Abstract
Genes coding for 10-deacetylbaccatin III-10-O-acetyl transferase and C-13 phenylpropanoid side chain-CoA acyltransferase were used as molecular markers for screening of Taxol-producing endophytic fungi. Using PCR, three out of 90 endophytic fungi, isolated from Taxus x media and Taxus yunnanensis, gave positive results. These 3 strains, when grown in 300 ml potato/dextrose liquid medium at 25°C for 10 days, contained 100–160 μg Taxol/g dry wt of mycelium.
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References
Jennewein S, Wildung MR, Chau M, Walker K, Croteau R (2004) Random sequencing of an induced Taxus cell cDNA library for identification of clones involved in Taxol biosynthesis. Proc Natl Acad Sci USA 101(24):9149–9154
Ji Y, Bi JN, Yan B, Zhu XD (2006) Taxol-producing fungi: a new approach to industrial production of taxol. Chin J Biotechnol 22:1–6
Kim W, Mauthe W, Hausner G, Klassen G (1990) Isolation of high-molecular-weight DNA and double-stranded RNAs from fungi. Can J Bot 68:1898–1902
Li J, Hu Y, Chen W, Lin Z (2006) Identification and pilot study of Taxus endophytic fungi’s taxol-producing correlation gene BAPT. Biotechnol Bull S1:356–371
Stierle A, Strobel G, Stierle D (1993) Taxol and taxane production by Taxomyces andreanae, an endophytic fungus of Pacific yew. Science 260:214–216
Strobel G, Yang X, Sears J, Kramer R, Sidhu RS, Hess WM (1996) Taxol from Pestalotiopsis microspora, and endophytic fungus from Taxus wallachiana. Microbiology 142:435–440
Walker K, Croteau R (2000) Molecular cloning of a 10-deacetylbaccatin III-10-O-acetyl transferase cDNA from Taxus and functional expression in Escherichia coli. Proc Natl Acad Sci USA 97(2):583–587
Walker K, Fujisaki S, Long R, Croteau R (2002) Molecular cloning and heterologous expression of the C-13 phenylpropanoid side chain-CoA acyltransferase that functions in Taxol biosynthesis. Proc Natl Acad Sci USA 99(20):12715–12720
Wang J, Li G, Lu H, Zheng Z, Huang Y, Su W (2000) Taxol from Tubercularia sp. strain TF5, an endophytic fungus of Taxus mairei. FEMS Microbiol Lett 193:249–253
Wildung MR, Croteau R (1996) A cDNA clone for taxadiene synthase, the diterpene cyclase that catalyzes the committed step of taxol biosynthesis. J Biol Chem 271(16):9201–9204
Zhou X, Wang Z, Jiang K, Wei Y, Lin J, Sun X, Tang K (2007) Screening of taxol-producing endophytic fungi from Taxus chinensis var. mairei. Appl Biochem Microbiol 43(4):439–443
Acknowledgements
This research is financially supported by National Natural Science Foundation of China (Grant 20776058) and New Century Talents Support Program by the Ministry of Education of China in 2006. We thank Mrs. Xiaoman Gu and Hong Cheng of the Analysis and Testing Center of Huazhong University of Science and Technology for mass spectroscopy analysis.
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Peng Zhang and Peng-peng Zhou contributed equally to this work.
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Zhang, P., Zhou, Pp., Jiang, C. et al. Screening of Taxol-producing fungi based on PCR amplification from Taxus . Biotechnol Lett 30, 2119–2123 (2008). https://doi.org/10.1007/s10529-008-9801-7
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DOI: https://doi.org/10.1007/s10529-008-9801-7