产紫杉醇内生真菌研究的回顾、进展及趋势

Progress and Trends on Researches of Taxol-producing Endophytic Fungi

自1993年在太平洋紫杉中发现产紫杉醇的内生真菌以来,其寄主植物除红豆杉属外,在罗汉松属、榧树和松柏科植物等26种植物中亦有分布。目前,发现的产紫杉醇的内生真菌有219株,广泛分布在55个属中。内生真菌的鉴定采用基于DNA条形码的分子鉴定为主,在此基础上进行形态学鉴定。紫杉醇的鉴定和产紫杉醇内生真菌的筛选方法从薄层层析分析,逐步发展为质谱和高效液相色谱为主,以及以核磁共振,基于紫杉醇合成途径关键基因的PCR法鉴定和细胞毒性分析为辅的综合性鉴定和筛选方法。紫杉醇含量分析主要应用高效液相色谱法,竞争性抑制酶免疫分析也可用于分析紫杉醇的含量,但其稳定性和可靠性需要进一步验证。通过产紫杉醇的内生真菌实现紫杉醇的规模化生产,除需要筛选高产菌株外,还需要运用代谢工程,基因组学和基因工程方法和技术。

Since the discovery of taxol-producing endophytic fungi was firstly except for Taxus sp. , the host plants of taxol-producing endophytic fungi were widely distributed in 26 species of plants such as Podocarpus sp. , Torreya grandifolia and Coniferav and etc. At p re sen t, 219 strains of taxol-produ-cing endophytic fungi had been found and widely distributed in 55 genus of fungi. As theods , the DNA barcode was applied to the identification of endophytic fungi and then the morphological identifica-tion was carried out based on the molecular identification. The identification of taxol and the ducing endophytic fungi were developed from thin layer chromatography to mass spectrum and high uid chromatography as the main methods , and the supplementary and comprehensive methods including the nuclear magnetic resonance , the screening for key genes in taxol synthesis routes and cytotoxic activity assay. As a mainmethod , the high performance liquid chromatography was applied to analyze the contents of taxol. The competitiveinhibition enzyme immunoassay could also be used to analyze the contents of taxol , but the stawere further needed to be verified. Large scale production of taxol by taxol-producing endophytic fungi were carried out through the high yield strain and the application of the method and technology of mics and gene engineering.