Design and production of genetically modified soybean protein with anti‐hypertensive activity by incorporating potent analogue of ovokinin(2–7)

FEBS Letters

Doyama, Naomi¹  Maruyama, Nobuyuki¹  Utsumi, Shigeru¹  Matoba, Nobuyuki¹  Yamada, Yuko¹  Yoshikawa, Masaaki¹ 

[1] Research Institute For Food Science, Kyoto University, Uji, Kyoto 611-0011, Japan

关键词: Ovokinin;    Anti-hypertensive peptide;    Soybean β-conglycinin;    Genetically modified food;    Spontaneously hypertensive rat;    APMSF;    (p-amidinophenyl)-methylsulfonyl fluoride;    GM;    genetically modified;    HPLC;    high performance liquid chromatography;    LB;    Luria–Bertani;    ODS;    octadecyl silica;    PAGE;    polyacrylamide gel electrophoresis;    SHR;    spontaneously hypertensive rat;    WKY;    Wistar–Kyoto rat;    TFA;    trifluoroacetic acid;   

DOI  :  10.1016/S0014-5793(01)02434-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The potent anti-hypertensive peptide, RPLKPW, has been designed based on the structure of ovokinin(2–7). The sequence encoding this peptide was introduced into three homologous sites in the gene for soybean β-conglycinin α′ subunit. The native α′ subunit as well as the modified, RPLKPW-containing α′ subunit were expressed in Escherichia coli, recovered from the soluble fraction and then purified by ion-exchange chromatography. The RPLKPW peptide was released from recombinant RPLKPW-containing α′ subunit after in vitro digestion by trypsin and chymotrypsin. Moreover, the undigested RPLKPW-containing α′ subunit given orally at a dose of 10 mg/kg exerted an anti-hypertensive effect in spontaneously hypertensive rats, unlike the native α′ subunit. These results provide evidence for the first time that a physiologically active peptide introduced into a food protein by site-directed mutagenesis could practically function in vivo even at a low dose.

【 授权许可】

Unknown