• Journal of Microbiology and Biotechnology
• /
• 제25권4호
• /
• pp.503-510
• /
• 2015

The iLOV protein belongs to a family of blue-light photoreceptor proteins containing a light-oxygen-voltage sensing domain with a noncovalently bound flavin mononucleotide (FMN) as its chromophore. Owing to advantages such as its small size, oxygen-independent nature, and pH stability, iLOV is an ideal candidate over other reporter fluorescent proteins such as GFP and DsRed. Here, for the first time, we describe the feasibility of applying LOV domain-based fluorescent iLOV as a metal sensor by measuring the fluorescence quenching of a protein with respect to the concentration of metal ions. In the present study, we demonstrated the inherent copper sensing property of the iLOV protein and identified the possible amino acids responsible for metal binding. The fluorescence quenching upon exposure to Cu²⁺ was highly sensitive and exhibited reversibility upon the addition of the metal chelator EDTA. The copper binding constant was found to be 4.72 ± 0.84 µM. In addition, Cu²⁺-bound iLOV showed high fluorescence quenching at near physiological pH. Further computational analysis yielded a better insight into understanding the possible amino acids responsible for Cu²⁺ binding with the iLOV protein.

• Bulletin of the Korean Chemical Society
• /
• 제17권9호
• /
• pp.808-813
• /
• 1996

To explore protein folding mechanism, we simulated a folding pathway in a simplified 3×3×3 cubic lattice. In the lattice folding Monte Carlo simulations, each of the 28 possible native packing pairs that exist in the native conformation was used as a conformational restraint. The native packing restraints in the lattice model could be considered as a disulfide linkage restraint in a real protein. The results suggest that proteins denatured with a small disulfide loop can, but not always, fold faster than proteins without any disulfide linkage and than proteins with a larger disulfide loop. The results also suggest that there is a rough correlation between loop size of the native packing restraint and folding time. That is, the order of native residue-residue packing interaction in protein folding is likely dependent on the residue-residue distance in primary sequence. The strength of monomer-monomer pairwise interaction is not important in the determination of the packing order in lattice folding. From the folding simulations of five strong folding lattice sequences, it was also found that the context encoded in the primary sequence, which we do not yet clearly understand, plays more crucial role in the determination of detailed folding kinetics. Our restrained lattice model approach would provide a useful strategy to the future protein folding experiments by suggesting a protein engineering for the fast or slow folding research.

• 한국미생물·생명공학회지
• /
• 제26권2호
• /
• pp.117-121
• /
• 1998

김치로부터 분리한 Lactobacillus plantarum bacteriophage SC 921은 M.O.I가 0.2일 경우 용균효과가 빠르게 진행됨을 알 수 있었고, SDS-PAGE를 실시하여 phage particle protein을 조사해 본 결과 4개의 major protein으로 구성되어 있는데 이들은 각각 48, 34, 32, 29 kDa으로 구성되어 있다. Exo III로 30분간 반응시킨 후 S1 nuclease를 처리하여 DNA의 형태를 조사해 본 결과 intact DNA는 linear form의 double strand를 유전전달 물질로 가지고 있었다. 제한효소에 대한 절단 효과를 조사한 결과, Sma I에 대해서 1개, Xba I, Cla I, Kpn I, EcoRI에 대해서 각각 2, 4, 5, 6개의 절단부위를 가지고 있으며, Hind III에 대해서는 절단부위가 매우 많은 것을 알 수 있었다. Hind III를 이용하여 intact DNA의 genome size를 측정해본 결과 약 66.5 kbp정도였다. 위의 실험결과와 restriction enzyme mapping을 통해 기존에 알려진 bacteriophage B2와 비교해본 결과 숙주 균주는 같으나 단백질적인 구조나 유전전달물질로 본 구조는 서로 다름을 알 수 있었다.

