Tetanus toxin production from Clostridium tetani, using a casein-based medium in a single-use bioreactor

Yong Ju Chung, Mi Young Jung, Jin A. Lee, Tae Yeon Kim, Yong Kyung Choe, Ik Hwan Kim

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)


Clostridium tetani, a spore-forming anaerobic bacterium, and a casein-based semisynthetic medium were used to produce tetanus toxin in this study. N-Z-Case TT (casein hydrolysate) solution and glucose stock media were mixed and autoclaved, which resulted in tetanus toxin expression. The toxin was expressed when the N-Z-Case TT solution reacted with the glucose stock at a high temperature, creating an adequate amount of Maillard reaction products (MRPs). After accumulating in C. tetani cells, tetanus toxin was secreted into the medium when cell lysis was induced by surface aeration. C. tetani was cultivated and tetanus toxin was expressed in a single-use bioreactor, which produced 80 Lf/mL of tetanus toxin in a medium with MRPs. While using the correct medium to induce tetanus toxin was important, other factors played a part in achieving the desired concentration of the toxin, including the medium processing and culture methods inside the bioreactor. Tetanus toxoid with a purity level greater than 2,500 Lf/mgPN was obtained by detoxifying and purifying the toxin recovered from the fermenter or single-use bioreactor. A single-use bioreactor could be used in a limited space without the need for constructing a large scale production facility, to produce the tetanus toxoid antigen for clinical trials.

Original languageEnglish
Pages (from-to)531-536
Number of pages6
JournalBiotechnology and Bioprocess Engineering
Issue number4
Publication statusPublished - 2016 Aug 1


  • Clostridium tetani
  • casein hydrolysate
  • semisynthetic medium
  • single-use bioreactor
  • tetanus toxin

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Biomedical Engineering


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