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Characterization of Pseudomonas sp. MN5 and Purification of Manganese Oxidizing Protein (Pseudomonas sp. MN5의 특성과 망간산화단백질 정제)

  • Lee, Seung-Hui;Park, Kyeong-Ryang
    • Journal of Life Science
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    • v.18 no.1
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    • pp.84-90
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    • 2008
  • Bacterial colonies which were able to oxidize the manganese were isolated from six soil samples in Byungchon area. Among them, one bacterial strain was selected for this study based on its high manganese oxidation activity. This selected bacterial strain was identified as Pseudomonas sp. MN5 through physiological-biochemical test and analysis of its 16s rRNA sequence. This selected bacterial strain was able to utilize fructose and maltose, but they doesn't utilizing various carbohydrates as a sole carbon source. Pseudomonas sp. MN5 showed a very sensitive to antibiotics such as kanamycin, chloramphenicol, streptomycin and tetracycline, but a high resistance up to mg/ml unit to heavy metals such as lithium, manganese and barium. Optimal manganese oxidation condition of Pseudomonas sp. MN5 was pH 7.5 and manganese oxidation activity was inhibited by proteinase K and boiling treatment. The manganese oxidizing protein produced by Pseudomonas sp. MN5 was purified by ammonium sulfate precipitation, HiTrap Q FF anion exchange chromatography and G3000sw $_{XL}$ gel filtration chromatography. By sodium dodecyl sulfate polyacrylamide gel electrophoresis, three manganese oxidizing protein with estimated molecular weights of 15 kDa, 46.7 kDa and 63.5 kDa were detected. Also, it was estimated that manganese oxidizing protein produced by Pseudomonas sp. MN5 were a kind of porin proteins through internal sequence and N-terminal sequence analysis.

The Effect of Potassium Phosphate as a pH Stabilizer on the Production of Gellan by Sphingomonas paucibilis NK-2000 (Sphingomonas paucibilis NK-2000에 의한 젤란의 생산에 미치는 pH 안정제로서의 인산칼슐의 영향)

  • Lee, Nam-Kyu;Jo, Young-Bae;Jin, Il-Hyuck;Son, Chang-Woo;Lee, Jin-Woo
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1033-1038
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    • 2009
  • Maximal productions of gellan by Sphingomnas paucibilis NK2000 from 20 g/l glucose and 10 g/l soybean pomace were 7.46 g/l in a flask and 7.35 g/l in a 7 l bioreactor, when the initial pH of media was 6.8. Maximal production of gellan in a 7 l bioreactor under pH control by sodium hydroxide was 8.42 g/l, whereas that under control by potassium phosphate was 8.50 g/l. The optimal concentration of potassium phosphate in a medium for production of gellan by S. paucibilis NK2000 was found to be 5.0 g/l. Maximal production of gellan in a medium containing 5.0 g/l potassium phosphate without pH control was 8.93 g/l in a 7 l bioreactor. In this study, a simple process without pH control was developed to enhance the production of gellan, with optimized concentration of potassium phosphate in the medium.

