Heterogeneity in the molecular species of heat-stable enterotoxin of Vibrio cholerae non-O1 expressed by Escherichia coli carrying the cloned toxin gene
Dohi, S.; Kasuga, H.; Nakao, H.; Ogawa, A.; Nair, G.B.; Takeda, T.
Fems Microbiology Letters 106(2): 223-227
1993
ISSN/ISBN: 0378-1097 PMID: 8454187 DOI: 10.1111/j.1574-6968.1993.tb05963.x
Accession: 008768414
The biological activity of the heat-stable enterotoxin of Vibrio cholerae non-O1 (NAG-ST) was found to be predominantly associated with the periplasmic extract (about four-fold higher than the culture supernatant) of a recombinant E. coli (JM109) strain carrying the NAG-ST toxin gene. Four molecular species of NAG-ST, two each from the periplasmic extract and culture supernatant of JM109, were purified. Amino acid sequence analysis of the four NAG-ST peptides isolated by HPLC revealed that they all differed from that of the mature 17-amino acid residue NAG-ST released by V. cholerae non-O1. The M-r-values of the peptides obtained from the periplasmic extract were 4331 and 2785, while those recovered from the culture supernatant were 3154 and 2785. It thus appears that V. cholerae NAG-ST is synthesized as larger molecules in the recombinant E. coli strain. The differences in sizes of the exported NAG-ST molecular could relate to differences in the enzyme cleavage system between E. coli and V. cholerae.