• Journal of Pharmaceutical Investigation
• /
• 제35권2호
• /
• pp.89-94
• /
• 2005

The micron or nano-sized lysozyme as a model protein drug was prepared using solution enhanced dispersion by supercritical fluid (SEDS) process at various conditions (e.g., solvent, temperature and pressure) to investigate the feasibility of pulmonary protein drug delivery. The lysozyme particles prepared were characterized by laser diffraction particle size analyzer, scanning electron microscopy (SEM) and powder X-ray diffractometry (PXRD). The biological activity of lysozyme particles after/before SEDS process was also examined. Lysozyme was precipitated as spherical particles. The precipitated particles consisted of 100 - 200 nm particles. Particle size showed the precipitates to be agglomerates with primary particles of size $1\;-\;5 \;{\mu}m$. The biological activity varied between 38 and 98% depending on the experimental conditions. There was no significant difference between untreated lysozyme and lysozyme after SEDS process in PXRD analysis. Therefore, the SEDS process could be a novel method to prepare micron or nano-sized lysozyme particles, with minimal loss of biological activity, for the pulmonary delivery of protein drug.

• Journal of Pharmaceutical Investigation
• /
• 제32권3호
• /
• pp.199-207
• /
• 2002

In order to develop a sustained release formulation of bovine somatotropin (BST), which has been used to increase the body weight of oxen or the milk production of dairy cows, poly(D,L-lactide-co-glyceride)(PLGA) microspheres were made by W/O/W multiple emulsification method and solvent extraction method. Physical properties including particle size, drug entrapment, drug release, protein denaturation, and in vivo body weight increase in rats were characterized. The size of the microspheres was increased as the molecular weight of PLGA increased. When Span 65 and stearic acid during preparation were added, the size was decreased but the amount of surface protein was increased, resulting in a high loading efficiency, with fast release of BST from the microspheres. Aggregation or fragmentation of BST by SDS-PAGE during microsphere preparation and drug release study was not observed. Body weight of Sprague-Dawley's male rats was significantly increased after subcutaneous administrations of BST-loaded PLGA microspheres. There was a good correlation between in vivo weight gain and in vitro release rate of microspheres. PLGA microspheres with a high surface protein ratio could be a good candidate for the sustained delivery of BST.

• Asian-Australasian Journal of Animal Sciences
• /
• 제12권3호
• /
• pp.388-394
• /
• 1999

Two experiments were conducted to subdivide isoleucine (exp. 1) and valine (exp. 2) requirements for maintenance from the requirements for growth of broilers aged 1 to 3 weeks. Purified diets were used, containing five graded levels of isoleucine and valine. Based on weight gain response, the isoleucine requirement for growth was 7.50 mg/g weight gain and the daily isoleucine need for maintenance (mg) was 0.044 per unit metabolic body size ($(Wg^{0.75})$). Based on the N gain response, the isoleucine requirement for growth was 0.317 mg/mg N gain and the daily isoleucine need for maintenance (mg) was 0.040 per unit metabolic body size $(Wg^{0.75})$. Based on weight gain and N gain response, the total isoleucine requirement was calculated 244 mg/day or 0.59% of the diet, 274 mg/day or 0.66% of the diet, respectively. From the relationship of weight gain and N gain, 5.07% of the retained protein was comprised of isoleucine; the reported isoleucine content of chick muscle was 4.42%. The valine requirement for growth was 9.84 mg/g weight gain and 0.36 mg/mg N gain whereas the maintenance requirement was 0.046 or 0.052 mg per unit of metabolic body size (Wgo.11. According to the model developed to estimate valine requirement, the total requirement was 319 mg/day or 0.77% of the diet, 315 mg/day or 0.76% of the diet, respectively. Previous reported valine requirements for growing chicks of 7~24 days old were in close agreement with these estimates. As a percentage of retained protein, valine was calculated to be 5.81% ; the reported valine concentration of crude protein of chicks' body including feathers was 6.72%.

