Antihypertensive activity of recombinant peptide IYPR expressed in Escherichia coli as inclusion bodies.

Abstract

To produce more angiotensin converting enzyme inhibitory peptides (ACEIP), we have established a high-efficiency Escherichia coli expression system. The DNA-coding sequence for the recombinant protein, which was subcloned into the vector pET-30a(+), has been expressed as inclusion bodies in E. coli BL21 (DE3). The influences of induction time and concentration of isopropyl-β-D-thiogalactopyranoside (IPTG) on the expression of recombinant protein were studied. The resulting expression level of the protein accounted for about 31% of cellular protein at a temperature of 37°C, IPTG concentration of 0.6mM and induction time of 7h. The inclusion bodies were washed, separated from the cells, and solubilized with urea. After purification by affinity chromatography, the recombinant protein was recovered with a high purity of about 90%. Molecular weight of the recombinant protein was measured using Tricine-SDS-PAGE. Peptide IYPR was obtained by cleavage of the recombinant protein with trypsin and the IC(50) value was 61 mg/L. The antihypertensive activity in spontaneously hypertensive rats (SHRs) was also investigated. Single oral administration of this peptide in 10-week old SHRs resulted in a significant reduction of systolic blood pressure to 50 mm Hg at 4 h. The data obtained provide a good reference for further development of peptide Ile-Tyr-Pro-Arg into an effective antihypertensive agent for prevention and treatment of hypertension.

• Authors: Liurong Huang, Haile Ma, Yunliang Li, Shuxiang Li
• Journal: Protein expression and purification (Protein Expr Purif) Vol. 83 Issue 1 Pg. 15-20 (May 2012) ISSN: 1096-0279 [Electronic] United States
• PMID: 22381462 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2012 Elsevier Inc. All rights reserved.

Chemical References

• Angiotensin-Converting Enzyme Inhibitors
• Antihypertensive Agents
• Oligopeptides
• Recombinant Fusion Proteins
• Peptidyl-Dipeptidase A

Topics

• Amino Acid Sequence
• Angiotensin-Converting Enzyme Inhibitors (chemistry, metabolism, pharmacology)
• Animals
• Antihypertensive Agents (chemistry, metabolism, pharmacology)
• Base Sequence
• Blood Pressure (drug effects)
• Escherichia coli (genetics)
• Inclusion Bodies (chemistry)
• Molecular Sequence Data
• Oligopeptides (biosynthesis, chemistry, genetics, pharmacology)
• Peptidyl-Dipeptidase A (metabolism)
• Rats
• Rats, Inbred SHR
• Recombinant Fusion Proteins (biosynthesis, chemistry, genetics, pharmacology)