Hollow-fire bioreactors compared to batch and chemostat culture for the production of a recombinant toxoid by a marine Iibrio.
Bioreactor selection is important for maximising the productivity of recombinant organisms. In this paper a comparison is made between growth and recombinant protein synthesis in three types of bioreactor containing a marine Vibrio capable of heterologous expression and secretion of the non-toxic B-subunit pentamer of Escherichia coli heat-labile enterotoxin, EtxB. The heterologous gene was located on the plasmid pMMB68. Resistance to carbenicillin was used to select for plasmid-containing cells. In batch and continuous culture. Volumetric productivities were highest when cells were grown in the presence of carbenicillin. Without antibiotic selection, the highest volumetric productivity (9.4 mg EtxB-1 h-1) was observed in hollow-fibre bioreactors, and the production phase could be maintained for over 50 h. The highest specific productivity under these conditions was found in batch culture, but the maximal production phase was only of 5 h duration. In hollow-fibre reactors the type of fibre used significantly affected productivity, both with regards to the maintenance of reactor integrity and by allowing passage of the recombinant toxoid through the selectively permeable membrane. Where contamination of the product with carbenicillin is to be avoided, these bioreactors are superior to batch or continuous culture.
Lloyd
J R
JR
School of Biological Sciences, University of Birmingham, Edgbaston, UK.J.R.Lloyd@bham.ac.uk
Hirst
T R
TR
Bunch
A W
AW
D001427:Bacterial Toxins / Q000096:biosynthesis*; D004768:Enterotoxins / Q000096:biosynthesis*; D004926:Escherichia coli / Q000472:pathogenicity*; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014121:Toxoids / Q000096:biosynthesis*; D014733:Vibrio / Q000378:metabolism*
D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D014121:Toxoids; C027538:heat-labile enterotoxin, E coli