• Environmental Analysis Health and Toxicology
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• 제29권
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• pp.7.1-7.8
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• 2014

Objectives The present study was designed to systematically characterize the denaturation and the renaturation of double stranded DNA (dsDNA), which is suitable for DNA hybridization. Methods A series of physical and chemical denaturation methods were implemented on well-defined 86-bp dsDNA fragment. The degree of each denaturation was measured and the most suitable denaturation method was determined. DNA renaturation tendency was also investigated for the suggested denaturation method. Results Heating, beads mill, and sonication bath did not show any denaturation for 30 minutes. However probe sonication fully denatured DNA in 5 minutes. 1 mol/L sodium hydroxide (alkaline treatment) and 60% dimethyl sulfoxide (DMSO) treatment fully denatured DNA in 2-5 minutes. Conclusions Among all the physical methods applied, the direct probe sonication was the most effective way to denature the DNA fragments. Among chemical methods, 60% DMSO was the most adequate denaturation method since it does not cause full renaturation during DNA hybridization.

• Preventive Nutrition and Food Science
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• 제2권4호
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• pp.281-284
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• 1997

Heat-induced gelation is an important functional property of whey proteins. Preheating of calcium reduced whey was reported to increase gel strength. 5% whey-protein solutions were preheated at pH7 and at various temperatures(60~8$0^{\circ}C$) for 15 minutes. The amount of soluble aggregates and denaturation enthalpy of preheated whey proteins were measured. Preheating temperature was negatively correlated with denaturation enthalpy($R^2$=0.857, P=0.08) and positive with the amount of soluble aggregates($R^2$=0.921, P=0.002). Denaturation enthalpy was negatively correlated with gel strength($R^2$=0.93, P=0.002). Soluble aggregates and gel strength were positively correlated($R^2$=0.972, P=0.0003). The formation of three dimensional gel network requires controlled protein denaturation and aggregation. Since preheating leads to the partial denaturation of proteins and the formation of soluble aggregates, preheated whey proteins have a higher gel strength than non-preheated one.

• Bulletin of the Korean Chemical Society
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• 제26권3호
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• pp.413-417
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• 2005

Fluorescence of green fluorescent protein mutant, 2-5 GFP is observed during denaturation by guanidine. The fluorescence intensity decreases exponentially but the fluorescence lifetime does not change during denaturation. The fluorescence lifetime of the denatured protein is shorter than that of native form. As the protein structure is modified by guanidine, solvent water molecules penetrate into the protein barrel and protonate the chromophore to quench fluorescence. Most fluorescence quenchers do not affect the fluorescence of native form but accelerate the fluorescence intensity decay during denaturation. Based on the observations, a simple model is suggested for the structural change of the protein molecule during denaturation.

• 한국안광학회지
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• 제12권1호
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• pp.9-15
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• 2007

본 연구는 UV-A 노출에 의해 유발된 안구 내에 존재하는 단백질 효소 중의 하나인 ribonuclease A(RNase A)의 변성을 차단할 수 있는 안경렌즈의 적절한 UV-A 차단율을 알아보기 위해 수행하였다. RNase A를 1, 3, 6, 24, 48, 72, 96시간 동안 365 nm의 UV-A에 노출시켜 노출시간에 따른 단백질의 변성 정도를 아크릴 아미드 겔 전기영동법으로 확인하였다. 또한, 20%, 50%, 80%, 99% UV 차단 효과를 가진 안경렌즈로 UV-A를 차단하였을 때 RNase A의 변성이 억제될 수 있는 지를 알아보았다. RNase의 변성은 1시간 동안의 UV 노출에 의해서도 유발되었으며, UV-A 노출시간이 길어질수록 그 정도는 심해졌다. 1시간의 UV-A 노출에 의해 유발된 미세한 RNase A의 변성은 20% 정도의 UV-A 차단으로 충분히 예방될 수 있었다. UV-A를 3시간 동안 노출하였을 때는 50%이상의 UV-A 차단율을 가진 렌즈가 RNase A의 변성을 막을 수 있었다. 6시간 동안 UV-A에 노출되었을 때에는 99%의 렌즈로 차단하였을 때조차도 RNase A의 변성이 완벽하게 차단되지 못했다. 그러나 96시간 동안 UV-A에 노출되었을 때 나타나는 심각한 단백질의 변성이 99%의 UV-A 차단 렌즈를 사용하였을 경우 크게 감소하였다.

• Journal of Photoscience
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• 제9권2호
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• pp.296-298
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• 2002

Irreversible photobleaching of bacteriorhodopsin (bR), namely denaturation induced by illumination of visible light, was investigated by absorption kinetic measurements. The denaturation kinetics revealed that light illumination significantly enhanced the structural decay of bR. The kinetic analyses showed that the molecular structure of bR denatures according to a single-exponential decay, whereas irreversible photobleaching has two decay components. The decay constant of the slow component of photobleaching is almost same as that in the dark. An Arrhenius plot of the denaturation kinetic constants for the fast and slow components showed similar activation energies of approximately 19 kcal/mol.

• BMB Reports
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• 제31권4호
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• pp.405-408
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• 1998

Endonuclease-sensitive sites detected by T4 endonuclease V or UvrABC nuclease treatments were compared in the dihydrofolate reductase gene of UV-irradiated Chinese hamster ovary B-11 cells. The number of endonuclease-sensitive sites detected by T4 endonuclease V treatment followed by NaOH denaturation was twice that of formamide denaturation. Repeated treatment of damaged genomic DNA with T4 endonuclease V resulted in no further increase in the number of endonuclease-sensitive sites detected. The numbers of endonuclease-sensitive sites detected by UvrABC nuclease using each denaturation condition were similar. Sequential treatment with the two endonucleases using formamide denaturation resulted in twice the number of endonuclease-sensitive sites detected by treatment of each nuclease alone. Due to a lack of AP endonuclease activity these results suggest the presence of T4 endonuclease V-sensitive sites which could be complemented by alkaline gel separation or by UvrABC nuclease treatment.

