Premium
Toxin production by Clostridium tetani in biphasic liquid cultures
Author(s) -
Puziss Milton,
Hedén C.G.
Publication year - 1965
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.260070305
Subject(s) - clostridium tetani , toxin , dextran , protoplast , microbiology and biotechnology , titration , peg ratio , bacillus anthracis , clostridium difficile toxin a , in vitro , chemistry , phase (matter) , chromatography , biology , bacteria , biochemistry , tetanus , clostridium difficile , immunology , organic chemistry , genetics , vaccination , finance , economics , antibiotics
An exploratory study is described on the use of a biphasic culture technique for production of toxin by C. tetani . The biphasic system used was a combination of dextran and poly(ethylene glycol) (PEG) in the presence of a protein‐free medium. The system supported good growth and gave a favorable phase ratio (top/bottom) of 15:1. The majority of the bacterial cells were in the bottom dextran phase, while the greater part of the toxin was in the upper PEG–medium phase. Toxin yields from a typical experiment were 10 9 mouse IP MLD/ml. of upper phase filtrate. The biphasic culture system was also adaptable to continuous culture, with approximately similar toxin levels found in the upper phase. The appearance of the clostridial cells in older cultures suggested the possibility of protoplast formation, perhaps stabilized by the polymers present. In vitro toxin titration (Lf) was inhibited by the presence of the polymers. Bacillus anthracis also grew well in the biphasic system and elaborated protective antigen, primarily into the upper phase.