• 생약학회지
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• 제24권4호
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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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• 제32권5호
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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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• 제37권2호
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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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• 제12권
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• pp.9-22
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• 1994

Soybean mosaic virus에 감수성 품종인 단경콩과 저항성 품종인 팔달콩의 cotyledon에 여러형태로 자극을 처리하였을때 유도되는 isoflavone aglycones(daidzein, genistein)와 이들 배당체들(daidzin, genistin)의 시간별 축적 정도를 조사하였다. UDP-glucose, $MgSO_4$, $HgCl_2$의 처리 및 Botrytis cinerea의 감염에 의하여 두 품종 모두 isoflavone aglycones와 이들 배당체들의 함량이 대조구(water 처리)와 비교하여 증가하였다. 그러나 citrate의 처리에 의해서는 isoflavone aglycones의 함량은 대조구와 큰 차이가 없었으나 isoflavone glucosides의 함량은 크게 증가하였다. UDP-glucose, $MgSO_4$ 및 $HgCl_2$의 처리에 의하여 isoflavone aglycones의 함량은 단경콩보다 팔달콩에서 크게 나타났다. 특히 daidzein의 경우, 단경콩보다 팔달콩에서 현저하게 크게 나타났다. 그러나 Botrytis cinerea의 감염에 의해서 daidzein의 최대 축적 양에는 두 품종 사이에 큰 차이가 없으나 최대 축적 시간은 팔달콩이 단경콩보다 빠르게 나타났다. 그리고 그 배당체인 daidzin과 geinstein의 축적은 단경콩보다 팔달콩에서 모든 자극처리에의해 높게 나타났다. UDP-glucose, $MgSO_4$, 및 $HgCl_2$의 처리에 의한 daidzein과 geinstein이 축적되는 양은 단경콩에서는 초기에는(24hr까지)에는 비슷하나, 48hr이후에는 daidzein의 축적이 geinstein의 함량보다 크게 나타났다. 그러나 팔달콩에서는 대체로 전 기간에서 daidzein의 함량이 geinstein의 함량보다 크게 나타났다. 병원균의 감염에 의해서는 두 품종에서 모두 daidzein의 양이 geinstein의 양보다 크게 나타났다.

• Journal of Microbiology and Biotechnology
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• 제22권11호
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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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• 제16권2호
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• pp.174-178
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• 2001

본 연구는 항산화작용, 항암작용, estrogen 유사작용, 항골다공증작용 등 다양한 생리적 기능을 가진 isoflavones의 추출조전을 추출용매획 농도, 추출온도, pH 및 추출시간으로 나누어 단계적으로 isoflavones을 추출함에 있어 최척의 추출 조건올 찾고자 하였으며, 보다 기능이 잘 알려진 4종 isoflavones 추출에 대한 결과를 분석하여 최적추출 초건을 선정하였다. 그 결과 75% ethanol, $80^{circ}C$, pH4, 3시간의 추출 조건에서 $4,024\mu\textrm{g}/mL$의 가장 높온 총 isofla vones 추출량을 나타내었고, 대두추출물 중의 단백질 제거를 위해 이용된 염화칼융의 농도가 증가할수록 보다 높은 함량의 isotla yones윤 얻을 수 있었다. 위의 결과로 얻은 최적추출조건에서의 대두추춤물에는 거의 대부분이 체내에 홉수되지 않는 daidzin과 genistin 같은 배당체의 형태로 존재해 있음을 확인하였다. 따라서 $beta$-glucosidase라는 효소를 이용하여 이러한 배당채 들흘 daidzein이나 genistein과 같이 체내여] 홉수 가능한 aglycones 형태로 전환시키는데 있어 효소의 농도, 반응온도 및 pH, 반웅시간의 조건에서 aglycon$\xi$s 형혜익 최적 전환수 율 조건을 각각 선정하였다. 그 결과 효소농도 8.7 units, 온도 $40^{circ}C$, pHS의 초건에서 4 40 분 풍안 반응시켰을 때 90%이상 전환된 aglycones 형태를 확인할 수 있었다.