Efficient production of casoxin D, a bradykinin agonist peptide derived from human casein, by Bacillus brevis.
We efficiently produced a small peptide by the host-vector system using Bacillus brevis as a host. DNA encoding the physiologically functional casoxin D, composed of seven amino acids, was ligated in tandem. An expression-secretion vector containing DNA, which codes for a fusion protein of epidermal growth factor-casoxin D pentamer, was constructed. B. brevis transformed with this plasmid produced about 0.5 g/liter of the fusion protein in the culture supernatant. The fusion protein was purified with ammonium sulfate fractionation from the supernatant and digested with two kinds of proteinases. A peptide well separated by high pressure liquid chromatography was identified as biologically active casoxin D.
Kato
M
M
Faculty of Agriculture, Nagoya University, Japan.
Fujiwara
Y
Y
Okamoto
A
A
Yoshikawa
M
M
Chiba
H
H
Udaka
S
S
D000595:Amino Acid Sequence; D001407:Bacillus / Q000235:genetics; D001483:Base Sequence; D001920:Bradykinin / Q000819:agonists*; D002364:Caseins / Q000737:chemistry*; D003001:Cloning, Molecular; D004815:Epidermal Growth Factor / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000235:genetics* / Q000494:pharmacology; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification
D002364:Caseins; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C097580:casoxin D; D004815:Epidermal Growth Factor; D001920:Bradykinin