Use of Bacillus brevis for synthesis and secretion of Des-B30 single-chain human insulin precursor.
A synthetic gene encoding a single chain human insulin precursor [B-chain (1-29)-A-chain] linked to the C-terminal lysine of human epidermal growth factor (1-28) (EGF-SCI) was constructed. This gene was expressed using Bacillus brevis. EGF-SCI was isolated from the supernatant of the culture broth. Treatment of EGF-SCI with lysyl endopeptidase resulted in the formation of des-B30 human insulin. The identification of the formed des-B30 human insulin was made by the measurement of molecular weight and amino acid analysis. The binding coefficient to anti-human insulin antibody was comparable to that of human insulin.
Koh
M
M
Institute for Applied Life Science, University of East Asia, Graduate School, Shimonoseki-shi, Yamaguchi, Japan.
Hanagata
H
H
Ebisu
S
S
Morihara
K
K
Takagi
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001407:Bacillus / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D006224:Cricetinae; D004815:Epidermal Growth Factor / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007328:Insulin / Q000031:analogs & derivatives* / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000078790:Insulin Secretion; D008969:Molecular Sequence Data; D011485:Protein Binding; D011498:Protein Precursors / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011972:Receptor, Insulin / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014169:Transformation, Bacterial
C414747:EGF-SCI protein, human; D007328:Insulin; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C045770:insulin, single chain des-(B30)-; D004815:Epidermal Growth Factor; D011972:Receptor, Insulin