An Escherichia coli host strain useful for efficient overproduction of secreted recombinant protein.
Periplasmic secretion of overexpressed Bacillus stearothermophilus alpha-amylase was analyzed in batch and fed-batch cultivations of Escherichia coli MG1655:pCSS4-p and the mutant strain CWML2:pCSS4-p. Under all conditions investigated, growth and product formation of MG1655:pCSS4-p were severely impaired by heterologous protein expression and/or processing, while E. coli CWML2:pCSS4-p was found to be more robust and to accumulate 2- to 3-fold higher maximum alpha-amylase levels. While this strain is itself potentially interesting for applications, its properties also illustrate the potential of the selection procedure that was employed to obtain it from its progenitor MG1655 (Weikert, C., Sauer, U., Bailey, J. E., 1997. Microbiol. 143: 1567-1574. Application of this procedure to existing industrial strains may lead to significantly improved process organisms.
Weikert
C
C
Institute of Biotechnology, ETH Zürich, CH-8093 Zürich, Switzerland.
Sauer
U
U
Bailey
J E
JE
D001709:Biotechnology / Q000295:instrumentation / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D001411:Geobacillus stearothermophilus / Q000201:enzymology* / Q000235:genetics; D007700:Kinetics; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D000516:alpha-Amylases / Q000096:biosynthesis* / Q000235:genetics
D011994:Recombinant Proteins; D000516:alpha-Amylases