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Studies on development of a new production method for anti-cancer agent, taxol by use of pestalotiopsis sp.

利用拟盘菌开发抗癌剂紫杉醇新方法的研究。

基本信息

批准号：
14560138
负责人：
TACHIBANA Sanro
金额：
$ 2.3万
依托单位：
Ehime University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

Using a taxol producing fungus isolated from Kyaraboku (Taxus cuspidata var. nana), improvement of taxol productivity by change of incubation conditons and othe methods by mutation and cell fusion of the fungus were conducted. And, taxol productivity was examined by the newly isolated fungi from inner bark of genus taxus trees. The following results were obtained.(1)The trial for increase in the taxol productivity : Culture methods were changed from standing cultures to shaking and immobilized cultures. However, the taxol productivity was about 1/10 times to the standing cultures. Effect of sugar as a biogenetic precursor on taxol production was examined by change the sugar composition and added amount of sugar in the medium. It was found that taxol productivity was increased 1.3 times when glucose and small amount of myoinositol was added to the culture. And, taxol productivity was tried to increase by addition of a biogenetic inhibitor. But, the productivity could not be increased. F … More urthermore, the effect of taxol productivity on change of added amount of nitrogen in the culture medium was examined. It was found that taxol productivity was increased 1.3 times when the amount of nitrogen was decreased 1/10 times. It was found that a kind of taxane compound (elucidated the structure) was produced except for taxol by the culture of the fungus.(2)The fungus was made to mutate by ultraviolet irradiation and chemical reagent. But, taxol productivity was not increased when each mutation fungus was incubated in the culture medium. And, the formation of fusants having higher taxol productivity was tried by protoplast fusion of the fungus. The taxol productivity by cultures of the fusants is being conducted.(3)Increase in taxol productivity : It was found that taxol productivity was drastically improved, when taxol production by fermentation of the fungus is used jointly with taxol production by immobilized cell cultures of Kyaraboku in a bioreactor. And, taxol productivity of some newly isolated fungi from genus taxus trees was examined. It was found that one newly fungus had higher taxol productivity than the fungus used in the research. And furthermore, it was found that antifungal activity of taxol produced by the fungus present in inner bark of Taxus cuspidata van nana are related to the antifungal activity of wood extracts. Less

利用从Kyaraboku（Taxus cuspidata var. nana）、改变培养条件提高紫杉醇产量以及通过诱变和细胞融合等方法提高紫杉醇产量。并对从红豆杉属树木内层树皮中分离到的真菌进行了紫杉醇产率测定。获得了以下结果。（1）提高紫杉醇产量的试验：将培养方法由静置培养改为振荡培养和固定化培养。但紫杉醇的产量约为静置培养的1/10。通过改变培养基中糖的组成和添加量，研究了糖作为生物遗传前体对紫杉醇合成的影响。结果发现，当向培养物中添加葡萄糖和少量肌醇时，紫杉醇的产量提高了1.3倍。并且，尝试通过添加生物遗传抑制剂来增加紫杉醇生产率。但是，生产率不能提高。F ...更多信息因此，研究了紫杉醇生产率对培养基中氮添加量变化的影响。结果表明，当氮素用量减少1/10时，紫杉醇产量增加1.3倍。结果表明，该菌在培养过程中除产生紫杉醇外，还产生一种紫杉烷类化合物（结构已阐明）。（2）采用紫外线诱变和化学诱变相结合的方法对该菌进行诱变。但是，紫杉醇的产量没有增加时，每一个突变真菌在培养基中孵育。并通过原生质体融合的方法尝试了紫杉醇高产融合子的形成。对融合子进行了紫杉醇生产能力的研究。（3）紫杉醇生产率的提高：发现当在生物反应器中通过真菌发酵生产紫杉醇与通过固定化Kyaraboku细胞培养物生产紫杉醇联合使用时，紫杉醇生产率显著提高。并对红豆杉属树木上新分离到的几种真菌产紫杉醇的能力进行了研究。结果表明，一株新的紫杉醇产生菌的紫杉醇产量高于本研究所用的菌株。此外，还发现东北红豆杉树皮真菌产生的紫杉醇的抑菌活性与木材提取物的抑菌活性有关。少