• Applied Biological Chemistry
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• 제27권3호
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• pp.180-186
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• 1984

비교적 높은 역가의 glucose oxidase(GOD)와 catalase(CAT)를 세포의 효소로 생산하는 균주인 Penicillium spp., PS-8을 선별배지를 사용하여 액체 배양하였으며, 그 결과 배양액 1ml당 2.7units의 glucose oxidase와 2.0units의 catalase를 얻었다. Glucose oxidase와 catalase를 분리하기 위하여 $60{\sim}90%\;(NH_4)_2SO_4$ 분별침전을 행한후 DEAE-cellulose column을 사용하여 두 효소를 완전히 분리하였으며, 이들 효소의 회수율은 54%와 34%이었다. 분리된 glucose oxidase와 catalase는 PS-8 균주를 효소 고정 매개체로 하여 2.5% glutaraldehyde를 가교제로 12시간 동안 처리함으로써 효소를 고정시켰다. 이들 고정화 효소는 CAT/GOD 값이 서로 다르게 동시 고정, 고정후 혼합, glucose oxidase만의 고정 등의 형태로 만들었다. pH에 따른 효소의 활성변화에서는 동시고정 및 고정후 혼합 방법이 수용성 효소보다 안정화됨을 보여 주었으며, 동시 고정에서는 CAT/GOD값이 높을수록 보다 완만한 pH 활성곡선을 나타내었다. GOD와 CAT/GOD=10의 Km' 값은 각각 $7.1{\times}10^{-2}$ 및 $5.1{\times}10^{-2}M$이었으며, 이들의 활성화 에너지값은 각각 3.97 및 2.98 kcal/mole/deg이었다.

• 한국식품과학회지
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• 제31권2호
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• pp.416-420
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• 1999

육계와 돼지의 혈장에서 포도당과 lysine이 축합하여 갈색의 물질을 생성하는 Maillard 반응을 억제시키기 위하여 혈장에 GOD 5 또는 10 units/g이나 제빵용 효모(0.3% w/w)를 첨가하여 탈당시켰다. GOD와 제빵용 효모의 첨가가 혈장 포도당 함량과 pH 변화에 미치는 영향을 조사했다. 초기 닭과 돼지 혈장의 평균 포도당 농도는 각각 $150\;mg/100\;cm^{3}$과 $143\;mg/100\;cm^{3}$이었다. GOD나 효모를 첨가한 혈장의 포도당 농도는 시간이 지남에 따라 감소되었다. 효모를 이용한 경우, 육계와 돼지의 혈장 포도당이 거의 없어지는데는 약 4시간 정도 걸렸다. 효모가 GOD 보다 혈장 포도당을 빨리 감소시켰고 GOD의 농도 10 units/g이 5 units/g보다 효과적이었다. 혈장 포도당을 제거함으로써 Maillard반응의 억제가 가능하리라 여겨진다. 혈장 포도당의 감소에 따라 pH도 같이 감소했다가 포도당 양이 안정된 후에는 pH 값이 약간 상승했다. 혈장의 pH를 측정함으로써 혈장의 탈당 정도를 간접적으로 알 수 있었다.

• 한국전기전자재료학회:학술대회논문집
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• 한국전기전자재료학회 1999년도 춘계학술대회 논문집
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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.

• BMB Reports
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• 제40권6호
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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.

• 한국미생물·생명공학회지
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• 제26권1호
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• pp.68-75
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• 1998

A. niger의 GOD(Glucose Oxidase) 대량생산과 효율적인 분비를 protein의 대량생산에 많이 사용되는 strain인 S. cerevisiae에서 시도하였다. S. cerevisiae의 ADH1과 GAL 10 promotor, 그리고 ${alpha}$-MF signal sequence와 A. oryzae의 ${alpha}$-amylase signal sequence 및 S. cerevisiae의 GAL7과 A. niger의 GOD terminator를 이용하여 4개의 expression vector를 합성한 후 S. cerevisiae 2805에 auxotroph 방법으로 형질변환시켰다. 변이체들을 배양하여 세포내와 세포외의 GOD활성도를 분석한 결과 GAL 10 promotor가 삽입된 pGAL변이체들이 ADH1 promotor가 삽입된 pADH 변이체들 보다 GOD 생산성이 높았다. GAL 10 promotor와 A. oryzae의 ${alpha}$-amylase signal sequence가 삽입된 pGALGO2에서 115시간 배양시 GOD의 생산이 가장 높았다($GOD_{total}$: 10.3 unit/mL, $GOD_{ex}$: 8.7 unit/mL). 이 수치는 같은 promotor인 GAL 10 promotor와 ${alpha}$-MF signal sequence가 삽입된 pGALGO1보다 3배정도 높다. 이 결과는 ADH 1 promotor를 사용하였을 경우에도 일치하였다. 또한 A. oryzae의 ${alpha}$-amylase signal sequence가 S. cerevisiae의 ${alpha}$-MF signal sequence보다 GOD를 더 효과적으로 분비시켰다. 상기 결과로 미루어 보면 signal sequence가 단백질의 분비 외에도 단백질 합성에도 많은 영향을 주는 것으로 추측된다. pGALGO1과 pGALGO2의 GOD분비효율은 각각 89%, 84%이었다. S. cerevisiae에서는 일반적으로 과당화가 일어나기 때문에 S. cerevisiae에서 합성된 재조합 GOD의 분자량은 250 kDa으로 A. niger의 GOD(170 kDa)보다 더 컸다.

• KSBB Journal
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• 제13권3호
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• pp.244-250
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• 1998

글루코스와 초산을 온라인 모니터링 하기위한 흐름주입분석기 술이 개발되었고 대장균발효공정에 이용되었다. 또한, Epoxy 고 분자 담체에 고정화된 GOD와 SOD 를 이용한 GOD-FIA와 SOD-FlA의 특성을 연구했다. 즉, FIA 의 조작온도, 운반용액 속의 첨 가제 (Triton, EDTA, natrium azid 등), 분석 시 료에 용 해된 신진대사물의 농도, pH, 초산측정시 사코진 농도 등에 따 라 고정화된 GOD와 SOD의 활성도, 즉 검출신호의 높이변화를 고찰했다. 최소배양액 과 복합배양액을 사용한 대장균 발효공정 에서 달루코스와 초산을 동시에 온라인 모니터링 하였으며 그결 과는 오프라인 분석결과와 잘 일치 하였다,

• Journal of Microbiology and Biotechnology
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• 제17권6호
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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.

• 센서학회지
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• 제27권3호
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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.

• Animal Bioscience
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• 제35권1호
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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.

• KSBB Journal
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• 제13권1호
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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.