• BMB Reports
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• v.38 no.6
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• pp.668-675
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• 2005

A full-length cDNA encoding taxadiene synthase (designated as TmTXS), which catalyzes the first committed step in the Taxol biosynthetic pathway, was isolated from young leaves of Taxus media by rapid amplification of cDNA ends (RACE). The full-length cDNA of TmTXS had a 2586 bp open reading frame (ORF) encoding a protein of 862 amino acid residues. The deduced protein had isoelectric point (pI) of 5.32 and a calculated molecular weight of about 98 kDa, similar to previously cloned diterpene cyclases from other Taxus species such as T. brevifolia and T. chinenisis. Sequence comparison analysis showed that TmTXS had high similarity with other members of terpene synthase family of plant origin. Tissue expression pattern analysis revealed that TmTXS expressed strongly in leaves, weak in stems and no expression could be detected in fruits. This is the first report on the mRNA expression profile of genes encoding key enzymes involved in Taxol biosynthetic pathway in different tissues of Taxus plants. Phylogenetic tree analysis showed that TmTXS had closest relationship with taxadiene synthase from T. baccata followed by those from T. chinenisis and T. brevifolia. Expression profiles revealed by RT-PCR under different chemical elicitor treatments such as methyl jasmonate (MJ), silver nitrate (SN) and ammonium ceric sulphate (ACS) were also compared for the first time, and the results revealed that expression of TmTXS was all induced by the tested three treatments and the induction effect by MJ was the strongest, implying that TmTXS was high elicitor responsive.

• Journal of Microbiology and Biotechnology
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• v.23 no.10
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• pp.1372-1380
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• 2013

Different endophytic fungi isolated from Himalayan Yew plants were tested for their ability to produce taxol. The BAPT gene (C-13 phenylpropanoid side chain-CoA acetyl transferase) involved in the taxol biosynthetic pathway was used as a molecular marker to screen taxol-producing endophytic fungi. Taxol extracted from fungal strain TBPJ-B was identified by HPLC and MS analysis. Strain TBPJ-B was identified as Fusarium redolens based on the morphology and internal transcribed spacer region of nrDNA analysis. HPLC quantification of fungal taxol showed that F. redolens was capable of producing $66{\mu}g/l$ of taxol in fermentation broth. The antitumour activity of the fungal taxol was tested by potato disc tumor induction assay using Agrobacterium tumefaciens as the tumor induction agent. The present study results showed that PCR amplification of genes involved in taxol biosynthesis is an efficient and reliable method for prescreening taxol-producing fungi. We are reporting for the first time the production of taxol by F. redolens from Taxus baccata L. subsp. wallichiana (Zucc.) Pilger. This study offers important information and a new source for the production of the important anticancer drug taxol by endophytic fungus fermentation.

• Journal of the Korean Wood Science and Technology
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• v.27 no.3
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• pp.91-98
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• 1999

The content of taxol and related compounds in needle and stem tissues of exotic yews growing in the Chollipo Arboretum which located in the West Sea of Korea were determined by HPLC. A considerable variation in contents of taxol and the related compounds within species and among cultivars were observed. The content of taxol was much higher in the needles than those in the young stem tissues except T. X media cv. Vermuleun, T. baccata cv. Washingtonii, and T. X cuspidata cv. Nana, and the content of 10-deacetyl baccatin III(10-DAB III) was also slightly higher in the needle. The needle of T. cuspidata cv. Nigra collected from April was contained the high amount of taxol(1,027 ${\mu}g$ per g dry weight) and also contained high quantity in 10-DAB III(267 ${\mu}g$ per g dry weight). Concentration of taxol in needle was significantly greater than those of meristemic and ovary tissues, Exotic yews growing at the Chollipo Arboretum containing T. cuspidata cv. Nigra could be suitable materials as a source for the tissue and cell cultures for production of taxol.