Cloning, Soluble Expression, and Production of Recombinant Antihypertensive Peptide Multimer (AHPM-2) in Escherichia coli for Bioactivity Identification

Title: Cloning, Soluble Expression, and Production of Recombinant Antihypertensive Peptide Multimer (AHPM-2) in Escherichia coli for Bioactivity Identification

Volume: 18 Issue: 7

Author(s): Shengqi Rao, Zhenzhen Xu, Yujie Su, Junhua Li, Jun Sun and Yanjun Yang

Affiliation:

Keywords: Antihypertensive peptide, antihypertensive peptide multimer, design, expression, Escherichia coliAntihypertensive peptide, antihypertensive peptide multimer, design, expression, Escherichia coli

Abstract: Recombinant antihypertensive peptide multimer (AHPM-2, 8kDa/68AA), a new designed polypeptide with potential antihypertensive effect in vivo, is composed of 15 low-molecular-weight antihypertensive peptides tandemly linked up according to the restriction sites of gastrointestinal proteases. After gene optimization, the DNA fragment encoding AHPM-2 was chemically synthesized, cloned into the pET32a, and successfully expressed in E.coli, above 90% in a soluble form. After chromatographic purification, the expressed fusion protein Trx-AHPM-2 was subject to the simulated gastrointestinal digestion, and the hydrolysate showed potent ACE inhibitory activity with an IC50 value of 4.5±0.3 ìg ml-1. The active fragments from the AHPM-2 were identified by UPLC-MS/MS. This method will be useful in obtaining an appreciable quantity of recombinant AHP at low cost, and the intact AHPM-2 is expected to be developed into functional food for preventing hypertension as well as for therapeutic.

Cite this article as:

Rao Shengqi, Xu Zhenzhen, Su Yujie, Li Junhua, Sun Jun and Yang Yanjun, Cloning, Soluble Expression, and Production of Recombinant Antihypertensive Peptide Multimer (AHPM-2) in Escherichia coli for Bioactivity Identification, Protein & Peptide Letters 2011; 18 (7) . https://dx.doi.org/10.2174/092986611795446067