• Title/Summary/Keyword: S-Layer protein

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Studies on the Composition of Protein and Lipid from Korean Walnut (Juglans regia L.) (한국산 호도의 단백질 및 지질의 조성에 관한 연구)

  • Choi, Cheong;Sung, Tae-Soo;Cha, Woen-Suep;Son, Cyu-Mok
    • Applied Biological Chemistry
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    • v.29 no.3
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    • pp.318-323
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    • 1986
  • We empolyed gel filtraction, polyacrylamide gel electrophoresis, amino acid autoanalyzer, thin layer chromatography for determining protein and lipid composition in walnut. The walnut contained 22.18% of crude protein and 64.23% of crude lipid. Glutamic acid (38.60%) was the major amino acid in soluble protein, followed by arginine and aspartic acid. The sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed 12 band in soluble protein of walnut, and collection rate of main protein fraction purified by Sephadex G-150 was 60.67%. The molecular weight for the main protein was estimated to be 43,000. The lipid fraction obtained by silicic acid column chromatography were mainly composed of about 93.05% neutral lipid, whereas compound lipid was only 7.0% level. Among the neutral lipid by thin layer chromatography, triglyceride was 82.05%, sterol ester and free fatty acid were 3.86% and 4.80%, repectively. The predominant fatty acids of total and neutral lipids were linoleic acid $(64.48{\sim}69.98%)$ and oleic acid $(13.89{\sim}15.36%)$. The major fatty acids of triglyceride separated from neutral lipid were linolenic acid (69.98%).

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Effect of Seaweeds Addition to Layer′s Rarion on Egg Productivity and Reduction of Calcium . Phosphorus Excretion (해조분의 첨가가 산란계의 생산성 및 칼슘 . 인의 배설량 저감에 미치는 영향)

  • 김동균;박정래;정인학
    • Journal of Animal Environmental Science
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    • v.6 no.3
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    • pp.141-151
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    • 2000
  • Over-grown seaweeds in Donghae(east sea of Korea peninsula) may impedes ocean environment, however, they can be a good nutrient resources for poultry feeds if they are utilized properly. In this study, seaweeds powder were tested for laying hens\` ration to investigate the effects on egg production rate, egg quality and calcium phosphorus excretion. One hundred 65wks-old brown layers were fed for 5 weeks alotted with seaweeds powder addition to experimental diet by 0(control), 0.5, 1.0 and 2.0%, respectively, and obtained following results; 1. Seaweeds proved a mid-protein low-energy feed resources with planty of K, Na, Ca, Mg, Sr and Fe. 2. Seaweeds addition by 0.5% and 1.0% improved egg production rate and egg-mass output markedly(p<0.05) than control. Seaweeds addition did not alter cholesterol level of yolk and yolk index, however egg shell thickness showed increasing trend by increasing seaweeds addition level. 3. Protein absorption and digestibility in seaweed addition treatments were significantly higher(p<0.05) than control group and protein contents of excreta in 0.5% and 1.0% treatments were reduced(p<0.05), which suggests effective protein metabolism for egg production. Increasing seaweeds addition reduces Ca and P contents in rectum and excreta, suggesting Ca and P utilization improvement in laying hens and lessening soil pollution. 4. In conclusion, seaweeds addition in layers' diet by 1.0% level improves egg-mass production and might be egg quality by increasing metabolism of protein, calcium and phosphorus.

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Calretinin-Containing Neurons in the Deeper Layers of the Hamster Superior Colliculus (햄스터 상구의 deeper layers에서 calretinin이 함유 신경세포)

