• Title/Summary/Keyword: Complex Enzyme

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Change of ROS Generation and Antioxidant Enzyme Activity of Flavonol Quercetin in the Presence of Vitamin E, L-Ascorbit acid, Reduced Glutathione on the B16F10 Murine Melanoma Cells (B16F10 세포에서 Quercetin과 Vitamin E, L-Ascorbic acid, 환원형 글루타치온과의 병용 투여에 의한 활성산소종 발생과 항산화 효소의 활성 변화)

  • 허정심;김안근
    • YAKHAK HOEJI
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    • v.47 no.6
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    • pp.432-437
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    • 2003
  • It has been known that quercetin, a bioflavonoid widely distributed in fruits and vegetables as dietary-derived flavonoid and exert significant multiple biological effects such as antioxidant and anti-inflammatory, anti-tumor effects. In addition, it has been shown to have a chemoprotective role in cancer, though complex effects on signal transduction involved in cell proliferation and angiogenesis. The present study investigated whether quercetin can enhance antioxidant enzyme activity (glutathione peroxidase: GPx, superoxide dismutase: SOD, catalase: CAT) and regulate the reactive oxygen species (ROS) generation in the presence of vitamin E, L-ascorbic acid, reduced glutathione (GSH) on B16F10 murine melanoma cells. After 48h treatment of cells with quercetin in the presence of vitamin E, L-ascorbic acid, GSH, we measured the cytotoxicities by MTT assay. The cells exhibited a dose-dependent inhibition in their proliferation in the presence of vitamin E, L-ascorbic acid, GSH respectively. We also investigated the effects of antioxidant enzyme activity and ROS generation. The antioxidant enzyme activity of quercetin in the presence of vitamin E was stronger than GSH, L-ascorbic acid, the same treatments decreased ROS generation in B16F10 murine melanoma cells. Taken together, these result demonstrate that the antioxidant effect of quercetin can enhanced in the presence of vitamin E and it might plays an important role in anti-oxidative effects.

Purification and Characterization of Extracellular $\beta$-Glucosidase from Sinorhizobium kostiense AFK-13 and Its Algal Lytic Effect on Anabaena flos-aquae

  • Kim, Jeong-Dong;Lee, Choul-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.745-752
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    • 2007
  • A $\beta$-glucosidase from the algal lytic bacterium Sinorhizobium kostiense AFK-13, grown in complex media containing cellobiose, was purified to homogeneity by successive ammonium sulfate precipitation, and anion-exchange and gel-filtration chromatographies. The enzyme was shown to be a monomeric protein with an apparent molecular mass of 52 kDa and isoelectric point of approximately 5.4. It was optimally active at pH 6.0 and $40^{\circ}C$ and possessed a specific activity of 260.4 U/mg of protein against $4-nitrophenyl-\beta-D-glucopyranoside$(pNPG). A temperature-stability analysis demonstrated that the enzyme was unstable at $50^{\circ}C$ and above. The enzyme did not require divalent cations for activity, and its activity was significantly suppressed by $Hg^{+2}\;and\;Ag^+$, whereas sodium dodecyl sulfate(SDS) and Triton X-100 moderately inhibited the enzyme to under 70% of its initial activity. In an algal lytic activity analysis, the growth of cyanobacteria, such as Anabaena flos-aquae, A. cylindrica, A. macrospora, Oscillatoria sancta, and Microcystis aeruginosa, was strongly inhibited by a treatment of 20 ppm/disc or 30 ppm/disc concentration of the enzyme.

Studies on the Cellulase. (V) -Fractionation of Cellulolytic Complex produced by Trichoderma $viride(O_2-1)$ (섬유소(纖維素) 분해효소(分解酵素)에 관(關)한 연구(硏究) (제5보(第5報)) -Trichoderma $(O_2-1)$가 생성(生成)하는 Cellulolytic Complex의 분별(分別)에 대(對)하여-)

