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Journal of Biological Sciences

Year: 2006 | Volume: 6 | Issue: 4 | Page No.: 651-658
DOI: 10.3923/jbs.2006.651.658
Molecular Cloning and Characterization of cDNA Encoding Taxane 2α-O-benzoyltransferase, Catalyzing Taxol Biosynthesis from Taxus media
Dongli Zhao, Qian Wang, Guoyin Kai, Xinglong Wang, Juan Lin, Yan Pi, Zhiqi Miao, Xiaofen Sun and Kexuan Tang

Abstract: By RACE-PCR cloning, a cDNA encoding taxane 2α-O-benzoyltransferase (designated as TmTBT) was isolated from Taxus media, which catalyzes the conversion of 2-debenzoyl-7,13-diacetylbaccatin III, a semisynthetic substrate, to 7,13-diacetylbaccatin III. The full-length cDNA of TmTBT was 1478 bp and contained a 1320 bp Open Reading Frame (ORF) encoding a protein of 440 amino acid residues with molecular weight of 50,090 Da and an isoelectric point (pI) of 6.25, similar to taxane 2α-O-benzoyltransferase from Taxus cuspidata. Sequence comparison analysis revealed that TmTBT had high similarity with other members of plant transferase family. Phylogenetic tree analysis showed that TmTBT had close relationship with taxane 2α-O-benzoyltransferase from T. cuspidata. Tissue expression pattern analysis revealed that TmTBT expressed only in leaves and no expression could be detected in fruits and stems, indicating that TmTBT was a tissue-specific gene.

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How to cite this article
Dongli Zhao, Qian Wang, Guoyin Kai, Xinglong Wang, Juan Lin, Yan Pi, Zhiqi Miao, Xiaofen Sun and Kexuan Tang, 2006. Molecular Cloning and Characterization of cDNA Encoding Taxane 2α-O-benzoyltransferase, Catalyzing Taxol Biosynthesis from Taxus media. Journal of Biological Sciences, 6: 651-658.

Keywords: Phylogenetic tree analysis, RACE, taxane 2a-O-benzoyltransferase, Taxus media and TmTBT

REFERENCES

  • Baloglu, E. and D.G.I. Kingston, 1999. The taxane diterpenoids. J. Nat. Prod., 62: 1448-1472.
    CrossRef    


  • Croom, E.M.J., 1995. Taxus for Toxol and Toxids. In: Taxol Science and Applications, Suffness, M. (Ed.). CRC Press, Boca Raton, pp: 37-70


  • Furmanowa, M. and K. Syklowska-Baranek, 2000. Hairy root cultures of Taxus media var. Hicksii Rehd. as a new source of paclitaxel and 10-deacetylbaccatin III. Biotechnol. Lett., 22: 683-686.
    CrossRef    


  • Geourjon, C. and G. Deleage, 1995. SOPMA: Significant improvement in protein secondary structure prediction by consensus prediction from multiple alignments. Cabios, 11: 681-684.
    Direct Link    


  • Han, K.H., P. Fleming, K. Walker, M. Loper and W.S. Chilton et al., 1994. Genetic transformation of mature Taxus: An approach to genetically control the in vitro production of the anticancer drug. Taxol. Plant Sci., 95: 187-196.
    Direct Link    


  • Hezari, M., N.G. Lewis and R. Croteau, 1995. Purification and characterization of taxa-4(5),11(12)-diene synthase from pacific yew (Taxus brevifolia) that catalyzes the first committed step of Taxol biosynthesis. Arch. Biochem. Biophys., 322: 437-444.
    PubMed    


  • Holton, R.A., C. Somoza, H.B. Kim, F. Liang and R.J. Biediger et al., 1995. The total synthesis of paclitaxel starting with camphor. ACS. Symp. Ser., 583: 288-301.


  • Jaakola, L., A.M. Pirtila, M. Halonen and A. Hohtola, 2001. Isolation of high quality RNA from bilberry (Vaccinium myrtillus L.) fruit. Mol. Biotechnol., 19: 210-213.
    CrossRef    


  • Jennewein, S. and R. Croteau, 2001. Taxol: biosynthesis, molecular genetics and biotechnological applications. Applied Microbiol. Biotechnol., 57: 13-19.
    CrossRef    


  • Kai, G., Miao, C. Qiu, L. Zhang, L. Zhao and Z. Li etal., 2004. Molecular cloning and characterization of a taxadienol acetyl transferase cDNA from Taxus media. Plant Sci., 167: 759-764.
    CrossRef    


  • Kohler, J. and B.R. Goldspiel, 1994. Evaluation of new drug Paclitaxel (Taxol). Pharmacotherapy, 14: 3-34.


  • Shin, S.W., Y.S. Kim and S. Lim, 2000. Elicitors for the regulation of baccatin III biosynthesis in plant cell culture system. Yakhak. Hoechi., 44: 60-65.


  • Thompson, J.D., D.G. Higgins and T.J. Gibson, 1994. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res., 22: 4673-4680.
    CrossRef    PubMed    Direct Link    


  • Walker, K. and R. Croteau, 2001. Molecules of interest: Taxol biosynthetic genes. Phytochemistry, 58: 1-7.
    Direct Link    


  • Yuan, Y.J., Z.J. Wei, Z.Q. Miao and J.C. Wu, 2002. Acting paths of elicitors on taxol biosynthesis pathway and their synergistic effect. Biochem. Eng. J., 10: 77-83.
    Direct Link    


  • Yukimune, Y., Y. Hara, E. Nomura, H. Seto and S. Yoshida, 2000. The configuration of methyl jasmonate affects paclitaxel and baccatin III production in Taxus cells. Phytochemistry, 54: 13-17.
    CrossRef    


  • Walker, K. and R. Croteau, 2000. Taxol biosynthesis-Molecular cloning of a benzoyl- CoA-taxane 2a-O-benzoyltransferase cDNA from Taxus and functional expression in Escherichia coli. Proc. Natl. Acad. Sci. USA., 97: 13591-13596.


  • Walker, K., A. Schoendorf and R. Croteau, 2000. Molecular cloning of a Taxa-4(20),11(12)-dien-5a-ol-O-Acetyl transferase cDNA from Taxus and functional expression in Escherichia coli. Arch. Biochem. Biophys., 374: 371-380.


  • Walker, K., R. Long and R. Croteau, 2002. The final acylation step in Taxol biosynthesis: Cloning of the taxoid C13-side-chain N-benzoyltransferase from Taxus. Proc. Natl. Acad. Sci. USA., 99: 9166-9171.


  • Wildung, M.R. and R. Croteau, 1996. A cDNA cloing for taxadiene synthase, the diterpene cyclase, that catalyzes the committed step of taxol biosynthesis. J. Biol. Chem., 271: 9201-9204.


  • Altschul, S.F., T.L. Madden, A.A. Schaffer, J. Zhang, Z. Zhang, W. Miller and D.J. Lipman, 1997. Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucl. Acids Res., 25: 3389-3402.
    CrossRef    PubMed    Direct Link    

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