High cell density fermentation of recombinant Vibrio cholerae for the production of B subunit of Escherichia coli enterotoxin |
| |
Authors: | Panda A K Ghorpade A Mukhopadhyay A Talwar G P Garg L C |
| |
Affiliation: | National institute of immunology, Aruna Asaf Ali Marg, New Delhi 110067, India. |
| |
Abstract: | High cell density fermentation studies were performed to produce the B subunit of Escherichia coli heat-labile enterotoxin (LTB) from a Vibrio cholerae culture that carries a recombinant plasmid with an ampicillin resistance gene, tac promoter, and the gene encoding LTB. Upon induction with isopropyl-beta-D-thiogalactopyranoside (IPTG) the culture secreted the protein into the extracellular milieu. Fed-batch fermentation with stepwise addition of a total of 5 mM of IPTG during the active growth phase of the organism resulted in the production of 400 mg/L of LTB in 9 h and a cell optical density (OD) of 24. The LTB was purified to homogeneity with 70% recovery from the fermentation broth and was found to be chemically and biologically identical to the native protein by N-terminal amino acid sequencing and receptor binding assay. (c) 1995 John Wiley & Sons, Inc. |
| |
Keywords: | Escherichia coli enterotoxin fed batch high cell density fermentation purification |
本文献已被 PubMed 等数据库收录! |