Overproduction of 15N-Labeled r-RGD-Hirudin in Pichia pastoris for NMR Studies
| Autor: | Jue Wang, Yanling Zhang, Wei Mo, Shiyun Li, Xingang Liu, Houyan Song, Min Yu, Linsen Dai, Xiaomin Yan |
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| Rok vydání: | 2010 |
| Předmět: |
Magnetic Resonance Spectroscopy
Nitrogen Isotopes Biological activity General Medicine Hirudins Biology biology.organism_classification Biochemistry Pichia Yeast Up-Regulation Pichia pastoris Thrombin Heteronuclear molecule Structural Biology Yield (chemistry) medicine Electrophoresis Polyacrylamide Gel Overproduction Heteronuclear single quantum coherence spectroscopy Biotechnology medicine.drug |
| Zdroj: | Protein & Peptide Letters. 17:1228-1233 |
| ISSN: | 0929-8665 |
| DOI: | 10.2174/092986610792231465 |
| Popis: | The novel recombinant hirudin, r-RGD-hirudin, inhibits thrombin and platelet aggregation. Here, we reported over-expression of (15)N-labeled r-RGD-hirudin by Pichia pastoris in minimal medium. After extensive optimization, the yield of active r-RGD-hirudin reached ≈600 mg/L when the yeast cells were culture in a fermenter. The purified (15)N-labeled r-RGD-hirudin retained full biological activity and was uniformly labeled. Heteronuclear NMR of the (15)N-labeled r-RGD-hirudin was performed for the first time, and all signals in the heteronuclear single quantum coherence (HSQC) spectrum were successfully assigned. |
| Databáze: | OpenAIRE |
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