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

Abstract

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 L L-conglycinin K KP P subunit. The native K KP P subunit as well as the modified, RPLKPW-containing K KP P 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 K KP P subunit after in vitro digestion by trypsin and chymotrypsin. Moreover, the undigested RPLKPW-containing K KP P subunit given orally at a dose of 10 mg/kg exerted an anti-hypertensive effect in spontaneously hypertensive rats, unlike the native K KP P 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. ß 2001 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

Improved Bile Acid-binding Ability of Soybean Glycinin A1a Polypeptide by the Introduction of a Bile Acid-binding Peptide (VAWWMY)

“…12,13) However, this study is the first report on the improvement of bile acidbinding ability of glycinin by the incorporation of the VAWWMY sequence.…”

Soybean Glycinin A1aB1b Subunit Has a Molecular Chaperone-like Function to Assist Folding of the Other Subunit Having Low Folding Ability

“…This modification allowed the RPLKPW sequences to be released by trypsin and chymotrypsin. Matoba et al 6) and Yamada et al 14) have designed soybean -conglycinin 0 subunit containing RPLKPW and confirmed that the introduced bioactive peptides were released by the action of trypsin and chymotrypsin and elicited high anti-hypertensive activity in vivo. These results suggest that RPLKPW could also be released from the modified versions of A3B4 by the action of trypsin and chymotrypsin and may exhibit high anti-hypertensive activity in vivo, since the positions of the modifications are in the disordered region on the molecular surface.…”

“…We also constructed expression plasmids for modified versions of A3B4. The DNA regions for two RPLKPW (strong anti-hypertensive peptide) 6) -like sequences of TPSYPP and RPSHGK corresponding to residues 268-273 and 275-280 of the hypervariable region in the A3B4 subunit were respectively changed to the DNA sequences for * Present address: Laboratory of Applied Molecular Design, Marine Biotechnology Institute, Kamaishi, Iwate 026-0001, Japan y To whom correspondence should be addressed. Fax: +81-774-38-3761; E-mail: sutsumi@kais.kyoto-u.ac.jp Biosci.…”

Isocaloric High-protein Diet Ameliorates Systolic Blood Pressure Increase and Cardiac Remodeling Caused by Maternal Caloric Restriction in Adult Mouse Offspring

“…SBP was measured with an indirect tail-cuff method by using MK-2000 (Muromachi Kikai Co. Ltd, Tokyo, Japan), as we [11] and others [20,21] described and validated. At 8 and 16 weeks of age (wks), SBP was measured at least five times in conscious mice (n=8-16 for each group).…”