首页|Human Proinsulin C-peptide from a Precursor Overexpressed in Pichia pastoris

Human Proinsulin C-peptide from a Precursor Overexpressed in Pichia pastoris

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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.

proinsulinC-peptidePichia pastoris

Yang-Bin HUANG、Jiang LI、Xin GAO、Jiu-Ru SUN、Yi LU、Tao FENG、Jian FEI、Da-Fu CUI、Qi-Chang XIA、Jun REN、You-Shang ZHANG

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Shanghai Newsummit Biopharma Company, Shanghai 200233, China

Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China

Zhongshan Hospital, Fudan University, Shanghai 200031, China

Shanghai Yizhong Biotechnology Company, Shanghai 201203, China

Shanghai Center of Research and Development of New Drugs, Shanghai 201203, China

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2006

生物化学与生物物理学报(英文版)
中国科学院上海生物化学研究所

生物化学与生物物理学报(英文版)

CSTPCDCSCD北大核心SCI
影响因子：0.772
ISSN：1672-9145
年,卷(期)：2006.38(8)
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