title: Expression and purification recombinant antihypertensive peptide ameliorates hypertension in rats with spontaneous hypertension.
abstract: A highly efficient Escherichia coli expression system was established to obtain an appreciable quantity of antihypertensive peptide. The DNA-coding sequence for the Gly-Val-Tyr-Pro-His-Lys peptide was chemically synthesized and linked to form a ten-copy in tandem. It was cloned into the vector pET-15b and expressed in E. coli BL21 (DE3). The optimal conditions for maximal expression were verified and included the induction time and the concentration of isopropyl-β-D-thiogalactopyranoside. The recombinant protein was purified by affinity chromatography to greater than 95% purity, and further purification was achieved by High-performance Liquid Chromatography after cleavage with trypsin. The product was identified by Electrospray Ionization-Mass Spectrometry. The antihypertensive effects of the recombinant AHP were investigated in spontaneously hypertensive rats. The in vivo results demonstrated that a single oral administration of this peptide in spontaneously hypertensive rats resulted in a significant reduction of systolic blood pressure at 2h. Systolic blood pressure was stabilized 4h later and remained at a low level for 24h. This study provides a practical method to develop the peptide into functional foods or drugs for the prevention and treatment of hypertension.
authors.affiliation: Life Science Institute, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China. Electronic address: xiaolitina@126.com.
authors.forename: Xiao-Li
authors.initials: XL
authors.lastname: Wang
authors.affiliation: Life Science Institute, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Shi-Nan
authors.initials: SN
authors.lastname: Ma
authors.affiliation: Life Science Institute, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Ya-Hong
authors.initials: YH
authors.lastname: Yuan
authors.affiliation: Life Science Institute, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Life Science Institute, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Dong-Sheng
authors.initials: DS
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000959:Antihypertensive Agents / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology*; D001483:Base Sequence; D001794:Blood Pressure / Q000187:drug effects; D006973:Hypertension / Q000503:physiopathology*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D051381:Rats; D011918:Rats, Inbred SHR; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology*
chemical_list: D000959:Antihypertensive Agents; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C085525:C111 peptide
keywords: Antihypertensive peptide; Expression; Hypertension; Purification; Tandem