• Title/Summary/Keyword: glucose oxidase(GO)

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Effects of Enzymes and Emulsifiers on the Loaf Volume and Crumb Hardness of Rice Breads (효소제 및 유화제의 첨가가 쌀빵의 부피와 경도에 미치는 영향)

  • Lee, Myung-Hee;Chang, Hak-Gil;Lee, Young-Tack
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.6
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    • pp.761-766
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    • 2008
  • The effects of various enzymes and emulsifiers on the loaf volume and crumb hardness of rice breads were studied. Four different enzymes [fungal ${\alpha}$-amylase (AMYL), maltogenic bacterial ${\alpha}$-amylase (NMYL), glucose oxidases (GO), and xylanase+hemicellulases (PTP)] and four emulsifiers [sorbitan monostearate (SMS), glycerol monostearate (GMS), sodium stearoyl lactylate (SSL), and glycerol ester+propylene glycol ester+sucrose ester+sorbitan ester (SP)] were supplemented to rice dough. The addition of AMYL, GO, and GO+AMYL increased loaf volume of rice breads. The highest loaf volume was observed in rice bread supplemented with AMYL. Rice breads supplemented with enzymes firmed at lower rates during storage, and AMYL, NMYL, and GO considerably decreased crumb hardness of rice breads, exhibiting a significant antistaling effect. The addition of emulsifiers produced rice breads with better specific loaf volume and crumb texture, and continuously retarded crumb hardness of rice breads during storage. Especially, rice bread supplemented with SSL demonstrated the highest loaf volume and the lowest crumb hardness during storage.

Effects of Heating on Hydroxyl Radical-Generated Toxicity in Mouse Forebrain Tissue Culture

  • Lee, Jeong-Chae;Lim, Kye-Taek
    • Toxicological Research
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    • v.14 no.3
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    • pp.301-306
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    • 1998
  • This experiment was carrid out to know the effects of heating and serum on hydroxyl radicals in embryonic mouse forebrain (cerebrum) culture. The heating to mouse embryonic cerebrum cells in culture was done in a water bath at 43${\circ}C$ for 60min. After that, two supernatants were prepared at 20 hrs and 48 hrs respectively after heat treatment to the brain cells. To find out the heating effects on neuron cells, mouse cerebrum cells (13 embryonic day) were cultured in hydroxyl radical generation system composed of 20mU/ml glucose oxidase (GO system), using condition of normal culture media (MEM, 5% serum, 5% $CO_2$or supernatant prepared after heating at 43${\circ}C$ for 60 min in a water bath. Supernatant prepared at 20 hrs after heat treatment had a greater protective effects against hydroxyl radical than supernatant prepared at 48 hrs after heat treatment . Otherwise, the protective effect of serum against hydroxyl radicals in the cultured brain cells is higher than that in the heat treatment. These results indicated that serum in culture media reduced cytotoxicity of hydroxyl radicals in mouse forebrain culture, also that heat treatment showed the protective effects against hydroxyl radicals generated with 20mU/ml GO system in mouse forebrain culture.

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Comparison with Some Antioxidants on Hydroxyl Radical in Mouse Whole Brain Culture

  • Lee, Jeong-Chae;Lim, Kye-Taek;Lee, Ki-Seoup;Jung, Hee-young
    • Toxicological Research
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    • v.14 no.4
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    • pp.541-545
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    • 1998
  • This experiment carried out to compare the protective effects of some antioxidants to hydroxyl radicals in embryonic mouse whole brain tissue culture. The ICR mouse whole brain (13 embryonic day) was cultured in hydroxyl radical system in which radicals were generated by 20 mU / ml glucose oxidase (GO). In this experiment, to make ferrous iron from ferric iron, iron as an accelerator, and ascorbic acid as a reductant were used. For comparison of the protective effects to hydroxyl radicals, antioxidants such as desferrioxamine (DFX), laccase. water or ethanol extracts from Rhus Vemiciflua Stokes (RVS), and $\alpha$-tocopherol were used, because they relate to metal ion. The results of this experiment showed that all antioxidants protected effectively the cytotoxicity from hydroxyl radicals in the brain cultures. More than 70% of cell viabilities among different antioxidants was at 1 mM DFX, 1.43 $\mu\textrm{m}$ laccase, 12.5 $\mu\textrm{m}$ water extract, 12.5 $\mu\textrm{m}$ ethanol extract and 50 $\mu\textrm{m}$ $\alpha$-tocopherol individually, compared with 20 mU/ml GO alone. In comparison to the antioxidative activities of antioxidants, laccase and extracts from RVS showed strong antioxidative effects even at low concentration.

