• KSBB Journal
• /
• v.18 no.1
• /
• pp.35-38
• /
• 2003

Analysis of proteome in amniotic fluid was performed by 2-D PAGE (polyacrylamide gel electrophoresis). Proteins in amniotic fluid were separated by centrifugation and solubilized in buffer solution for IEF, using an IPG strip of pH 4-7L. Both a homogeneous slab gel of 12.5% and a gradient gel of 8-18%, were used. After 2-D PAGE, spots were stained with silver nitrate and picked up for in-gel digestion. Digested peptides were analyzed by MALDI-TOF and proteins were further identifical. More protein spots were detected in the gradient gels and a protein not previously reported was identified.

• KSBB Journal
• /
• v.20 no.2 s.91
• /
• pp.71-75
• /
• 2005

Alignment of 2D-gel images of biological samples can visualize the difference of expression profiles and also inform us candidates of protein spots to be further analyzed. However, comparison of two proteome images between the case and control does not always successfully identify differentially expressed proteins because of sample-to-sample variation, poor reproducibility of 2D-gel electrophoresis and inconsistent electrophoresis conditions. Multiple alignment of 2D-gel image must be preceded before visualizing the difference of expression profiles or clustering proteome images. Thus, a software for the alignment of multiple 2D-Gel images and their clustering was developed by applying various algorithms and statistical methods. Microsoft Visual C++ was used to implement the algorithms in this work. Multiresoultion-multilevel algorithm was found out to be suitable for fast alignment and for largely distorted images. Clustering of 10 different proteome images of Fetal Alcohol Syndrome, was carried out by implementing a k-means algorithm and it gave a phylogenetic tree of proteomic distance map of the samples. However, the phylogenetic tree does not discriminate the case and control. The whole image clustering shows that the proteomic distance is more dependent to age and sex.

• 한국생물공학회:학술대회논문집
• /
• 2003.10a
• /
• pp.746-749
• /
• 2003

Alignment of 2D-gel images of biological samples can visualize the difference of expression profiles and also inform us candidates of protein spots to be further analyzed. However, comparison of two proteome images between case and control does not always successfully identify differentially expressed proteins due to sample-to-sample variation. Because of poor reproducibility of 2D-gel electrophoresis, sample-by-sample variations and inconsistent electrophoresis conditions, multiple number of 2D-gel image must be processed to align each other to visualize the difference of expression profiles and to deduce the protein spots differentially expressed with reliability. Alignment of multiple 2D-Gel images and their clustering were carried out by applying various algorithms and statistical methods. In order to align multiple images, multiresolution-multilevel algorithm was found out to be suitable for fast alignment and for distorted images. Clustering of 12 different images implementing a k-means algorithm gives a phylogenetic tree of distance map of the proteomes. Microsoft Visual C++ was used to implement the algorithms in this work.

• BMB Reports
• /
• v.37 no.3
• /
• pp.298-303
• /
• 2004

In general, a SYPRO Ruby dye is well known as a sensitive fluorescence-based method for detecting proteins by one-or two-dimensional SDS-PAGE (1-DE or 2-DE). Based on the SYPRO Ruby dye system, the combined two-dimensional fibrin zymography (2-D FZ) with SYPRO Ruby staining was newly developed to identify the Bacillus sp. proteases. Namely, complex protein mixtures from Bacillus sp. DJ-4, which were screened from Doen-Jang (Korean traditional fermented food), showed activity on the zymogram gel. The gel spots on the SYPRO Ruby gel, which corresponded to the active spots showing on the 2-D FZ gel, were analyzed by a matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometric analysis. Five intracellular fibrinolytic enzymes of Bacillus sp. DJ-4 were detected through 2-D FZ. The gel spots on the SYPRO Ruby dye stained 2-D gel corresponding to 2-D FZ were then analyzed by MALID TOF MS. Three of the five gel spots proved to be quite similar to the ATP-dependent protease, extracellular neutral metalloprotease, and protease of Bacillus subtilis. Also, the extracellular proteases of Bacillus sp. DJ-4 employing this combined system were identified on three gels (e.g., casein, fibrin, and gelatin) and the proteolytic maps were established. This combined system of 2-D zymography and SYPRO Ruby dye should be useful for searching the specific protease from complex protein mixtures of many other sources (e.g., yeast and cancer cell lines).

