• 한국식품영양과학회지
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• 제22권2호
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• pp.161-164
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• 1993

본 연구에서는 Kjeldahl 방법으로 시유의 질소분획물의 함량과 유청단백질 변성정도를 측정함으로써 이들이 시유의 열처리 정도를 비교할 수 있는 검색의 지표로서 타당하는지 알아보았다. 각 열처리법에 따른 질소분획물들의 함량을 보면, 원유에서는 시료 100g당 총질소가 431.3mg, 케이신질소가 341.0mg, 비케이신질소가 90.3mg, 이 중에서 비단백태질소가 31.6mg그리고 유청단백질소는 58.8mg을 보인 반면, 시유인 저온살균유, 고온순간살균유, 초고온처 리유의 질소분획물들의 함량이 다르게 나타났다. 유청단백질 변성정도는 저온살균유가 26.7%, 고온순간살균유가 32.9%, 초고온살균유가 60.7%, 그리고 초고온멸균유가 38.4%를 보여 열처리 정도가 높아질수록 변성정도도 높게 나타남으로써 열처리법에 따라 유청단백질의 변성정도의 구분이 뚜렷하였다.

• 한국안광학회지
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• 제18권3호
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• pp.253-260
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• 2013

목적: 본 연구에서는 안구 내에 존재하는 효소들의 변성을 차단할 수 있는 안경렌즈의 UV-B 차단율을 밝히고자 하였다. 방법: RNase A와 catalase, superoxide dismutase(SOD)를 1, 3, 6, 24, 96시간 동안 312 nm의 UV-B에 노출시킨 후 아크릴아미드 겔 전기영동법으로 손상 정도를 확인하였다. 또한, 50%, 80%, 95%, 99%의 UV-B 차단율을 가진 안경렌즈의 효소 손상 억제 효과를 알아보았다. 결과: RNase A는 1시간 동안 UV-B의 노출에서 손상이 유발되었으며 1시간 이상 6시간 이하의 노출에 의한 효소의 손상을 막기 위해서는 95%, 24시간 이상 96시간 이하의 노출에는 99% UV-B 차단 렌즈가 효소의 손상을 완벽하게 억제하였다. Catalase는 UV-B에 대한 1시간 이하의 노출에는 영향을 받지 않아 아무런 변성이 발생하지 않았고, 3시간 이상 6시간 이하의 단시간 노출에서는 50% 이상의 UV-B 차단 렌즈, 24시간 이상 96시간 이하의 장시간 노출에는 95% 이상 UV-B 차단 렌즈로 효소의 변성을 완벽하게 억제할 수 있었다. SOD는 6시간 이하의 노출에는 손상되지 않았고, 24시간 정도의 노출에서도 50% 차단 렌즈로 효소 손상을 억제할 수 있었다. 그러나 96시간 UV-B에 노출될 경우는 95% 이상의 차단율을 가진 안경렌즈에 의해 SOD의 손상이 완벽하게 억제되었다. 결론: 본 연구에서는 UV-B 노출에 의한 효소의 변성을 억제하기에 적절한 안경렌즈 차단율은 각 효소마다 상이하였으며 일정 수준 이상의 UV-B 차단율을 가진 경우에만 효과가 있음을 밝혔다.

• 한국축산식품학회지
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• 제21권3호
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• pp.265-271
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• 2001

We investigated changes of immunoglobulin G (IgG) concentrations by heating and drying condition. Also it is performed to group for commercial product by promoting of IgG preservation and reducing of protein denaturation. The result was that content of IgG in colostrum was higher than normal milk. Especially, IgG content of colostrum within 12 hrs after parturition was over 44.67mg/ml and it is 60 times of normal milk. IgG contents was reduced rapidly according as passage of the time. IgG content of the sample heating at 30min at 65$^{\circ}C$ was still a little higher that heating for 10sec at 72$^{\circ}C$. IgG denaturation of heat treatment at 100$^{\circ}C$ for 10sec was lower than at 85$^{\circ}C$ for 30min. We investigated the changes of IgG concentrations of kinds of market milk different with heating processing. This result showed that IgG denaturation ratio by ultra high temperature pasteurization (UHT) was higher than long time low temperature pasteurization (LTLT). On the other hands, IgG content by spray drying was 14.5mg/g and freezing drying was 10.8mg/g. It showed that denaturation of protein content by freezing drying was more than spray drying.

