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Journal of microbiology and biotechnology 36건

  1. [국내논문]   An Efficient and Stable Method for the Transformation of Heterogeneous Genes into Cephalosporium acremonium Mediated by Agrobacterium tumefaciens   피인용횟수: 3

    XU WEI (Department of Pharmaceutical Engineering, Shenyang Pharmaceutical University ) , ZHU CHUNBAO (Department of Biopharmacevtics, Shanghai Institute of Pharmaceutical Industry(SIPI) ) , ZHU BAOQUAN (Department of Biopharmacevtics, Shanghai Institute of Pharmaceutical Industry(SIPI))
    Journal of microbiology and biotechnology v.15 no.4 ,pp. 683 - 688 , 2005 , 1017-7825 ,

    초록

    A transformation system mediated by Agrobacterium tumefaciens is routinely used for the genetic engineering of plants. Here, we report an efficient and stable method for transformation of heterogeneous genes into an industrial Cephalosporium acremonium by using a similar transformation system established in plants. Both the phleomycin-resistant gene and vgb gene were used as screening markers to confirm the success of transformation by either Southern hybridization or PCR amplification. It was found that acetosyringone (AS) was necessary only for protoplast transformation and the heterogeneous genes transferred were integrated into the genome of C. acremonium. The transformation efficiency obtained with this system was much higher than the conventional techniques used for transformation of C. acremonium.

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  2. [국내논문]   Partial Purification and Characterization of Thermostable Alkaline $\beta$-Mannanase from Bacillus sp. JB-99 Suitable for Pulp Bleaching   피인용횟수: 4

    VIRUPAKSHI S. (Department of Biotechnology, Gulbarga University ) , BABU K. GlREESH (Department of Biotechnology, Gulbarga University ) , NAIK GAJANAN R. (Department of Biotechnology, Gulbarga University)
    Journal of microbiology and biotechnology v.15 no.4 ,pp. 689 - 693 , 2005 , 1017-7825 ,

    초록

    Bacillus sp. JB-99, when grown in a chemically defined medium containing lactose as a carbon source, yielded 3,860 U/ml extracellular $\beta$ -mannanase, which was high compared to other examined carbon sources. Among the nitrogen sources, yeast extract enhanced the enzyme activity. The enzyme production was growth-associated. The enzyme was optimally active at $65^{\circ}C$ , pH 10, and had a half-life of 190 min at $65^{\circ}C$ . N-Bromosuccinamide and $AgNO_3,\;CuSO_4$ , and $HgCl_2$ strongly inhibited the enzyme, whereas $Ca^{2+}$ stimulated the enzyme activity. The $\alpha$ -galactosidase enzyme production was not found in any of the enzyme assays.

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  3. [국내논문]   Functional Characterization of the Madlp, a Spindle Checkpoint Protein in Fission Yeast  

    Kim, In-Gyu (School of Life Science and Biotechnology, Korea University ) , Rhee, Dong-Keun (School of Life Science and Biotechnology, Korea University ) , Lee, Hee-Cheul (School of Life Science and Biotechnology, Korea University ) , Lee, Joo (Department of Biology, Yonsei University ) , Kim, Hyong-Bai (Department of Biotechnology and Bioinformatics, Korea University)
    Journal of microbiology and biotechnology v.15 no.4 ,pp. 694 - 700 , 2005 , 1017-7825 ,

    초록

    Defects in the mitotic spindle or in the attachment of chromosomes to the spindle are believed to release an activated form of spindle checkpoint complex that inhibits APC-dependent ubiquitination and subsequently arrests the cell cycle at metaphase. When the spindle assembly is disrupted, the fission yeast mitotic arrest deficient (mad) mutants fail to arrest and rapidly lose viability. To enhance our understanding of the molecular mechanisms for the pathway of checkpoint function, the functional characterizations of Mad 1 p from Schizosaccharomyces pombe involved in this process have been carried out. Yeast two-hybrid and various deletion analyses of S. pombe Mad1 p reveal that the C terminus of Mad1p is critical for the binding of Mad2p and maintenance of Mad 1 p-Mad2p interaction. In addition, it was found. that the Mad1p region (residues 206-356) is essential for Mad1p-other checkpoint components. Mad1p truncating this region is sufficient to bind Mad2p but abolishes the checkpoint function, indicating that the checkpoint function is necessary for interaction of Mad 1 p-other checkpoint components. The possible functions of S. pombe Mad1p at the cell cycle checkpoint are discussed.

