• 폴리머
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• 제32권3호
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• pp.246-250
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• 2008

본 연구에서는 분해성 고분자의 종류와 제조조건에 따른 나노입자의 분산 안정성을 평가하기 위하여 광산란 입도 분석기를 이용하여 입자 크기 변화를 조사하였다. 이를 위해 생분해성 고분자인 폴리(D,L-락타이드-co-글리콜라이드) 단독과 폴리(L-락타이드)와의 블렌드, 그리고 폴리(L-락타이em)-g-폴리(에틸렌 글리콜)과의 블렌드, 세 종류를 개선된 자발적 에멀션/용매 확산 방법을 이용하여 나노입자를 제조하였다. 제조조건의 영향을 파악하기 위하여서는 나노입자의 제조시에 대표적인 분산 안정제인 폴리(비닐 알코올)(PVA)의 사용 유무의 영향을 조사하였다. 시간이 진행함에 따라 분산 안정제가 적용되지 않은 경우에서는 나노입자의 분해와 분해를 통해 붕괴된 입자의 응집에 의하여 입자의 크기가 bimodal 분포를 나타내었다. 나노입자가 분산된 수용액에 단백질 용액을 투입한 경우에는 분산 안정제인 PVA의 사용 유무에 의해 극명한 차이를 나타내었으며 PVA가 분산 안정성에 크게 기여함을 확인할 수 있었다. 또한 낮은 분자량의 폴리(에틸렌 글리콜)이 그래프트 고분자를 미량 블렌드함에 따라 입자의 단백질 흡착에 대한 분산 안정성은 침전이 일어나지 않고 1일 정도 분산 상태를 유지하는 것으로부터 일부 향상됨을 확인하였다.

• Bulletin of the Korean Chemical Society
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• 제33권8호
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• pp.2508-2512
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• 2012

Cold shock proteins (CSPs) are a family of proteins induced at low temperatures. CSPs bind to single-stranded nucleic acids through the ribonucleoprotein 1 and 2 (RNP 1 and 2) binding motifs. CSPs play an essential role in cold adaptation by regulating transcription and translation via molecular chaperones. The solution nuclear magnetic resonance (NMR) or X-ray crystal structures of several CSPs from various microorganisms have been determined, but structural characteristics of psychrophilic CSPs have not been studied. Therefore, we optimized the purification process to obtain highly pure Lm-Csp and determined the three-dimensional structure model of Lm-Csp by comparative homology modeling using MODELLER on the basis of the solution NMR structure of Bs-CspB. Lm-Csp consists of a ${\beta}$-barrel structure, which includes antiparallel ${\beta}$ strands (G4-N10, F15-I18, V26-H29, A46-D50, and P58-Q64). The template protein, Bs-CspB, shares a similar ${\beta}$ sheet structure and an identical chain fold to Lm-Csp. However, the sheets in Lm-Csp were much shorter than those of Bs-CspB. The Lm-Csp side chains, E2 and R20 form a salt bridge, thus, stabilizing the Lm-Csp structure. To evaluate the contribution of this ionic interaction as well as that of the hydrophobic patch on protein stability, we investigated the secondary structures of wild type and mutant protein (W8, F15, and R20) of Lm-Csp using circular dichroism (CD) spectroscopy. The results showed that solvent-exposed aromatic side chains as well as residues participating in ionic interactions are very important for structural stability. Further studies on the three-dimensional structure and dynamics of Lm-Csp using NMR spectroscopy are required.