• Fisheries and Aquatic Sciences
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• 제2권1호
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• pp.12-16
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• 1999

Denaturation of actomyosin from the obliquely striated mantle muscle of squids (Todarodes pacificus) was studied by measuring the changes in $Ca^{2+}$-ATPase activity, relative viscosity, and solubility during frozen storage at three different temperature zones of maximum ice crystal formation $(-3^{\circ}C,\;-\;-5^{\circ}C)$, the eutectic point $(-11^{\circ}C)$, and $-20^{\circ}C$. The logarithms of $Ca^{2+}$-ATPase activity, relative viscosity and solubility of the actomyosin solutions (0.6 M KCl) and suspensions (0.05 M KCl) tended to decrease during frozen storage. The denaturation of squid actomyosin at the zone of maximum ice crystal formation significantly differed by only two degree of temperature difference between $-3^{\circ}C$ and $-5^{\circ}C$, and it (0.05 M KCl) at $-3^{\circ}C$ was less than those of other temperature. The denaturation at $-11^{\circ}C$ was more rapid than at $-5^{\circ}C$. The logarithms of $Ca^{2+}$ -ATPase activity, relative viscosity, and solubility were changed slower in the suspensions (0.05 M KCl) than the solutions (0.6 M KCl) at all experimental temperatures.

• Journal of Pharmaceutical Investigation
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• 제31권3호
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• pp.191-196
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• 2001

Biodegradable polymeric microspheres were studied for their usefulness as carriers for the delivery of vaccine antigens. However, protein antigen could be denatured during microencapsulation processes due to the exposure to the organic phase and stress condition of cavitation and shear force. Therefore this study was carried out to re-evaluate the degree of protein denaturation during microencapsulation with poly(lactide-co-glycolide) (PLGA) copolymer. PLGA microspheres containing ovalbumin (OVA), prepared by W/O/W multiple emulsification method, were suspended in pH 7.4 PBS and incubated with shaking at $37.5^{\circ}C$. Drug released medium was collected periodically and analyzed for protein contents by micro-BCA protein assay. In order to evaluate the protein integrity, release medium was subjected to the analyses of SDS-PAGE and size exclusion chromatography (SEC). And enzyme-linked immunosorbent assay (ELISA) was introduced to measure the immunoreactivity of entrapped OVA and to get an insight into the three-dimensional structure of epitope. The structures of entrapped protein were not affected significantly by the results of SDS-PAGE and SEC. However, immunoreactivity of released antigen was varied, revealing the possibility of protein denaturation in some microspheres when it was evaluate by ELISA method. Therefore, in order to express the degree of protein denaturation, antigenicity ratio (AR) was obtained as follows: amount of immunoreactivity of OVA/total amount of OVA released ${\times}100(%)$. ELISA method was an efficient tool to detect a protein denaturation during microencapsulation and the comparison of AR values resulted in more accurate evaluation for immunoreactivity of entrapped protein.

• 한국축산식품학회지
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• 제20권2호
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• pp.146-151
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• 2000

The objective of this study was to investigate the characteristics on the thermal denaturation of free drip released from pork loin during chilled storage using DSC(differential scanning calorimetry). DSC thermogram of drip released from normal pork(NORD) was characterized by a minor peak and two major peaks with temperature maxima at $61.5^{\circ}C$, $71.7^{\circ}C$ (associated with sarcoplasmic proteins) and $84.3^{\circ}C$ (associated with protein-protein interaction and aggregation). In the denaturation temperature of drip released from PSE pork (PSED), the peak(Tmax) at $59.0^{\circ}C$ was reduced by $2.5^{\circ}C$. When the thermograms were divided into segments correponding to the three peaks, $\Delta$H2 was shown to be reduced by 10% in PSED as compared to NORD. With the decrease in the solubility of sarcoplasmic proteins in PSE muscle, there was a corresponding increase the drip loss during the storage.

• 한국안광학회지
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• 제20권1호
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• pp.75-81
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• 2015

