从洛氏拟杆菌培养基中纯化神经氨酸酶及其性质

Purification and properties of neuraminidase from the culture medium of Bacteroides loescheii.

作者信息

Takeshita T, Takagaki M, Amano A, Murakami Y, Nagata H, Shizukuishi S

机构信息

Osaka University Faculty of Dentistry, Japan.

出版信息

Oral Microbiol Immunol. 1991 Oct;6(5):316-9. doi: 10.1111/j.1399-302x.1991.tb00501.x.

DOI:10.1111/j.1399-302x.1991.tb00501.x

PMID:1820573

Abstract

Neuraminidase was purified from the culture medium of Bacteroides loescheii ATCC 15930 by ultrafiltration followed by a DEAE-Sephacel anion exchange chromatography, fast-protein liquid chromatography using Mono P column and high-performance liquid chromatography using a Shim-pack Diol-300 column. The molecular weight of the purified enzyme was measured to be approximately 87 kDa and the optimal pH was at 4.8. Although the enzyme was able to hydrolyze the substrates with alpha,2-3, alpha,2-6, and alpha,2-8 linkages of N-acetylneuraminic acid, the rate of hydrolysis of N-acetylneuraminosyl-alpha,2-3-lactose was greater than that of the alpha,2-6-isomer.

摘要

通过超滤，随后进行DEAE-葡聚糖凝胶阴离子交换色谱、使用Mono P柱的快速蛋白质液相色谱以及使用Shim-pack Diol-300柱的高效液相色谱，从洛氏拟杆菌ATCC 15930的培养基中纯化出神经氨酸酶。测得纯化酶的分子量约为87 kDa，最适pH为4.8。尽管该酶能够水解具有N-乙酰神经氨酸α,2-3、α,2-6和α,2-8连接的底物，但N-乙酰神经氨糖基-α,2-3-乳糖的水解速率大于α,2-6-异构体的水解速率。