• 한국축산식품학회지
• /
• 제36권2호
• /
• pp.267-274
• /
• 2016

The purposes of this study were to investigate the impacts of concentration levels of whey protein isolate (WPI) and inulin on the formation and physicochemical properties of WPI/inulin nano complexes and to evaluate their potential prebiotic effects. WPI/inulin nano complexes were produced using the internal gelation method. Transmission electron microscopy (TEM) and particle size analyzer were used to assess the morphological and physicochemical characterizations of nano complexes, respectively. The encapsulation efficiency of resveratrol in nano complexes was studied using HPLC while the potential prebiotic effects were investigated by measuring the viability of probiotics. In TEM micrographs, the globular forms of nano complexes in the range of 10 and 100 nm were successfully manufactured. An increase in WPI concentration level from 1 to 3% (w/v) resulted in a significant (p<0.05) decrease in the size of nano complexs while inulin concentration level did not affect the size of nano complexes. The polydispersity index of nano complexes was below 0.3 in all cases while the zeta-potential values in the range of -2 and -12 mV were observed. The encapsulation efficiency of resveratrol was significantly (p<0.05) increased as WPI and inulin concentration levels were increased from 1 to 3% (w/v). During incubation at 37℃ for 24 h, WPI/inulin nano complexes exhibited similar viability of probiotics with free inulin and had significantly (p<0.05) higher viability than negative control. In conclusions, WPI and inulin concentration levels were key factors affecting the physicochemical properties of WPI/inulin nano complexes and had potential prebiotic effect.

• 산업식품공학
• /
• 제21권4호
• /
• pp.391-402
• /
• 2017

본 연구에서는 건조조건과 분쇄조건을 달리하거나 추출한 식물류의 영양성분 함량과 영양성분의 보존율을 비교분석하였다. 동결분쇄한 식물류는 일반분쇄 하거나 추출한 식물류에 비하여 영양성분의 보존율이 높은 것으로 나타났다. 인공생체막에서 영양성분의 투과도를 분석한 결과 동결분쇄한 식물류의 영양성분이 잘 투과하였다. 분쇄과정에서 열발생을 최소화하고 입도를 작게 하는 동결분쇄 기술이 식물류 원료를 우수한 식물소재로 가공할 수 있는 기술임을 확인하였다.

• 동아시아식생활학회지
• /
• 제20권1호
• /
• pp.187-192
• /
• 2010

The study investigated the effect of rice particle size on the physicochemical properties of soybean porridge prepared at different pH. Redness, yellowness and viscosity of soybean porridge increased according to the rice particle size. Protein content of soybean porridge was increased 2-fold, compared to that of rice porridges. Total amino acid content of soybean porridge was 1,821.2 mg/100 g and that of rice was 1,719.1 mg/100 g. Lysine and threonine contents of limited amino acid of soybean porridge were increased. Half grain rice soybean porridge exhibited the highest sensory scores. The results indicated that half grain size soybean porridge is preferred, both physicochemically and nutritionally.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제44권4호
• /
• pp.174-181
• /
• 2018

Objectives: The number of patients with medication-related osteonecrosis of the jaw (MRONJ) is increasing, but treatment remains controversial. Published papers and systematic reviews have suggested that surgical treatment is effective in patients with MRONJ. The purpose of this study was to determine whether preoperative University of Connecticut Osteonecrosis Numerical Scale (UCONNS), other serologic biomarkers, and size of necrosis are prognostic factors for outcome of surgical treatment in MRONJ. Materials and Methods: From January 2008 to December 2016, 65 patients diagnosed with MRONJ at the Department of Oral and Maxillofacial Surgery in College of Dentistry, Dankook University who required hospitalization and surgical treatment were investigated. Patient information, systemic factors, and UCONNS were investigated. In addition, several serologic values were examined through blood tests one week before surgery. The size of osteolysis was measured by panoramic view and cone-beam computed tomography in all patients. With this information, multivariate logistic regression analysis with backward elimination was used to examine factors affecting postoperative outcome. Results: In multivariate logistic analysis, higher UCONNS, higher C-reactive protein (CRP), larger size of osteolysis, and lower serum alkaline phosphate were associated with higher incidence of incomplete recovery after operation. This shows that UCONNS, CRP, serum alkaline phosphate, and size of osteolysis were statistically significant as factors for predicting postoperative prognosis. Conclusion: This study demonstrated that CRP, UCONNS, serum alkaline phosphate, and size of osteolysis were statistically significant factors in predicting the prognosis of surgical outcome of MRONJ. Among these factors, UCONNS can predict the prognosis of MRONJ surgery as a scale that includes various influencing factors, and UCONNS should be used first as a predictor. More aggressive surgical treatment and more definite surgical margins are needed when the prognosis is poor.