Increased production of low molecular weight recombinant proteins in Escherichia coli.
A general method for obtaining high-level production of low molecular weight proteins in Escherichia coli is described. This method is based on the use of a novel Met-Xaa-protein construction which is formed by insertion of a single amino acid residue (preferably Arginine or Lysine) between the N-terminal methionine and the protein of interest. The utility of this method is illustrated by examples for achieving high-level production of human insulin-like growth factor-1, human proinsulin, and their analogs. Furthermore, highly produced insulin-like growth factor-1 derivatives and human proinsulin analogs are converted to their natural sequences by removal of dipeptides with cathepsin C.
Belagaje
R M
RM
Department of Biotechnology, Eli Lilly and Company, Indianapolis, Indiana 46285, USA. BELAGAJE_RAMA_M@LILLY.COM
Reams
S G
SG
Ly
S C
SC
Prouty
W F
WF
D000595:Amino Acid Sequence; D001120:Arginine; D001483:Base Sequence; D020789:Cathepsin C; D003001:Cloning, Molecular; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D008239:Lysine; D008715:Methionine; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D011384:Proinsulin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*
D010446:Peptide Fragments; D011994:Recombinant Proteins; D007334:Insulin-Like Growth Factor I; D011384:Proinsulin; D001120:Arginine; D008715:Methionine; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D020789:Cathepsin C; D008239:Lysine