Design and production of genetically modified soybean protein with anti-hypertensive activity by incorporating potent analogue of ovokinin(2-7).
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 beta-conglycinin alpha' subunit. The native alpha' subunit as well as the modified, RPLKPW-containing alpha' 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 alpha' subunit after in vitro digestion by trypsin and chymotrypsin. Moreover, the undigested RPLKPW-containing alpha' subunit given orally at a dose of 10 mg/kg exerted an anti-hypertensive effect in spontaneously hypertensive rats, unlike the native alpha' 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.
Matoba
N
N
Research Institute For Food Science, Kyoto University, Uji, 611-0011, Kyoto, Japan.
Doyama
N
N
Yamada
Y
Y
Maruyama
N
N
Utsumi
S
S
Yoshikawa
M
M
D000284:Administration, Oral; D020816:Amino Acid Motifs / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D052179:Antigens, Plant; D000959:Antihypertensive Agents / Q000008:administration & dosage* / Q000302:isolation & purification / Q000378:metabolism; D001794:Blood Pressure / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D004527:Egg Proteins / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005916:Globulins / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000235:genetics*; D011355:Prodrugs / Q000008:administration & dosage* / Q000302:isolation & purification / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods; D021122:Protein Subunits; D051381:Rats; D011918:Rats, Inbred SHR; D011994:Recombinant Proteins / Q000008:administration & dosage* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D055314:Seed Storage Proteins; D030262:Soybean Proteins / Q000235:genetics; D013025:Glycine max; D014357:Trypsin / Q000378:metabolism
D052179:Antigens, Plant; D000959:Antihypertensive Agents; D004527:Egg Proteins; D005916:Globulins; D010446:Peptide Fragments; D011355:Prodrugs; D021122:Protein Subunits; D011994:Recombinant Proteins; D055314:Seed Storage Proteins; D030262:Soybean Proteins; C013894:beta-conglycinin protein, Glycine max; C096585:ovokinin; D002918:Chymotrypsin; D014357:Trypsin