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KMID : 1094720160210040537
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Biotechnology and Bioprocess Engineering
2016 Volume.21 No. 4 p.537 ~ p.543
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Optimization of diphtheria toxin production by Corynebacterium diphtheriae using a casein-based medium in a fermenter
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Chung Yong-Ju
Lee Jin-A
Jung Mi-Young
Lee Sang-Mi
Kim Tae-Yeon
Choe Yong-Kyung
Kim Ik-Hwan
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Abstract
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The Td-based combined vaccine contains only small amounts of the diphtheria toxoid antigen. However, a high level of purity is necessary for this antigen. The diphtheria toxin is produced by growing Corynebacterium diphtheriae in a semisynthetic, casein-based medium in a fermenter. In order to obtain a highly pure diphtheria toxoid, the optimal conditions to express the toxin at 300 Lf/mL in a fermenter culture were determined. When C. diphtheriae was cultivated in a fermenter and a high concentration of toxin was obtained, specific patterns for the pH and dissolved oxygen levels identified. Overall, the fermenter cultivation process was divided into four stages according to variations in the pH. A specific range of K La in the fermenter (0.0092 ~ 0.0093/sec) was required to produce high level expression of diphtheria toxin. The amount of toxin expression varied significantly according to culture conditions. Agitation and aeration in the fermenter affected toxin expression, even when the optimal K La value for toxin production was maintained. A previous study has reported that the amounts of agitation and aeration are important factors when cultivating fungus in the fermenter to produce chitinolytic enzyme. A mass production of diphtheria toxoid with a purity level greater than 2,500 Lf/ mgPN was obtained through purification and detoxification from this optimized toxin production.
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KEYWORD
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casein hydrolysate, diphtheria toxin, fermenter, semisynthetic media, Td vaccine
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