• Title/Summary/Keyword: species characterization

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Purification and Characterization of Transglutaminase from a Newly Isolated Streptomyces platensis YK-2 (토양 방선균 Streptomyces platensis YK-2가 생산하는 Transglutaminase의 정제 및 효소학적 특성)

  • Ko, Hee-Sun;Kim, Hyun-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.6
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    • pp.801-806
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    • 2009
  • A species producing transglutaminase (EC 2.3.2.13) was isolated from forest soil and identified as Streptomyces platensis YK-2. The transglutaminase was purified from culture broth by 50% methanol precipitation, followed by successive chromatography on DEAE-Sephadex. The yield and purification-fold was 63.4% and 2.2-fold, respectively. The purified microbial transglutaminase (MTG) migrated as a single band of approximately 45 kDa upon sodium dodecyl sulfate polyacrylamide gel eletrophoresis. The isoelectric point determined by multichambered electrofocusing was pH $6.0{\sim}7.0$. The enzyme was strongly inhibited by $Hg^{++}$, but was activated by $Cd^{++}$, $Mg^{++}$, $Mn^{++}$, $Pb^{++}$ and reducing agents such as dithiothreitol and mercaptoethanol.

Production and Characterization of Monoclonal Antibodies to Glutamate Dehydrogenase from Thermophile Sulfolobus solfataricus

  • Cho, Sung-Woo;Ahn, Jee-Yin;Bahn, Jae-Hoon;Jeon, Seong-Gyu;Park, Jin-Seu;Lee, Kil-Soo;Choi, Soo-Young
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.587-594
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    • 2000
  • Monoclonal antibodies against glutamate dehydrogenase (GDH) from Sulfolobus solfataricus were produced and characterized using epitope mapping and biosensor technology, Five monoclonal antibodies raised against S. solfataricus GDH were each identified as a single protein band that comigrated with purified S. solfataricus GDH on the SDS-polyacrylamide gel electrophoresis and immunoblot. Epitope mapping analysis showed that only one subgroup among the antibodies tested recognized the same peptide fragments of GDH. Using the anti-S. solfataricus GDH antibodies as probes, the cross-reactivities of GDHs from various sources were investigated and it was found that the mammalian GDH is not immunologically related to S. solfataricus GDH. The structural differences between the microbial and mammalian GDHs were further investigated using biosensor technology (Pharmacia BIAcore) and monoclonal antibodies against S. solfataricus and bovine brain. The binding affinity of S. solfataricus glutamate dehydrogenase anti-S. solfataricus for GDH ($K_D$=11 nM) was much tighter than that of anti-bovine for GDH ($K_D$=450 nM). These results, together with the epitope mapping analysis, suggest that there may be structural differences between the two GDH species, in addition to their different biochemical properties.

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Characterization of Water Quality and Microbial Communities in Rivers in Changwon city (창원시 하천의 수질 및 미생물상 분석)

  • Kim Sun-A;Kim Chung-Hye;Lim Byung-Ran;Cho Kwang-Hyun;Park Hee-Chang;Joo Woo Hong
    • Journal of Life Science
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    • v.16 no.1
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    • pp.148-155
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    • 2006
  • The diversity of bacterial populations in rivers flowing through Changwon City, was investigated using quinone profiling. The physicochemical properties such as temperature, pH, dissolved oxygen(DO), dissolved organic carbon (DOC) and biochemical oxygen demand (BOD) were also measured in this study. Ubiquinone (UQ)-8, UQ-9 and UQ-10 were observed in all samples for the sites investigated. UQ-8 was the -predominant quinone species in rivers except for Namch'on downstream, T'owolch'on, and Kaumchongch'on in autumn, while UQ-8 was also found as major quinones in the sample except for Hanamch'on, T'owolch'on, Kaumchongch'on, and Namsanch'on in winter. A higher concentration of DOC in rivers yield high concentration of plastoquinone (PQ-9) in autumn and those of total quinones in winter, respectively. Correlation analysis also indicate that BOD is considered to be a major factor controlling the concentration of PQ in rivers.

