دورية أكاديمية
Glycosylation analysis of recombinant neutral protease I from Aspergillus oryzae expressed in Pichia pastoris.
| العنوان: | Glycosylation analysis of recombinant neutral protease I from Aspergillus oryzae expressed in Pichia pastoris. |
|---|---|
| المؤلفون: | Lei D, Xu Y, He Q, Pang Y, Chen B, Xiong L, Li Y |
| المصدر: | Biotechnology letters [Biotechnol Lett] 2013 Dec; Vol. 35 (12), pp. 2121-7. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Kluwer Academic Publishers Country of Publication: Netherlands NLM ID: 8008051 Publication Model: Print Cited Medium: Internet ISSN: 1573-6776 (Electronic) Linking ISSN: 01415492 NLM ISO Abbreviation: Biotechnol Lett Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 1999- : Dordrecht : Kluwer Academic Publishers Original Publication: [Kew, Eng., Science and Technology Letters] |
| مواضيع طبية MeSH: | Aspergillus oryzae/*enzymology , Fungal Proteins/*chemistry , Metalloendopeptidases/*chemistry , Pichia/*genetics , Recombinant Proteins/*chemistry, Amino Acid Sequence ; Aspergillus oryzae/genetics ; Chromatography, Liquid ; Fungal Proteins/genetics ; Fungal Proteins/metabolism ; Glycosylation ; Metalloendopeptidases/genetics ; Metalloendopeptidases/metabolism ; Models, Molecular ; Molecular Sequence Data ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Tandem Mass Spectrometry |
| مستخلص: | Neutral protease I from Aspergillus oryzae 3.042 was expressed in Pichia pastoris and its N-glycosylation properties were analyzed. After purification by nickel-affinity chromatography column, the recombinant neutral protease (rNPI) was confirmed to be N-glycosylated by periodicacid/Schiff's base staining and Endo H digestion. Moreover, the deglycosylated protein's molecular weight decreased to 43.3 kDa from 54.5 kDa analyzed by SDS-PAGE and MALDI-TOF-MS, and the hyperglycosylation extent was 21 %. The N-glycosylation site of rNPI was analyzed by nano LC-MS/MS after digesting by trypsin and Glu-C, and the unique potential site Asn41 of mature peptide was found to be glycosylated. Homology modeling of the 3D structure of rNPI indicated that the attached N-glycans hardly affected neutral protease's activity due to the great distance away from the active site of the enzyme. |
| المشرفين على المادة: | 0 (Fungal Proteins) 0 (Recombinant Proteins) EC 3.4.24.- (Metalloendopeptidases) |
| تواريخ الأحداث: | Date Created: 20131001 Date Completed: 20140528 Latest Revision: 20131217 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1007/s10529-013-1314-3 |
| PMID: | 24078118 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Full Text Finder | تدمد: | 1573-6776 |
|---|---|
| DOI: | 10.1007/s10529-013-1314-3 |