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‐te...

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Bibliographic Details
Published in:Protein science 1997-09, Vol.6 (9), p.1953-1962
Main Authors: Belagaje, Rama M., Reams, Stephen G., Ly, Stan C., Prouty, Walter F.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Arginine
Base Sequence
Cathepsin C
Cloning, Molecular
Dipeptidyl-Peptidases and Tripeptidyl-Peptidases - metabolism
Escherichia coli - genetics
Escherichia coli - metabolism
Gene Expression
high‐pressure liquid chromatography
human proinsulin
Humans
Insulin-Like Growth Factor I - biosynthesis
Insulin-Like Growth Factor I - chemistry
Insulin-Like Growth Factor I - genetics
insulin‐like growth factor‐1
Lysine
Methionine
Molecular Sequence Data
Molecular Weight
Peptide Fragments - chemistry
Peptide Fragments - genetics
polyacrylamide gel electrophoresis
Proinsulin - biosynthesis
Proinsulin - chemistry
Proinsulin - genetics
protein production
Recombinant Proteins - biosynthesis
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Summary: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.
ISSN:0961-8368
1469-896X
DOI:10.1002/pro.5560060916