Study on production of biologically active substances by immobilized cells

采用固定化细胞制备了红豆杉（Taxus cuspidata var. nana）和银杏（Icho）的紫杉醇和银杏内酯。此外，还研究了从枇杷内皮分离的拟盘菌生产紫杉醇的方法。细胞悬浮培养制备紫杉醇。紫杉醇的产量约为完整植株的一半。然而，通过在细胞培养中添加激发子和生物遗传前体，产量增加了约10%。孵育后的细胞可以用海藻酸盐固定。固定细胞释放紫杉醇到培养基中。释放到培养基中的紫杉醇量约为细胞中产生的紫杉醇的60 ~ 80%。固定细胞连续释放紫杉醇4周。紫杉醇释放量为0.3 ~ 0.6mg/L。认识到利用生物反应器生产紫杉醇的可能性。从枇杷内皮中分离到部分拟盘菌。该真菌可生产紫杉醇等紫杉醇类化合物，但其产量尚在研究中。以银杏种子为原料，采用悬浮培养法制备银杏内酯细胞。银杏内酯的产量几乎与完整植株相同。在培养物中加入激发子和生物遗传前体后，银杏内酯的产量增加了约5倍。孵育后的细胞可以用海藻酸盐固定。固定化细胞释放银杏内酯到时间介质中。释放到培养基中的银杏内酯量约为细胞产生银杏内酯的20%。认识到利用生物反应器生产紫杉醇的可能性。

Production of taxol and ginkgolides by immobilized cells from Kyaraboku (Taxus cuspidata var. nana) and Icho (Ginkgo biloba) was conducted. And, production of taxol by pestalotia fungi isolated from inner bark of Kyaraboku was also conducted.1. Cells induced from Kyaraboku stems were produced taxol by cell suspension cultures. The amount of taxol production was about a half compared to those of the intact plant. However, the amount of the production was increased about 10-holds by addition of an elicitor and a biogenetic precursor to the cell cultures.2. Incubated cells could be immobilized with alaginate. The immobilized cells released taxol into medium. The amount of taxol released into the medium was about 60 to 80% of the taxol produced in the cells. The immobilized cells released continually taxol for 4 weeks. The amount of taxol released into the medium was 0.3 to 0.6mg/L.A possiblity for production of taxol by use of a bioreactor was recognized.3. Some pestalotia fungi were isolated from inner bark of Kyaraboku. The fungi could be produced taxanes including taxol, however, the amount of taxol production are under investigation.4. Cells from Ginkgo seeds were produced ginkgolides by cell suspension cultures. The amount of ginkgolides production was almost same as those of the intact plant. The amount of ginkgolides production was increased about 5 times by addition of an elicitor and a biogenetic precursor to the cultures.5. Incubated cells could be immobilized with alaginate. The immobilized cells released ginkgolides into time medium. The amount of ginkgolides released into the medium was about 20% of ginkgolides produced in the cells. A possiblity for production of taxol by use of a bioreactor was recognized.