Purification and Properties of Neuraminidase Isozymes in Arthrobacter ureafaciens Mutant

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Abstract
An Arthrobacter ureafaciens mutant (M1057) capable of producing neuraminidase con-stitutively was isolated by NTG mutagenesis from A. ureafaciens KMS 3663. Four molecular species (L, Ml, M2, and S) of neuraminidase isozymes were homogeneously purified from the mutant and parent strains by means of DEAE-cellulose, affinity chro-matography, ammonium sulfate precipitation, chromatofocusing, and Ultrogel AcA44 gel filtration. The molecular weights of L, Ml, 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 iV-acetylneuraminosyl-a, (2–6)-lactose were 4.5–5.5 and 0.6–0.8 mM. Neuraminidase L, Ml, M2, and S were able to hydrolyze oligosaccharides, glycoproteins and gangliosides containing a, (2–3)-, a, (2–6)-, and a, (2–8)-linked N-acetylneuraminic acid. Among these isozymes isolated, isozyme S was most active on colominic acid.