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Increased production of low molecular weight recombinant proteins in Escherichia coli
Author(s) -
Belagaje Rama M.,
Reams Stephen G.,
Ly Stan C.,
Prouty Walter F.
Publication year - 1997
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560060916
Subject(s) - proinsulin , escherichia coli , biochemistry , recombinant dna , methionine , lysine , chemistry , arginine , residue (chemistry) , amino acid , insulin , biology , microbiology and biotechnology , gene
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‐X aa ‐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.