title: Cloning, soluble expression, and production of recombinant antihypertensive peptide multimer (AHPM-2) in Escherichia coli for bioactivity identification.
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 μg 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.
authors.affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
authors.forename: Shengqi
authors.initials: S
authors.lastname: Rao
authors.forename: Zhenzhen
authors.initials: Z
authors.lastname: Xu
authors.forename: Yujie
authors.initials: Y
authors.lastname: Su
authors.forename: Junhua
authors.initials: J
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Sun
authors.forename: Yanjun
authors.initials: Y
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D000959:Antihypertensive Agents / Q000737:chemistry* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D053719:Tandem Mass Spectrometry
chemical_list: D000806:Angiotensin-Converting Enzyme Inhibitors; D000959:Antihypertensive Agents; D010455:Peptides; D011993:Recombinant Fusion Proteins