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 journalArticle

1 Citation (Scopus)

Abstract

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
Volume21
Issue number4
DOIs
Publication statusPublished - 2016 Aug 1

Fingerprint

Tetanus Toxin
Casein
Clostridium
Bioreactors
Caseins
Clostridium tetani
Reaction products
Glucose
Maillard Reaction
Tetanus Toxoid
Fermenters
Antigens
Bacteria
Anaerobic Bacteria
Spores
Culture Media
Processing
Clinical Trials
Temperature

Keywords

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

ASJC Scopus subject areas

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

Cite this

Tetanus toxin production from Clostridium tetani, using a casein-based medium in a single-use bioreactor. / Chung, Yong Ju; Jung, Mi Young; Lee, Jin A.; Kim, Tae Yeon; Choe, Yong Kyung; Kim, Ik Hwan.

In: Biotechnology and Bioprocess Engineering, Vol. 21, No. 4, 01.08.2016, p. 531-536.

Research output: Contribution to journalArticle

Chung, Yong Ju ; Jung, Mi Young ; Lee, Jin A. ; Kim, Tae Yeon ; Choe, Yong Kyung ; Kim, Ik Hwan. / Tetanus toxin production from Clostridium tetani, using a casein-based medium in a single-use bioreactor. In: Biotechnology and Bioprocess Engineering. 2016 ; Vol. 21, No. 4. pp. 531-536.
@article{18383af222e94f3fb35b3262ed559fde,
title = "Tetanus toxin production from Clostridium tetani, using a casein-based medium in a single-use bioreactor",
abstract = "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.",
keywords = "casein hydrolysate, Clostridium tetani, semisynthetic medium, single-use bioreactor, tetanus toxin",
author = "Chung, {Yong Ju} and Jung, {Mi Young} and Lee, {Jin A.} and Kim, {Tae Yeon} and Choe, {Yong Kyung} and Kim, {Ik Hwan}",
year = "2016",
month = "8",
day = "1",
doi = "10.1007/s12257-016-0355-6",
language = "English",
volume = "21",
pages = "531--536",
journal = "Biotechnology and Bioprocess Engineering",
issn = "1226-8372",
publisher = "Korean Society for Biotechnology and Bioengineering",
number = "4",

}

TY - JOUR

T1 - Tetanus toxin production from Clostridium tetani, using a casein-based medium in a single-use bioreactor

AU - Chung, Yong Ju

AU - Jung, Mi Young

AU - Lee, Jin A.

AU - Kim, Tae Yeon

AU - Choe, Yong Kyung

AU - Kim, Ik Hwan

PY - 2016/8/1

Y1 - 2016/8/1

N2 - 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.

AB - 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.

KW - casein hydrolysate

KW - Clostridium tetani

KW - semisynthetic medium

KW - single-use bioreactor

KW - tetanus toxin

UR - http://www.scopus.com/inward/record.url?scp=84990821998&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84990821998&partnerID=8YFLogxK

U2 - 10.1007/s12257-016-0355-6

DO - 10.1007/s12257-016-0355-6

M3 - Article

VL - 21

SP - 531

EP - 536

JO - Biotechnology and Bioprocess Engineering

JF - Biotechnology and Bioprocess Engineering

SN - 1226-8372

IS - 4

ER -