Painful Channels in Sensory Neurons

  • Lee, Yunjong;Lee, Chang-Hun;Oh, Uhtaek
    • Molecules and Cells
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    • v.20 no.3
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    • pp.315-324
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    • 2005
  • Pain is an unpleasant sensation experienced when tissues are damaged. Thus, pain sensation in some way protects body from imminent threat or injury. Peripheral sensory nerves innervated to peripheral tissues initially respond to multiple forms of noxious or strong stimuli, such as heat, mechanical and chemical stimuli. In response to these stimuli, electrical signals for conducting the nociceptive neural signals through axons are generated. These action potentials are then conveyed to specific areas in the spinal cord and in the brain. Sensory afferent fibers are heterogeneous in many aspects. For example, sensory nerves are classified as $A{\alpha}$, $-{\beta}$, $-{\delta}$ and C-fibers according to their diameter and degree of myelination. It is widely accepted that small sensory fibers tend to respond to vigorous or noxious stimuli and related to nociception. Thus these fibers are specifically called nociceptors. Most of nociceptors respond to noxious mechanical stimuli and heat. In addition, these sensory fibers also respond to chemical stimuli [Davis et al. (1993)] such as capsaicin. Thus, nociceptors are considered polymodal. Recent advance in research on ion channels in sensory neurons reveals molecular mechanisms underlying how various types of stimuli can be transduced to neural signals transmitted to the brain for pain perception. In particular, electrophysiological studies on ion channels characterize biophysical properties of ion channels in sensory neurons. Furthermore, molecular biology leads to identification of genetic structures as well as molecular properties of ion channels in sensory neurons. These ion channels are expressed in axon terminals as well as in cell soma. When these channels are activated, inward currents or outward currents are generated, which will lead to depolarization or hyperpolarization of the membrane causing increased or decreased excitability of sensory neurons. In order to depolarize the membrane of nerve terminals, either inward currents should be generated or outward currents should be inhibited. So far, many cationic channels that are responsible for the excitation of sensory neurons are introduced recently. Activation of these channels in sensory neurons is evidently critical to the generation of nociceptive signals. The main channels responsible for inward membrane currents in nociceptors are voltage-activated sodium and calcium channels, while outward current is carried mainly by potassium ions. In addition, activation of non-selective cation channels is also responsible for the excitation of sensory neurons. Thus, excitability of neurons can be controlled by regulating expression or by modulating activity of these channels.

Bonding Quality of Adhesives Formulated with Okara Hydrolyzates and Phenol-formaldehyde Resins for Bonding Fancy Veneer onto High-density Fiberboard (두부비지 가수분해물과 페놀수지로 조제한 마루판 화장용 접착제의 접착성능)

  • Yang, In;Ahn, Sye-Hee;Choi, In-Gyu;Choi, Won-Sil;Kim, Sam-Sung;Oh, Sei-Chang
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.4
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    • pp.388-396
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    • 2009
  • In our study, the potential of okara as an ingredient of new bio-based adhesives was investigated for the production of fancy-veneered flooring boards. Okara was hydrolyzed by 1% sulfuric acid solution (AC) and 1% sodium hydroxide solution (AK). Phenol formaldehyde (PF) prepolymers were prepared as a cross-linker of okara hydrolyzates. Then, okara-based adhesive resins were formulated with 35% AC, 35% AK and 30% PF prepolymer on solid content basis. The adhesive resins were applied on high-density fiberboards (HDF) with the spread rate of $300g/m^2$. After that, oak fancy veneers are covered on the HDF, and then pressed with the pressure of $7kg/m^2$ at $120^{\circ}C$. The experimental variables were three mole ratios of formaldehyde to phenol (1.8, 2.1, 2.4), three assembly time (0, 10, 20 min), and two press time (90 sec, 120 sec), respectively. The fancy-veneered high-density fiberboards were tested by dry tensile strength, glueline failure by wetting and formaldehyde emission. Tensile strength of the boards exceeded the requirement of KS standard. The formaldehyde emissions were approached at the E0 level specified in KS standard. Based on these results, okara can be used as an ingredient of environmentally friendly adhesive resin systems for the production of flooring boards.