  • Kim, Ye-Eun;Choi, Jae-Sik;Kim, Hye-Hyun;Yeo, Jin-Yeon;Jeon, Chang-Jin
    • Journal of Life Science
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    • v.16 no.5
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    • pp.750-758
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    • 2006
  • Calcium-binding protein calretinin is thought to play important roles in calcium buffering. Recently, we reported on the distribution, morphology of calretinin-immunoreactive (IR) neurons and the effects of eye enucleation on the immunoreactivity of calretinin in the superficial layers of the hamster superior colliculus (SC). In the present study, we describe the distributions and types of labeled cells and effects of enucleation in the deeper layers by immunocytochemistry. We also compare this labeling to that of GABA, the major inhibitory neurotransmitter in the central nervous system. In contrast to the superficial layers, the deeper layers contained many calretinin-IR neurons which formed two tiers. The first tier, which was very distinctive, was found within the intermediate gray layer. The second tier was found in the deep gray layer. Labeled neurons varied dramatically in morphology and included vertical fusiform, stellate, round/oval, and horizontal neurons. In contrast to the superficial layers, enucleation appeared to have no effect on the distribution of calretinin immunoreactivity in the deeper layers. Two-color immunofluorescence revealed that none of calretinin-IR neurons were labeled with an antibody to GABA. The present results demonstrate that calretinin identifies unique neuronal sublaminar organizations in the hamster SC. The present results also demonstrate that none of the calretinin-IR neurons in the hamster SC is GABAergic interneurons. As many calretinin-IR cells are GABAergic interneurons in most other brain areas, this phenomenon in hamster SC is exceptional.

중성자 산란을 이용한 생체물질의 구조 연구 : 단백질의 생체유사막의 흡착

  • Sin, Gwan-U;Rafailovich, M.H.;Sokolov, J.;Pernodet, N.;Satija, S.K.
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.30-33
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    • 2002
  • We have shown that it is possible to form a fibrilar network of fibronectin on a polyelectrolyte polymer film whose dimensions are similar to those reported on the extra cellular matrix. The fibronectin network was observed to form only when the charge density of the polymer was in excess of the natural charge density of the cell wall. Furthermore, the self-organized fibronectin layer was much thicker than the polymer film, indicating that long ranged interaction may playa key role in the assembly process. It is therefore important to understand the structure of the polymer layer/protein interface. Here we report on a neutron reflectivity study where we explore the structure of the polyelectrolyte layer, in this case sulfonated polystyrene (PSSx,), with varying degree of sulfonation (x<30%), as a function of sulfur content and counter ion concentration. These results are then correlated with systemic study of the adsorption and the multilayer formation of fibronectin as a function of incubation time for various sulfonation levels of $PSSx.^1$

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Detection of Anticancer Activity from the Root of Angelica gigas In Vitro

  • Ahn, Kyung-Seop;Sim, Woong-Seop;Kim, Ik-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.5 no.2
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    • pp.105-109
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    • 1995
  • Anticancer activity of a fraction of the ethanol extract from the root of Korean angelica (Angelica gigas Nakai) was recognized in human cancer cell lines HeLa $S_3$, K-562, and Hep $G_2$. The extract blocked the phorbol ester-inducing megakaryocytic differentiation of K-562 cells, which indicated the modification of protein kinase C (PKC) activity. In vitro assay showed the activation of PKC by the extract. An effective fraction of the Angelica gigas extract, of which $R_f$ value was 0.64 in a thin layer chromatography, was a different component from those of European angelicas. The $ED_50$ value of the fraction was 8, 9, and $16\;\mu\textrm{m}/ml$ against HeLa $S_3\;Hep\;G_2$, and K-562 cells, respectively, while the fraction showed higher $ED_50$ values against normal cell lines.

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Ribonucleotide Reductase Activity of Schizosaccbarornyces pombe Is Inhibited by Escherichia coli RecA Antibody (Schizosaccbarornyces pombe에서 Escherichia coli RecA 항체에 의한 Ribonucleotide Reductase 효소활성 저해)

