• Title/Summary/Keyword: Glucose oxidase (GOD)

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Preparation and Properties of Coimmobilized Glucose Oxidase-Catalase (Glucose Oxidase와 Catalase의 동시 고정화 제품과 성질)

  • Lee, Suk-Hee;Uhm, Tai-Boong;Cho, Sook-Ja;Byun, Si-Myung
    • Applied Biological Chemistry
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    • v.27 no.3
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    • pp.180-186
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    • 1984
  • For the study of glucose oxidase(GOD) and catalase(CAT) coimmobilization system, the enzymes were obtained from Penicillium spp., PS-8, and the strain itself was used as an immobilizing matrix. To separate glucose oxidase and catalase after the ammonium sulfate fractionation of the culture broth, DEAF-cellulose column was used and its activity yield was 54 and 34%, respectively. Both enzymes were immobilized on the cell matrix, followed crosslinking with 2.5% glutaraldehyde for 12hr. In the determination of efficiencies of GOD and CAT of dual, mixed and soluble enzyme systems, the dual immobilized one w-as superior to those of the soluble or mixed ones. In the comparison of pH profiles, the dual and mixed types showed broader maximum pH ranges than the soluble type. Varying CAT/GOD ratio of the dual system, the higher the ratio showed the broader activity profile. In the comparison of apparent $K_m$ of GOD only and CAT/GOD=10, they were $7.1{\times}10^{-2}$ and $5.1{\times}10^{-2}M$. Their activation energies showed 3.98kcal/mole/deg for GOD only and 2.98kcal/mole/deg for CAT/GOD=10.

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Glucose Content and pH of Broiler and Porcine Blood Plasma by Glucose Oxidase or Baker's Yeast Addition (Glucose Oxidase 및 제빵용 효모 첨가에 따른 육계와 돼지의 혈장 포도당과 pH 변화)

  • Lee, Jae-Jun;Yi, Young-Hyoun
    • Korean Journal of Food Science and Technology
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    • v.31 no.2
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    • pp.416-420
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    • 1999
  • The effects of GOD and yeast on glucose content and pH of broiler and porcine blood plasma were investigated. The initial glucose concentration of broiler and porcine blood plasma were $150mg/100cm^{3}\;and\;143mg/100cm^{3}$, respectively. Addition of GOD and yeast decreased glucose contents in broiler and porcine plasma. As expected, plasma glucose content decreased as incubation time increased. While 1080 and 1110 min were required to remove glucose from both broiler and porcine plasma at GOD 5 units/g and 480 and 1020 min were required at GOD 10 units/g, respectively; both required 240 min at 0.3% yeast (w/w). The Maillard reaction can be prevented by desugarization. During the removal of glucose, pH of the plasma decreased. As glucose content in plasma leveled off, the pH value of plasma increased. Therefore, pH may be used as an index of desugarization.

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Electrochemical Properties of Polypyrrole/ Glucose Oxidase Enzyme Electrode (Polypyrrole/Glucose Oxidase 효소전극의 전기화학적 특성)

  • 김현철;구할본
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 1999.05a
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    • pp.357-361
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    • 1999
  • GOD electrochemically immobilized in PPy/GOD complex have an effect on redox properties of the complex. In the cyclicvoltammetry, GOD shows the redox reaction at the potential below -0.6Y vs. Ag/AgCI. That leads to new peaks in the cyclicvoltammograms in additional to typical PPy peaks. The pH of electrolyte solution during potential swing decreased to 4.4, and then increased to 10. That suggests the redox of GOD for the cycling. As the concentration of GOD was increased, the anodic wave of the new peaks was strong as much as increased. GOD obstructs the diffusion of electrolyte anion because of its net chain. Insulating property of GOD is cause that it made the faradic impedance of complex large in charge transfer. It suggests that increase of the concentration of GOD be against electrochemical coupling. Therefore, the concentration of GOD and electrochemical coupling should be dealt with each other. The apparent Michaelis-lenten constant ( K\`$_{M}$ ) was determined by 30.7 mmol d $m^{-3}$ fur the PPy/GOD complex. The value is of the same order of magnitude as that for soluble glucose oxidase from Aspergillus Niger.r.

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Cloning and Characterization of a Single Chain Antibody to Glucose Oxidase from a Murine Hybridoma

  • Sellrie, Frank;Schenk, Jorg A.;Behrsing, Olaf;Drechsel, Oliver;Micheel, Burkhard
    • BMB Reports
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    • v.40 no.6
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    • pp.875-880
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    • 2007
  • Glucose oxidase (GOD) is an oxidoreductase catalyzing the reaction of glucose and oxygen to peroxide and gluconolacton (EC 1.1.3.4.). GOD is a widely used enzyme in biotechnology. Therefore the production of monoclonal antibodies and antibody fragments to GOD are of interest in bioanalytics and even tumor therapy. We describe here the generation of a panel of monoclonal antibodies to native and heat inactivated GOD. One of the hybridomas, E13BC8, was used for cloning of a single chain antibody(scFv). This scFv was expressed in Escherichia coli XL1-blue with the help of the vector system pOPE101. The scFv was isolated from the periplasmic fraction and detected by western blotting. It reacts specifically with soluble active GOD but does not recognize denatured GOD adsorbed to the solid phase. The same binding properties were also found for the monoclonal antibody E13BC8.