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  4. [국내논문]   Effective Screening of Antagonist for the Biological Control of Soilborne Infectious Disease (Damping-Off)   피인용횟수: 4

    LEE BAEK-SEOK (Department of Biological Engineering, Inha University ) , LEE HYANG-BOK (Department of Biological Engineering, Inha University ) , CHOI SUNG-WON (Greenbiotech. Co. Ltd. ) , YUN HYUN-SHIK (Department of Biological Engineering, Inha University ) , KIM EUN-KI (Department of Biological Engineering, Inha University)
    Journal of microbiology and biotechnology v.15 no.4 ,pp. 701 - 709 , 2005 , 1017-7825 ,

    초록

    An efficient method of selecting an antagonistic strain for use as a biological control agent strain was developed. In this improved method, the surface tension reduction potential of an isolate was included in the 'decision factor,' in addition to two other factors; the growth rate and pathogen inhibition. By using a statistically designed method, an isolate from the soil was selected and identified as Bacillus sp. GB 16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth were observed when the Bacillus sp. GB 16 was used. The action of the surface tension reducing component was assumed to enhance the wetting, spreading, and residing of the antagonistic strain in the rhizosphere. This result showed that the improved selection method was quite effective in selecting the best antagonistic strain for the biological control of soilborne infectious plant pathogens.

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  5. [국내논문]   Characterization of an Improved Recombinant Baculovirus Producing Polyhedra that Contain Bacillus thuringiensis Cry1Ac Crystal Protein   피인용횟수: 10

    Kim Jae-Su (School of Agricultural Biotechnology, College of Agriculture & Life Science, Seoul National University, Dongbu Hannong Chemical Co., Ltd., Agricultural Technology Research Institute (ATRI) ) , Cho Jae-Young (School of Agricultural Biotechnology, College of Agriculture & Life Science, Seoul National University ) , Chang Jin-Hee (School of Agricultural Biotechnology, College of Agriculture & Life Science, Seoul National University ) , Shim Hee-Jin (School of Agricultural Biotechnology, College of Agriculture & Life Science, Seoul National University ) , Roh Jong-Yul (School of Agricultural Biotechnology, College of Agriculture & Life Science, Seoul National University ) , Jin Byung-Ae (College of Natural Resources and Life Sciences, Dong-A University ) , Je Yeon-Ho (School of Agricultural Biotechnology, College of Agriculture & Life Science, Seoul National University)
    Journal of microbiology and biotechnology v.15 no.4 ,pp. 710 - 715 , 2005 , 1017-7825 ,

    초록

    A novel recombinant baculovirus, Bactrus, was constructed by the insertion of the Bacillus thuringiensis cry1Ac gene between two polyhedrin genes of Autographa californica nucleopolyhedrovirus (AcNPV) under the control of the polyhedrin gene promoter. Polyhedra produced by Bactrus in insect cells were incorporated with 130 kDa of polyhedrin-Cry1Ac-polyhedrin fusion protein, and 30 kDa of intact polyhedrin, resulting from a homologous recombination between two polyhedrin genes, was also expressed. The insecticidal activity of Bactrus against Spodoptera exigua larvae was similar to that of AcNPV, but it showed significantly higher toxicity towards Plutella xylostella larvae in comparison with that of AcNPV. The expression level of fusion protein and the insecticidal activity of recombinant polyhedra produced by the Bactrus against P. xylostella larvae were decreased after serial passages. In conclusion, the Bactrus had improved insecticidal activity and returned to wild-type AcNPV after several passages.

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  6. [국내논문]   Characteristics of Sulfur Oxidation by a Newly Isolated Burkholderia spp.   피인용횟수: 5

    JUNG JE, SUNG (Department of Food Science and Technology, Kyung Hee University ) , JANG KI-HYO (Department of Food and Nutrition, Samcheok National University ) , SIHN EON-HWAN (Department of Hotel Culinary Arts, Ulsan College ) , PARK SEUNG-KOOK (School of Agricultural Biotechnology, College of Agriculture & Life Science, Seoul National University ) , PARK CHANG-HO (Industrial Liaison Research Institute, Kyung Hee University, Department of Chemical Engineering, Kyung Hee University)
    Journal of microbiology and biotechnology v.15 no.4 ,pp. 716 - 721 , 2005 , 1017-7825 ,