  • Lee, Jung-Sup;Chun, Min-Suck;Kim, Ok-Bong;Park, Jong-Kun;Kim, Si-Wouk;Park, Yeal;Yang, Young-Ki;Hong, Seung-Hwan;Park, Sang-Dai
    • The Korean Journal of Zoology
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    • v.38 no.4
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    • pp.565-569
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    • 1995
  • We have previoosly demonstrated that the RecA-like protein of Schizosaccharomyces pombe (S. pombe) is immunologically related to Escherichia coil (E. coil) RecA protein and that the cellular level of the protein is significantly increased by inhibitors of nucleotide pool-forming enzymes such as hydroxyurea (HU) and methotrexate (MTX) (lee and Park, 1994; lee et al., 1994). In this study, we report that the ribonudeotide redudase activity of S. pombe is inhibited by E. coil RecA antibody, as determined by thin layer chromatography using [5-$^3$H]CDP as a substrate. The relative activity of ribonucleotide reductase was dramatically inhibited by 100 mM of flu (26.4% reduction) in in vitro assay, compared to that of non-treated control. The ribonucleotide reductase activity was also inhibited by immunoprecipitation with E. coil RecA antibody (43.3% reduction). These results indicate that the strudure of S. pombe ribonucleotide reductase is in part similar to that of E. coil RecA protein.

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Study of Efficiency with Rose, Lavender essential oil about Skin Inflammation Iinduced by Surfactant (계면활성제에 의해 유도된 피부염증에 있어서 Rose, Lavender essential oil 의 유효성 연구)

  • Choi, Jeung-Sook;Nam, Jeung-Hae
    • Fashion & Textile Research Journal
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    • v.8 no.4
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    • pp.471-475
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    • 2006
  • The summary of a study of efficacy of Rose essential oil and Lavender essential oil against skin-inflammation caused to surfactant is as follows. As a result of protein analysis, although Rose essential oil and Lavender essential oil stimulates skin, there is no much alteration of protein. The alteration of protein's bend to be under part of 43Kd is appeared in the group treated Rose essential oil and Lavender essential oil. The stratum of the group treated by surfactant is more, the stratum of the group treated by surfactant is more thick. As the observed result of alteration of basal layer, it is seen that the group treated Rose essential oil and Lavender essential oil takes the shape of cell layer constant, and the group treated Rose essential oil is made the basal cell layer of normal structure than that of Lavender essential oil. As the observed result of alteration of Mast cell group treated by Rose essential oil and Lavender essential oil for 3 weeks Luna's stain(${\times}20$), (${\times}100$), it is observed that the damaged group treated by Lavender essential oil is numerous in alteration of mast cell's number, and Mast cell's size is larger than the damaged group.

Cloning and Characterization of a Multidomain GH10 Xylanase from Paenibacillus sp. DG-22

  • Lee, Sun Hwa;Lee, Yong-Eok
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1525-1535
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    • 2014
  • The xynC gene, which encodes high molecular weight xylanase from Paenibacillus sp. DG-22, was cloned and expressed in Escherichia coli, and its nucleotide sequence was determined. The xynC gene comprised a 4,419bp open reading frame encoding 1,472 amino acid residues, including a 27 amino acid signal sequence. Sequence analysis indicated that XynC is a multidomain enzyme composed of two family 4_9 carbohydrate-binding modules (CBMs), a catalytic domain of family 10 glycosyl hydrolases, a family 9 CBM, and three S-layer homologous domains. Recombinant XynC was purified to homogeneity by heat treatment, followed by Avicel affinity chromatography. SDS-PAGE and zymogram analysis of the purified enzyme identified three active truncated xylanase species. Protein sequencing of these truncated proteins showed that all had identical N-terminal sequences. In the protein characterization, recombinant XynC exhibited optimal activity at pH 6.5 and $65^{\circ}C$ and remained stable at neutral to alkaline pH (pH 6.0-10.0). The xylanase activity of recombinant XynC was strongly inhibited by 1 mM $Cu^{2+}$ and $Hg^{2+}$, whereas it was noticeably enhanced by 10 mM dithiothreitol. The enzyme exhibited strong activity towards xylans, including beechwood xylan and arabinoxylan, whereas it showed no cellulase activity. The hydrolyzed product patterns of birchwood xylan and xylooligosaccharides by thin-layer chromatography confirmed XynC as an endoxylanase.

