• BMB Reports
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• 제33권4호
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• pp.321-325
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• 2000

Quinone reductase was purified to homogeneity from bovine liver by using ammonium sulfate fractionation, ionexchange chromatography, and gel filtration chromatography. The enzyme utilized either NADH or NADPH as the electron donor. The enzyme catalyzed the reduction of several quinones and other artificial electron acceptors. Furthermore, the enzyme catalyzed NAD(P)H-dependent reduction of azobenzene. The apparent Km for 1,4-benzoquinone and azobenzene was 1.64 mM and 0.524 mM, respectively. The reduction of azobenzene by quinone reductase was almost entirely inhibited by dicumarol or Cibacron blue 3GA, potent inhibitors of the mammalian quinone reductase. In the presence of 1.0${\mu}M$ Cibacron blue 3GA, azoreductase activity was lowered by 45%, and almost complete inhibition was seen above 2.0 ${\mu}M$ Cibacron blue 3GA.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권1호
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• pp.84-87
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• 2006

Batch binding experiments were performed to assess the recovery performance of glyceraldehyde 3-phosphate dehydrogenase (G3PDH) bound to the unshielded and polymer (polyvinyl pyrrolidone. PVP)-shielded dye-ligand (Cibacron Blue 3GA) adsorbent. The adoption of a polymer-shielded, dye-ligand technique facilitated the elution efficiency of bound G3PDH. It was demonstrated that the recovery of G3PDH using polymer-shielded dye-ligand adsorption yielded higher elution efficiency, at 60.5% and a specific activity of 42.3 IU/mg, after a low ionic strength elution (0.15 M NaCl). The unshielded dye-ligand yielded lower elution efficiency. at 6.5% and a specific activity of 10.2 IU/mg.

• 생명과학회지
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• 제12권3호
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• pp.281-287
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• 2002

소 간으로부터 퀴논 환원효소를 정제하였으며 정제된 효소는 벤조퀴논과 나프토퀴논 뿐만 아니라 페난트렌 퀴논의 환원도 촉매하였다. 소 간으로부터 정제된 퀴논 환원 효소는 아릴니트로소 화합물의 생변환을 촉매하였으며 반응 생성물은 TLC, GC, GC-MS, NMR을 사용하여 확인되었다. 이 반응은 포유동물 퀴논 환원효소의 강력한 저해제인 Cibacron blue 3GA나 dicumarol에 의하여 크게 저해되었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.638-639
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• 2000

Quinone reductase was purified to electrophoretic homogeneity from bovine liver by using ammonium sulfate fractionation, ion-exchange chromatography, and gel filtration chromatography. The enzyme utilized either NADH or NADPH as the electron donor. The optimum pH of the enzyme was pH 8.5, and the activity of the enzyme was greatly inhibited by $Cu^{2+}$ and $Hg^{2+}$ ions, dicumarol and cibacron blue 3GA. The enzyme catalyzed the reduction of several quinones and other artificial electron acceptors. Furthermore, the enzyme catalyzed NAD(P)H-dependent reduction of azobenzene or 4-nitroso-N,N-dimethylaniline. The apparent $K_m$ for 1,4-benzoquinone, azobenzene, and 4-nitroso-N,N-dimethylaniline was 1.64mM, 0.524mM and 0.225mM, respectively. The reduction of azobenzene or 4-nitroso-N,N-dimethylaniline by quinone reductase was strongly inhibited by dicumarol or cibacron blue 3GA, potent inhibitors of quinone reductase.

• 한국동물학회지
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• 제29권4호
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• pp.283-293
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• 1986

사람의 alpha-fetoprotein(AFP)에 대한 모노클론 항체의 생산 및 분석을 위하여, 태아조직을 재료로 추출법, DEAE-cellulose 및 concanavalin A-Sepharose, Cibacron blue F3GA-agarose, immunoadsorbent column등의 흡착크로마토그라피법에 의해 AFP를 분리하였다. 총 534g의 태아조직에서 AFP의 量은 8.76 mg으로서 순수분리되었음을 확인하였다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.552-555
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• 2005

The influence of whole yeast cells $(0{\sim}15%\;w/v)$ on the protein adsorption performance in dye-ligand chromatography was explored. The adsorption of a model protein, bovine serum albumin (BSA), was selected to demonstrate this approach. The UpFront adsorbent $(p=1.5\;g/cm^3)$ derivatised with Cibacron Blue 3GA and a commercially available expanded bed column (20 mm i.d.) from UpFront Chromatography, Denmark, were employed in the batch binding and expanded bed operation. The BSA binding capacity was demonstrated to not be adversely affected by the presence of yeast cells. The dynamic binding capacity of BSA at a $C/C_0=0.1$ biomass concentration of 5, 10, 15% w/v were 9, 8, and 7.5mg/mL of settled adsorbent, respectively.