Metabolic engineering of Vit C: Biofortification of potato

  • Upadhyaya, Chandrama P.;Park, Se-Won
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.10a
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    • pp.14-14
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    • 2010
  • Vitamin C (ascorbic acid) is an essential component for collagen biosynthesis and also for the proper functioning of the cardiovascular system in humans. Unlike most of the animals, humans lack the ability to synthesize ascorbic acid on their own due to a mutation in the gene encoding the last enzyme of ascorbate biosynthesis. As a result, vitamin C must be obtained from dietary sources like plants. In this study, we have developed two different kinds of transgenic potato plants (Solanumtuberosum L. cv. Taedong Valley) overexpressing strawberry GalUR and mouse GLoase gene under the control of CaMV 35S promoter with increased ascorbic acid levels. Integration of the these genes in the plant genome was confirmed by PCR and Southern blotting. Ascorbic acid(AsA) levels in transgenic tubers were determined by high-performance liquid chromatography(HPLC). The over-expression of these genes resulted in 2-4 folds increase in AsA intransgenic potato and the levels of AsA were positively correlated with increased geneactivity. The transgenic lines with enhanced vitamin C content showed enhanced tolerance to abiotic stresses induced by methyl viologen(MV), NaCl or mannitol as compared to untransformed control plants. The leaf disc senescence assay showed better tolerance in transgenic lines by retaining higher chlorophyll as compared to the untransformed control plants. Present study demonstrated that the over-expression of these gene enhanced the level of AsA in potato tubers and these transgenics performed better under different abiotic stresses as compared to untransformed control. We have also investigated the mechanism of the abiotic stress tolerance upon enhancing the level of the ascorbate in transgenic potato. The transgenic potato plants overexpressing GalUR gene with enhanced accumulation of ascorbate were investigated to analyze the antioxidants activity of enzymes involved in the ascorbate-glutathione cycle and their tolerance mechanism against different abiotic stresses under invitro conditions. Transformed potato tubers subjected to various abiotic stresses induced by methyl viologen, sodium chloride and zinc chloride showed significant increase in the activities of superoxide dismutase(SOD, EC 1.15.1.1), catalase, enzymes of ascorbate-glutathione cycle enzymes such as ascorbate peroxidase(APX, EC 1.11.1.11), dehydroascorbate reductase(DHAR, EC 1.8.5.1), and glutathione reductase(GR, EC 1.8.1.7) as well as the levels of ascorbate, GSH and proline when compared to the untransformed tubers. The increased enzyme activities correlated with their mRNA transcript accumulation in the stressed transgenic tubers. Pronounced differences in redox status were also observed in stressed transgenic potato tubers that showed more tolerance to abiotic stresses when compared to untransformed tubers. From the present study, it is evident that improved to lerance against abiotic stresses in transgenic tubers is due to the increased activity of enzymes involved in the antioxidant system together with enhanced ascorbate accumulated in transformed tubers when compared to untransformed tubers. At moment we also investigating the role of enhanced reduced glutathione level for the maintenance of the methylglyoxal level as it is evident that methylglyoxal is a potent cytotoxic compound produced under the abiotic stress and the maintenance of the methylglyoxal level is important to survive the plant under stress conditions.

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The Study on Emulsifying and Foaming Properties of Buckwheat Protein Isolate (분리 메밀 단백질의 유화 및 기포특성에 관한 연구)