  • Sung, Nack-Kie
    • Applied Biological Chemistry
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    • v.12
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    • pp.99-105
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    • 1969
  • The yield of cellulase derived from Trichoderma $(O_2-1)$ was remarkably varied with various concentration of ethanol and acetone in purification of the enzyme. In the purification with ethanol of ${\beta}-glucosidase$, the best result was obtained in the concentration of 60% and, of CMCase and of filter paper disintegrating enzyme 80%. And in the purification with acetone of ${\beta}-glucosidase$, filter paper disintegrating enzyme, and CMCase, in the concentration of 60%, 80%, and 90% respectively, was shown the best yield. The activities of crude Cellulase preparation could be seperated into few of fractions by column chromatography with Silica gel, Cellulose powder, and gauze. Most of CMCase, avicelase, and ${\beta}-glucosidase$ were eluted, but most of filter paper disintegrating enzyme and the rest of enzymes mentioned the above were absorbed, and were eluted with water. Therefore, it was considered that CMCase is different from filter paper disintegrating enzyme in properties. The relative activity of CMCase was different from that of avicelase in the peak of elusion part. And it was considered that filter paper disintegrating enzyme and cellulose powder saccharifying enzyme was seperated respectively as absorption part and non absorption part. The auther came to the conclusion that at least there were more than three sorts of cellulase in Trichoderma $(O_2-1)$ cellulase preparation.

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Growth Inhibition and Apoptosis Induction of Gastric Cancer Cells by Copper (II) Glycinate Complex

  • JE CHUL LEE;JEONG, YONG WOOK;KISUNG KIM;JAE YOUNG OH;JONG CHUN PARK;JUNG HWAN BANG;ANG WON CHOI
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.394-399
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    • 2003
  • The in vitro cytotoxic effects of newly synthesized copper (II) glycinate complex were investigated in two gastric cancer cell lines of SNU484 and SNU638 cells. The complex inhibited the growth and decreased the viability of both gastric cancer cells in a dose-dependent manner. Gastric cancer tells treated with the complex exhibited the features of apoptosis, as demonstrated by fragmentation of chromosomal DNA, activation of caspase-3-like enzyme, and cleavage of poly[ADP-ribose] polymerase (PARP). With the treatment of copper (II) glycinate complex, the active form of caspase-3 was observed in SNU484 cells, but not in SNU638 cells, indicating that an alternative pathway of apoptosis might have been triggered in SNU638 cells. In conclusion, copper (II) glycinate complex induces apoptosis of SNU484 and SNU638 gastric cancer cells, and it is suggested that novel copper (II) glycinate complex is highly active against human gastric cancer cells.

Enhancement of ${\beta}$-Glucan Content in the Cultivation of Cauliflower Mushroom (Sparassis latifolia) by Elicitation

  • Park, Hyun;Ka, Kang-Hyeon;Ryu, Sung-Ryul
    • Mycobiology
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    • v.42 no.1
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    • pp.41-45
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    • 2014
  • The effectiveness of three kinds of enzymes (chitinase, ${\beta}$-glucuronidase, and lysing enzyme complex), employed as elicitors to enhance the ${\beta}$-glucan content in the sawdust-based cultivation of cauliflower mushroom (Sparassis latifolia), was examined. The elicitors were applied to the cauliflower mushroom after primordium formation, by spraying the enzyme solutions at three different levels on the sawdust-based medium. Mycelial growth was fully accomplished by the treatments, but the metabolic process during the growth of fruiting bodies was affected. The application of a lysing enzyme resulted in an increase in the ${\beta}$-glucan concentration by up to 31% compared to that of the control. However, the treatment resulted in a decrease in mushroom yield, which necessitated the need to evaluate its economic efficiency. Although we still need to develop a more efficient way for using elicitors to enhance functional metabolites in mushroom cultivation, the results indicate that the elicitation technique can be applied in the cultivation of medicinal/edible mushrooms.

