• Title/Summary/Keyword: Aglycon

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Enzymatic Biotransformation of Ginsenoside Rb1 and Gypenoside XVII into Ginsenosides Rd and F2 by Recombinant β-glucosidase from Flavobacterium johnsoniae

  • Hong, Hao;Cui, Chang-Hao;Kim, Jin-Kwang;Jin, Feng-Xie;Kim, Sun-Chang;Im, Wan-Taek
    • Journal of Ginseng Research
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    • v.36 no.4
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    • pp.418-424
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    • 2012
  • This study focused on the enzymatic biotransformation of the major ginsenoside Rb1 into Rd for the mass production of minor ginsenosides using a novel recombinant ${\beta}$-glucosidase from Flavobacterium johnsoniae. The gene (bglF3) consisting of 2,235 bp (744 amino acid residues) was cloned and the recombinant enzyme overexpressed in Escherichia coli BL21(DE3) was characterized. This enzyme could transform ginsenoside Rb1 and gypenoside XVII to the ginsenosides Rd and F2, respectively. The glutathione S-transferase (GST) fused BglF3 was purified with GST-bind agarose resin and characterized. The kinetic parameters for ${\beta}$-glucosidase had apparent $K_m$ values of $0.91{\pm}0.02$ and $2.84{\pm}0.05$ mM and $V_{max}$ values of $5.75{\pm}0.12$ and $0.71{\pm}0.01{\mu}mol{\cdot}min^{-1}{\cdot}mg$ of $protein^{-1}$ against p-nitrophenyl-${\beta}$-D-glucopyranoside and Rb1, respectively. At optimal conditions of pH 6.0 and $37^{\circ}C$, BglF3 could only hydrolyze the outer glucose moiety of ginsenoside Rb1 and gypenoside XVII at the C-20 position of aglycon into ginsenosides Rd and F2, respectively. These results indicate that the recombinant BglF3 could be useful for the mass production of ginsenosides Rd and F2 in the pharmaceutical or cosmetic industry.

Optimum Conversion to the Aglycone Form Using $\beta$-glucosidase and Isoflavone Extraction from Soybean (대두로부터 Isoflavone추출 및 $\beta$-glucosidase를 이용한 Aglycone 형태로의 전환 최적 조건)

  • 김기욱;전병수
    • 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.

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Purification of Antimicrobial Compounds and Antimicrobial Effects of Schima wallichii subsp. liukiuensis against Candida sp. (Schima wallichii subsp. liukiuensis의 Candida종에 대한 항균효과 및 항균물질의 분리정제)

  • Choi, Mynug-Suk;Shin, Kuem;Yang, Jae-Kyung;Ahan, Jin-Kwon;Kwon, Oh-Woong;Lee, Yi-Young
    • KSBB Journal
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    • v.16 no.3
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    • pp.269-273
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    • 2001
  • To develop natural antimicrobial substances from Theaceae, Schima wallichii subsp. liukiuensis was selected from 218 woody plants, and antimicrobial compounds against bacteria, fungi, and yeast were isolated. The antimicrobial activity of ethanol extracts proved higher than those of other organic solvents. The antimicrobial activity of S. liukiuensis extract showed no differences in sesonal variation, but, that of plant part was high in bark at autumn. An antimicrobial substance was isolated from the extract of Schima using column chromatography packed with silica gel and sephadex LH-20, and then a purified antimicrobial substance (Compound I) was obtained using HPLC analysis. The Compound I in the analysis of UV, IR, and GC-MS presumed a triterpene or steroidal saponin, ${\alpha}$-sitisterol as aglycon combined three sugars. The minimal inhibitory concentration (MIC) of the Compound I against a bacteria, fungi, and yeast were 1.25 g/L, 5.0 g/L, and 0.040 g/L, respectively. This is much lower than the MIC of hinokitiol, an natural antimicrobial compound used commercially, which suggests that Compound I could be developed as a natural preservative and pharmaceuticals.

