• Title/Summary/Keyword: ${\beta},D-glucosidase$

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Isolation, Identification and Mutant Development of Butanol Tolerance Bacterium (부탄올 내성 미생물의 분리, 동정 및 변이주의 개발)

  • Jung, Hyesook;Lee, Jinho
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.26-32
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    • 2013
  • Butanol-resistant bacteria were isolated from butanol solvent. The cell growth of isolated strains declined with increasing concentrations of butanol, and isolated strain BRS02 displayed more resistance to 12.5 g/L of butanol than other isolated strains. In addition, strain BRS251, which was resistant to even higher concentrations of butanol, was developed by the mutation of BRS02 using UV. BRS251 could grow in LB medium containing up to 17.5 g/L of butanol, 32.5 g/L of propanol, or 6 g/L of pentanol, whereas the control strain Escherichia coli was found to be tolerant to 7.5 g/L of butanol, 20 g/L of propanol, or 2 g/L of pentanol. The isolated BRS02, a Gram(+) bacterium seen to have a cocci form under the microscope, grew in 6.5% NaCl. According to biochemical tests, BRS02 can metabolize and produce acid with D-galactose, D-maltose, D-mannitol, D-mannose, methyl-${\beta}$-Dglucopyranoside, D-ribose, sucrose, or D-trehalose, as carbon sources. Also, this strain showed resistance to bacitracin, vibriostatic agent O/129, and optochin, alongside positive activities for arginine dihydrolase, ${\alpha}$-glucosidase, and urease. The BRS02 strain was identified as Staphylococcus sp. by analyses of the 16S rRNA gene, phylogenetic tree, and biochemical tests.

Proteomic Identification of Differentially Expressed Proteins in Arabidopsis Mutant ntm1-D with Disturbed Cell Division

  • Lee, Kyung Hyeon;Kim, Youn-Sung;Park, Chung-Mo;Kim, Hie-Joon
    • Molecules and Cells
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    • v.25 no.1
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    • pp.70-77
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    • 2008
  • Proteome analysis was performed to identify proteins differentially expressed in an Arabidopsis mutant, ntm1-D. In this mutant the NAC transcription factor NTM1 is constitutively expressed and the resultant phenotypic changes include dwarfism, serrated leaves, and altered floral structures, probably due to reduced cell division. Marked elevation of proteins mediating environmental stress responses, including annexin, vegetative storage proteins, beta-glucosidase homolog 1, and glutathione transferases was observed. Overexpression of annexin was confirmed by RT-PCR and Western blotting. These observations suggest that the reduced growth observed in the ntm1-D mutant is caused by enhancement of its stress responses, possibly resulting in a cost in fitness.

Characteristics of Enzymatic Hydrolysis of Sodium Hydroxide pretreated Suwon Poplar (NaOH 전처리된 현사시나무의 효소가수분해 특성)

  • 박영기;오정수
    • Journal of Korea Foresty Energy
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    • v.20 no.2
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    • pp.20-27
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    • 2001
  • An effective method for production of glucose was developed using enzymatic hydrolysis of Suwon poplar by the cellulase. Enzymatic hydrolysis of wood is the reaction to produce glucose from wood using enzyme which derives from microorganism. Glucose can be transferred easily to ethanol by fermentation. Ethanol is the starting material for producing acetone, butanol, citric acid and lactic acid. The mechanism of the enzymatic hydrolysis of cellulose are reasonably explained in terms of the sequential action of three different types of enzymes, endo-cellulase, ex-cellulase, and $\beta$ -glucosidase. The goal of this work was to investigate the cellulose hydrolysis pretreated polar with various concentration NaOH, the crystallinity of cellulose, lignin contents and the degree of hydrolysis.

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Apoptosis Induction of Persicae Semen Extract in Human Promyelocytic Leukemia (HL-60) Cells

  • Kwon, Hee-Young;Hong, Seon-Pyo;Hahn, Dong-Hoon;Kim, Jeong-Hee
    • Archives of Pharmacal Research
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    • v.26 no.2
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    • pp.157-161
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    • 2003
  • The major ingredient of Persicae Semen is a cynogenic compound, amygdalin (D-mandelonitrile-$\beta$-gentiobioside). Controversial results on the anticancer activity of amygdalin were reported due to its conversion to its inactive isomer, neoamygdalin. In order to inhibit the epimerization of amygdalin, we used newly developed simple acid boiling method in preparation of Persicae Semen extract. HPLC analysis revealed most of amygdalin in Persicae Semen extract was active D-form. Persicae Semen extract was used to analyze its effect on cell proliferation and induction of apoptosis in human promyelocytic leukemia (HL-60) cells. Persicae Semen extract was cytotoxic to HL-60 cells with $IC_{50}$ of 6.4 mg/mL in the presence of 250 nM of $\beta$-glucosidase. The antiproliferative effects of Persicae Semen extract appear to be attributable to its induction of apoptotic cell death, as Persicae Semen extract induced nuclear morphology changes and internucleosomal DNA fragmentation.