Optimum Reaction Condition of Enzymatic Hydrolysis for Production of Reducing Sugar from Enteromorpha intestinalis (창자파래로부터 환원당 생산을 위한 효소가수분해의 최적 반응조건)

  • Kim, A-Ram;Kim, Dong-Hyun;Jeong, Gwi-Taek
    • KSBB Journal
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    • v.30 no.2
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    • pp.53-57
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    • 2015
  • In this study, the production of total reducing sugar from macro green-algae Enteromorpha intestinalis by enzymatic hydrolysis was investigated. As a result of enzymatic hydrolysis using 13 kind commercial enzymes, the highest yield of 8.75% was obtained from Viscozyme L, which is multi-enzyme complex such as cellulase, arabanase, beta-glucanase, hemicellulase and xylanase. As a control, only 0.33% and 0.27% yield were obtained from 1% sulfuric acid and 0.05 M citrate buffer (pH 4.8), respectively. In the case of enzyme mixture, the mixture of $Viscozyme^{(R)}$ L and $Cellic^{(R)}$ CTec2 (1:1) was presented the highest yield of 10.67%. Finally, the 14.99% yield was obtained at 36 hr under the condition of 10% biomass and 30% enzyme mixture.

3,4-Dihydroxyphenyl-L-alanine의 효소적 생산에 대한 반응첨가물의 영향

  • Lee, Seung-Goo;Ro, Hyeon-Su;Hong, Seung-Pyo;Sung, Moon-Hee
    • Microbiology and Biotechnology Letters
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    • v.24 no.2
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    • pp.222-226
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    • 1996
  • The enzymatic synthesis of 3, 4-dihydroxyphenyl-L-alanine (L-DOPA) was examined for the effects of the reaction additives such as sodium borate, alcohol, and organic solvents. The enzyme used was tyrosine phenol-lyase of Citrobacter freundii KCTC 2006 produced in Escherichia coli. The amounts of tyrosine phenol-lyase and pyridoxal-5-phosphate were optimized to 2.0 units/ml and 0.1 mM, respectively, for the synthetic reaction. Sodium borate, a substance that forms a complex with pyrocatechol, reduced the enzyme deactivation by pyrocatechol although it seriously inhibited the enzyme activity. Among the organic solvents tested, dimethylsulfoxide, dimethylformamide, and alcohol increased the productivity of the L-DOPA synthesis. In a reaction system with 5% methanol, L-DOPA concentration increased up to 210 mM after 24 hours, and 77.1% of which was separated as precipitates. The L-DOPA was purified to 99.96% by solubilizing and recrystallyzing the precipitates.

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Cell Biological Studies on Growth and Development Effect of polyamine and $Ca^{2+}$ on D-glucose-6-phosphate cyclohydrolase activity in carrot root protoplast (생체생장에 관한 세포생물학적 연구 당근 뿌리의 원형질체에서 D-glucose-6-phosphate cyclohydrolase 활성도에 미치는 polyamine과 $Ca^{2+}$의 영향)

  • 이순희
    • Journal of Plant Biology
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    • v.30 no.4
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    • pp.249-255
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    • 1987
  • The effect of polyamine and Ca2+ on D-glucose-6-phosphate cyclohydrolase activity was studied in Daucus carota root. The enzyme activity was reduced in response to increase in concentration of Ca2+, not the Ca2+-calmodulin complex. The inhibition effect due to Ca2+ was reversed by polyamine, especially remarkable at low concentration of Ca2+. The effect of the Ca2+ on the enzyme seemed to compete with polyamine according to the Lineweaver-Burk plot. The enzyme activity from carrot root protoplast cultured in the prescence of verapamil was higher than that of the control. Such cumulative results suggest that the inhibition by the Ca2+ and enhancement or reversal by polyamine could regulate the biosynthesis of pectin and hemicellulose to some extent.

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Effects of Dietary Carbohydrase Enzyme Complex and Microbial Phytase Supplementation on Productivity and Nutrient Digestibility in Growing Pigs (탄수화물 분해 복합효소제와 미생물 파이테이즈의 첨가가 육성돈의 생산성 및 영양소 소화율에 미치는 영향)

