Details

Autor(en) / Beteiligte

• Huang, Liurong
• Ma, Haile
• Li, Yunliang
• Li, Shuxiang

Titel

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

Ist Teil von

• Protein expression and purification, 2012-05, Vol.83 (1), p.15-20

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2012

Quelle

MEDLINE

Beschreibungen/Notizen

• ► Construction of a novel recombinant plasmid to produce ACEIP. ► Factors such as induction time, IPTG concentration were examined. ► Recombinant protein was purified. ► Antihypertensive activity of the inhibitor was investigated. 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 IC50 value was 61mg/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 50mmHg at 4h. 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.