N-linked glycosylation influences on the catalytic and biochemical properties of Penicillium purpurogenum beta-D-glucuronidase
文献类型：期刊论文
| 作者 | Zou, Shuping2; Xie, Luping1; Liu, Yanli1; Kaleem, Imdad1; Zhang, Guifeng3; Li, Chun1 |
| 刊名 | JOURNAL OF BIOTECHNOLOGY
 |
| 出版日期 | 2012-02-10 |
| 卷号 | 157期号:3页码:399-404 |
| ISSN号 | 0168-1656 |
| 关键词 | beta-D-Glucuronidase Penicillium purpurogenum N-glycosylation Thermal stability Conformation |
| 通讯作者 | Li, C |
| 英文摘要 | To study the influence of N-linked carbohydrate moiety on the catalytic and biochemical properties of glycosylated enzyme, a recombinant beta-D-glucuronidase (PGUS-P) from Penicillium purpurogenum as a model glycoprotein, was deglycosylated with peptide-N-glycosidase F (PNGase-F) under native conditions. The enzymatic deglycosylation procedure resulted in the complete removal of carbohydrate moiety. Compared with the glycosylated PGUS-P, the deglycosylated PGUS-P exhibited 20-70% higher activity (p < 0.05) within pH 6-9, but 15-45% lower activity (p < 0.05) at 45-70 degrees C. The apparent decrease in the thermal stability of the deglycosylated enzyme was reflected by a decrease in the denaturation temperature (T-d) values determined by differential scanning calorimetry (DSC). The removal of N-linked glycans also reduced enzyme's sensitivity to certain metal ions. The deglycosylated PGUS-P displayed lower K-m vaules, but higher k(cat)/K-m ratios than the glycosylated isoform towards glycyrrhizin. The consequent conformational changes were also determined by circular dichroism (CD) and fluorescence spectroscopy which revealed no significant difference in the secondary but a slight dissimilarity between the tertiary structures of both isoforms of PGUS-P. (C) 2011 Elsevier B. V. All rights reserved. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biotechnology & Applied Microbiology |
| 研究领域[WOS] | Biotechnology & Applied Microbiology |
| 关键词[WOS] | SACCHAROMYCES-CEREVISIAE ; MONO-GLUCURONIDE ; PICHIA-PASTORIS ; GLYCYRRHIZIN ; STABILITY ; ENZYME ; DEGLYCOSYLATION ; KINETICS ; BIOSYNTHESIS ; METABOLISM |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000300451800006 |
| 公开日期 | 2013-10-26 |
| 版本 | 出版稿 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/4294]  |
| 专题 | 过程工程研究所_生化工程国家重点实验室 |
| 作者单位 | 1.Beijing Inst Technol, Sch Life Sci & Technol, Beijing 100081, Peoples R China 2.Zhejiang Univ Technol, Inst Bioengn, Hangzhou 310014, Zhejiang, Peoples R China 3.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing 100080, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zou, Shuping,Xie, Luping,Liu, Yanli,et al. N-linked glycosylation influences on the catalytic and biochemical properties of Penicillium purpurogenum beta-D-glucuronidase[J]. JOURNAL OF BIOTECHNOLOGY,2012,157(3):399-404. |
| APA | Zou, Shuping,Xie, Luping,Liu, Yanli,Kaleem, Imdad,Zhang, Guifeng,&Li, Chun.(2012).N-linked glycosylation influences on the catalytic and biochemical properties of Penicillium purpurogenum beta-D-glucuronidase.JOURNAL OF BIOTECHNOLOGY,157(3),399-404. |
| MLA | Zou, Shuping,et al."N-linked glycosylation influences on the catalytic and biochemical properties of Penicillium purpurogenum beta-D-glucuronidase".JOURNAL OF BIOTECHNOLOGY 157.3(2012):399-404. |
入库方式： OAI收割
来源：过程工程研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。