A Fermented Ginseng Extract, BST204, Inhibits Proliferation and Motility of Human Colon Cancer Cells

  • Park, Jong-Woo;Lee, Jae-Cheol;Ann, So-Ra;Seo, Dong-Wan;Choi, Wahn-Soo;Yoo, Young-Hyo;Park, Sun-Kyu;Choi, Jung-Young;Um, Sung-Hee;Ahn, Seong-Hoon;Han, Jeung-Whan
    • Biomolecules & Therapeutics
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    • v.19 no.2
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    • pp.211-217
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    • 2011
  • Panax ginseng CA Meyer, a herb from the Araliaceae, has traditionally been used as a medicinal plant in Asian countries. Ginseng extract fermented by ginsenoside-${\beta}$-glucosidase treatment is enriched in ginsenosides such as Rh2 and Rg3. Here we show that a fermented ginseng extract, BST204, has anti-proliferative and anti-invasive effects on HT-29 human colon cancer cells. Treatment of HT-29 cells with BST204 induced cell cycle arrest at $G_1$ phase without progression to apoptosis. This cell cycle arrest was accompanied by up-regulation of tumor suppressor proteins, p53 and p21$^{WAF1/Cip1}$, down-regulation of the cyclin-dependent kinase/cyclins, Cdk2, cyclin E, and cyclin D1 involved in $G_1$ or $G_1/S$ transition, and decrease in the phosphorylated form of retinoblastoma protein. In addition, BST204 suppressed the migration of HT-29 cells induced by 12-O-tetradecanoylphorbol-13-acetate, which correlated with the inhibition of metalloproteinase-9 activity and extracellular signal-regulated kinase activity. The effects of BST204 on the proliferation and the invasiveness of HT-29 cells were similar to those of Rh2. Taken together, the results suggest that fermentation of ginseng extract with ginsenoside-${\beta}$-glucosidase enhanced the anti-proliferative and the anti-invasive activity against human colon cancer cells and these anti-tumor effects of BST204 might be mediated in part by enriched Rh2.

Gram-Scale Production of Ginsenoside F1 Using a Recombinant Bacterial β-Glucosidase

  • An, Dong-Shan;Cui, Chang-Hao;Siddiqi, Muhammad Zubair;Yu, Hong Shan;Jin, Feng-Xie;Kim, Song-Gun;Im, Wan-Taek
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1559-1565
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    • 2017
  • Naturally occurring ginsenoside F1 (20-O-${\beta}$-$\text\tiny{D}$-glucopyranosyl-20(S)-protopanaxatriol) is rare. Here, we produced gram-scale quantities of ginsenoside F1 from a crude protopanaxatriol saponin mixture comprised mainly of Re and Rg1 through enzyme-mediated biotransformation using recombinant ${\beta}$-glucosidase (BgpA) cloned from a soil bacterium, Terrabacter ginsenosidimutans Gsoil $3082^T$. In a systematic step-by-step process, the concentrations of substrate, enzyme, and NaCl were determined for maximal production of F1. At an optimized NaCl concentration of 200 mM, the protopanaxatriol saponin mixture (25 mg/ml) was incubated with recombinant BgpA (20 mg/ml) for 3 days in a 2.4 L reaction. Following octadecylsilyl silica gel column chromatography, 9.6 g of F1 was obtained from 60 g of substrate mixture at 95% purity, as assessed by chromatography. These results represent the first report of gram-scale F1 production via recombinant enzyme-mediated biotransformation.

Antidiabetic Activity and Enzymatic Activity of Commercial Doenjang Certified for Traditional Foods (전통식품 품질인증 일부 시판 된장의 효소활성 및 항당뇨 활성)

  • Lee, So-Young;Kim, In-Sun;Park, So-Lim;Lim, Seong-Il;Choi, Hye-Sun;Choi, Shin-Yang
    • KSBB Journal
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    • v.27 no.6
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    • pp.361-366
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    • 2012
  • We investigated the anti-diabetic activity and enzymatic activity of 24 commercial doenjang samples certified for traditional foods. Twenty four doenjang samples showed the wide ranges in enzymatic activities (protease activities 0-50.45 unit/g, ${\alpha}$-amylase activities 0-675.9 unit/g, ${\beta}$-amylase 13.6-308.6 unit/g), and there were no difference in enzymatic activity by the producing region. To evaluate the potential anti-diabetic activity of 24 doenjang samples, we examined the effect of doenjang methanol extract (DME) on 2-[n-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl) amyno]-2-deoxy-d-glucose (2-NBDG) uptake. Ten samples among 24 samples significantly stimulated the uptake of 2-NBDG. When the cells were treated with DME at 400 ug/mL, No. 17 and 23 specially stimulated 2-NBDG uptake by 1.23-fold and 1.25-fold, respectively, compared with untreated control cell. And there were no cytotoxicity in the C2C12 cells treated with DME at concentration of 500 ug/mL. Among 24 samples, No. 6, 7, 12, 21 and 24 showed the ${\alpha}$-glucosidase inhibitor activity at concentration of 10 mg/mL; however, they were less effective than acarbose which is a commercial ${\alpha}$-glucosidase inhibitor.

