Zhang Peng, Zhou Peng-Peng, Yu Long-Jiang
Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.
Curr Microbiol. 2009 Sep;59(3):227-32. doi: 10.1007/s00284-008-9270-1. Epub 2009 May 30.
Fermentation processes using taxol-producing fungi other than Taxus spp. may be an alternative way to produce taxol, which is an important antitumor agent used widely in the clinic setting. In this study, a taxol-producing endophytic fungus strain MD2 was isolated from the inner bark of Taxus media. Strain MD2 produced taxol when grown in potato dextrose liquid medium. The fungal taxol-which was analyzed by ultraviolet, high-performance liquid chromatography and mass spectrometry-was shown to be identical to authentic taxol and 10-deacetylbaccatin III. Further analysis with nuclear magnetic resonance (NMR) spectroscopy to show the chemical structure of the fungal taxol indicated that the fungal taxol produced an NMR spectrum identical to that of authentic taxol. Strain MD2 was identified as Cladosporium cladosporioides according to morphology of the fungal culture, characteristics of the spores, and analysis of 18S rDNA sequence. In addition, 10-deacetylbaccatin III-10-O-acetyl transferase gene of C. cladosporioides MD2 was cloned for the first time and was shown to share 99% identity with that of T. x media and 97% identity with that of T. wallichiana var. mairei.
使用除红豆杉属植物之外的产紫杉醇真菌进行发酵过程，可能是生产紫杉醇的一种替代方法，紫杉醇是一种在临床环境中广泛使用的重要抗肿瘤药物。在本研究中，从南方红豆杉的内皮中分离出一株产紫杉醇的内生真菌菌株MD2。菌株MD2在马铃薯葡萄糖液体培养基中生长时会产生紫杉醇。通过紫外、高效液相色谱和质谱分析的真菌紫杉醇，显示与正品紫杉醇和10 - 去乙酰巴卡亭III相同。用核磁共振（NMR）光谱进一步分析以显示真菌紫杉醇的化学结构，表明真菌紫杉醇产生的NMR光谱与正品紫杉醇的光谱相同。根据真菌培养的形态、孢子特征以及18S rDNA序列分析，菌株MD2被鉴定为枝孢霉。此外，首次克隆了枝孢霉MD2的10 - 去乙酰巴卡亭III - 10 - O - 乙酰转移酶基因，显示与南方红豆杉的该基因有99%的同一性，与云南红豆杉的该基因有97%的同一性。