• Proceedings of the Korean Society for Bioinformatics Conference
• /
• 2002.06a
• /
• pp.35-47
• /
• 2002

2D-PAGE/MALDI-TOF는 프로-테옴 연구의 중요한 실험 기법중의 하나이다. 이는 단백질의 발현 분석을 위한 방법으로, 2D-PAGE 결과로 얻어진 영상 데이터의 분석에 대한 정확도가 단백질 발현에 대한 분석 결과의 질을 결정하는 중요한 요인으로 작용한다. 2D Electrophoresis에 의한 Gel Protein Database는 현재 많은 연구자들에 의해 생산되고 있으며, 대단히 많은 데이터들이 인터넷을 통하여 접근이 가능하다. 이러한 대량 정보의 Database 활용이 가능한 상황은 2D-PAGE에 의해 생산된 Gel Image의 상호 비교에 대한 요구를 도출하였다. 본 발표에서는 영상처리 및 형태인식 기술과 2D-PAGE 연구의 결합을 주제로 하여, 2D-PAGE Gel 영상 처리 및 비교에 관련되는 전처리 (preprocessing), spot detection, feature extraction, spot matching 및 image comparison 기술을 소개한다.

• Journal of dental hygiene science
• /
• v.10 no.2
• /
• pp.79-83
• /
• 2010

This study manufactured hydrogel, which was contained NSAIDs(non-steroidal anti-inflammatory drugs) Ketorolac tromethamine and hydrolyzed products of gardeniae fructus extract, and experimented viscosity, surface tension, tensile strength and bio-adhesiveness by using hairless mouse. Thus, it was performed in expectation for being probably able to develop as effective auxiliary agent of periodontal disease after non-surgical or surgical periodontal treatment. As a result, the following conclusions were obtained. 1. Out of KGE and KGH gel materials, the content of ketorolac tromethamine was 1.02~0.97%. The content of geniposide was 0.34% in KGE gel A and C. However, it got lower to 0.11% in KGH gel B and D. The content of genipin wasn't shown in KGE gel A and C, but was shown with 0.13% in KGH gel B and D. 2. As for viscosity according to temperature in gel material, the gel, which used independently Carbopol 940 as gel inoculant, maintained the higher viscosity than the gel, which added Poloxamer 407. The surface tension in each material showed 34.77~40.58 dyne/cm at 37. As for tensile strength in material, KGH gel B was shown the higher tensile strength in about 3.5 times compared to the control group. 3. As for bio-adhesiveness, the back-skin upper part(epidermis) and abdomen skin were shown to be 50.62 N in KGH gel B, thereby having indicated higher value in about 5 times compared to control group. The back-skin lower part(dermis) and abdomen skin were shown to be 35.93 N in KGH gel B, thereby having indicated higher value in about 3.5 times compared to control group.

• Journal of Applied Biological Chemistry
• /
• v.51 no.3
• /
• pp.88-94
• /
• 2008

Three different protein extraction methods-phenol extraction, trichloroacetic acid (TCA) precipitation, and desalting/TCA precipitation-were compared to determine the optimal reproducible high resolution 2-dimensional (2-D) electrophoresis for each chungkugjang, doenjang, and kochujang samples. The soluble proteins from Chungkugjang extracted by phenol were separated with high reproducibility and resolution, and gained 1.75- to 3-fold more protein spots on 2-D gel than those from the other methods. On the contrary, the extracted proteins from doenjang and kochujang treated by desalting/TCA precipitation method showed about 1.5- to 3.3-fold more protein spots on 2-D gel. Using the established methods, the changes in the protein profiles of the fermented soy foods were monitored during the fermentation period by 2-DE. One of the major proteins in soy, $\beta$-conglycinin $\alpha$-subuint, and some proteins with unknown functions were localized on 2-D gel as the protease-resistant proteins throughout the fermentation period of doenjang. Changes in the protein profile monitored by the established methods can provide basic information on unfolding the mechanisms of the generation of biofunctional activity in the fermented soy foods.