  • 손경희;최희선
    • Korean journal of food and cookery science
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    • v.9 no.1
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    • pp.43-51
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    • 1993
  • Buckwheat protein isolate was tested for the effects of pH, addition of sodium chloride and heat treatment on solubility, emulsion capacities, emulsion stability, surface hydrophobicity, foam capacities and foam stability. The solubility of buckwheat protein isolate was affected by pH and showed the lowest value at pH 4.5, the isoelectric point of buckwheat protein isolate. The solubility significantly as the pH value reached closer to either ends of the pH, i.e., pH 1.0 and 11.0. The effects of NaCl concentration on solubility were as follows; at pH 2.0, the solubility significantly decreased when NaCl was added; at pH 4.5, it increased above 0.6 M; at pH 7.0 it increased; and at pH 9.0 it decreased. The solubility increased above $80^{\circ}C$, at all pH ranges. The emulsion capacity was the lowest at pH 4.5. It significantly increased as the pH approached higher acidic or alkalic regions. At pH 2.0, when NaCl was added, the emulsion capacity decreased, but it increased at pH 4.5 and showed the maximum value at pH 7.0 and 9.0 with 0.6 M and 0.8 M NaCl concentrations. Upon heating, the emulsion capacity decreased at acidic pH's but was maximised at pH 7.0 and 9.0 on $60^{\circ}C$ heat treatment. The emulsion stability was the lowest at pH 4.5 but increased with heat treatment. At acidic pH, the emulsion stability increased with the increase in NaCl concentration but decreased at pH 7.0 and 9.0. Generally, at other pH ranges, the emulsion stability was decreased with increased heating temperature. The surface hydrophobicity showed the highest value at pH 2.0 and the lowest value at pH 11.0. As NaCl concentrationed, the surface hydrophobicity decreased at acidic pH. The NaCl concentration had no significant effects on surface hydrophobicity at pH 7.0, 9.0 except for the highest value observed at 0.8 M and 0.4 M. At all pH ranges, the surface hydrophobicity was increased, when the temperature increased. The foam capacity decreased, with increased in pH value. At acidic pH, the foam capacity was decreased with the increased in NaCl concentration. The highest value was observed upon adding 0.2 M or 0.4 M NaCl at pH 7.0 and 9.0. Heat treatments of $60^{\circ}C$ and $40^{\circ}C$ showed the highest foam capacity values at pH 2.0 and 4.5, respectively. At pH 7.0 and 9.0, the foam capacity decreased with the increased in temperature. The foam stability was not significantly related to different pH values. The addition of 0.4 M NaCl at pH 2.0, 7.0 and 9.0 showed the highest stability and the addition of 1.0 M at pH 4.5 showed the lowest. The higher the heating temperature, the lower the foam stability at pH 2.0 and 9.0. However, the foam stability increased at pH 4.5 and 7.0 before reaching $80^{\circ}C$.

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Electrophoretic Characterization of Job′s tears (Yulmoo: Coix lachryma-jobi L. var. Ma-yuen stapf. & Yeomjoo: Coix lachryma-jobi L.) proteins (율무와 염주 단백질의 전기영동 특성)

  • 우자원
    • Korean journal of food and cookery science
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    • v.7 no.3
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    • pp.13-20
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    • 1991
  • This study was performed to examine the electrophoretic properties of Job's tears (Yulmoo: Coix lachryma-jobi L. var. Ma-yuen Stapf. & Yeomjoo: Coix lachryma-jobi L.) proteins. Albumins, globulins, gliadins and glutelins were extracted from the polished Yulmoo and brown Yeomjoo by the modified Osborne method. For a comparison, rice proteins were extracted and fractionated by the same method. The relative proportions of protein fractions were 17.4 : 19.6 : 55.2 : 7.7% in polished Yulmoo, 12.6 : 62.2 : 4.2 :21.0% in brown Yeomjoo and 14.2 : 57 4 : 0.77 : 27.8% in rice, in the order of albumis, globulins, gliadins and glutelins. Polyacrylamide gel electrophoresis (PAGE) and SDS-polyacrylamide gel electrophoresis (SDS-PAGE) were peformed to identify the subfractions of each protein fraction extracted from polished Yulmoo, brown Yeomjoo and rice. The electro-phoregrams of polyacrylamide gel electrophoresis showed that the same fractions of both polished Yulmoo protein and brown Yeomjoo protein had very similar electrophoretic patterns to each other respectively, but there were significant differences in the patterns between Job's tears proteins and rice proteins.

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Studies on the Regulation for Use, Metabolism, Intake, and Safety of Sodium Nitrite in Meat Products (육가공품에 사용되는 아질산염의 사용기준, 대사, 섭취량과 안전성에 대한 조사 연구)

