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 短叶红豆杉(Shvifolia Pacific Yew)，显示出相当大的抗癌前景 毒品。然而，没有可靠的大规模紫杉醇供应。 红豆杉的组织培养可以提供一种替代供应。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 紫杉醇高产微生物培养系统的研制 可能需要区分细胞类型，从而能够将 有毒化合物，远离活跃分裂的细胞。为了实现这一目标，我们 建议在已完成工作的基础上开发红豆杉根瘤培养系统 与其他木本植物一起生长。结节是密集的细胞团，形成、生长 并作为一个有凝聚力的单位进行繁殖。结核特别吸引人 紫杉醇的产生，因为它们表现出高度的细胞和组织 差异化。从理论上讲，根瘤培养可以作为一种产品 紫杉醇系统，因为外部快速分裂的细胞层保持 生物量增殖所需的生长，而内部分化 细胞为紫杉醇的生产提供了一个孤立的场所。根瘤培养 也将作为紫杉醇生物合成研究的可操作模型。 我们将调查影响结核发育、生长的条件 和组织高效液相色谱紫杉烷图谱。了解紫杉醇的要求 紫杉烷在完整植物中的生产、免疫细胞学定位 并将确定季节性环境影响。是这样的 信息将被用来推断最大值所需的条件 紫杉醇在根瘤培养中的合成。分批和灌装的影响 还将确定生物反应器系统的生产情况。一旦 用于提高紫杉醇产量的红豆杉根瘤系统的开发 完成后，才可以用来将研究延伸到阐释 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.