Immunohistochemical and Immunocytochemical Study about the Glial Fibrillary Acidic Protein in the Tanycytes of the Area Postrema of Bat (박쥐 맨아래구역 띠뇌실막세포의 Glial Fibrillary Acidic Protein에 대한 면역조직화학 및 면역세포화학적 연구)

  • Yang, Young-Chul;Cho, Byung-Pil;Kang, Ho-Suck
    • Applied Microscopy
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    • v.30 no.4
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    • pp.377-387
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    • 2000
  • There are a few tanycytes between the general ependymal cells lining the ependymal layer of the brain ventricle. These cells are considered as modified ependymal cells which possess a long basal process. Tanycytes are known to have an ability to communicate by absorbing substances from cerebrospinal fluid and transporting them to the blood vessels and/or to the neurons in the CNS. The third and fourth ventricular tanycytes were mainly studied as subjects but it's rare to find reports about the tanycytes of the area postrema. Glial fibrillary acidic protein is an intermediate filament protein that is expressed especially in astrocytes of the CNS. But GFAP is also found in filament of the tanycytes and its process. Therefore this study was carried out for the examination of the GFAP immunoreactive tanycytes lining the area postrema of the bat, and we also examined the ultrastructure of tanycytes using electron microscope. GFAP immunoreactive tanycytes were located in the caudal portion of the fourth ventricle, and especially mainly in the transitional zone between the floor of the caudal fourth ventricle and ependymal layer lining the area postrema. A few GFAP immunoreactive tanycytes were also found in the ependymal layer lining the area postrema, and some groups of tanycytes were found in the ependymal layer of the area postrema near the floor of the caudal fourth ventricle , The processes of tanycytes were stained deeply with anti-GFAP antibody. Especially the GFAP immunoreactive tanycytes lining the area postrema had very long processes that cross the whole width of the area postrema. In the electron microscope, the cell body of ependymal tanycyte was located on the ependymal layer and had a long basal process. Intermediate filaments were observed around the nucleus and well developed in the process of tanycrte. Longitudinal oriented long mitochondria and a few lipid droplets were also found in this process. After immunocytocheical staining, the gold particles were found only in the intermediate filaments. We could not determine the function of the tanycytes in the area postrema. Thus, further investigation is required to determine the functional relationship between the tanycytes and the area postrema in hibernating animal, the bat.

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Sequence Analysis of a Cryptic Plasmid pKW2124 from Weissella cibaria KLC140 and Construction of a Surface Display Vector

  • Kim, Soo Young;Oh, Chang Geun;Lee, Young Joo;Choi, Kyu Ha;Shin, Doo Sik;Lee, Si Kyung;Park, Kab Joo;Shin, Hakdong;Park, Myeong Soo;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.545-554
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    • 2013
  • Plasmid isolation of kimchi-derived Weissella cibaria KLC140 revealed six different plasmids. The smallest plasmid, pKW2124, was DNA sequenced and characterized, showing 2,126 bp with a GC content of 36.39% and five putative open reading frames (ORFs). In silico analysis of these ORFs showed ORF1 encodes a putative replication protein similar to rolling circular replication proteins from other lactic acid bacteria. However, a single-stranded intermediate was not detected when S1 nuclease was treated, suggesting it may follow theta replication. Interestingly, the replication initiation site of this plasmid is 100% identical to other plasmids from lactic acid bacteria, suggesting it may function for replication initiation. To construct a surface layer expression vector, pTSLGFP, slpA encoding the surface layer protein from Lactobacillus acidophilus was PCR amplified and fused with the gfp gene, forming a SLGFP fused gene. The plasmid pKW2124 was cloned into the XbaI site of pUC19, forming an Weissella-E. coli shuttle vector pKUW22. NheI-linearized pTSLGFP was ligated into pKUWCAT containing pKUW22 and the chloramphenicol acetyltransferase gene from pEK104, resulting in an 8.6 kb pKWCSLGFP surface layer expression vector. After transformation of this vector into W. cibaria KLC140, a GFP fluorescence signal was detected on the surface of the transformant, substantiating production of SLGFP fused protein and its secretion. This is the first report for construction of a Weissella surface layer expression vector, which may be useful for surface layer production of beneficial proteins in Weissella.