A型产气荚膜梭菌合成培养基使用参数及培养毒素浓缩工艺研究

The research between the application parameter in the synthesized culture medium of Clostridium perfringens A type and the ultrafiltration of cultured toxin

关键词:  A型产气荚膜梭菌  合成培养基  使用参数  浓缩工艺

Key words:  Clostridium perfringens A type  synthesized culture medium  application parameter  ultrafiltration

摘要:

经过对A型产气荚膜梭菌合成培养基pH值、灭菌方式和配制用水进行优化，确定最适pH值为8.0～8.5，灭菌方式为116℃30min，配制用水为去离子水。合成培养基培养产气荚膜梭菌CVCC37株6小时后毒力达50～100MLD/ml，随着时间的延长，毒力不再增强；培养温度为36℃或37℃时，产毒效果最佳。比较不同截留分子量的超滤膜浓缩效果，10Ku截留分子量的收获率为40%，其透出液静脉接种0.2ml,小鼠2/2死亡；而8Ku收获率达75%，其透出液静脉接种0.2ml,小鼠0/2死亡。因此，8Ku截留分子量膜包适宜对A型菌培养毒素的浓缩。

Abstract:

The optimal conditions for the synthesized culture medium of Clostridium perfringens A type is confirmed that the most suitable pH is about 8.0～8.5;the method of sterilisation in autoclave is 116℃30min;the used water is deionic water 。For the strain(CVCC37) of Clostridium perfringens, the toxin activity attains 50～100MLD/ml after growing in optimized medium for 6 hours, the toxic activity does not increase with even longer growth time ;when compared with different culture temperature，the highest level of toxin activity at 36℃ or 37℃ is researched. Ultrafiltration with different membrane by MWCO(Molecular Weight Cutoff) is used to concentrate the culture supernatant,the harvest ratio of the 10Ku membrane by MWCO is 40%, and the discarded effluent results in 2/2 lethal ratio on rats injected by 0.2ml in vein;while the harvest ratio of the 8Ku membrane by MWCO is 75% and the discarded effluent has no lethal activity；Therefore, the ultrafiltration with 8Ku membrane by MWCO is suitable for the ultrafiltration of cultured toxin produced by Clostridium perfringens A type。