• 한국식품과학회지
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• 제24권3호
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• pp.294-299
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• 1992

본 연구에서는 분리대두단백질에 단백질 가수분해효소를 처리하여 대두단백질에 가수분해적 변성을 야기하여 그 기능성을 변화 고찰하고 실제 식품에의 이용방법으로 효소처리된 두유로 두부를 만들어 물성실험과 관능평가를 실시하였다. 장엽에서 SPI를 제조한 후 15분간 bromelain으로 처리하여 2.7% 가수분해된 MSPI를 만들어 기능특성을 측정 비교하였다. MSPI는 전 pH범위에서 용해도가 증가하였고, 특히 등전점에서 15% 정도 증가하였다. 유화형성력과 기포팽창력은 증가하였으나 이들의 안정성은 감소하였는데, 특히 기포안정성은 알칼리쪽에서 급격히 감소하였고, 등전점에서는 가장 큰 것으로 나타났다. 표준두유와 변형두유를 혼합하여 제조한 두부로 물성실험과 관능평가를 실시하였을 때 압착실험에서는 변형두부I이 연하고 탄력성 있는 두부를 형성하는 것으로 나타났다. 각 두부의 물성모형은 spring 한개와 Maxwell 모형 세개를 가진 7요소모형으로 동일한 물성모형을 나타내었다. 관능평가에서는 표준두유와 변형두유를 3 : 1의 비율로 혼합하여 제조한 두부의 품질이 가장 우수한 것으로 평가되었다.

• Journal of Microbiology and Biotechnology
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• 제25권6호
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• pp.782-787
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• 2015

In this study, we developed an assay system for missense mutations in human phenylalanine hydroxylases (hPAHs). To demonstrate the reliability of the system, eight mutant proteins (F39L, K42I, L48S, I65T, R252Q, L255V, S349L, and R408W) were expressed in a mutant strain (pah^-) of Dictyostelium discoideum Ax2 disrupted in the indigenous gene encoding PAH. The transformed pah - cells grown in FM minimal medium were measured for growth rate and PAH activity to reveal a positive correlation between them. The protein level of hPAH was also determined by western blotting to show the impact of each mutation on protein stability and catalytic activity. The result was highly compatible with the previous ones obtained from other expression systems, suggesting that Dictyostelium is a dependable alternative to other expression systems. Furthermore, we found that both the protein level and activity of S349L and R408W, which were impaired severely in protein stability, were rescued in HL5 nutrient medium. Although the responsible component(s) remains unidentified, this unexpected finding showed an important advantage of our expression system for studying unstable proteins. As an economic and stable cell-based expression system, our development will contribute to mass-screening of pharmacological chaperones for missense PAH mutations as well as to the in-depth characterization of individual mutations.

• 미생물학회지
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• 제38권4호
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• pp.218-218
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• 2002

Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAPl) of Candide albicans. Three intracellular forms of SAPI were detected by immunoblotting using menoclonal antibody (MAb) CAPl. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAPl and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAPI. These results show that SAPI is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor farm undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretary vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAPl was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in parti be related to enzyme stability and activity.

• Journal of Microbiology
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• 제38권4호
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• pp.218-223
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• 2000

Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAPl) of Candide albicans. Three intracellular forms of SAPI were detected by immunoblotting using menoclonal antibody (MAb) CAPl. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAPl and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAPI. These results show that SAPI is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor farm undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretary vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAPl was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in parti be related to enzyme stability and activity.

• 한국재료학회지
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• 제28권3호
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• pp.182-188
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• 2018

Nitrogen (N)-doped protein-based carbon as platinum (Pt) catalyst supports from tofu for oxygen reduction reactions are synthesized using a carbonization and reduction method. We successfully prepare 5 wt% Pt@N-doped protein-based carbon, 10 wt% Pt@N-doped protein-based carbon, and 20 wt% Pt@N-doped protein-based carbon. The morphology and structure of the samples are characterized by field emission scanning electron microscopy and transmission electron micro scopy, and crystllinities and chemical bonding are identified using X-ray diffraction and X-ray photoelectron spectroscopy. The oxygen reduction reaction are measured using a linear sweep voltammogram and cyclic voltammetry. Among the samples, 10 wt% Pt@N-doped protein-based carbon exhibits exellent electrochemical performance with a high onset potential of 0.62 V, a high $E_{1/2}$ of 0.55 V, and a low ${\Delta}E_{1/2}=0.32mV$. Specifically, as compared to the commercial Pt/C, the 10 wt% Pt@N-doped protein-based carbon had a similar oxygen reduction reaction perfomance and improved electrochemical stability.