Overproduction and High Level Secretion of Glucose Oxidase in Saccharomyces cerevisiae (Glucose Oxidase의 Saccharomyces cerevisiae에서의 대량생산 및 고효율 분비)

  • 홍성용;최희경;이영호;백운화;정준기
    • Microbiology and Biotechnology Letters
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    • v.26 no.1
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    • pp.68-75
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    • 1998
  • The overproduction and high level secretion of Glucose Oxidase (GOD) from A. niger in S. cerevisiae was carried out by cloning GOD gene. For this purpose, using two different strong promoters (ADH1 promoter, GAL10 promoter) and signal sequences (${alpha}$-MF signal sequence of S. cerevisiae and ${alpha}$-amylase signal sequence of A. oryzae) and GAL7- and GOD terminator, four expression vectors were constructed. All the expression vectors were transformed in S. cerevisiae 2805 using auxotroph method. By the flask culture, transformants of pGAL expression vector series containing GAL 10 promotor showed much higher GOD productivity than transformants of pADH expression vector series containing ADH1 promoter Transformants of pGALGO2 containing GAL10 promotor and ${alpha}$-amylase signal sequence has shown the best productivity of GOD ($GOD_{total}$: 10.3 unit/mL, $GOD_{ex}$: 8.7 unit/mL) at 115 hr. This value was three fold higher than that of pGALGO1 containing GAL 10 promotor and ${alpha}$-MF signal sequence, even if the same promotor was involved. Through the ${alpha}$-amylase signal sequence of A. oryzae, GOD was secreted much more than the case of ${alpha}$-MF signal sequence from S. cerevisiae. These results suggest that signal sequence may play a important roles in not only the secretion but also the overproduction of foreign protein. Secretion rate of GOD in pGALGO1 and pGALGO2 was 89% and 84%, respectively, Because of the overglycosylation in S. cerevisiae the molecular weight of recombinant GOD in S. cerevisiae was much larger (250 kDa) than that of nature GOD in A. niger (170 kDa).

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On-line Monitoring of Glucose and Acetate by Flow-Injection Analysis in Escherichia coli Fermentation Process (대장균 발효공정에서 흐름주입분석을 이용한 글루코스와 초산의 온라인 모니터링)

  • 이종일
    • KSBB Journal
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    • v.13 no.3
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    • pp.244-250
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    • 1998
  • Flow-injection analysis (FIA) for on-line monitoring of glucose and acetate are described and employed in E.coli fermentation process. Glucose oxidase (GOD) for the detection of glucose is immobilized on epoxy polymer support, which is packed in a small cartirdge, and applied to a GOD-FIA system. The detection of acetate is based on the inhibition of acetate to the oxidation of sarcosine by sarcosine oxidase (SOD). SOD is also immobilized on epoxy polymer support and used for a SOD-FIA system. GOD-FIA system is characterized as well as SOD-FIA system by the investigation of the effects of pH, temperature and metabolites in samples on the peak height. GOD-FIA and SOD-FIA systems were also applied for on-line On-line measurements buy FIA measurements by FIA were in god agreement with off-line measurements.

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Simultaneous and Sequential Co-Immobilization of Glucose Oxidase and Catalase onto Florisil

  • Gul, Ozyilmaz;Tukel, S. Seyhan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.960-967
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    • 2007
  • The co-immobilization of Aspergillus niger glucose oxidase (GOD) with bovine liver catalase (CAT) onto florisil (magnesium silicate-based porous carrier) was investigated to improve the catalytic efficiency of GOD against $H_2O2$ inactivation. The effect of the amount of bound CAT on the GOD activity was also studied for 12 different initial combinations of GOD and CAT, using simultaneous and sequential coupling. The sequentially co-immobilized GOD-CAT showed a higher efficiency than the simultaneously co-immobilized GOD-CAT in terms of the GOD activity and economic costs. The highest activity was shown by the sequentially co-immobilized GOD-CAT when the initial amounts of GOD and CAT were 10 mg and 5 mg per gram of carrier. The optimum pH, buffer concentration, and temperature for GOD activity for the same co-immobilized GOD-CAT sample were then determined as pH 6.5, 50 mM, and $30^{\circ}C$, respectively. When compared with the individually immobilized GOD, the catalytic activity of the co-immobilized GOD-CAT was 70% higher, plus the reusability was more than two-fold. The storage stability of the co-immobilized GOD-CAT was also found to be higher than that of the free form at both $5^{\circ}C\;and\;25^{\circ}C$. The increased GOD activity and reusability resulting from the co-immobilization process may have been due to CAT protecting GOD from inactivation by $H_2O2$ and supplying additional $O_2$ to the reaction system.

