• Journal of Microbiology and Biotechnology
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• v.18 no.2
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• pp.248-254
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• 2008

Candida magnoliae, an osmotolerant and erythritol producing yeast, prefers D-fructose to D-glucose as carbon sources. For the investigation of the fructophilic characteristics with respect to sugar transportation, a sequential extraction method using various detergents and ultracentrifugation was developed to isolate cellular membrane proteins in C. magnoliae. Immunoblot analysis with the Pma1 antibody and two-dimensional electrophoresis analysis coupled with MS showed that the fraction II was enriched with membrane proteins. Eighteen proteins out of 36 spots were identified as membrane or membrane-associated proteins involved in sugar uptake, stress response, carbon metabolism, and so on. Among them, three proteins were significantly upregulated under the fructose supplying conditions. The hexose transporter was highly homologous to Ght6p in Schizosaccharomyces pombe, which was known as a predominant transporter for the fructose uptake of S. pombe because it exhibited higher affinity to D-fructose than D-glucose. The physicochemical properties of the ATP-binding cassette transporter and inorganic transporter explained their direct or indirect associations with the fructophilic behavior of C. magnoliae. The identification and characterization of membrane proteins involved in sugar uptake might contribute to the elucidation of the selective utilization of fructose to glucose by C. magnoliae at a molecular level.

• Proceedings of the Korean Biophysical Society Conference
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• 2002.06b
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• pp.56-56
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• 2002

Muscle contraction and relaxation are regulated by the sarcoplasmic reticulum (SR)-mediated $Ca^{2＋}$ release and $Ca^{2＋}$ uptake. The SR functions are closely related with the proteins residing in the SR such as ryanodine receptor, $Ca^{2＋}$-ATpase, calsequestrin, triadin and junctin. In an effort to further identify important functional SR proteins, experiments of sucrose-density gradient of SR fractionation, concanavalin A treatment, 2D gel electrophoresis, $^{45}$ Ca$^{2+}$ overlay, Strains-all staining, and peptide finger printing (PFP) were carried out.(omitted)d)

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

• Preventive Nutrition and Food Science
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• v.15 no.2
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• pp.143-146
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• 2010

Organic zinc was included in the diet of broiler chickens to examine its effect on the skin characteristics and the expression level of skin proteins. Broiler chicks (Ross$\times$Ross) were fed a corn-wheat-soybean meal basal diet, either as control or containing an additional 80 ppm of zinc proteinate for 4 weeks, and then five broilers from each treatment were selected randomly, slaughtered, and their skin characteristics were examined. There were significant increases (p<0.05) in thigh skin epidermis and dermis thickness in the chicks fed organic zinc. Collagen content in the skin of broilers was also increased by the addition of organic zinc to the diet. 2D-gel electrophoresis patterns indicated that expression levels of the three proteins, glyoxylase 1, hypothetical protein, and dispersin B were affected by zinc feeding. These results suggest that adding organic zinc to the chicken's feed may contribute to decreased skin tearing.

• Reproductive and Developmental Biology
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• v.33 no.4
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• pp.237-242
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• 2009

Pregnancy is a unique event in which a fetus develops in the uterus despite being genetically and immunologically different from the mother, and the underlying mechanisms remain poorly understood. To analyze the differential gene expression profiles in nonpregnant and 7 days post coitus (dpc) pregnant uterus of mice, we performed a global proteomic study by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. The uterine proteins were separated using 2-DE, Approximately 1,000 spots were detected on staining with Coomassie brilliant blue. An image analysis using Melanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between pregnant and nonpregnant uterus. Twenty-one spots were identified as differentially expressed proteins, of which 10 were up-regulated proteins such as alpha-fetoprotein, chloride intracellular channel 1, transgelin, heat-shock protein beta-1, and carbonic anhydrase II, while 11 were down-regulated proteins such as X-box binding protein, glutathione S-transferase omega 1, olfactory receptor Olfr204, and metalloproteinase-disintegrin domain containing protein TECADAM. Most of the identified proteins appeared to be related with catabolism, cell growth, metabolism, regulation, cell protection, protein repair, or protection. Our results uncovered key proteins of mouse uterus involved in pregnancy.

• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1222-1228
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• 2006

Two kinds of Haematococcus pluvialis cells (green vegetative cells cultivated under optimal cell culture conditions and red cyst cells maintained under high light stress conditions to induce astaxanthin production) were used to investigate the protein expression profiles by two-dimensional electrophoresis, image analysis, and peptide mass fingerprinting. The cellular accumulation of astaxanthin was evident after exposure to high light intensity and reached the maximum cellular level after 78 h of high light stress. In a 2-D electrophoresis analysis, 22 proteins were upregulated over 2-fold in the red cyst cells when compared with the green vegetative cells and selected for further analysis by chemically assisted fragmentation (CAF)-MALDI-TOF sequencing to identify the protein functions. Among 22 different spots, several key enzymes specific to the carotenoid pathway, including isopentenyl pyrophosphate isomerase (IPP) and lycopene $\beta$-cyclase, appeared in H. pluvialis after exposure to high light intensity. Therefore, IPP and lycopene $\beta$-cyclase would appear to be involved with carotenoid accumulation in the cytoplasm, as these peptides were preferentially upregulated by high light intensity preceding an increase in carotenoid, and only these forms were detected in the red cyst cells.

• Plant Resources
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• v.6 no.1
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• pp.81-88
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• 2003

Higenamine [1-(4-hydroxy-6, 7-dihydroxy-l, 2, 3, 4-tetrahydroisoquinoline) is a cardiotonic constituent of Aconiti tuber, one of the most widely prescribed oriental medicines. S-(-)higenamine was reported to have a stronger cardiotonic activity than R-(+)-higenamine and known as a central intermediate in the biosynthesis of various benzyl isoquionoline alkaloids in plants. The separation of higenamine enantiomers has been accomplished with capillary electrophoresis using cyclodextrins (CDs) as chiral selectors. Good resolution of this enantiomers was obtained using a 50 mM sodium phosphate buffer containing hydroxypropyl $\beta$-CDs using 27 cm fused silica capillary (50${\mu}{\textrm}{m}$ i.d., 20 cm to detector) at 25 $^{\circ}C$. With the electric field of 340 V/cm, the separation time of higenamine enantiomers was less than 6 min. Under this optimum conditions, the relative standard deviations of migration time and peak area were less than 1.6% and 3.2%. A 512-channel diode array detector was confirmed for the higenamine. The detection limits (S/N = 3) of these enantiomers are $1.5mutextrm{m}$/mL. We confirmed the chiral form of higenamine in medicinal plants.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.525.1-525
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• 1986

To clone ${\beta}$-glucosidase gene from Cellulomonas sp. a gene library was constructed using E. coli JM83 pUC9. Among 2,500 pseudotransformants obtained, 20 clones developed yellow color on the p-nitrophenyl- -D-glucopyranoside filter paper These 20 clones were classified into three groups based on the results of activity staining using nondenaturating polyacrylamide gel electrophoresis and restriction enzyme digestions. Among the three groups, only one group containing pCEl plasmid has specificity for cellobiose.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2004.10a
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• pp.62-62
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• 2004

• Proceedings of the KAIS Fall Conference
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• 2009.05a
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• pp.3-4
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• 2009

한국인 아동의 우식치아로부터 S. mutans를 분리하고, acid stress하에서 분리한 S. mutans의 내산성 능력과 관련된 단백질을 규명하고자 하였다. S. mutans의 16S rDNA 유전자 핵산염기서열을 바탕으로 설계된 프라이머 쌍을 이용하여 중합효소연쇄반응을 실시하고 16S rDNA sequencing을 실시한 결과, 한국인 아동으로부터 분리 된 S. mutans K7은 기존에 보고된 표준균주와 99.9% 일치하는 결과를 나타내 S. mutans로 판명되었다. 또한 2D gel electrophoresis 를 수행한 결과, acid stress에 관여하는 것으로 추정되는 단백질들을 확인 할 수 있었다.