Protein & Peptide Letters

Prof. Ben M. Dunn  
Department of Biochemistry and Molecular Biology
University of Florida
College of Medicine
P.O. Box 100245
Gainesville, FL
USA
Email: bdunn@ufl.edu

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

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

Affiliation: 1800 Lihu Avenue, Wuxi, Jiangsu Province, China 214122

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.

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

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Article Details

VOLUME: 18
ISSUE: 7
Page: [699 - 706]
Pages: 8
DOI: 10.2174/092986611795446067
Price: $58