• 한국생물정보학회:학술대회논문집
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• 한국생물정보시스템생물학회 2004년도 The 3rd Annual Conference for The Korean Society for Bioinformatics Association of Asian Societies for Bioinformatics 2004 Symposium
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• pp.265-271
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• 2004

The interaction network of protein -protein plays an important role to understand the various biological functions of cells. Currently, the high -throughput experimental techniques (two -dimensional gel electrophoresis, mass spectroscopy, yeast two -hybrid assay) provide us with the vast amount of data for protein-protein interaction at the proteome scale. In order to recognize the role of each protein in their network, the efficient bioinformatical and computational analysis methods are required. We propose a systematic and mathematical method which can analyze the protein -protein interaction network rigorously and enable us to capture the biological and physical essence of a topological character and stability of protein -protein network, and sensitivity of each protein along the biological pathway of their network. We set up a Laplacian matrix of spectral graph theory based on the protein-protein network of yeast proteome, and perform an eigenvalue analysis and apply a perturbation method on a Laplacian matrix, which result in recognizing the center of protein cluster, the identity of hub proteins around it and their relative sensitivities. Identifying the topology of protein -protein network via a Laplacian matrix, we can recognize the important relation between the biological pathway of yeast proteome and the formalism of master equation. The results of our systematic and mathematical analysis agree well with the experimental findings of yeast proteome. The biological function and meaning of each protein cluster can be explained easily. Our rigorous analysis method is robust for understanding various kinds of networks whether they are biological, social, economical...etc

• 한국응용과학기술학회지
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• 제17권3호
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• pp.158-166
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• 2000

A conductimetric study of foam formed from mixture of the protein, ${\beta}-lactoglobulin$, and the nonioinc surfactant, SML, revealed that their stability was reduced at concentrations of SML in the range $3{\sim}10mM$. The interaction of SML with ${\beta}-lactoglobulin$ was investigated by fluorimetry and a dissociation constant of $0.2{\mu}M$ was calculated for the complex. Surface tension studies confirmed the presence of interaction between the two components and provided evidence for the progressive displacement of ${\beta}-lactogloblin$ from the air/water interface with increasing SML concentration. Experiments using air-suspended microscopic thin liquid films revealed transitions in the chainage characteristics and thickness of the film at SML concentrations below that which resulted in destabilization of the foam. However, measurements of surface mobility of fluorescent-labeled ${\beta}-lactoglobulin$ by a photobleaching method identified that a transition to a mobile system occurred at a SML concentration which correlated with the onset of instability in the disperse phase. The results would indicate that maintenance of the viscoelastic properties of the surface is paramount importance in determining the stability of interfaces comprising mixtures of protein and surfactant.

• 생명과학회지
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• 제20권11호
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• pp.1656-1661
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• 2010

난황단백질 가수분해를 위한 알칼리성 단백질분해효소를 5가지 담체 Duolite A568, Celite R640, Dowex-1, Dowex 50W 그리고 Silica gel R60 에 고정화하였다. Duolite A568의 경우에 24.7%의 최대 고정화 효율을 나타내었다. 자유 효소와 고정화 효소에 대한 최적의 pH는 각각 8과 9였고, 최적의 pH는 고정화에 의해 염기성으로 1만큼 증가하였다. 그러나 최적 온도는 자유 효소와 고정화 효소 모두 $50^{\circ}C$로 같았다. 고정화 효소가 자유 효소에 비해 높은 열 안정성을 보였다. 재사용 회분식 공정에서 10 cycle 동안 효소활성은 초기 활성의 86%를 유지하였다. 연속 공정을 위한 충진층 반응기에서 여러 유속에 대한 장기 조업에서 효소 활성의 안정성 평가하였는데 낮은 유속일수록 높은 활성을 유지하였다. 연속 조업에서 casein과 난황 단백질을 사용하여 원료에 대한 고정화 효소의 활성에 대한 영향을 조사하였다. 96시간 연속 조업에서 casein의 경우는 초기 활성의 83%를 유지하였고 난황 단백질의 경우는 초기 활성의 61%를 유지하였다.