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High cell density fermentation of recombinant Vibrio cholerae for the production of B subunit of Escherichia coli enterotoxin
Author(s) -
Panda Amulya K.,
Ghorpade Anuja,
Mukhopadhyay Asok,
Talwar G. P.,
Garg L. C.
Publication year - 1995
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260450309
Subject(s) - lac operon , enterotoxin , fermentation , escherichia coli , vibrio cholerae , heat labile enterotoxin , recombinant dna , extracellular , protein subunit , plasmid , biochemistry , heat stable enterotoxin , biology , chemistry , microbiology and biotechnology , gene , bacteria , genetics
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‐β‐ D ‐thiogalactopyranoside (IPTG) the culture secreted the protein into the extracellular milieu. Fed‐batch fermentation with stepwise addition of a total of 5 m M 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. © 1995 John Wiley & Sons, Inc.