• 멤브레인
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• 제8권1호
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• pp.50-58
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• 1998

Polysulfone 재질의 다공성 평판막 및 실관막에 키토산 피막층을 형성시킨 후 반응성 염료인 Cibacron Blue 3GA를 고정화시켜 human serum albumin(HSA)의 결합용량이 최대 70 $\mu{g/cm}^2$인 단백질 친화성 막을 제조하였다. 친화성 평판막 모듈을 대상으로 HSA에 대한 용출 크로마토그래피 실험을 수행하여 eluent 용액의 최적 환경조건을 결정하였는바, 1M KCl이 첨가된 농도 0.06 M, pH 10의 universal buffer를 eluent로 사용했을 때 리간드와 결합된 단백질의 용출이 가장 우수하였다. 친화성 평판막 및 실관막 모듈을 대상으로 HSA의 전열 크로마토그래피 실험을 수행하여 단백질에 대한 동적 결합용량을 측정하였다. 이 결과 동적 결합용량은 평판막 모듈의 경우에는 loading 용액의 유량과 HSA의 농도가 증가함에 따라 평형 결합용량 값으로부터 크게 감소하였으나, 실관막 모듈의 경우에는 loading 용액의 유량과 HSA의 농도에 관계없이 항상 평형 결합용량 수준을 유지하였는바, 따라서 실관막 모듈이 평판막 모듈보다 단백질 친화성 크로마토그래피 분리관으로서 더 효과적이었다.

• BMB Reports
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• 제32권2호
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• pp.127-132
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• 1999

The NAD(P)H-quinone oxidoreductase (EC 1. 6. 99. 2) was purified from S. cerevisiae. The native molecular weight of the enzyme is approximately 111 kDa and is composed of five identical subunits with molecular weights of 22 kDa each. The optimum pH of the enzyme is pH 6.0 with 1,4-benzoquinone as a substrate. The apparent $k_m$ for 1,4-benzoquinone and 1,4- naphthoquinone are 1.3 mM and $14.3\;{\mu}M$, respectively. Its activity is greatly inhibited by $Cu^{2+}$ and $Hg^{2+}$ ions, nitrofurantoin, dicumarol, and Cibacron blue 3GA. The purified NAD(P)H-quinone oxidoreductase was found capable of reducing aromatic nitroso compounds as well as a variety of quinones, and can utilize either NADH or NADPH as a source of reducing equivalents. The nitroso reductase activity of the purified NAD(P)H-quinone oxidoreductase is strongly inhibited by dicumarol.

• BMB Reports
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• 제34권3호
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• pp.225-229
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• 2001

Besides a variety of quinones, purified bovine liver quinone reductase catalyzed the reduction of N,N-p-nitrosoaniline to N,N-dimethyl-p-phenylenediamine. The formation of N,N-dimethyl-p-phenylenediamine was identified by TLC, GC, GC-MS and NMR. Quinone reductase can utilize either NADH or NADPH as a source of reducing equivalents. The apparent Km for 1,4-benzoquinone and N,N-dimethyl-p-nitrosoaniline was 1.64 mM and 0.22 mM, respectively The reduction of N,N-dimethyl-p-nitrosoaniline was almost entirely hampered by dicumarol or Cibacron blue 3GA, potent inhibitors of mammalian quinone reductase. During the bovine liver quinone reductase-catalyzed reduction of N,N-dimethyl-p-nitrosoaniline, benzoquinonediiminium ion was produced.

• KSBB Journal
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• 제13권2호
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• pp.125-132
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• 1998

Protein affinity membrane was prepared via the coating of chitosan gel on the porous flat polysulfone membrane surface, followed by the immobilization f the reactive dye (Cibacron Blue 3GA) to the chitonsan gel. The maximum protein binding capacity of affinity membrane was about 70${\mu}g/cm^2$ determined by the batch adsorption experiments of human serum albumin (HSA). Using module of this membrane, the characteristics of protein chromatography were investigated through the experiments of elution and frontal chromatography of HSA. This membrane module promises as a chromatography column, since it represented a lower pressure drop and a greater reproducibility. The protein separation ratio was significantly influenced by the flow rate of mobile phase and the injection quantity of HSA. The dynamic protein binding capacity of module decreased from the equilibrium binding capacity with increasing flow rate and approached the value of 15 - 20 ${\mu}g/cm^2$ for flow rates above 6 mL/min.