TAXOL PRODUCTION UTILIZING A NODULE CULTURE SYSTEM

紫杉醇，一种从生长缓慢的红豆杉组织中提取的二萜类化合物 短叶太平洋紫杉）树，显示出相当大的希望作为抗癌 药 然而，没有可靠的大规模紫杉醇供应。 红豆杉属植物的组织培养可能提供一种替代的来源。 不幸的是， 紫杉醇通过与微管结合，可能对大多数细胞有毒;因此， 紫杉醇高产微培养系统的建立 可能需要分化细胞类型，能够分配 有毒化合物远离活跃分裂的细胞。 因此我们 在此基础上，提出了红豆杉根瘤培养体系的研究建议 与其他木本植物。 结节是致密的细胞簇， 并作为一个内聚单元倍增。 结核对人类特别有吸引力， 因为它们在细胞和组织中都表现出高度紫杉醇生产 分化 诺依文化理论上可以作为 由于外部快速分裂的细胞层维持了紫杉醇的细胞系统， 生长所需的生物量增殖，而内部分化 细胞为紫杉醇的生产提供了一个隔离的场所。 结节培养物 也将作为紫杉醇生物合成研究的可操作模型。 我们将研究影响根瘤发育、生长 和组织HPLC紫杉烷谱。 了解紫杉醇的要求 完整植物中紫杉烷的产生、免疫细胞学定位 并确定季节性环境影响。 等 信息将被用来推断最大化所需的条件。 根瘤培养物中紫杉醇合成。 批次和灌注的影响 还将确定生物反应器系统对生产的影响。 一旦 开发一个红豆杉根瘤系统以提高紫杉醇的产量， 完成，然后它可以用来扩展研究的说明 参与紫杉醇合成的酶和基因。

Taxol, a diterpenoid extracted from tissues of the slow growing Taxus brevifolia Pacific yew) tree, shows considerable promise as an anticancer drug. However, a reliable large scale supply of taxol is not available. Tissue culture of Taxus may provide an alternative supply. Unfortunately, taxol, by binding to microtubules, is likely toxic to most cells; thus the development of a microculture system for the high level production of taxol may require the differentiation of cell types capable of partitioning the toxic compound away from actively dividing cells. To achieve this, we propose to develop a nodule culture system for Taxus based on work done with other woody plants. Nodules are dense cell clusters which form, grow and multiply as a cohesive unit. Nodules are particularly attractive for taxol production as they display a high degree of both cellular and tissue differentiation. Nodule cultures can theoretically serve as a production system for taxol since the outer rapidly dividing cell layers maintain the growth needed for biomass proliferation while the inner differentiated cells provide an isolated location for taxol production. Nodule cultures will also serve as a manipulatable model for studies of taxol biosynthesis. We will investigate the conditions which affect nodule development, growth and tissue HPLC taxane profiles. To understand the requirements taxol production, immunocytological localization of taxanes within intact plants and the seasonal environmental influences will be determined. Such information will be used to deduce the conditions required for maximal taxol synthesis in nodule cultures. The influence of batch and perfused bioreactor systems on production will also be determined. Once the development of a Taxus nodule system for enhanced taxol production is accomplished, then it can be used to extend the research to the elucidation of the enzymes and genes involved in taxol synthesis.