• Korean Journal of Pharmacognosy
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• v.24 no.4
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• pp.309-312
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• 1993

A new method for separation and quantitative determination of isoflavone glucosides in rhizomes of Belamcanda chinensis (Iridaceae) by high performance liquid chromatography was elaborated. A reverse-phase system with a Spheri-5 RP-18 column using MeOH:HOAc:$H_2O$(24 : 5 : 71) as a mobile phase was developed. The isoflavonoids were detected at 268 nm and the analysis was successfully carried out within 15 min.

• Journal of Microbiology and Biotechnology
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• v.32 no.5
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• pp.657-662
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• 2022

Glycosyltransferase (GT)-specific degenerate PCR screening followed by in silico sequence analyses of the target clone was used to isolate a member of family1 GT-encoding genes from the established fosmid libraries of soil actinomycetes Micromonospora echinospora ATCC 27932. A recombinant MeUGT1 was heterologously expressed as a His-tagged protein in E. coli, and its enzymatic reaction with semi-synthetic phenoxodiol isoflavene (as a glycosyl acceptor) and uridine diphosphate-glucose (as a glycosyl donor) created two different glycol-attached products, thus revealing that MeUGT1 functions as an isoflavonoid glycosyltransferase with regional flexibility. Chromatographic separation of product glycosides followed by the instrumental analyses, clearly confirmed these previously unprecedented glycosides as phenoxodiol-4'-α-O-glucoside and phenoxodiol-7-α-O-glucoside, respectively. The antioxidant activities of the above glycosides are almost the same as that of parental phenoxodiol, whereas their anti-proliferative activities are all superior to that of cisplatin (the most common platinum chemotherapy drug) against two human carcinoma cells, ovarian SKOV-3 and prostate DU-145. In addition, they are more water-soluble than their parental aglycone, as well as remaining intractable to the simulated in vitro digestion test, hence demonstrating the pharmacological potential for the enhanced bio-accessibility of phenoxodiol glycosides. This is the first report on the microbial enzymatic biosynthesis of phenoxodiol glucosides.

• Molecules and Cells
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• v.37 no.2
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• pp.172-177
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• 2014

A glycosyltransferase, YjiC, from Bacillus licheniformis has been used for the modification of the commercially available isoflavonoids genistein, daidzein, biochanin A and formononetin. The in vitro glycosylation reaction, using UDP-${\alpha}$-D-glucose as a donor for the glucose moiety and aforementioned four acceptor molecules, showed the prominent glycosylation at 4' and 7 hydroxyl groups, but not at the $5^{th}$ hydroxyl group of the A-ring, resulting in the production of genistein 4'-O-${\beta}$-D-glucoside, genistein 7-O-${\beta}$-D-glucoside (genistin), genistein 4',7-O-${\beta}$-D-diglucoside, biochanin A-7-O-${\beta}$-D-glucoside (sissotrin), daidzein 4'-O-${\beta}$-D-glucoside, daidzein 7-O-${\beta}$-D-glucoside (daidzin), daidzein 4', 7-O-${\beta}$-D-diglucoside, and formononetin 7-O-${\beta}$-D-glucoside (ononin). The structures of all the products were elucidated using high performance liquid chromatography-photo diode array and high resolution quadrupole time-of-flight electrospray ionization mass spectrometry (HR QTOF-ESI/MS) analysis, and were compared with commercially available standard compounds. Significantly higher bioconversion rates of all four isoflavonoids was observed in both in vitro as well as in vivo bioconversion reactions. The in vivo fermentation of the isoflavonoids by applying engineered E. coli $BL21(DE3)/{\Delta}pgi{\Delta}zwf{\Delta}ushA$ overexpressing phosphoglucomutase (pgm) and glucose 1-phosphate uridyltransferase (galU), along with YjiC, found more than 60% average conversion of $200{\mu}M$ of supplemented isoflavonoids, without any additional UDP-${\alpha}$-D-glucose added in fermentation medium, which could be very beneficial to large scale industrial production of isoflavonoid glucosides.

• Current Research on Agriculture and Life Sciences
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• v.12
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• pp.9-22
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• 1994

The changes of the time-dependent accumulation of isoflavone aglycones(daidzein, genistein) and their glucosides(daidzin, genistin) by various stress-inducing treatment on cotyledon of soybeans(Dankyungkong and Paldalkong) were investigated. Levels of isoflavone aglycones and their glucosides in soybeans treated with UDP-glucose, $MgSO_4$ and $HgCl_2$ and infected with phytopathogen appeared to be higher than those of treatment with distilled water. When compared with data for control, the levels of isoflavone glucosides in citrate-treated soybeans were increased but those of their aglycones did not appear appreciable differences. In Paldalkong treated with UDP-glucose, $MgSO_4$ and $HgCl_2$ the levels of isoflavone aglycones and their glucosides were higher than in Dankyungkong. In particular, the accumulation of daidzein in Paldalkong was significantly higher than in Dankyungkong. By infection with Botrytis cinerea, the maximum amount of accumulation of daidzein in two cultivar did not to be a large different, but accumulation time appeared more rapidly in Paldalkong than Dankyungkong. The accumulation amount of daidzein and genistein in Dankyungkong treated with elicitors appeared to be similar at initial stage, but the level of daidzein after 48hours appeared to be higher than that of genistein. In Paldalkong, the level of daidzein was higher than that of genistein at all stage. The level of daidzein in soybeans infected with phytopathogen appeared to be higher than that of genestein.

• Journal of Microbiology and Biotechnology
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• v.22 no.11
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• pp.1523-1531
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• 2012

The metabolite profile of meju during fermentation was analyzed using mass spectrometry techniques, including GC-MS and LC-MS, and the bacterial diversity was characterized. The relative proportions of bacterial strains indicated that lactic acid bacteria, such as Enterococcus faecium and Leuconostoc lactis, were the dominant species. In partial least-squares discriminate analysis (PLS-DA), the componential changes, which depended on fermentation, proceeded gradually in both the GC-MS and LC-MS data sets. During fermentation, lactic acid, amino acids, monosaccharides, sugar alcohols, and isoflavonoid aglycones (daidzein and genistein) increased, whereas citric acid, glucosides, and disaccharides decreased. MS-based metabolite profiling and bacterial diversity characterization of meju demonstrated the changes in metabolites according to the fermentation period and provided a better understanding of the correlation between metabolites and bacterial diversity.

• KSBB Journal
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• v.16 no.2
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• pp.174-178
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• 2001

Soybeans contain the phytoestrogens genistein and daidzein, their glucosides genistin and daidzin and coumesterol. These isoflavonoid compounds are capable of producing an estrogenic response in a number of diverse species. This study determined optimum conditions for the extraction of the main isoflavones(daidzin, genistin, daidzein, genistein) in defatted soybean meal using high-performance liquid chromatography. The best optimum extraction was achieved at 75% ethanol, $80^{circ}C$, pH4 and a three hour contact time. In addition, isoflavones with high purity were separated by adding up to 4%(w/v) of calcium chloride dihydrate. Most soybean extracts were composed of $beta$-glucosidic conjugate(daidzin, genistin) which is difficult to adsorb in body. Therefore, $beta$-glucosidase was used to convert as conjugate to aglycone form (daidzein, genistein) which is easy to adsorb. The optimal conditions of enzyme reaction involved to be 8.4 units of enzyme concentration, pH5.0, $40^{circ}C$ and 40 minutes.