Optimal Conditions and Substrate Specificity for Trehalose Production by Resting Cells of Arthrobacter crystallopoietes N-08

  • Seo, Yi-Seul;Shin, Kwang-Soon
    • Preventive Nutrition and Food Science
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    • v.16 no.4
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    • pp.357-363
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    • 2011
  • Recently, we found that Arthrobacter crystallopoietes N-08 isolated from soil directly produces trehalose from maltose by a resting cell reaction. In this study, the optimal set of conditions and substrate specificity for the trehalose production using resting cells was investigated. Optimum temperature and pH of the resting cell reaction were $55^{\circ}C$ and pH 5.5, respectively, and the reaction was stable for two hours at $37{\sim}55^{\circ}C$ and for one hour at the wide pH ranges of 3~9. Various disaccharide substrates with different glycosidic linkages, such as maltose, isomaltose, cellobiose, nigerose, sophorose, and laminaribiose, were converted into trehalose-like spots in thin layer chromatography (TLC). These results indicated broad substrate specificity of this reaction and the possibility that cellobiose could be converted into other trehalose anomers such as ${\alpha},{\beta}$- and ${\beta},{\beta}$-trehalose. Therefore, the product after the resting cell reaction with cellobiose was purified by ${\beta}$-glucosidase treatment and Dowex-1 ($OH^-$) column chromatography and its structure was analyzed. Component sugar and methylation analyses indicated that this cellobiose-conversion product was composed of only non-reducing terminal glucopyranoside. MALDI-TOF and ESI-MS/MS analyses suggested that this oligosaccharide contained a non-reducing disaccharide unit with a 1,1-glucosidic linkage. When this disaccharide was analyzed by $^1H$-NMR and $^{13}C$-NMR, it gave the same signals with ${\alpha}$-D-glucopyranosyl-(1,1)-${\alpha}$-D-glucopyranoside. These results suggest that cellobiose can be converted to ${\alpha},{\alpha}$-trehalose by the resting cells of A. crystallopoietes N-08.

Cellulose Degradation and Extracellulat Enzymatic Activity of the Mud Flat in Sunchon Bay (순천만 갯벌 토양의 섬유소 분해능 및 체외효소 활성)

  • 백근식;최지혁;성치남
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.130-135
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    • 2000
  • Decomposition rate of organic matiter in the mud flat of Sunchon Bay was estimated. Physicochemical parameters, cellulose degradation rate. distribution of heterotrophic bacteria, and extracellular enzymatic activities were measured from August 1997 to July 1998. Soil temperatures, water contents, concentration of $PO_4$-P and organic matter were -1-~$30^{\circ}C$, 42.1-53.1%, 0.0779-0.1961 mgig and 1.99-7.64%, respectively. Decomposition rate of cellulose film ranged from 7.7 to 100%imonth, high in summer and low in winter. The number of heterotrophic bacteria ranged from $0.87{\times}10^6 to 3.6{\times}10^7 $CUFsIg dq soil. Enzymatic activities of phosphatase, $\alpha$-D-gluEosidase, $\beta$-D-glucosidase and cellobiohydrolase, which were measured as decomposition rate of methylumbelliferyl(MLiF)-substrate, were 152.23-1779.80 nMIhr, 2.67-202.18 nM/hr, 5.03-258.26 M h r and 3.42-63.07 nM/hr, respectively Cellulose degradaaon rate and extracellular extracellular enzymatic activities were conelated with each other, and showed high correlation coefticiency with soil temperature.

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Isolation of Stenotrophomonas rhizopilae Strain GFC09 with Ginsenoside Converting Activity and Anti-wrinkle Effects of Converted Ginsenosides (사포닌 전환 활성 Stenotrophomonas rhizopilae Strain GFC09 균주의 분리 동정 및 전환 사포닌의 주름 개선 효과)

  • Min, Jin Woo;Kim, Hye-Jin;Joo, Kwang-Sik;Kang, Hee-Cheol
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.4
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    • pp.375-382
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    • 2015
  • Ginsenosides (ginseng saponin) as the one of important pharmaceutical compounds of ginseng and is responsible for the pharmacological and biological activities. These ginsenoside produces diverse small molecules ginsenoside which have more pharmacological activities including anti-wrinkle, anti-cancer and anti-oxidant effects. In the present study, we isolated bacteria using esculin agar, to produce ${\beta}$-glucosidase, and we focused on the bio-transformation of ginsenoside. Phylogenetic tree analysis was performed by comparing the 16S rRNA sequences; we identified the strain as Stenotrophomonas rhizopilae strain GFC09. In order to determine the optimal conditions for enzyme activity, the crude enzyme was incubated with 1 mM ginsenoside $Rb_1$. Bioconversion of ginsenoside $Rb_1$ were analyzed using TLC and HPLC. The crude enzyme hydrolyzed the ginsenoside $Rb_1$ along the following pathway: LB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$ into compound K, TSB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$. The structure of the hydrolyzed metabolites were identified by NMR. The activity screening tests showed that the conversion product induced the production of type I procollagen in a dose-dependent manner. These results suggested that hydrolyzed ginseng product containing the ginsenoside $F_2$ and compound K could be useful as an active ingredient for wrinkle-care cosmetics.