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An Assay of L-Ascorbic Acid with Crude Enzyme from Cucumber(Cucumis sativus L.) (오이(Cucumis sativus L.)에서 추출한 조효소 용액을 이용한 L-Ascorbic Acid의 정량분석)

  • Lee, Suk-Yeong;Kim, Hong-Seop;Go, Dae-Hui
    • The Korean Journal of Food And Nutrition
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    • v.8 no.3
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    • pp.159-164
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    • 1995
  • The test possibility of L-ascorbic acid(AsA) assay by ascorbate oxidase(A.O) solution obtained from cucumber was estimated. The results obtained were summarized as follow. The difference of absorbance before and after oxidation of AsA by A.O solution was proportional to the AsA concentration. Iso- AsA compared to same concentration of AsA was 97% response. Therefore, the 0 specificity on Iso- AsA a was deficient. On the sucrose, glucose, fructose were coexisted from 5 times, 500 times of AsA concentration, none of them were affected by any level of AsA concentration. In the case of EDTA, it was not nearly affected that EDTA was coexisted 50 times of AsA concentration, but EDTA Inhibited considerably from 500 times of AsA concentration. The effect of citric acid on the AsA concentration was not in the same level of AsA, however, it was slightly inhibited from 5 times, 50 times of AsA concentration and 500 times of AsA inhibited remarkably As known L-AsA was catalycally autooxidated by Cu2+ and Fe2+. They were not nearly affected In the above 0.01M level on the Ash assay. However, Fe3+ ion was neither nearly affected in the 0.001M level nor In the above 0.01M level on the AsA assay.

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Identification of the Genes Involved in the Fruiting Body Production and Cordycepin Formation of Cordyceps militaris Fungus

  • Zheng, Zhuang-Li;Qiu, Xue-Hong;Han, Ri-Chou
    • Mycobiology
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    • v.43 no.1
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    • pp.37-42
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    • 2015
  • A mutant library of Cordyceps militaris was constructed by improved Agrobacterium tumefaciens-mediated transformation and screened for degradation features. Six mutants with altered characters in in vitro and in vivo fruiting body production, and cordycepin formation were found to contain a single copy T-DNA. T-DNA flanking sequences of these mutants were identified by thermal asymmetric interlaced-PCR approach. ATP-dependent helicase, cytochrome oxidase subunit I and ubiquitin-like activating enzyme were involved in in vitro fruiting body production, serine/threonine phosphatase involved in in vivo fruiting body production, while glucose-methanol-choline oxidoreductase and telomerase reverse transcriptase involved in cordycepin formation. These genes were analyzed by bioinformatics methods, and their molecular function and biology process were speculated by Gene Ontology (GO) analysis. The results provided useful information for the control of culture degeneration in commercial production of C. militaris.

Antioxidative Effects of Ethanol Extracts from Rhus Verniciflua Stokes (RVS) on Mouse Whole Brain Cells (옻나무 에탄올 추출물의 쥐 뇌세포에 대한 항산화효과)

  • Lim, Kye-Taek;Shim, Jae-Han
    • Korean Journal of Food Science and Technology
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    • v.29 no.6
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    • pp.1248-1254
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    • 1997
  • To measure antioxidative activities, the various extracts from RVS (Rhus Verniciflua Stokes) were tried out with either DPPH or thiocyanate method. Also we used the GO (Glucose Oxidase) 20 mU/mL hydroxyl radical system in mouse whole brain cell culture. Chloroform, n-hexane or ethanol were used as extract solutions which had different polarity respectively. In DPPH and thiocyanate method, the antioxidative activities of the crude ethanol extracts were stronger than other extracts. The crude ethanol extracts were fractionated 5 peaks by glass column. Among of them, antioxidative activity of peak II $(P_{II})$ was shown stronger than other fractions, a little for peak III $(P_{III})$ and peak IV $(P_{IV})$, and none for peak I $(P_I)$ and Peak V $(P_V)$. In the antioxidative effects of crude ethanol extracts (30 mg/mL), cell viabilities were evaluated $1\;{\mu}L\;(297\;{\mu}g/mL)$, $2\;{\mu}L\;(588\;{\mu}g/mL)$ of crude ethanol extracts 59%, 68% respectively. $10\;{\mu}L\;(2,727\;{\mu}g/mL)$ addition of crude ethanol extracts had 95% cell viabilities, 0.01% significant, comparing control. In addition, the compounds related to antioxidative effect of crude ethanol extract might be glycoproteins by means of SDS-PAGE. Comparison to antioxidative effects between several antioxidants (ascorbic acid, ${\alpha}-tocopherol$, catalase) $273\;{\mu}L/mL$ addition of crude ethanol extracts corresponds to $1\;{\mu}g/mL$ catalase in antioxidative effects.

