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

摘要

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 ℃,30 min;the used water is deionic water.For the strain(CVCC37) of Clostridium perfringens,the toxin activity attains 50~100 MLD/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 10 ku membrane by MWCO is 40%,and the discarded effluent results in 2/2 lethal ratio on rats injected by 0.2 mL in vein;while the harvest ratio of the 8 ku membrane by MWCO is 75% and the discarded effluent has no lethal activity;Therefore,the ultrafiltration with 8 ku membrane by MWCO is suitable for the ultrafiltration of cultured toxin produced by Clostridium perfringens A type.%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℃30 min,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 h后毒力达50～100 MLD/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,10 ku截留分子量的收获率为40%,其透出液静脉接种0.2 mL,小鼠2/2死亡;而8 ku收获率达75%,其透出液静脉接种0.2 mL,小鼠0/2死亡。因此,8 ku截留分子量膜包适宜对A型菌培养毒素的浓缩。