    초록

    The role of an effective microbial species is critical to the successful application of biological processes to remove sulfur compounds. A bacterial strain was isolated from the soil of a malodorous site and identified as Burkholderia spp. This isolate was able to oxidize thiosulfate to sulfate, with simultaneous pH decrease and accumulation of elemental sulfur. The specific growth rate and the sulfate oxidation rate using the thiosulfate basal medium were $0.003 h^{-1}\;and\;3.7 h^{-1}$ , respectively. The isolated strain was mixotrophic, and supplementation of $0.2\%$ (w/v) of yeast extract to the thiosulfate-basal medium increased the specific growth rate by 50-fold. However, the rate of sulfate oxidation was more than ten times higher without yeast extract. The isolate grew best at pH 7.0 and $30^{\circ}C$ , and the sulfate oxidation rate was the highest at 0.12 M sodium thiosulfate. In an upflow biofilter, the isolated strain was able to degrade $H_2S\;with\;88\%$ efficiency at 8 ppm and 121/h of incoming gas concentration and flow rate, respectively. The cell density at the bottom of the column reached $3.2{\times}10^8$ CFU/(g bead) at a gas flow rate of 121/h.

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  7. [국내논문]   Cloning of Human Liver Cytosolic Sialidase from Genomic DNA Using Splicing by Overlap Extension and Its Characterization  

    HA KI-TAE (National Research Laboratory for Glycobiology and Department of Biochemistry and Molecular Biology, College of Oriental Medicine, Dongguk University ) , CHO SEUNG-HAK (Division of Intestinal Bacteriology, National Institute of Health ) , KANG SUNG-KOO (National Research Laboratory for Glycobiology and Department of Biochemistry and Molecular Biology, College of Oriental Medicine, Dongguk University ) , KIM YEON-KYE (National Fisheries Research Institute ) , KIM JUNE-KI (Division of Intestinal Bacteriology, National Institute of Health ) , KIM CHEORL-HO (National Research Laboratory for Glycobiology and Department of Biochemistry and Molecular Biology, College of Oriental Medicine, Dongguk University)
    Journal of microbiology and biotechnology v.15 no.4 ,pp. 722 - 727 , 2005 , 1017-7825 ,

    초록

    Cytosolic sialidase (Neu2), a member of the sialidase family that is responsible for hydrolysis of sialic acid from the terminal position of sialoglycoconjugates, is poorly expressed in skeletal muscle and not detected in any other adult tissues. Thus, we isolated Neu2 cDNA using splicing by overlap extension (SOEing). In order to further characterize this enzyme, a His-tagged derivative was expressed in the bacterial expression system and purified by $Ni^{2+}$ -affinity chromatography. A recombinant product of approximately 42 kDa had sialidase activity toward 4-methyl-umbelliferyl- $\alpha$ -D-N-acetylneuraminic acid (4MU-NeuAc). The optimal pH and temperature of the recombinant Neu2 for 4MU-NeuAc was 6.0 and $37.5^{\circ}C$ , respectively. The metal ions, such as $Cu^{2+}\;and\;Cd^{2+}$ , showed strong inhibitory effect on the activity of the enzyme. The enzyme efficiently hydrolyzed the gangliosides GM3 and GD3 and had relatively low activities on ganglioside GD1a and GD1b, $\alpha$ 2-3 sialyllactose, and sialylated glycoproteins such as fetuin, transferrin, and orsomucoid, but had hardly any activities on $\alpha$ 2-6 sialyllactose and ganglioside GM1 and GM2. We concluded that the recombinant Neu2 has a sialidase activity toward glycoproteins as well as gangliosides.

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  8. [국내논문]   Comparison of Antibiotic Resistance of Blood Culture Strains and Saprophytic Isolates in the Presence of Biofilms, Formed by the Intercellular Adhesion (ica) Gene Cluster in Staphylococcus epidermidis   피인용횟수: 7

    CHO BONG-GUM (Laboratory of Enteric Infections, Department of Microbiology, National Institute of Health, Department of Applied Biochemistry, College of Natural Sciences, Konkuk University ) , KIM CHEORL-HO (National Research Laboratory for Glycobiology, Ministry of Science and Technology of Korean Government and Department of Biochemistry and Molecular Biology, Dongguk University COM ) , LEE BOK KWON (Laboratory of Enteric Infections, Department of Microbiology, National Institute of Health ) , CHO SEUNG-HAK (Laboratory of Enteric Infections, Department of Microbiology, National Institute of Health)
    Journal of microbiology and biotechnology v.15 no.4 ,pp. 728 - 733 , 2005 , 1017-7825 ,

