Isolation and characterisation of sialidase from a strain of Streptococcus oralis

被引:34
作者
Byers, HL [1 ]
Tarelli, E [1 ]
Homer, KA [1 ]
Beighton, D [1 ]
机构
[1] GKT Dent Inst, Joint Microbiol Res Unit, London SE5 9RW, England
关键词
D O I
10.1099/0022-1317-49-3-235
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Streptococcus oralis, the most virulent of the viridans streptococci, produces a sialidase and this exo-glycosidase has been implicated in the disease process of a number of pathogens. The sialidase of S. oralis strain AR3 was purified in order to understand the characteristics of this putative virulence determinant. The enzyme isolated as a high mol. wt aggregate (c. 325 kDa) was purified 4520-fold from late exponential phase cultures by a combination of ultrafiltration, ammonium sulphate precipitation, ion-exchange and gel filtration chromatography. The sialidase component had a mol. wt of 144 kDa as determined by SDS-PAGE analysis. The purified sialidase released N-acetylneuraminic acid from a range of sialoglycoconjugate including human al-acid glycoprotein, bovine submaxillary mucin, colominic acid and sialyl-alpha 2,3- and sialyl-alpha 2,6-lactose. Also, N-glycolylneuraminic acid was cleaved from bovine submaxillary mucin. The sialidase had a K-m of 11.8 mu M for alpha(1)-acid glycoprotein, was active over a broad pH range with a pH optimum of 6.0 and cleaved alpha 2,3-, alpha 2,6- and alpha 2-8-sialyl glycosidic linkages with a marked preference for alpha 2,3-linkages. The enzyme was competitively inhibited by the sialic acid derivative, 2,3-dehydro-N-acetylneuraminic acid, with a K-IC of 1.2 mu M. The characteristics of the purified sialidase would support a nutritional role for this enzyme that may be significant in the proliferation of this organism in the oral cavity and at extra-oral sites in association with life-threatening infections.
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页码:235 / 244
页数:10
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