목적: 본 연구는 안구에 존재하는 항산화효소인 superoxide dismutase(SOD)와 catalase(CAT)가 UV-A에 반복적으로 노출되었을 때 이들의 구조 및 활성의 변화가 유발되는지 알아보고 이들의 상관관계를 밝히고자 수행되었다. 방법: SOD와 CAT의 표준품으로 각각의 효소용액을 제조하고 하루 30분, 1시간 및 2시간씩 365 nm의 UV-A에 노출시키는 조건으로 1, 2, 3, 4 및 5일 동안 UV-A에 반복적으로 노출시켰다. UV-A 반복노출에 따른 SOD와 CAT의 구조변성은 전기영동분석으로 확인하였으며, 이들 효소의 활성은 분석키트를 이용하여 비색분석법으로 측정하였다. 결과: UV-A에 반복노출된 SOD는 일일 1시간 이상 조건으로 반복노출되었을 때 전기영동분석에서 효소의 다중화(polymerization)가 관찰되었으나 활성의 변화는 12% 이내로 나타났다. 반면 UV-A에 반복노출된 CAT는 전기영동 시 효소의 밴드크기가 감소하여 구조변성이 나타났음을 알 수 있었으며, 효소활성 또한 유의하게 감소됨을 확인하였다. 반복노출시간이 긴 경우 CAT은 전기영동분석에서는 효소밴드를 보임에도 불구하고 그 활성은 완전히 소실됨을 알 수 있었다. 결론: 이상의 결과로 UV-A 반복노출에 따른 항산화효소의 구조변성은 효소의 종류에 따라 그 정도와 양상이 다르게 나타나며, 구조변성이 효소활성의 감소정도와 반드시 일치하는 것은 아님을 알 수 있었다.

• 한국식품과학회지
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• 제19권2호
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• pp.89-96
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• 1987

방어와 갈색띠 매물고둥에서 myosin과 paramyosin 을 추출하고, 이들 단백질의 가열중에 일어나는 변성기구를 아미노산 잔기와 SH기의 변화 및 단백질간의 상호작용 등을 측정하므로서 분석하였다. 각 단백질을 이루는 구성아미노산의 측쇄중 소수성 잔기의 유리정도는 가열온도 $65^{\circ}C$까지는 증가하였으나, 그 이상의 가열온도에서는 감소하는 경향을 보였다. 유리 소수성 잔기가 증가하여 감에 따라, 단백질간 상호작용도 활발하여 갔으며, 소수성 잔기의 유리정도가 감소하는 가열온도$65^{\circ}C$부터는 단백질의 응집이 일어나기 시작 하였다. 단백질간의 상호작용을 탁도로써 분석하여 Arrhenius식으로 해석한 결과, 방어 myosin은 3단계이상의 변성과정으로 구분할 수 있었으며, 갈색띠 매물고둥 paramyos은 2단계의 변성과정으로 구분할 수 있었다. 이들 두 단백질 소수성, 용해도, 유리 SH기의 수 및 단백질간의 상호작용 등은 온도함수와 밀접한 상관관계를 보였다.

• Archives of Pharmacal Research
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• 제24권2호
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• pp.150-158
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• 2001

The activity of nonsteroidal antiinflammatory drugs (NSAIDs) in rheumatoid arthritis is not only due to the inhibition of the production of prostaglandins, which can even have beneficial immunosuppressive effects in chronic inflammatory processes. Since we speculated that these drugs could also act by protecting endogenous proteins against denaturation, we evaluated their effect on heat-induced denaturation human serum albumin (HSA) in comparison with several fatty acids which are known to be potent stabilizers of this protein. By the Mizushimas assay and a recently developed HPLC assays we observed that NSAIDs were slightly less active [$EC_{50}~10^{-5}-10^{-4}$ M] than FA and that the HPLC method was less sensitive but more selective than the turbidimetric assay, i.e. it was capable of distinguishing true antiaggregant agents like FA and NSAIDs from substances capable of inhibiting the precipitation of denatured protein aggregates. In conclusion, this survey could be useful for the development of more effective agents in protein condensation diseases like rheumatic disorders, cataract and Alzheimers disease.

• Molecules and Cells
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• 제26권1호
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• pp.61-66
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• 2008

Cyclic AMP receptor protein (CRP) is allosterically activated by cAMP and functions as a global transcription regulator in enteric bacteria. Structural information on CRP in the absence of cAMP (apo-CRP) is essential to fully understand its allosteric behavior. In this study we demonstrated interdomain interactions in apo-CRP, using a comparative thermodynamic approach to the intact protein and its isolated domains, which were prepared either by limited proteolysis or using recombinant DNA. Thermal denaturation of the intact apo-CRP, monitored by differential scanning calorimetry, revealed an apparently single cooperative transition with a slight asymmetry. Combined with circular dichroism and fluorescence analysis, the thermal denaturation of apo-CRP could be interpreted as a coupled process involving two individual transitions, each attributable to a structural domain. When isolated individually, both of the domains exhibited significantly altered thermal behavior, thus pointing to the existence of non-covalent interdomain interactions in the intact apo-CRP. These observations suggest that the allosteric conformational change of CRP upon binding to cAMP is achieved by perturbing or modifying pre-existing interdomain interactions. They also underline the effectiveness of a comparative approach using calorimetric and structural probes for studying the thermodynamics of a protein.