【作者】 张彩琴；

【导师】 王远亮；

【作者基本信息】 重庆大学 ， 药物化学， 2006， 硕士

【摘要】 埃博霉素B是一种新型抗癌剂,来源于纤维堆囊菌代谢产物之一。由于它具有比紫杉醇更高的抗癌活性,近来受到了极大的关注。本文以纤维堆囊菌Soce90为菌种,进行液体发酵培养,获得其最佳的生长条件。研究了该菌种的生长周期,并以此为基础,探究了发酵时,菌龄对发酵的影响。以及比较了一次填加培养基和间隔填加培养基发酵、基本培养基发酵和基本培养基中添加微量元素发酵、基本发酵和基本培养基发酵中pH调节发酵的不同,确定了最佳的发酵条件。另外,对Soce90菌种进行了固定化培养,通过比较了海藻酸钠、聚乙烯醇、玉米芯及丝瓜布的发酵结果,确定了最好的固定化材料—海藻酸钠,并确定了固定化时海藻酸钠用量,浓度及细菌浓度等参数。同时,通过浸提、萃取、层析等方法,获取了发酵液中的抗肿瘤活性物质—埃博霉素B。主要结论如下:1. Soce90菌种生长周期的结果为,第1天细菌数达到21.3×106个/mL,第2天细菌数达到最大,为27.1×106个/mL,第3天稍有所下降,这段时间为它的稳定期。此后细菌数越来越少,直到第9天,细菌数只有14.0×106个/mL。2.不同菌龄的发酵结果为,第2天的种子菌发酵后细菌干重为0.101g/tube,第3天的种子菌发酵后细菌干重为0.074g/tube;间隔添加培养基发酵后细菌的干重为0.304g/tube;一次添加培养基发酵后细菌的干重为0.088g/tube;基本培养基发酵的干重为0.088 g/tube;基本培养基pH调节发酵后细菌干重为0.272 g/tube;基本培养基中添加微量元素发酵的细菌干重为0.138g/tube。3.固定化Soce90细菌的最佳材料为海藻酸钠,固定化时,海藻酸钠的浓度为4%,用量为10mL,菌液量为8mL。固定化技术的长处在于它是一种良好发酵和分离耦合的生物反应器,能提高微生物的耐性和利用率,能减少后期分离的工艺过程。4.游离细菌发酵液通过初分,层析,HPLC-MS检测之后,鉴定在发酵液中含有埃博霉素B,其含量为22.5mg/L.更多还原

【Abstract】 The epothilone B is a vital anticancer agent. It is one of the secongd-metabolics from Sorangium cellulosum. It has got a great of attention in the world Recently.Fermention was conducted by Sorangium cellulosum 90,and got the best condition of its growth.The cycle of its growth was studied ,and on the basis of it,the fermentive effect was explored about ages of seeds .The fermentive results were compared between intermittent fed-batch process and fed-batch process、between ordinary fermention and fermention added microelements、between ordinary fermention and fermention modulated by pH. At the same time , the epothilone B was obtained by the methods of mention, extraction, separation and LC-MS.In addition, Soce90 was immobilizated by sodium polymannuronate、PVA、loofa and corncob.The best immobilizated material was sodium polymannuronate.And the parameters were ensured about dosage,concentration of sodium polymannuronate and the concentration of So ce90 during the immobilization. Some importants results are as the follows:1. The cycle of Soce90 growth is about 9 days.On the first day, the number of Soce90 reaches 21.3×106/mL.The peak of its number is obtainted about 27.1×106/mL on the second day.Its number is down from the third day.At last,it is only 14.0×106/mL。2. The fermentive results of the difference ages of seeds are as the follows:The fermentive dry weight of two-days old seeds is 0.101g/tube.Another fermentive dry weight of three-days old seeds is 0.074g/tube.The fermentive dry weight of intermittent fed-batch process is 0.304g/tube. The fermentive dry weight of fed-batch process is 0.088g/tube. Another weight of ordinary fermention is 0.088 g/tube. The third weight of fermention modulated by pH is 0.272 g/tube.The fourth weight of fermention added microelements is 0.138g/tube.3. The epothiloneB is checked up by mention, extraction, separation and LC-MS. The content is 22.5mg/L.4. The best immobilizated material is sodium polymannuronate. The dosage, concentration of sodium polymannuronate is 4%,and 10mL.The volume of Soce90 during the immobilization is 10mL.更多还原