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Identification of an UDP-glucose: Flavonol 3-O-glucosyl-transferase from cell suspension cultures of soybean (Glycine max L.)

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Abstract

A glucosyltransferase, which catalyses the glucosylation of flavonols, using uridine diphosphate-D-glucose as glucose donor, has been isolated and purified about 5–10 fold from cell suspension cultures of soybean (Glycine max L., var. Mandarin). The pH optimum for this reaction was ca. 8.5 in glycine-NaOH buffer, and no additional cofactors were required. The enzyme glucosylated the following flavonols predominantly at the 3-position: quercetin (Km 126 μM), kaempferol (Km 172 μM), isorhamnetin (Km 200 μM) and fisetin (Km 270 μM). With quercetin as substrate, the apparent Km value for uridine diphosphate-D-glucose was 0.3 M. Glucosylation of flavonols and flavones by this preparation occurred weakly also at the 7-position. No activity was found with dihydroquercetin, naringenin, 4,2′,4′-trihydroxychalcone, daidzein or texasin. The enzyme was specific for flavonoid compounds, since no activity was observed towards cinnamic acids or simple phenols. However, the preparation was contaminated by a vanillic acid glucosyltransferase, from which it could be partially separated by ionexchange chromatography. The specific activity of the flavonol 3-O-glucosyltransferase increased with age of the culture, reaching a maximum late in the growth cycle of the culture.

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Abbreviations

SAM:

S-adenosyl-L-methionine

CMT, SAM:

caffeate 3-O-methyltransferase

FMT, SAM:

flavonoid O-methyltransferase

UDP-glucose:

uridine diphosphate-D-glucose

PAL:

phenylalanine ammonia-lyase

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Author notes

  1. J. E. Poulton

    Present address: Department of Biochemistry and Biophysics, University of California, 95616, Davis, California, USA

Authors and Affiliations

  1. Lehrstuhl für Biochemie der Pflanzen am Biologischen Institut II der Universität Freiburg, Schänzlestr. 1, D-7800, Freiburg, Federal Republic of Germany

    J. E. Poulton & Margit Kauer

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  1. J. E. Poulton
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  2. Margit Kauer
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Poulton, J.E., Kauer, M. Identification of an UDP-glucose: Flavonol 3-O-glucosyl-transferase from cell suspension cultures of soybean (Glycine max L.). Planta 136, 53–59 (1977). https://doi.org/10.1007/BF00387925

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  • DOI: https://doi.org/10.1007/BF00387925

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