Open AccessProduction of diphtheria toxin CRM228 in B. subtilis
Author(s) -
Hemilä Harri,
Glode L.Michael,
Palva Ilkka
Publication year - 1989
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1989.tb03621.x
Subject(s) - diphtheria toxin , diphtheria , toxin , microbiology and biotechnology , corynebacterium diphtheriae , bacillus subtilis , chemistry , biology , virology , bacteria , vaccination , genetics
The gene coding for a nontoxic diptheria toxin (DT), tox 228, was isolated from lysogenic Corynebacterium diphtheriae and cloned into pBR322. A mature form of the tox 228 gene, lacking its signal sequence, was expressed in Bacillus subtilis using a B. amyloliquefaciens α‐amylase secretion vector. To test the possibility of producing partially deleted DT molecules, which could be used for cell‐directed toxin conjugates, a truncated form lacking 151 amino acids from the C‐terminus of the DT was generated by oligonucleotide mutagenesis. Both the truncated and intact DT were efficiently secreted into the culture medium. During prolonged cultivation, the truncated form was less stable than the intact DT molecule.