  • Shim, Y.H.;Chae, B.J.;Lee, J.H.
    • Journal of Animal Science and Technology
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    • v.45 no.4
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    • pp.569-576
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    • 2003
  • An experiment was conducted to investigate the effect of microbial phytase (Natuphos$^{\circledR}$) supplementation, individually and in combination with carbohydrase enzyme complex (composed of enzymes targeted to SBM dietary components such as $\alpha$-galactosides and galactomannans; ENDO-POWER$^{\circledR}$) to corn-soy basis diet with low nutrient levels on growth performance and nutrient digestibility of growing pigs. A total of 48 crossbred weaned pigs (Landrace${\times}$Yorkshire${\times}$Duroc), 29.1$\pm$0.14 kg of initial body weight, were randomly allotted to four dietary treatments, based on weight and age, according to a Randomized Complete Block Design. There were three pens per treatment and 4 pigs per pen. The dietary treatments were 1) CON (control diet with 3,380 kcal/kg of metabolizable energy, 18.96% of crude protein, 1.10% of lysine, 0.75% of calcium and 0.35% of available phosphorus), 2) LP+NTPS (CON diet with 0.15% unit lower available P levels+0.1% phytase (500 FTU/kg; Natuphos$^{\circledR}$)), 3) LEL+ENP (CON diet with 3.0% unit lower ME and lysine levels + 0.1% carbohydrase enzyme complex (ENDO-POWER$^{\circledR}$), and 4) LPEL+ENZ (CON diet with 0.15% unit lower available P levels and 3.0% unit lower ME and lysine levels+0.1% ENDO-POWER$^{\circledR}$ and 0.1% Natuphos$^{\circledR}$ (500 FTU/kg). There was no significant difference (p〉0.05) in average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) among dietary treatments during the whole experimental period (0 to 4 weeks). Apparent digestibility of gross energy was greater in LP+NTPS and LPEL+ENZ groups than in the LEL+ENP (p<0.05). Apparent digestibility of phosphorus was greater in LP+NTPS than in LEL+ENP (p<0.05). Dry matter excretion was lowest in LPEL+ENZ and phosphorus excretion was lowest in LP+NTPS (p<0.05). Overall, pigs fed on LPEL+ENZ group tended to have better nutrient digestibility (dry matter, gross energy, crude protein and phosphorus) than pigs fed on control group. All dietary enzyme treatment groups showed lower feed cost/body weight gain of pigs than control group. In conclusion, the results from the present study suggest that the simultaneous inclusion of phytase and carbohydrase enzyme complex to diets is advantageous with respect to reducing nutrient excretion of growing pigs and may contribute to increased economic return when added to corn-soy based growing pig diets.

Processing of Sardine Sauce from Sardine Scrap (정어리잔사를 이용한 정어리간장의 제조)

  • LEE Eung-Ho;CHO Soon-Yeong;HA Jae-Ho;OH Kwang-Soo;KIM Chang-Yang
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.2
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    • pp.117-124
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    • 1984
  • Sarine scrap usually comprises about $40\%$ of the raw fish in processing. The purpose of this study is to establish the desirable methods for proteinaceous materials from the sardine scrap through autolysis or enzymatic digestion and to convert them into useful by-products such as sardine sauce. Sardine scrap was chopped and mixed with equal weight of water, and be hydrolyzed them by autolysis and/or by addition of commercial proteolytic enzyme and various concentrations of sodium chloride. The optimal conditions for hydrolysis of sardine scrap were revealed in temperature at $55^{\circ}C$ and 4 hours digestion with bromelain($0.4\%$) and commercial complex enzyme ($6.0\%$), and those conditions were also applicated in autolysis. The maximum hydrolyzing rate of protein and released amino nitrogen were $82.5\%,\;5.2\%$ through autolysis, $84.3\%,\;5.8\%$ by bromelain digestion and $92.5\%,\;5.9\%$ by complex enzyme, respectively. In the products prepared from sardine scrap through autolysis or bromelain digestion, hypoxanthine was dominant, as $17.4 {\mu}mole/g$, dry matter for autolysis and $16.0 {\mu}mole/g$, dry matter, for bromelain digestion among the nucleotidcs and their related compounds, respectively. The abundant free amino acids were leucine, glutamic acid, lysnie, valine and alanine. The contents of those amino acids were $51.3\%,\;48.3\%$ of the total free amino acids, respectively. And the contents of 5'-IMP and TMAO were negligible but total creatinine was developed in value from $9.2\%\;to\;10\%$ of total extracted nitrogen. The flavor of sardine sauce prepared from sardine scrap by autolysis or enzyme digestion were not inferior to that of traditional Korean soy sauce by sensory evaluation.

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