CAJ | 학술논문

确定C型产气英膜梭菌合成培养基使用参数及建立配套的毒素浓缩工艺。比较不同pH值、灭菌温度、配制用水的合成培养基及其不同培养时间和温度下培养产气荚膜梭菌CVCC60102株的毒力；按毒素的分子量及超滤膜的截留分子量设计并比较2种不同浓缩工艺的毒素收获率及透出液的毒力。结果表明，pH值为8．0～8．4、灭菌方式为116℃30min、配制用水为去离子水时产毒最佳，毒力达到500—1000MLD／mL；合成培养基培养产气英膜梭菌CVCC60102株18h后毒力达500—1000MLD／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；不同截留分子量的超滤膜浓缩，10ku截留分子量的收获率为68％，其透出液静脉接种0．2mL，小鼠2／2死亡；而8ku收获率达80％，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C型菌培养毒素的浓缩。上述结果为C型产气荚膜梭菌合成培养基的应用提供了数据支撑。
학정C형산기영막사균합성배양기사용삼수급건립배투적독소농축공예。비교불동pH치、멸균온도、배제용수적합성배양기급기불동배양시간화온도하배양산기협막사균CVCC60102주적독력；안독소적분자량급초려막적절류분자량설계병비교2충불동농축공예적독소수획솔급투출액적독력。결과표명，pH치위8．0～8．4、멸균방식위116℃30min、배제용수위거리자수시산독최가，독력체도500—1000MLD／mL；합성배양기배양산기영막사균CVCC60102주18h후독력체500—1000MLD／mL，수착시간적연장，독력불재증강；배양온도위36℃혹37℃시，산독효과최가；불동절류분자량적초려막농축，10ku절류분자량적수획솔위68％，기투출액정맥접충0．2mL，소서2／2사망；이8ku수획솔체80％，기투출액정맥접충0．2mL，소서0／2사망。인차8ku절류분자량막포괄의대C형균배양독소적농축。상술결과위C형산기협막사균합성배양기적응용제공료수거지탱。
To define the parameters in the synthesized culture medium for Clostridum peofringens C type and the ultrafiltration of cultured toxin, the experiment compared the toxin activity of Clostridum peofringens CVCC60102 cultured in the synthesized medium prepared with different pH, sterilisation temperature, water type, also under condition of different culture time and temperature, and designed two different methods to concentrate the toxin according to the molecular weight and the MWCO ( Molecular Weight Cut Off) of the uhrafihration membrane, and compared the harvest ratio and activity of toxin obtained by the previous two methods. Results showed that the optimal synthesized culture medium should be prepared with deionic water at the most suitable pH of 8.0 ~ 8.4 and autoclaved for 30 min at 116 ~C. For the strain (CVCC60102) of Clostridum perfringens, the toxic activity reached 500- 1000 MLD/mL after growth for 18 hours. The toxicity did not increase with even longer growth time. The highest level of toxicity would be reached when cultured at 36 or 37 ℃. When concentrated by the 10 ku MWCO membrane, the harvest ratio of toxin was 68%, and the discarded effluent resulted in 2/2 lethal ratio on rat injected by 0.2 mL in vein. While by 8 ku MWCO, the harvest ratio was 80% and the discarded effluent had no lethal activity. Therefore, the ultrafiltration with membrane of 8 ku MWCO was suitable for the concentration of the cultured toxin. The above results provided the data support for the application of the synthesized culture medium for Clostridum pefringens C type, and these would be consulted for the manufactural application of corporations.