title: Refolding and activation of human prorenin expressed in Escherichia coli: application of recombinant human renin for inhibitor screening.
abstract: Human prorenin was expressed in Escherichia coli as a fusion protein of thioredoxin. The chimeric protein, which accumulated insoluble inclusion bodies, was solubilized in 4 M guanidine-HCl and refolded by an arginine-detergent buffer system and by systematic dialysis. The refolded fusion prorenin was activated by trypsin. The antiserum against human kidney renin specifically inhibited the recombinant human renin activity. Using the recombinant human renin, we screened its inhibitory activity in fermented soybean paste (miso) and demonstrated that miso contained renin inhibitory activity derived from soybean. The IC(50) values for soybean and steamed soybean extracts were determined to be 1.9 and 1.6 mg/ml, respectively. This is the first demonstration of renin inhibitory activity in miso and soybean.
authors.affiliation: Akita Research Institute for Food and Brewing, Akita. saori@arif.pref.akita.jp
authors.forename: Saori
authors.initials: S
authors.lastname: Takahashi
authors.forename: Hironobu
authors.initials: H
authors.lastname: Ogasawara
authors.forename: Takayuki
authors.initials: T
authors.lastname: Watanabe
authors.forename: Masanori
authors.initials: M
authors.lastname: Kumagai
authors.forename: Hiroyasu
authors.initials: H
authors.lastname: Inoue
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Hori
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D017510:Protein Folding; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D012083:Renin / Q000378:metabolism*; D000092502:Renin Inhibitors
chemical_list: D017931:DNA Primers; D011994:Recombinant Proteins; D012083:Renin; D000092502:Renin Inhibitors