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Effect of Nitrogen Fertilization levels on Growth and Isoflavone Content in Soybean (질소시비수준이 콩의 생육 및 isoflavone 함량에 미치는 영향)

  • Lee Mi-Ja;Park Jong-Chul;Oh Young-Jin;Kim Kyong-Ho;Kim Hyung-Soon;Lee Sang-Bok;Kim Jung-Gon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.5
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    • pp.445-450
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    • 2006
  • The nitrogen fertilization effect on growth characteristics and isoflavone content was investigated in this study, and isoflavone analyzed by HPLC with photodiode array (PDA) detector and reverse-phase $C_{18}$ column. Fertilization levels were no-fertilization, no nitrogen, 50% decreased in nitrogen, standard and 50% increased in nitrogen fertilization. The 50% increased nitrogen fertilization showed the highest growth characteristics then other fertilization level and the number of pod and seed showed maximum value 31.9 and 72.3, respectively, and seed yield was 2,460 kg/ha. During growth stages, isoflavone content in leaf, stem and root of soybean plants decreased to R5 stage then increased to R7 stage. Isoflavone content according to various nitrogen fertilization condition, in case of none fertilization, no nitrogen, 50% decreased nitrogen fertilization showed higher value than those of standard and 50% increased nitrogen fertilization levels. Aglycon content among the isoflavone isomers showed much higher in plant than in seed. The highest isoflavone content was found in the root of soybean plant parts. Isoflavone content of seed was higher in none, no nitrogen, 50% decreased nitrogen than those of standard and 50% increased nitrogen fertilization.

Glucosylation of Resveratrol Improves its Immunomodulating Activity and the Viability of Murine Macrophage RAW 264.7 Cells (당화된 레스베라트롤의 대식세포 RAW 264.7세포의 생존능력과 레스베라트롤의 면역제어 활성을 증가)

  • Pandey, Ramesh Prasad;Lee, Jisun;Park, Yong Il;Sohng, Jae Kyung
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.19-26
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    • 2017
  • Effects of resveratrol glucosylation on the immunomodulation properties of resveratrol and on the viability of macrophage cells have been studied by using murine macrophage RAW 264.7 cells. Nitric oxide (NO) and interleukin 6 (IL-6) expression in macrophages in vitro were studied after treatment with different concentrations of (E)-resveratrol, (E)-resveratrol 3-O-${\beta}$-${\small{D}}$-glucoside (R-3-G), or (E)-resveratrol 4'-O-${\beta}$-${\small{D}}$-glucoside (R-4'-G). In vitro viability of RAW 264.7 cells after treatment with the aforementioned three compounds was also studied. As demonstrated by macrophage cell viability assays, two different resveratrol monoglucosides, R-3-G and R-4'-G, exhibited 50-80% reduced cytotoxicity in comparison to (E)-resveratrol in A549 and HepG2 cells. Compared to the resveratrol aglycon, both glucosylated resveratrol derivatives positively modulated NO and IL-6 production in macrophages positively via transcriptionally up-regulating IL-6 and iNOS expression. Conjugation of a glucose moiety on resveratrol was found to enhance the immunomodulating activity of resveratrol and the viability of RAW 264.7 cells.

Analysis of Bioconversion Components of Fermentation Hwangryunhaedok-tang (발효 황련해독탕의 생물 전환 성분분석)

  • Lee, Kwang Jin;Lee, BoHyoung;Jung, Pil Mun;Lian, Chun;Ma, Jin Yeul
    • YAKHAK HOEJI
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    • v.57 no.4
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    • pp.293-298
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    • 2013
  • Hwangryunhaedok-tang (HRT) is a traditional herbal medicine, which has been known as a useful prescription for anti-biotic, anti-inflammatory, anti-oxidative and immunosuppressive activity. In this study, the variation in the amount of eight bioactive components of Hwangryunhaedok-tang (HRT) and its fermentation HRT with Lactobacillus casei KFRI 127, Lactobacillus curvatus KFRI 166 and Lactobacillus confuses KFRI 227 was investigated via high-performance liquid chromatography coupled with diode array detection (HPLC-DAD). Simultaneous qualitative and quantitative analysis of eight bioactive components; geniposide, genipin, baicalin, wogonoside, palmatine, berberine, baicalein and wogonin was achieved by comparing their retention times ($t_R$) and UV spectra with those of the standard components. All calibration curve of standard components showed good linearity ($r^2$ >0.979). As a result, the geniposide amount was $15.52{\pm}0.19{\mu}/mg$ that as a main components in HRT. The wogonoside was decreased by 29.28~58.35% with Lactobacillus casei KFRI 127 and L. confuses KFRI 227 ($3.17{\pm}0.31{\mu}g/mg$ and $3.55{\pm}0.13{\mu}g/mg$) compared with the original HRT ($5.02{\pm}0.14{\mu}g/mg$). Otherwise wogonin was increased by 16.28~41.86% with Lactobacillus casei KFRI 127 and L. confuses KFRI 227 ($0.61{\pm}0.01{\mu}g/mg$ and $0.50{\pm}0.02{\mu}g/mg$) compared with the original HRT($0.43{\pm}0.00{\mu}g/mg$). HRT fermented with L. casei KFRI 127 and L. confuses KFRI 227 were evaluated as creating the changes in wogonoside to that aglycon wogonine. In the fermented HRT using Lactobacillus acidophilus KFRI 166, the genipin was only detected, among 3 species of fermentation strains. Thus, these results considered that the strains 166 were exhibited the remarkable changes in genipin.