Fabrication and Improved Sensitivity with Surface Treatment of TiO2/GOD Mixture based Glucose Biosensor (TiO2/GOD 혼합물 기반의 글루코스 바이오 센서의 제작과 표면 처리를 통한 감도개선)

  • Lee, Junyeop;Jung, Dong Geon;Lee, Jae Yong;Kim, Jae Keon;Jung, Daewoong;Kong, Seong Ho
    • Journal of Sensor Science and Technology
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    • v.27 no.3
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    • pp.170-174
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    • 2018
  • In this paper, the $TiO_2$/glucose oxidase (GOD) mixture has synthesized through simple and low-cost fabrication methods. The physical properties of the mixture were proved using an FT-IR/NIR spectrometer, an X-Ray diffractometer, and a Raman spectrometer. GOD maintained its bioactivity during all fabrication process. The current characteristics of the glucose biosensor were proportional to the glucose concentration and effective surface area of square pyramid on a silicon substrate. The maximum current change was measured in a pH 7.0 buffer solution. The simple and low-cost fabrication process and surface treatment can be used widely in previous research for improvements in effective surface area.

Effects of dietary supplementation of glucose oxidase, catalase, or both on reproductive performance, oxidative stress, fecal microflora and apoptosis in multiparous sows

  • Sun, Xiaojiao;Piao, Longguo;Jin, Haifeng;Nogoy, K. Margarette C.;Zhang, Junfang;Sun, Bin;Jin, Yi;Lee, Dong Hoon;Choi, Seong-Ho;Smith, Stephen B;Li, Xiangzi
    • Animal Bioscience
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    • v.35 no.1
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    • pp.75-86
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    • 2022
  • Objective: The objective of this experiment was to investigate the effect of dietary glucose oxidase (GOD), catalase (CAT), or both supplementation on reproductive performance, oxidative stress, and apoptosis in sows. Methods: A total of 104 multiparous sows were randomly assigned to four groups (n = 26) with each group given a basal diet, basal diet plus GOD at 60 U/kg, basal diet plus CAT at 75 U/kg, and basal diet plus GOD at 60 U/kg and CAT at 75 U/kg. Sows were fed the experimental diets throughout gestation and lactation. Results: Dietary GOD supplementation increased average daily feed intake of sows and litter weight at weaning (p<0.05). Dietary CAT supplementation reduced the duration of parturition, stillbirth, and piglet mortality and increased growth performance of weaned piglets (p<0.05). Dietary GOD and CAT supplementation enhanced antioxidant enzyme activities and lessened oxidative stress product levels in plasma of sows and elevated antioxidant capacity of 14-day milk and plasma in weaned piglets (p<0.05). Dietary GOD supplementation increased fecal Lactobacillus counts and reduced Escherichia coli counts of sows (p<0.05). Compared with the basal diet, the GOD diet reduced fecal Escherichia coli counts of sows, but the addition of CAT did not reduce Escherichia coli counts in the GOD diet. Dietary GOD and CAT supplementation reduced the apoptosis rate of the liver, endometrium, and ovarian granulosa cells in sows (p<0.05). In the liver, uterus, and ovary of sows, the mRNA expression of caspase-3 and caspase-9 was downregulated by dietary GOD and CAT supplementation (p<0.05). Conclusion: Dietary GOD and CAT supplementation could improve the antioxidant capacity of sows and weaned piglets, and alleviate hepatic, ovarian and uterine apoptosis by weakening apoptosis-related gene expression. Glucose oxidase regulated fecal microflora of sows, but supplementation of CAT to GOD could weaken the inhibitory effect of GOD on fecal Escherichia coli.

Electrochemical Determination of Immobilization Technique for Glucose Sensor Fabrication (포도당 센서의 제작을 위한 고정화 방법의 전기화학적 결정)

  • 정태훈;홍석인;노봉수;정용섭;윤정원;김태진
    • KSBB Journal
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    • v.13 no.1
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    • pp.52-57
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    • 1998
  • The present work proposes a simple electrochemical method applicable to any immobilization processes of oxidase using a Clark type oxygen electrode as a base transducer. The present work suggests an optimal immobilization technique among three different methods of glucose oxidase(GOD) onto one side of $37[\mu}$mthick blend membranes, composed o 80% of cellulose triacetate and 20% of polycaprolactone, on the basis of the maximum Michaelis-Menten parameter(Vm) determined by either steady state or transient analyses. The electrode system was made of disk type gold cathode(4mm diameter) and Ag/AgCl anode. One side of the blend membrane was in contact with the cathode surface while the other side was immobilized with GOD either in covalent-bond or cross-linked forms, the latter being covered by $25{\mu}$m thick dialysis membrane of cellulose acetate. The resultant current density was on-line monitored by a potentiostat while glucose level was varied from 1 to 20 mM. The present study shows that direct cross-linking of GOD with glutaraldehyde was mostly preferred for fabrication of glucose sensor, on the basis of resultant kinetic parameters from either steady state or transient analyses.

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