  • Lee Keun-Taik;Kang Jong-Ok;Kim Cheon-Jei;Lee Mooha;Lee Sung Ki;Lee Joo-Yeon;Lee Ju-Woon;Cho Soo-Hyun;Joo Seon-Tea;Chin Koo B.;Choi Sung-Hee
    • Food Science of Animal Resources
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    • v.25 no.1
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    • pp.103-120
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    • 2005
  • During the 1970s, concern arose that cured meats contained high levels of residual nitrite and preformed nitrosamines. Therefore, the search for alternatives and alternative approaches to the use of nitrite have been still continued, however no complete alternative for nitrite has yet been identified. Recently, it was publicized in Korea that nitrite-containing meat products would be detrimental to health, about which consumers have been seriously concerned. Therefore, this study was carried out to inform the consumer of the safety status of nitrite and thereby to lead proper consumption of meat products. For assessing the safety of nitrite, data regarding the regulation for use, metabolism in human body, and dietary intake amounts of nitrite were collected and analyzed. The mean intake level of nitrite for Korean per capita was recently reported to be not more than 1% of ADI set by JECFA. On the contrary, a calculation indicated that the daily nitrite intake per capita from saliva by ingestion of vegetables in Korea would be about 300-fold higher than that from cured meats. In consideration of the low consumption amount of meat products per capita of Korean, that is, at least one fifth, compared to European and American, there is no particular reason to concern about the impairment of health by nitrite intake from meat products for Korean. However, any effort for the reduction of residual nitrite content in cured meats should be given with an idea to minimize the intake of nitrite even from the minor source.

Effect of Activated Carbon and Fish Oil Addition on the Physico-Chemical Characteristics in Chicken Meat (활성탄과 어유의 첨가가 계육의 이화학적 특성에 미치는 영향)

  • 박창일;김영직;김덕진;안종호;김영길
    • Food Science of Animal Resources
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    • v.22 no.3
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    • pp.206-211
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    • 2002
  • This study was conducted to investigate the influence of dietary activated carbon(0.9%) and fish oil(0, 1, 2, 4%) addition on the feed efficiency, blood-cholesterol, proximate composition, pH and minerals in breast and thigh of chicken meat. Broilers were randomly assigned to one of the five dietary treatment: 1) Control (commercial feed) 2) T1(commercial feed supplemented with 0.9% activated carbon) 3) T2 (commercial feed with 0.9% activated carbon and 1% fish oil) 4) T3 (commercial feed with 0.9% activated carbon and 2% fish oil) 5) T4 (commercial feed with 0.9% activated carbon and 4% fish oil). They were fed with one of the experimental diets for five weeks and slaughtered. After that, the meat samples were vacuum packaged and stored over a period of 10 days at 4$\pm$1$\^{C}$. When broilers were fed with dietary activated carbon and fish oil, the feed efficiency of birds were higher compared with that of control diet. The blood cholesterol was tended to decrease in dietary activated carbon and fish oil(p<0.05). However, effects of diets containing graded levels of activated carbon and fish oil on proximate composition were not found(p>0.05). The pH of all treatments significantly increased during the storage periods. The activated carbon and fish oil diet increased the calcium, potassium and sodium content of chicken meat, and tended to increase total mineral contents.

Purification and Characterization of a Keratinase from Bacillus licheniformis Strain for Degradation of Egg Shell Membrane (낙각막 분해를 위한 Bacillus licheniformis로 부터 Keratinase의 정제 및 특성)

  • 전태욱;박기문
    • Food Science of Animal Resources
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    • v.22 no.3
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    • pp.259-266
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    • 2002
  • The egg shell membrane degrading isolated from soil was identified as Bacillus licheniformis by 16S rDNA identification method. A keratinase was isolated from the Baciilu licheniformis culture. DEAE-cellulose ion-exchange and Sephadex C-75 gel chromatograhies were used to purify the enzyme. The specific activity was increased 17.3-fold by the purification procedures. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and Sephadex G-75 chromatography indicated that the purified keratinase was monomeric and had a molecular weight of 65 kDa. The enzyme showed optimum activity at pH 9.0, and was stable above pH 9.0. The optimum temperature was 50$\^{C}$ and the enzyme was stable in the temperature ranges from 20$\^{C}$ to 50t. By the addition of 1 mM and 10 mM FeSO4, the activities of the enzyme were increased to 111$\pm$4.6% and 133$\pm$3.79%, respectively. The keratinase was an alkaline serine pretense because it was inhibited only by phenylmethylsulfonylfluorice (PMSF).