Kumaran Rangarajulu Senthil, Hur Byung-Ki
Department of Biological Engineering, Institute of Industrial Biotechnology, Inha University, Yonghyun-Dong, Nam-Gu, Incheon, South Korea.
Biotechnol Appl Biochem. 2009 Jul 6;54(1):21-30. doi: 10.1042/BA20080110.
Three different strains of the endophytic fungus Phomopsis were isolated from the healthy leaves of Taxus cuspidata (Japanese yew), Ginkgo biloba (ginkgo or maidenhair tree) and Larix leptolepis (Japanese larch) and screened for the production of taxol on a modified liquid medium for the first time. The presence of taxol was confirmed by spectroscopic and chromatographic analyses. The amount of taxol produced by this fungus was quantified by HPLC. The maximum amount of fungal taxol production was recorded as 418 microg/litre in the strain BKH 27. The yield was increased to 8360-fold that found for the fungus Taxomyces andreanae reported previously [Stierle, Strobel and Stierle (1993) Science 260, 214-216]. The fungal taxol extracted also showed a strong cytotoxicity towards the human cancer cells in an apoptosis assay. All the three isolates showed positive sign towards PCR for the conserved sequence of the taxadiene synthase gene. The results suggest that Phomopsis could be an excellent alternative source for taxol and may serve as a potential genetic-engineered species for the enhanced production of taxol.
从东北红豆杉（日本红豆杉）、银杏（银杏或白果树）和日本落叶松的健康叶片中首次分离出三种不同的内生真菌拟茎点霉，并在改良液体培养基上筛选其紫杉醇的产生情况。通过光谱和色谱分析确认了紫杉醇的存在。用高效液相色谱法对该真菌产生的紫杉醇量进行了定量。在菌株BKH 27中，真菌紫杉醇的最大产量记录为418微克/升。产量比先前报道的安德列紫杉菌[Stierle、Strobel和Stierle（1993年）《科学》260，214 - 216]提高到了8360倍。在凋亡检测中，提取的真菌紫杉醇对人类癌细胞也显示出很强的细胞毒性。所有三种分离物在紫杉二烯合酶基因保守序列的聚合酶链反应中均呈阳性。结果表明，拟茎点霉可能是紫杉醇的一个极好的替代来源，并可能作为一种潜在的基因工程物种用于提高紫杉醇的产量。
