oligosaccharides, glycoproteins and gangliosides containing alpha,(2-3)-, alpha,(2-6)-, and alpha,(2-8)-linked N-acetylneuraminic acid. Among these isozymes isolated, isozyme S was most active on colominic acid.
authors.affiliation: Kyoto Research Laboratories, Marukin Shoyu Co., Ltd.
authors.forename: Y
authors.initials: Y
authors.lastname: Ohta
authors.forename: Y
authors.initials: Y
authors.lastname: Tsukada
authors.forename: T
authors.initials: T
authors.lastname: Sugimori
mesh_terms: D001173:Arthrobacter / Q000201:enzymology* / Q000235:genetics; D004355:Drug Stability; D004790:Enzyme Induction; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D009154:Mutation; D009439:Neuraminidase / Q000302:isolation & purification* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D007527:Isoenzymes; D009439:Neuraminidase