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Molecular cloning and characterization of an isoflavone 7-O-glucosyltransferase from Pueraria lobata

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Abstract

Key message

A novel isoflavone 7- O -glucosyltransferase PlUGT1 was isolated from Pueraria lobata . PlUGT1 could convert daidzein to daidzin, genistein to genistin as well as formononetin to ononin.

Abstract

Pueraria lobata roots are traditionally consumed as a rich source of isoflavone glycosides that have various human health benefits. However, to date, the genes encoding isoflavone UDP-glycosyltransferases (UGTs) have only been isolated from the roots of soybean seedlings (GmIF7GT), soybean seeds (UGT73F2) and Glycyrrhiza echinata cell suspension cultures (GeIF7GT). To investigate the isoflavone metabolism in P. lobata, 40 types of partial UGT cDNAs were isolated from P. lobata, and seven full-length UGT candidates with preferential expression in roots were identified. Functional assays in yeast (Saccharomyces cerevisiae) revealed that one of these UGT candidates, designated PlUGT1 (official UGT designation UGT88E12), efficiently glycosylated isoflavone aglycones at the 7-hydroxy group. Recombinant PlUGT1 purified from Escherichia coli cells was characterized and shown to be relatively specific for isoflavone aglycones, while flavonoid substrates were poorly accepted. The biochemical results suggested that PlUGT1 was an isoflavone 7-O-glucosyltransferase. The deduced amino acid sequence of PlUGT1 shared only 26 % identity with GeIF7GT, 27 % with UGT73F2 and 63 % with GmIF7GT. The PlUGT1 gene was highly expressed in P. lobata roots relative to other organs and strongly induced by methyl jasmonate signal in P. lobata cell suspension culture. The transcript abundance of PlUGT1 was correlated with the accumulation pattern of isoflavone glycosides such as daidzin in P. lobata plants or in cell suspension culture. The biochemical properties and gene expression profile supported the idea that PlUGT1 could play a role in isoflavone glycosylation in P. lobata.

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Abbreviations

cDNAs:

Complementary DNAs

HID:

Trihydroxyisoflavanone dehydratase

HPLC:

High-performance liquid chromatography

IFS:

Isoflavone synthase

IPTG:

Isopropyl-D-thiogalactopyranoside

LC–MS:

Liquid chromatography–mass spectrometry

MeJA:

Methyl jasmonate

MS:

Murashige and Skoog

NAA:

Naphthalene acetic acid

PSPG:

Putative secondary plant glycosyltransferase

qRT-PCR:

Quantitative RT-PCR

RACE:

Rapid-amplification of cDNA ends

UGTs:

UDP-glycosyltransferases

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Acknowledgments

This project was partially supported by the National Natural Science Foundation of China (Project No. 31170284), the Grant for One Hundred Talents Program of the Chinese Academy of Sciences, China (Project No. Y129441R01) and the Innovation Project of Chinese Academy of Science (Project No. KSCX2-EW-J-20).

Conflict of interest

The authors declare that they have no conflict of interests.

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Authors and Affiliations

  1. CAS Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan, 430074, China

    Jia Li, Zhaobo Li, Changfu Li, Junbo Gou & Yansheng Zhang

  2. University of Chinese Academy of Sciences, Beijing, 100049, China

    Zhaobo Li & Junbo Gou

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  1. Jia Li
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  2. Zhaobo Li
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  3. Changfu Li
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  4. Junbo Gou
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  5. Yansheng Zhang
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Corresponding author

Correspondence to Yansheng Zhang.

Additional information

Communicated by C. F. Quiros.

J. Li and Z. Li contributed equally to this work.

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Li, J., Li, Z., Li, C. et al. Molecular cloning and characterization of an isoflavone 7-O-glucosyltransferase from Pueraria lobata . Plant Cell Rep 33, 1173–1185 (2014). https://doi.org/10.1007/s00299-014-1606-7

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