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The Effect of Nitric Oxide Donor or Nitric Oxide Synthase Inhibitor on Oxidant Injury to Cultured Rat Lung Microvascular Endothelial Cells (산화질소 공여물과 산화질소 합성효소 길항제가 백서 폐미세혈관 내피세포 산화제 손상에 미치는 영향)

  • Chang, Joon;Michael, John R.;Kim, Se-Kyu;Kim, Sung-Kyu;Lee, Won-Young;Kang, Kyung-Ho;Yoo, Se-Hwa;Chae, Yang-Seok
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.6
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    • pp.1265-1276
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    • 1998
  • Background : Nitric oxide(NO) is an endogenously produced free radical that plays an important role in regulating vascular tone, inhibition of platelet aggregation and white blood cell adhesion to endothelial cells, and host defense against infection. The highly reactive nature of NO with oxygen radicals suggests that it may either promote or reduce oxidant-induced cell injury in several biological pathways. Oxidant injury and interactions between pulmonary vascular endothelium and leukocytes are important in the pathogenesis of acute lung injury, including acute respiratory distress syndrome(ARDS). In ARDS, therapeutic administration of NO is a clinical condition providing exogenous NO in oxidant-induced endothelial injury. The role of exogenous NO from NO donor or the suppression of endogenous NO production was evaluated in oxidant-induced endothelial injury. Method : The oxidant injury in cultured rat lung microvascular endothelial cells(RLMVC) was induced by hydrogen peroxide generated from glucose oxidase(GO). Cell injury was evaluated by $^{51}$chromium($^{51}Cr$) release technique. NO donor, such as S-nitroso-N-acetylpenicillamine(SNAP) or sodium nitroprusside(SNP), was added to the endothelial cells as a source of exogenous NO. Endogenous production of NO was suppressed with N-monomethyl-L-arginine(L-NMMA) which is an NO synthase inhibitor. L-NMMA was also used in increased endogenous NO production induced by combined stimulation with interferon-$\gamma$(INF-$\gamma$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), and lipopolysaccharide(LPS). NO generation from NO donor or from the endothelial cells was evaluated by measuring nitrite concentration. Result : $^{51}Cr$ release was $8.7{\pm}0.5%$ in GO 5 mU/ml, $14.4{\pm}2.9%$ in GO 10 mU/ml, $32.3{\pm}2.9%$ in GO 15 mU/ml, $55.5{\pm}0.3%$ in GO 20 mU/ml and $67.8{\pm}0.9%$ in GO 30 mU/ml ; it was significantly increased in GO 15 mU/ml or higher concentrations when compared with $9.6{\pm}0.7%$ in control(p < 0.05; n=6). L-NMMA(0.5 mM) did not affect the $^{51}Cr$ release by GO. Nitrite concentration was increased to $3.9{\pm}0.3\;{\mu}M$ in culture media of RLMVC treated with INF-$\gamma$ (500 U/ml), TNF-$\alpha$(150 U/ml) and LPS($1\;{\mu}g/ml$) for 24 hours ; it was significantly suppressed by the addition of L-NMMA. The presence of L-NMMA did not affect $^{51}Cr$ release induced by GO in RLMVC pretreated with INF-$\gamma$, TNF-$\alpha$ and LPS. The increase of $^{51}Cr$ release with GO(20 mU/ml) was prevented completely by adding 100 ${\mu}M$ SNAP. But the add of SNP, potassium ferrocyanate or potassium ferricyanate did not protect the oxidant injury. Nitrite accumulation was $23{\pm}1.0\;{\mu}M$ from 100 ${\mu}M$ SNAP at 4 hours in phenol red free Hanks' balanced salt solution. But nitrite was not detectable from SNP upto 1 mM The presence of SNAP did not affect the time dependent generation of hydrogen peroxide by GO in phenol red free Hanks' balanced salt solution. Conclusion : Hydrogen peroxide generated by GO causes oxidant injury in RLMVC. Exogenous NO from NO donor prevents oxidant injury, and the protective effect may be related to the ability to release NO. These results suggest that the exogenous NO may be protective on oxidant injury to the endothelium.