• Food Engineering Progress
• /
• v.21 no.1
• /
• pp.36-41
• /
• 2017

The release profiles of nisin from nisin-incorporating gel foods and the antimicrobial activities of the gels on the growth of Brochothrix thermosphacta in an aqueous system containing the gels have been investigated. A linear regression model was applied to determine the diffusion coefficient (D) for the diffusion of nisin in the gel. The antimicrobial activities of nisin released from 1, 2 and 3% (w/v) agar gels on the growth of B. thermosphacta in a broth medium with and without nisin were investigated. The D decreased from $1.2{\times}10^{-2}$ to $8.2{\times}10^{-3}$ and $6.4{\times}10^{-3}cm^2/s$ as the agar concentration in the gel increased from 1 to 2 and 3% (w/v), demonstrating the diffusion rate in the gels can be controlled by the agar concentration in gel. The agar gel incorporating nisin inhibited the B. thermosphacta growth in the broth medium by prolonging the lag phase. The growth inhibition was enhanced by the addition of nisin in the medium. The results of this study exhibited that the gel food is a feasible nisin delivery system with a controlled release achieved by the adjustment of agar concentration in the system, demonstrating the potential of nisin-incorporating gel for preserving particulate-containing drinks.

• Bulletin of the Korean Chemical Society
• /
• v.33 no.8
• /
• pp.2489-2493
• /
• 2012

A new fluorescent system (acridine/RTIL hybrid gel) confined in the 3D micro-structure of a poly(lactic acid) membrane were prepared from 1-butyl-3-methylimidazolium-based ionic liquids ([bmim]X (X = $SbF_6$, $NTf_2$, Cl); RTILs), poly(lactic acid) (PLA), and acridine via the sol-gel route. SEM images showed that, in the presence of [bmim]$SbF_6$ and [bmim]$NTf_2$, 3D-ly paticulated structures were created inside the PLA membranes and acridine/RTIL hybrid gels were confined in gabs of particulates. However, the use of [bmim]Cl induced the formation of a 3D-ly porous structure containing the hybrid gel of acridine/[bmimCl in the micropores. The three fluorescent systems exhibited different fluorescence behaviors (fluorescence maximum and intensity) depending on the C2-H acidity scale of the RTILs (or their anion type). Acridine gels hybridized with [bmim]$SbF_6$ and [bmim]$NTf_2$ showed blue fluorescence with relative high intensity, whereas the hybrid gel with [bmim]Cl exhibited almost no fluorescence under dry conditions. However, the acridine/[bmim]Cl hybrid system in the micro-porous PLA membrane started to emit fluorescent light under humid conditions and showed a possible response, indicating that it could be applied as a humidity sensor.

• Korean Journal of Microbiology
• /
• v.26 no.4
• /
• pp.324-331
• /
• 1988

Ascorbate oxidizing enzyme from the crude extract of Pleurotus ostreatus was purified by ammonium sulfate precipitation, preparative polyacrylamide gel electrophoresis, DEAE Sepharose CL-6B ion exchange chromatography and Sephadex G-150 gel filtration chromatography. The molecular weight of the enzyme estimated by Sephadex G-150 gel filtration chromatography was 140,000 and that of its subunit by SDS-polyacrylamide gel electrophoresis 66,000. The optimum pH for the maximum activity of the enzyme was 5.2 and the isoelectric point of the enzyme was 6.0 Km values for L-ascorbic acid and D-isoascorbic acid were both 2.2.$\mu$M, which indicates that the enzyme has the asme affinity towards both substrates.