• BMB Reports
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• 제36권4호
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• pp.367-372
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• 2003

The stability of four hemoglobins (Hb) in dimer forms (low concentration) were investigated by the kinetics of denaturation. The rate constants of denaturation were obtained by variation of 280 nm absorption versus time in 10 mM Tris-HCl, 10 mM EDTA, pH 8.0 at $45^{\circ}C$ in the absence and presence of 0.5 M ethanol, dimethyl sulfoxide (DMSO), formamide, and glycerol. The results show the trend of rate constants in different co-solvents in the following order: chicken hemolysate < human hemolysate and chicken Hb D < chicken Hb A. The buried surface area was calculated for Hb samples in the absence of cosolvents. Accordingly, the trend points out that: chicken Hb D > chicken Hb A > human Hb A. These results suggest that both chicken hemolysate and chicken Hb D are relatively more stable than human and chicken Hb A, respectively. However, the denaturation rate constants of Hb in different co-solvents have designated the following order: ethanol > DMSO > formamide > glycerol. As a matter of fact, this phenomenon is an indication of an increase in the denaturation capacity (DC) and hydrophobicity, and a decrease in the surface tension of the solution in the preceding co-solvents.

• 한국축산식품학회지
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• 제37권1호
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• pp.44-51
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• 2017

Heat treatment of milk aims to inhibit the growth of microbes, extend the shelf-life of products and improve the quality of the products. Heat treatment also leads to denaturation of whey protein and the formation of whey protein-casein polymer, which has negative effects on milk product. Hence the milk heat treatment conditions should be controlled in milk processing. In this study, the denaturation degree of whey protein and the combination degree of whey protein and casein when undergoing heat treatment were also determined by using the Native-PAGE and SDS-PAGE analysis. The results showed that the denaturation degree of whey protein and the combination degree of whey protein with casein extended with the increase of the heat-treated temperature and time. The effects of the heat-treated temperature and heat-treated time on the denaturation degree of whey protein and on the combination degree of whey protein and casein were well described using the quadratic regression equation. The analysis strategy used in this study reveals an intuitive and effective measure of the denaturation degree of whey protein, and the changes of milk protein under different heat treatment conditions efficiently and accurately in the dairy industry. It can be of great significance for dairy product proteins following processing treatments applied for dairy product manufacturing.

• 한국안광학회지
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• 제12권3호
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• pp.97-103
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• 2007

본 연구는 UV-A 노출에 의해 유발된 안구 내에 존재하는 항산화효소인 catalase와 superoxide dismutase(SOD)의 변성을 차단할 수 있는 안경렌즈의 적절한 UV-A 차단율을 알아보기 위해 수행하였다. Catalase와 SOD를 1, 3, 6, 24, 96시간 동안 365 nm의 UV-A에 노출시켜 노출시간에 따른 단백질의 변성 정도를 아크릴아미드 겔 전기영동법으로 확인하였다. 또한, 20, 50, 80 및 99% UV 차단 효과를 가진 안경렌즈로 UV-A를 차단하였을 때 catalase와 SOD의 변성이 억제될 수 있는 지를 알아보았다. Catalase는 3시간 이상의 UV-A 노출에 의해 변성되기 시작하였다. 그러나, SOD는 6시간 이상 노출시에 변성이 되기 시작하였다. 99% 차단렌즈에 의해 3시간 동안의 UV-A 노출에 의한 catalase 변성을 완전히 억제할 수 있었다. 그러나 노출시간이 3시간보다 길어지거나 차단율이 99%보다 낮은 렌즈를 사용한 경우, 어느 정도까지는 변성 억제작용은 있었으나 완벽하게 변성을 억제하지는 못하였다. 비록 50% 차단율을 가진 렌즈는 UV-A에 대한 SOD의 변성에 대해 부분적인 억제 작용을 나타냈으나, 80% 및 99%의 차단율을 가진 안경렌즈는 UV-A에 의한 SOD의 변성을 완전히 억제하였다.

• 한국축산식품학회:학술대회논문집
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• 한국축산식품학회 2004년도 정기총회 및 제33차 춘계 학술대회
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• pp.198-201
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• 2004

To investigate hydrostatic pressure (HP) effect on myofibrillar protein (Mf) extracted from bovine Semitendinosus muscle, Ca- and Mg-ATPase activities to evaluate denaturation of myosin and actin, and soluble protein contents were observed. In Mf treated with 100 MPa for 5 min was not observed denaturation of myosin and actin. In Mf treated with 200 MPa for 5 min, denaturation of myosin and actin were observed but inactivation rate was low (0.0136 $min^{-1}$). Inactivation rate of myosin and actin was dramatically increased above 300 MPa treatment. However denaturation of myosin and actin was not that critical with duration time. By increasing pressure size, the amount of myosin and actin in soluble protein eluted in 20 mM potassium phosphate buffer (pH 7.0) containing 0.6 M NaCl were decreased. SDS-PAGE of soluble protein released from Mf suspension in 0.1 M NaCl buffer (pH 7.0) showed that low molecular weight proteins (15${\sim}$36 KDa) were released by HP treatment above 200 MPa. From the results, denaturation of myosin and actin, and release of light molecule proteins of Mf were observed by HP treatment over 200 MPa.