PURIFICATION AND PROPERTIES OF NEURAMINIDASE ISOZYMES IN ARTHROBACTER-UREAFACIENS MUTANT
PURIFICATION AND PROPERTIES OF NEURAMINIDASE ISOZYMES IN ARTHROBACTER-UREAFACIENS MUTANT
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DOI：
10.1093/oxfordjournals.jbchem.a122969
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发表时间：
1989-12-01
影响因子：
2.7
通讯作者：
SUGIMORI, T
中科院分区：
文献类型：
--
作者：
OHTA, Y;TSUKADA, Y;SUGIMORI, T
利用NTG诱变技术从产脲节杆菌（Arthrobacter ureafaciens）中分离到一株组成型产神经氨酸酶突变株（M1057）。产脲菌KMS 3663。通过DEAE-纤维素、亲和层析、硫酸铵沉淀、层析聚焦和Ultrogel AcA 44凝胶过滤，从突变株和亲本菌株中均匀纯化了四种分子种类（L、M1、M2和S）的神经氨酸酶同工酶。L、M1、M2和S同工酶的分子量分别约为88，000、66，000、66，000和52，000。这些同工酶对N-乙酰神经氨酰-α，神经氨酸酶L、M1、M2和S能够水解寡糖、糖蛋白和神经节苷脂，（2-3）-，α，（2-6）-，和α，（2-8）-连接的N-乙酰神经氨酸。在分离的同工酶中，同工酶S对Colominic酸的活性最高。
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.