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| Content Provider | Springer Nature Link |
|---|---|
| Author | Du, Cuihong Yi, Xiaoping Zhang, Yuanxing |
| Copyright Year | 2010 |
| Abstract | Endostatin, a 20 kDa C-terminal fragment of collagen XVIII, is a specific inhibitor of endothelial cell proliferation and angiogenesis. In the present study, we produced soluble and biologically active recombinant human endostatin (rhEndostatin) in Escherichia coli by expressing via fusion with solubility-promoting peptides and optimizing the expression conditions. The rhEndostatin was expressed via fusion with glutathione S-transferase (GST) and NusA protein, respectively. It revealed that NusA protein enhanced the production of soluble rhEndostatin; but GST didn’t. By optimizing the expression conditions, the production of soluble NusA-rhEndostatin fusion protein was about 50% of total cellular proteins and about 90% of the products appeared in the cellular supernatant fraction. The soluble NusA-rhEndostatin fusion protein was purified by one-step hydrophobic interaction chromatography and NusA was removed by thrombin. Then rhEndostatin was purified by affinity chromatography and gel filtration chromatography. As a result, a simple and economical purification procedure for rhEndostatin isolation was obtained. The biological activity of the rhEndostatin was demonstrated in vitro using a human vascular endothelial cells (HuVECs) proliferation assay. Our study provides a feasible and convenient approach to produce soluble and biologically active rhEndostatin. |
| Starting Page | 229 |
| Ending Page | 235 |
| Page Count | 7 |
| File Format | |
| ISSN | 12268372 |
| Journal | Biotechnology and Bioprocess Engineering |
| Volume Number | 15 |
| Issue Number | 2 |
| e-ISSN | 19763816 |
| Language | English |
| Publisher | The Korean Society for Biotechnology and Bioengineering |
| Publisher Date | 2010-05-09 |
| Publisher Place | Heidelberg |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Human endostatin soluble expression Escherichia coli purification procedure Industrial and Production Engineering Biotechnology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Bioengineering Biomedical Engineering Applied Microbiology and Biotechnology Biotechnology |
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