Human proinsulin C-peptide from a precursor overexpressed in Pichia pastoris
文献类型：期刊论文
| 作者 | Huang, YB; Li, J; Gao, X; Sun, JR; Lu, Y; Feng, T; Fei, J; Cui, DF; Xia, QC; Ren, J |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 2006 |
| 卷号 | 38期号:8页码:586-592 |
| 关键词 | proinsulin C-peptide Pichia pastoris |
| 通讯作者 | Ren, J (reprint author), Shanghai Newsummit Biopharma Co, Shanghai 200233, Peoples R China.,renjun@newsummibio.com ; yszhang@sibs.ac.cn |
| 英文摘要 | In this article we report the production of human proinsulin C-peptide with 31 amino acid residues from a precursor overexpressed in Pichia pastoris. A C-peptide precursor expression plasmid containing nine C-peptide genes in tandem was constructed and used to transform P. pastoris. Transformants with a high copy number of the C-peptide precursor gene integrated into the chromosome of P. pastoris were selected. In high-density fermentation in a 300 liter fermentor using a simple culture medium composed mainly of salt and methanol, the C-peptide precursor was overexpressed to a level of 2.28 g per liter. A simple procedure was established to purify the expression product from the culture medium. The purified C-peptide precursor was converted into C-peptide by trypsin and carboxypeptidase B joint digestion. The yield of C-peptide with a purity of 96% was 730 mg per liter of culture. The purified C-peptide was characterized by mass spectrometry, N- and C-terminal amino acid sequencing, and sodium dodecylsulfate-polyacrylamide gel electrophoresis. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | MULTIPLE JOINED GENES ; ESCHERICHIA-COLI ; DIABETIC-RATS ; DYSFUNCTION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000239784200010 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/1750]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Huang, YB,Li, J,Gao, X,et al. Human proinsulin C-peptide from a precursor overexpressed in Pichia pastoris[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2006,38(8):586-592. |
| APA | Huang, YB.,Li, J.,Gao, X.,Sun, JR.,Lu, Y.,...&Zhang, YS.(2006).Human proinsulin C-peptide from a precursor overexpressed in Pichia pastoris.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,38(8),586-592. |
| MLA | Huang, YB,et al."Human proinsulin C-peptide from a precursor overexpressed in Pichia pastoris".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 38.8(2006):586-592. |
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