    초록

    To elucidate the question of whether biofilm formed by the intercellular adhesion (ica) gene cluster has influences on antibiotic resistance in Staphylococcus epidermidis, we compared 124 skin strains with strains isolated from 50 blood cultures that cause septicemic diseases. The results revealed that the blood culture isolates were more resistant to the antibiotics tested than the saprophytic isolates. Moreover, antibiotic multiresistance was more prevalent in the clinical isolates. In the blood culture isolates, $46\%$ of the strains were resistant to three or more antibiotics, whereas only $12\%$ of the saprophytic isolates were resistant to three or more antibiotics. Interestingly, these characteristics were highly correlated with the biofilm formed by the ica gene cluster. In biofilm-producing strains, $84\%$ of the blood culture isolates and $44\%$ of the saprophytic isolates were antibiotic multiresistant, whereas only $22\%=;and\;9\%$ , respectively, were antibiotic multiresistant in biofilm-nonproducing strains. Additionally, in the biofilm-producing ica-positive strains, $89\%$ of the blood culture isolates and $57\%$ of the saprophytic isolates were antibiotic multiresistant. However, the rate of the antibiotic multiresistance in the ica-negative strains was very low, thus indicating that the biofim formed by the lea gene cluster in S. epidermidis is an important pathogenic factor in association with the antibiotic multiresistance.

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  9. [국내논문]   Purification and Characterization of CDMHK, a Growth Inhibitory Molecule Against Cancer Cell Lines, from Myxobacterium sp. HK1 Isolated from Korean Soil   피인용횟수: 3

    LEE HAN-KI (Division of Bioscience and Bioinformatics, Myongji University ) , LEE IN-HYE (Division of Bioscience and Bioinformatics, Myongji University ) , YIM JEE-SUN (Division of Bioscience and Bioinformatics, Myongji University ) , KIM YONG-HO (Division of Bioscience and Bioinformatics, Myongji University ) , LEE SANG-HEE (Division of Bioscience and Bioinformatics, Myongji University ) , LEE KISAY (Department of Environmental Engineering and Biotechnology, Myongji University ) , KOO YOON-MO (Department of Biological Engineering, Inha University ) , KIM SANG-JIN (Microbiology Laboratory, Korea Ocean Research and Development Institute ) , JEONG BYEONG-CHUL (Division of Bioscience and Bioinformatics, Myongji University)
    Journal of microbiology and biotechnology v.15 no.4 ,pp. 734 - 739 , 2005 , 1017-7825 ,

    초록

    Myxobacterium sp. HK1, isolated from Korean soil, degrades cellulose, differentiates to fruiting body, and its 16s rDNA has $95\%$ similarity to Polyangium sp. An anticancer molecule, CDMHK, was identified from culture broth of Myxobacterium sp. HK1, and purified by Diaion HP20, Silica gel, Sephadex LH-20 chromatography, and preparative HPLC using an YMC OSD-A C18 column. The molecular structure and formula were determined to be $C_{l2}H_{l9}N_3O_2$ (M.W 237) by MS spectrometry, 300 MHz $^{1}H\;and\;^{13}C$ NMR. The CDMHK was not active against Escherichia coli, Staphylococcus aureus, and Candida albicans. However, this molecule inhibited the growth of various cancer cell lines. The $ED_{50}$ values of CDMHK were determined to be 0.147, 0.086, 0.18, 0.166, and 0.142 $\mu$ g/ml against A549, SK-OV-3, SK-MEL-2, VF498, and HCTl5 cancer cell lines, respectively. In addition, the CDMHK was able to induce apoptosis of the CCRF-CEM cancer cell line, evidenced by DNA fragmentation assay and DAPI staining.

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  10. [국내논문]   Effect of Bifidobacterium Cell Fractions on IL-6 Production in RAW 264.7 Macrophage Cells   피인용횟수: 7

    Lee, Byung-Hee (Department of Food Science and Nutrition, Seoul National University ) , Ji, Geun-Eog (Research Center, BIFIDO Co., LTD.)
    Journal of microbiology and biotechnology v.15 no.4 ,pp. 740 - 744 , 2005 , 1017-7825 ,

    초록

    Bifidobacterium has been previously shown to potentiate immune function, which was mediated through the stimulation of cytokine production by macrophage. This study was performed to further characterize the effective component of Bifidobacterium by measuring the level of interleukin (IL)-6 cytokine using the RAW 264.7 murine cell line as a macrophage model. RAW 264.7 cells were cultured for 24 h in the presence of whole cells (WCs), cell walls (CWs), and cell-free extracts (CFEs) from various strains of Bifidobacterium and other lactic acid bacteria at various concentrations. The most effective component was different depending on the strains and the concentrations used. When tested with each cell fraction from Bifidobacterium sp. BGN4, heat treatment of the cell fractions lowered the production of IL-6. Synergistic effect was obtained, especially when CWs and CFEs were combined. Sonicated WCs stimulated IL-6 production more than intact WCs. The in vitro approaches employed here should be useful in further characterization of the effects of Bifidobacterium on gastrointestinal and systemic immunity.

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