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

使用从Kyaraboku(Taxus cupidata var.)分离的紫杉醇产生真菌。Nana)，通过改变培养条件等方法提高紫杉醇的产量。同时，对新分离的红豆杉属树皮内真菌进行了紫杉醇产量的测定。主要研究结果如下：(1)提高紫杉醇产量的试验：将培养方法由静置培养改为摇瓶固定化培养。而紫杉醇的产量约为常温培养的1/10。通过改变发酵液中糖的组成和加入量，考察了糖作为生物合成前体对紫杉醇生产的影响。结果表明，当添加葡萄糖和少量肌醇时，紫杉醇产量提高1.3倍。此外，还试图通过添加生物遗传抑制物来提高紫杉醇的产量。但是，生产率并没有提高。F…此外，还考察了紫杉醇产量对培养基氮素添加量变化的影响。结果表明，氮素用量减少1/10，紫杉醇产量提高1.3倍。通过对该真菌的培养发现，除紫杉醇外，还产生了一种紫杉烷类化合物(阐明了紫杉醇的结构)。(2)利用紫外线照射和化学试剂对该真菌进行诱变。但是，每种诱变真菌在培养基中培养时，紫杉醇的产量并没有增加。并通过原生质体融合的方法，尝试了紫杉醇产量较高的融合子的形成。通过融合子培养生产紫杉醇的研究正在进行中。(3)紫杉醇产量的提高：研究发现，将真菌发酵生产紫杉醇与固定化Kyaraboku细胞培养在生物反应器中生产紫杉醇相结合，可以显著提高紫杉醇产量。并对几种新分离的红豆杉属真菌的紫杉醇产量进行了测定。结果发现，一种新的真菌比研究中使用的真菌具有更高的紫杉醇产量。此外，还发现东北红豆杉内皮真菌产生的紫杉醇的抗真菌活性与木材提取物的抗真菌活性有关。较少

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