• BMB Reports
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• 제38권6호
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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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• 제23권10호
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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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• 제27권3호
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• pp.91-98
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• 1999

우리나라에 식재된 외래주목의 taxol 및 전구체인 10-deacetyl baccatin III(10-DAB III)의 함랑을 조사하기 위해 천리포수목원에 식재된 외래주목의 잎과 줄기로부터 추출하여 HPLC로 정량분석하였다. 외래주목의 잎과 줄기의 taxol 함량은 종간 및 종내, 그리고 부위에 따라 상당한 변이를 보였다. Taxol 함량은 T. X media cv. Vcrmulcun, T. baccata cv. Washingtonii, T. X cuspidata cv. Nana을 제외하고 잎에서 놓았으며, 전구체인 10-deacetyl baccatin III(10-DAB III)는 모든 종의 잎에서 높았다. 그 중 4월에 채집한 T. cuspidata cv. Nigra의 잎의 taxol 함량은 건물중 당 1,027 ${\mu}g$, 10-DAB III 의 함량은 건물중 당 267 ${\mu}g$로 보고된 태평양주목의 함량보다 월등히 높았다. 이종의 부위별 tax이 함량은 생장점 및 자방 조직보다 잎에서 높았으며, 이 결과들은 조직 및 세포배양의 재료로 널리 이용할 수 있을 것이다.