N-glycosylation is necessary for enzymatic activity of a beetle (Apriona germari) cellulase

文献类型: 外文期刊

第一作者: Wei, YD

作者: Wei, YD;Lee, SJ;Lee, KS;Gui, ZZ;Yoon, HJ;Kim, I;Je, YH;Guo, XJ;Sohn, HD;Jin, BR

作者机构:

关键词: Apriona germari;baculovirus;beetle;cellulase;N-glycosylation;endoglucanase;enzyme;insect cells

期刊名称:BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ( 影响因子:3.575; 五年影响因子:3.381 )

ISSN: 0006-291X

年卷期: 2005 年 329 卷 1 期

页码:

收录情况: SCI

摘要: We previously reported that the beta-1,4-endoglucanase (EGase) belonging to glycoside hydrolase family 45 cloned from the mulberry longicorn beetle, Apriona germari (Ag-EGase I), is composed of 237 amino acid residues and has a potential N-glycosylation site at 97-100 arnino acid residues (NSTF). We here describe the N-glycosylation and its role for enzymatic activity of the Ag-EGase I. The N-glycosvlation of Ag-EGase I was revealed by the treatment of tunicamycin to the recombinant virus-infected insect Sf9 cells and by endoglycosidase F to the purified recombinant Ag-EGase I, demonstrating that the carbohydrate moieties are not necessary for secretion but essential for Ag-EGase I enzyme activity. To further elucidate the functional role of the N-glycosylation in Ag-EGase I, we have assayed the cellulase enzyme activity in Thr99Gln mutant. Lack of N-glycosylation in Ag-EGase I showed no substantial enzyme activity. This result demonstrates that N-glycosylation at site 97-100 amino acid residues (NSTF) is essential for enzyme activity. (C) 2005 Elscvier Inc. All rialits reserved.

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