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Effects of Daehwanggamchoeumja and its component groups on diabetes, free radical and antioxidative defense system in Alloxan-induced diabetic rats (대황감초음자(大黃甘草飮子)와 그 구성약물군(構成藥物群)이 Alloxan 유도 당뇨(糖尿) 백서(白鼠)의 혈청 조성 및 항산화 효과에 미치는 영향)

  • Go Won-Do;Gwak Dong-Gul;Shin Hwa-Seog;Choi Oi-Chul;Park Sun-Dong
    • Herbal Formula Science
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    • v.10 no.2
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    • pp.159-188
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    • 2002
  • The purpose of this study was to reseach the effect of Daehwanggamchoeumja(大黃甘草飮子) and its component groups on diabetes, free radicals, and antioxidants system in Alloxan-induced diabetic rats. The experimental group was divided into three groups: Daehwanggamchoeumja(DG), and its components groups, Gamdutang (Gamcho&Daedu; DG-I) and Daehwanggamchotang(DG-2). The results were obtained as follows: 1. In the study of effect on diabetic metabolic dysfunction(Glucose, Triglyceride, Total Cholesterol, HDL Cholesterol, Total Protein, Albumin, Creatine, BUN), only DG has a significant effect. 2. In the study on free radical scavenging effect in vitro(the suppressing effect on peroxidation of linoleic acid on concentration, the scavenging effect of DPPH radical, inhibitory effect of superoxide in xanthine-xanthine oxidase system, inhibitory effect on lipid peroxidation reaction by hydroxy radical in $H_2O_2Fe^{2-}$system, and the effect on Nitrate reductase activity), DG and DG-2 have more effect than DG-l relatively. 3. In the study on antioxidants system in vivo(The level of serum LPO, The level of hepatic LPO, Catalase, GSH, GST), only DG has a significant effect. These results suggest that Daehwanggamchoeumja(大黃甘草飮子) has an effect on diabetes, peroxidative damage by free radical, so it seems to be useful to prevent and treat diabetes. The mechanisms of these are supposed to be involved in antioxidant and three drugs' cooperative synergy effect.

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Combined Effects of Vital Gluten, Gum, Emulsifier, and Enzyme on the Properties of Rice Bread (활성글루텐, 검, 유화제 및 효소제의 복합첨가에 따른 쌀빵의 품질특성)

  • Kim, Kyung-Eun;Lee, Young-Tack
    • Food Engineering Progress
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    • v.13 no.4
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    • pp.320-325
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    • 2009
  • The effects of adding additives such as vital gluten, gum, emulsifier, and enzyme to rice flour on baking quality were examined. The effects of different gums on the pasting and dough properties of rice flour containing vital gluten were studied using a Rapid Visco Analyzer (RVA) and a Brabender farinograph. The RVA peak, breakdown, and final viscosities decreased with the addition of gums, while setback viscosity increased. The farinogram showed that rice flour supplemented with gums such as tara gum, guar gum, and locust bean gum (LBG) increased water absorption and dough stability, yielding strengthened dough similar to wheat flour dough. The addition of guar or tara gum/sodium stearoyl lactylate (SSL)/fungal $\alpha$-amylase (AMYL) or glucose oxidase (GO) blend improved the volume and reduced the crumb firmness of rice bread prepared from rice flour containing 14% vital gluten. Therefore, the combined addition of gum, emulsifier and enzyme into rice flour significantly improved the rice bread quality, allowing the decrease of the vital gluten level in rice bread formula.

Enhanced and Targeted Expression of Fungal Phytase in Saccharomyces cerevisiae

  • LIM, YOUNG-YI;EUN-HA PARK;JI-HYE KIM;SEUNG-MOON PARK;HYO-SANG JANG;YOUN-JE PARK;SEWANG YOON;MOON-SIK YANG;DAE-HYUK KIM
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.915-921
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    • 2001
  • Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. In order to express a high level of fungal phytase in Saccharomyces cerevisiae, various expression vectors were constructed with different combinations of promoters, translation enhancers, signal peptides, and terminator. Three different promoters fused to the phytase gene (phyA) from Aspergillus niger were tested: a galactokinase (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, and yeast hybrid ADH2-GPD promoter consisting of alcohol dehydrogenase II (ADH2) and a GPD promoter. The signal peptides of phytase, glucose oxidase (GO), and rice amylase 1A(RAmy1A) were included. Plus, the translation enhancers of the ${\Omega}$ sequence and UTR70 from the tobacco mosaic virus (TMV) and spinach, respectively, were also tested. Among the recombinant vectors, pGphyA06 containing the GPD promoter, the ${\Omega}$ sequence, RAmy1A, and GAL7 terminator expressed the highest phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase was also performed by inserting an endoplasmic reticulum (ER) retention signal, KDEL sequence, into the C-terminus of the phytase within the vector pHphyA-6. It appeared that the KDEL sequence directed most of the early expression of phytase into the intracellular compartment yet more than $60\%$ of the total phytase activity was still retained within the cell even after the prolonged (>3 days) incubation of the transformant. However, the intracellular enzyme activity of the transformant without a KDEL sequence was as high as that of the extracellular one, thereby strongly suggesting that the secretion of phytase in S. cerevisiae appeared to be the rate-limiting step for the expression of a large amount of extracellular recombinant phytase, when compared with other yeasts.

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