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cDNA cloning and expression of isoflavonoid-specific glucosyltransferase from Glycyrrhiza echinata cell-suspension cultures

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Abstract

A cDNA encoding UDP-glucose: formononetin 7-O-glucosyltransferase, designated UGT73F1, was cloned from yeast extract-treated Glycyrrhiza echinata L. cell-suspension cultures using probes from Scutellaria baicalensis UDP-glucose: flavonoid 7-O-glucosyltransferase. The open reading frame of the UGT73F1 cDNA encodes a 441-amino-acid protein with a predicted molecular mass of 48.7 kDa. The deduced amino acid sequence showed that the protein is related to the stress-inducible glucosyltransferases. UGT73F1 mRNA was not detected in untreated G. echinata cultures but was transiently induced by treatment with yeast extract. Recombinant UGT73F1 was expressed as a histidine-tag fusion protein in Escherichia coli and purified to near homogeneity by nickel chelate chromatography. The purified recombinant enzyme was selective for isoflavonoid, formononetin and daidzein as substrates, while flavonoids and various tested non-flavonoid compounds were poor substrates.

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Abbreviations

GT:

UDP-glycosyltransferase

rUGT73F1:

recombinant UGT73F1

UBGT: UDP-glucose::

baicalein 7-O-glucosyltransferase

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Acknowledgments

The authors thank Prof. S. Ayabe for supplying the G. echinata callus and the cDNA library. This research was funded by a Sasakawa Scientific Research Grant from the Japan Science Society and by a Kitasato University Research Grant for Young Researchers.

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Authors and Affiliations

  1. School of Pharmaceutical Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, 108-8641, Tokyo, Japan

    Shigeyuki Nagashima, Rieko Inagaki, Akiko Kubo, Masao Hirotani & Takafumi Yoshikawa

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  1. Shigeyuki Nagashima
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  2. Rieko Inagaki
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  3. Akiko Kubo
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  4. Masao Hirotani
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  5. Takafumi Yoshikawa
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Corresponding author

Correspondence to Takafumi Yoshikawa.

Additional information

The nucleotide sequence data reported in this paper will appear in the DDBJ/EMBL/GenBank nucleotide sequence databases with the accession number AB098614.

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Nagashima, S., Inagaki, R., Kubo, A. et al. cDNA cloning and expression of isoflavonoid-specific glucosyltransferase from Glycyrrhiza echinata cell-suspension cultures. Planta 218, 456–459 (2004). https://doi.org/10.1007/s00425-003-1118-0

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  • DOI: https://doi.org/10.1007/s00425-003-1118-0

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