Expression and purification of human urodilatin by small ubiquitin-related modifier fusion in Escherichia coli

Applied Microbiology and Biotechnology
Ziyong SunJian-Ning Liu

Abstract

To prevent in vivo degradation, small peptides are usually expressed in fusion proteins from which target peptides can be released by proteolytic or chemical reagents. In this report, small ubiquitin-related modifier (SUMO) linked with a hexa-histidine tag was used as a fusion partner for the production of recombinant human urodilatin, a hormone for the treatment of acute decompensated heart failure. The fusion protein, which was overexpressed mainly as inclusion bodies in Escherichia coli, constituted about 25% of the total cell proteins. After purification by Ni-sepharose affinity chromatography and renaturation in refolding buffer, the fusion protein was cleaved with SUMO protease 1. Urodilatin was separated from the fusion partner by the subtractive chromatography using Ni-sepharose once again, and then further purified with reverse-phase high performance liquid chromatography. In vitro activity assay demonstrated that the recombinant urodilatin had a potent vasodilatory effect on rabbit aortic strips with an EC50 of 1.77+/-0.53 microg/ml, which was similar to that of the synthetic urodilatin standard. The expression strategy presented in this study allows convenient high yield and easy purification of small recombinant pep...Continue Reading

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Citations

Jun 24, 2009·Applied Microbiology and Biotechnology·Xianglei GaoSheng Xiong
Jul 8, 2009·Applied Microbiology and Biotechnology·Jian Feng LiShuang Quan Zhang
Oct 15, 2013·Applied Microbiology and Biotechnology·Chao LuanYi Zhen Wang
May 19, 2011·Protein Expression and Purification·Makkapati Satakarni, Robin Curtis
Jan 27, 2010·Journal of Biotechnology·Santosh Kumar UpadhyayRakesh Tuli
Jan 21, 2009·Protein Expression and Purification·Ziyong SunJian-Ning Liu
Jul 7, 2009·Biotechnology and Applied Biochemistry·Yifeng Li
Feb 24, 2015·Journal of Biotechnology·Jiaxin ZhangQiang Zhuge
Aug 20, 2016·Applied Microbiology and Biotechnology·Haiquan YangFei Xu
Nov 26, 2015·Biotechnology and Applied Biochemistry·Chunlan XiaoJing Zhang
Jun 8, 2010·The Protein Journal·Jian Feng LiShuang Quan Zhang

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