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Zingiber zerumbet CYP71BA1 catalyzes the conversion of α-humulene to 8-hydroxy-α-humulene in zerumbone biosynthesis

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Abstract

Plant cytochrome P450s are involved in the biosynthesis of various classes of secondary metabolites. To elucidate the biosynthesis of zerumbone, a sesquiterpenoid with multiple potential anticancer properties, a family of P450 genes expressed in rhizomes of Zingiber zerumbet Smith, were cloned using a PCR-based cloning strategy. After functional expression in yeast, one of these P450s was found to convert α-humulene into 8-hydroxy-α-humulene, a proposed intermediate of zerumbone biosynthesis. This P450 has been designated CYP71BA1, a new member of the CYP71 family. CYP71BA1 transcripts were detected almost exclusively in rhizomes and showed a similar expression pattern to ZSS1 transcripts during rhizome development. Coexpression of a gene cluster encoding four enzymes of the mevalonate pathway with CYP71BA1 and ZSS1 in Escherichia coli leads to the production of 8-hydroxy-α-humulene in the presence of mevalonate, suggesting the possibility of microbial production of this zerumbone intermediate from a relatively simple carbon source by metabolic engineering.

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Abbreviations

ATR2:

Arabidopsis P450 reductase 2 gene

MK:

Mevalonate kinase

PMK:

Phosphomevalonate kinase

MPPD:

Mevalonate diphosphate decarboxylase

IPPI:

Isopentenyl diphosphate isomerase

IPP:

Isopentenyl diphosphate

DMAPP:

Dimethylallyl diphosphate

CPR:

Cytochrome P450 reductase

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Acknowledgments

We thank Mr. T. Ishida, Ms. R. Sawa, and Mr. H. Miyawaki for providing the ginger plants. We are also grateful to Dr. D. Pompon (CNRS, Gif-sur-Yvette, France) for providing the yeast expression system. This work was supported in part by the Research and Development Program for New Bio-industry Initiatives (2006–2010) from the Bio-oriented Technology Research Advancement Institution (BRAIN), the “Academic Frontier” Project for Private Universities: Matching fund subsidy from the Ministry of Education, Culture, Sports, Science and Technology (2004–2008), and the Sasagawa Scientific Research Grant from the Japan Science Society.

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Authors and Affiliations

  1. Department of Bioscience, Graduate School of Agriculture, Kinki University, Nakamachi, Nara, 631-8505, Japan

    Fengnian Yu, Sho Okamoto, Kazuhisa Yamasaki & Ryutaro Utsumi

  2. Central Laboratories for Frontier Technology, Kirin Holdings Co., Ltd, i-BIRD 3-570, Suematsu, Nonoichi, Ishikawa, 921-8836, Japan

    Hisashi Harada & Norihiko Misawa

Authors
  1. Fengnian Yu
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  2. Sho Okamoto
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  3. Hisashi Harada
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  4. Kazuhisa Yamasaki
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  5. Norihiko Misawa
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  6. Ryutaro Utsumi
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Corresponding author

Correspondence to Ryutaro Utsumi.

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F. Yu and S. Okamoto contributed equally to this work.

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Yu, F., Okamoto, S., Harada, H. et al. Zingiber zerumbet CYP71BA1 catalyzes the conversion of α-humulene to 8-hydroxy-α-humulene in zerumbone biosynthesis. Cell. Mol. Life Sci. 68, 1033–1040 (2011). https://doi.org/10.1007/s00018-010-0506-4

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  • DOI: https://doi.org/10.1007/s00018-010-0506-4

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