Fate of 14C - Carobofuran in Rice Plant and Paddy Soil (수도체(水稻體) 및 담수토양중(湛水土壤中) $^{14}C$-Carbofuran의 행적(行跡)에 관한 연구(硏究))

  • Lee, Young-Deuk;Lee, Kyung-Hwi;Park, Chang-Kyu
    • Korean Journal of Environmental Agriculture
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    • v.6 no.1
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    • pp.7-16
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    • 1987
  • A study was undertaken to investigate the absorption, translocation and metabolism of carbofuran in rice paddies. Ring-3-$^{14}C-carbofuran$ applied onto the paddy soil surface was rapidly absorbed and translocated into rice plants. Within 2 days after treatment, it was observed that carbofuran reached shoot tips and accumulated. More than 15% of total radioactivity was recovered in rice plant from 3 to 20 days after treatment. In organic soluble fraction of rice plant extract, 3-hydroxycarbofuran was the major metabolite recording 43% and 4% of total organic soluble radioactivity in shoot and root at 20 days respectively. 3-Ketocarbofuran and phenolic metabolites including carbofuran phenol, 3-hydroxycarbofuran phenol and 3-ketocarbofuran phenol were also detected in the organic soluble fractions. Some glycosidic conjugates of carbofuran metabolites were found in water soluble fraction of rice plant extract and 3-hydroxycarbofuran was the most abundant aglycone. Radioactivity in paddy soil was rapidly decreased until 3 days after treatment and then maintained almost constant level. A significant portion (42∼56 %) of the total radioactivity remained in soil as nonextractable residue from 5 to 20 days after treatment. The nonextractable radioactivity was mainly located in soil organic matter distributing in humin, fulvic acid and humic acid fractions with the decreasing order. Evolution of $^{14}CO_2$ from ring cleavage of $3-^{14}C-carbofuran$ was negligible recording only 1.8% of total radioactivity during 20 days after treatment.

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Studies on Potato Glycoalkaloid Determination by Acid-hydrolysis Method (산 가수분해 방법에 의한 감자 glycoalkaloid성분의 정량성 검토)

  • Yoon, Kyung-Soon;Byun, Gwang-In
    • Journal of the Korean Society of Food Culture
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    • v.24 no.1
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    • pp.84-89
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    • 2009
  • This paper was conducted to evaluate aglycones and carbohydrates produced by acid hydrolysis of three potato glycoalkaloids [(PGA); ${\alpha}$-chaconine, ${\alpha}$-solanine, and demissine] in potatoes. Standard solanidine and demissidine were dissolved in 1N HCl and then heated at $100^{\circ}C$ for 10-120 min. Solanidine was rapidly decomposed during acid hydrolysis and one peak that was identified as solantherene ($M^+$=379) by GC-MS was detected. The transformation solanidine to solanthrene was approximately 50% complete after 10 min, approximately 90% complete after 60 min and 100% complete after 120 min. Demissidine was hydrolyzed using the same method that was used to hydrolyze the solanidine. However, demissidine produced only one peak upon GC-MS ($M^+$=399) analysis and was found to be very stable at increased temperatures. Acidy hydrolysis of ${\alpha}$-chaconine, ${\alpha}$-solanine and demissine resulted in the decomposition of ${\alpha}$-chaconine and ${\alpha}$-solanine to solanidine and solanthrene, respectively. Therefore, this hydrolysis method should not be utilized to produce PGA combining with solanidine as aglycone. The individual carbohydrates produced by the two PGAs by hydrolysis were very stable at increased temperatures; therefore, it was possible to quantify these PGAs based on calculation of the individual carbohydrate content. Conversely, because demissidine produced by the hydrolysis of demissine was extremely stable at increased temperatures, it was possible to quantify the PGA based on the aglycone produced by hydrolysis.

