The expression of 1-deoxy-d-xylulose synthase and geraniol-10-hydroxylase or anthranilate synthase increases terpenoid indole alkaloid accumulation in Catharanthus roseus hairy roots

Christie A.M. Peebles; Guy W. Sander; Erik H. Hughes; Ryan Peacock; Jacqueline V. Shanks; Ka Yiu San

doi:10.1016/j.ymben.2010.11.005

The expression of 1-deoxy-d-xylulose synthase and geraniol-10-hydroxylase or anthranilate synthase increases terpenoid indole alkaloid accumulation in Catharanthus roseus hairy roots

Christie A.M. Peebles, Guy W. Sander, Erik H. Hughes, Ryan Peacock, Jacqueline V. Shanks, Ka Yiu San

Chemical Engineering

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84 Scopus citations

Abstract

The terpenoid indole alkaloid (TIA) pathway in Catharanthus roseus produces two important anticancer drugs, vinblastine and vincristine, in very low yields. This study focuses on overexpressing several key genes in the upper part of the TIA pathway in order to increase flux toward downstream metabolites within hairy root cultures. Specifically, we constructed hairy root lines with inducible overexpression of 1-deoxy- D-xylulose synthase (DXS) or geraniol-10-hydroxylase (G10H). We also constructed hairy root lines with inducible expression of DXS and anthranilate synthase α subunit (ASA) or DXS and G10H. DXS overexpression resulted in a significant increase in ajmalicine by 67%, serpentine by 26% and lochnericine by 49% and a significant decrease in tabersonine by 66% and hörhammericine by 54%. Co-overexpression of DXS and G10H caused a significant increase in ajmalicine by 16%, lochnericine by 31% and tabersonine by 13%. Likewise, DXS and ASA overexpression displayed a significant increase in hörhammericine by 30%, lochnericine by 27% and tabersonine by 34%. These results point to the need for overexpressing multiple genes within the pathway to increase the flux toward vinblastine and vincristine.

Original language	English (US)
Pages (from-to)	234-240
Number of pages	7
Journal	Metabolic Engineering
Volume	13
Issue number	2
DOIs	https://doi.org/10.1016/j.ymben.2010.11.005
State	Published - Mar 2011

Bibliographical note

Funding Information:
The authors would like to thank Dr. Nam-Hai Chua at the Rockefeller University for providing the inducible promoter plasmid (pTA7002), Dr. Eugene Nester at the University of Washington for providing the A. rhizogenes 15834 strain used, Dr. Johan Memelink at the University of Leiden for providing us with the geraniol-10-hydroxylase sequence prior to publication, and Dr. Patricia Leon from the Universidad Nacional Autonoma de Mexico for providing the cDNA clone of Arabidopsis DXS. This work was supported by funding from the National Science Foundation (Grant numbers BES-0003730 , BES-9906978 , BES0224593 , BES0224600 , BES0420840 , and CBET0729622 ). Erik Hughes was supported by a training grant from the National Institutes of Health ( T32-GM08362 ). Christie Peebles was supported by the NSF Graduate Fellowship .

Keywords

Madagascar periwinkle
Methyl-erythritol phosphate pathway
Plant metabolic engineering
Terpenoid indole alkaloid
Tissue culture

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10.1016/j.ymben.2010.11.005

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The expression of 1-deoxy-d-xylulose synthase and geraniol-10-hydroxylase or anthranilate synthase increases terpenoid indole alkaloid accumulation in Catharanthus roseus hairy roots. / Peebles, Christie A.M.; Sander, Guy W.; Hughes, Erik H. et al.

In: Metabolic Engineering, Vol. 13, No. 2, 03.2011, p. 234-240.

Research output: Contribution to journal › Article › peer-review

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abstract = "The terpenoid indole alkaloid (TIA) pathway in Catharanthus roseus produces two important anticancer drugs, vinblastine and vincristine, in very low yields. This study focuses on overexpressing several key genes in the upper part of the TIA pathway in order to increase flux toward downstream metabolites within hairy root cultures. Specifically, we constructed hairy root lines with inducible overexpression of 1-deoxy- D-xylulose synthase (DXS) or geraniol-10-hydroxylase (G10H). We also constructed hairy root lines with inducible expression of DXS and anthranilate synthase α subunit (ASA) or DXS and G10H. DXS overexpression resulted in a significant increase in ajmalicine by 67%, serpentine by 26% and lochnericine by 49% and a significant decrease in tabersonine by 66% and h{\"o}rhammericine by 54%. Co-overexpression of DXS and G10H caused a significant increase in ajmalicine by 16%, lochnericine by 31% and tabersonine by 13%. Likewise, DXS and ASA overexpression displayed a significant increase in h{\"o}rhammericine by 30%, lochnericine by 27% and tabersonine by 34%. These results point to the need for overexpressing multiple genes within the pathway to increase the flux toward vinblastine and vincristine.",

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note = "Funding Information: The authors would like to thank Dr. Nam-Hai Chua at the Rockefeller University for providing the inducible promoter plasmid (pTA7002), Dr. Eugene Nester at the University of Washington for providing the A. rhizogenes 15834 strain used, Dr. Johan Memelink at the University of Leiden for providing us with the geraniol-10-hydroxylase sequence prior to publication, and Dr. Patricia Leon from the Universidad Nacional Autonoma de Mexico for providing the cDNA clone of Arabidopsis DXS. This work was supported by funding from the National Science Foundation (Grant numbers BES-0003730 , BES-9906978 , BES0224593 , BES0224600 , BES0420840 , and CBET0729622 ). Erik Hughes was supported by a training grant from the National Institutes of Health ( T32-GM08362 ). Christie Peebles was supported by the NSF Graduate Fellowship . ",

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The expression of 1-deoxy-d-xylulose synthase and geraniol-10-hydroxylase or anthranilate synthase increases terpenoid indole alkaloid accumulation in Catharanthus roseus hairy roots

Abstract

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