Antioxidant Activity and Whitening Effect of Cedrela sinensis A. Juss Shoots Extracts (참죽나무 새순 추출물의 항산화 활성과 미백 작용에 관한 연구)

  • Kim, Sun-Young;Kim, Chae-Rin;Kim, Hyun-Min;Kong, Myung;Lee, Ji-Hee;Lee, Hyun-Jun;Lim, Myoung-Sun;Jo, Na-Rae;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.3
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    • pp.175-182
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    • 2010
  • In this study, the antioxidative effects, inhibitory effects on tyrosinase of Cedrela sinensis extracts were investigated. The ethyl acetate fraction of extract ($3.54\;{\mu}g/mL$) and aglycone fraction of extract ($2.15\;{\mu}g/mL$) showed more excellent free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) than the activity of (+)-$\alpha$-tocopherol ($8.98\;{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of Cedrela sinensis extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction of extract ($0.15\;{\mu}g/mL$) and aglycone fraction of extract ($0.12\;{\mu}g/mL$) showed 10 times more excellent ROS scavenging activity than activity of L-ascorbic acid ($1.50\;{\mu}g/mL$). The protective effects of fractions of Cedrela sinensis on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethyl acetate fraction of extract and aglycone fraction of extracts suppressed photohemolysis in a concentration dependent manner ($5{\sim}25\;{\mu}g/mL$). The inhibitory effect of Cedrela sinensis extracts on tyrosinase was investigated to assess their whitening efficacy. Inhibitory effects ($IC_{50}$) on tyrosinase were determined with ethyl acetate fraction of Cedrela sinensis extract ($48.00\;{\mu}g/mL$) and aglycone fraction of extract ($5.88\;{\mu}g/mL$). The aglycone fraction showed 40 times more remarkable tyrosinase inhibitory effect than whitening agent, arbutin ($226.88\;{\mu}g/mL$) These results indicate that fractions of Cedrela sinensis can be used as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. The fractions of Cedrela sinensis can be applicable to new functional cosmetics for antioxidant and whitening.

Physicochemical components of Astragalus membranaceus fermented with mushroom mycelia (담자균 균사체가 배양된 황기의 이화학적 성분분석)

  • Jang, Yeon-Jeong;Lee, Yun-Hye;Lee, Chan-Jung;Kim, Jae-Hyeon;Kim, Eun-Ju;Ji, Soo-Jeong;Park, Shin-Young
    • Food Science and Preservation
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    • v.23 no.1
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    • pp.49-56
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    • 2016
  • This study analyzed the physicochemical characteristics of Astragalus membranaceus (AM) fermented with seven different mushroom mycelia. Physicochemical characteristics, such as contents of moisture, pH, total reducing sugars, free sugar, and isoflavonoid, were investigated. The moisture content was increased in most of the samples. The pH values of AM fermented with Phellinus linteus and Flammulina velutipes were increased, while the pH of other samples were similar to that of non-fermented AM. The reducing sugar content was in the range of 211.69~391.74 mg/100 g. The extraction yield using water was higher than that when extracted with 80% ethanol. The free sugar content was increased through fermentation with mushroom mycelia. However, the glucose contents of the 80% ethanol and water extracts were decreased. Finally, the calycosin and formononetin contents in 80% ethanol and water extracts of AM fermented with Phellinus linteus were 2,549.24 mg/g, and 827.66 mg/g for calycosin, and 1,366.69 mg/g and 221.28 mg/g for formononetin, respectively. These results suggest that fermentation with mushroom mycelia could be used to increase the bioactivity of AM. The mycelium-fermented AM might be a valuable source of functional material and edible resource for industry.