Isolation and Characterization of an Agarase-Producing Bacterial Strain, Alteromonas sp. GNUM-1, from the West Sea, Korea

  • Kim, Jonghee;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1621-1628
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    • 2012
  • The agar-degrading bacterium GNUM-1 was isolated from the brown algal species Sargassum serratifolium, which was obtained from the West Sea of Korea, by using the selective artificial seawater agar plate. The cells were Gram-negative, $0.5-0.6{\mu}m$ wide and $2.0-2.5{\mu}m$ long curved rods with a single polar flagellum, forming nonpigmented, circular, smooth colonies. Cells grew at $20^{\circ}C-37^{\circ}C$, between pH 5.0 and 9.0, and at 1-10% (w/v) NaCl. The DNA G+C content of the GNUM-1 strain was 45.5 mol%. The 16S rRNA sequence of the GNUM-1 was very similar to those of Alteromonas stellipolaris LMG 21861 (99.86% sequence homology) and Alteromonas addita $R10SW13^T$(99.64% sequence homology), which led us to assign it to the genus Alteromonas. It showed positive activities for agarase, amylase, gelatinase, alkaline phosphatase, esterase (C8), lipase (C14), leucine arylamidase, valine arylamidase, ${\alpha}$-chymotrypsin, acid phosphatase, naphthol-AS-BI-phosphohydrolase, ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\beta}$-glucosidase, catalase, and urease. It can utilize citrate, malic acid, and trisodium citrate. The major fatty acids were summed feature 3 (21.5%, comprising $C_{16:1}{\omega}7c/iso-C_{15:0}$ 2-OH) and C16:0 (15.04%). On the basis of the variations in many biochemical characteristics, GNUM-1 was considered as unique and thus was named Alteromonas sp. GNUM-1. It produced the highest agarase activity in modified ASW medium containing 0.4% sucrose, but lower activity in rich media despite superior growth, implying that agarase production is tightly regulated and repressed in a rich nutrient condition. The 30 kDa protein with agarase activity was identified by zymography, and this report serves as the very first account of such a protein in the genus Alteromonas.

Debaryomyces hansenii Strains from Valle De Los Pedroches Iberian Dry Meat Products: Isolation, Identification, Characterization, and Selection for Starter Cultures

  • Ramos, Jose;Melero, Yessica;Ramos-Moreno, Laura;Michan, Carmen;Cabezas, Lourdes
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1576-1585
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    • 2017
  • Yeasts, filamentous fungi, and bacteria colonize the surface of fermented sausages during the ripening process. The source of this microbiota is their surrounding environment, and is influenced by the maturing conditions and starter cultures. Debaryomyces hansenii was previously isolated from several dry-cured meat products and associated with the lipolytic and proteolytic changes that occur in these products, influencing their taste and flavor. Therefore, this study isolated the yeast microbiota present in the casing from different meat products ("lomo," "chorizo," and "$salchich{\acute{o}}n$") from the Valle de los Pedroches region in southern Spain. D. hansenii was by far the most abundant species in each product, as all 22 selected isolates were identified as D. hansenii by biochemical and/or molecular methods. In contrast, no yeasts were found in the meat batter. These data constitute the first study of the yeasts present in "lomo" sausages and particularly the highly appreciated Valle de los Pedroches "lomo" sausages. Furthermore, the resistance of these isolates to different pHs, temperatures, and saline stress was studied, together with their catabolic characteristics. Based on the results, certain isolates are proposed as valuable candidate starter cultures that could improve both the manufacture and the flavor of such dry-cured meat products, and provide an understanding of new mechanisms involved in stress tolerance. Applied medium-scale industrial tests are currently in progress.

Preparation and Characterization of Ni-Co Bimetallic Catalyst for Methanation (메탄화 반응을 위한 Ni-Co 이원 금속 촉매의 제조와 특성 분석)

  • Yia, Jong-Heop;Kanga, Mi-Yeong;Kim, Woo-Young;Cho, Won-Jun
    • Journal of the Korean Institute of Gas
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    • v.13 no.5
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    • pp.33-38
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    • 2009
  • Synthetic natural gas was producd by the reaction of carbon monoxide and hydrogen via methanation. Ni-Co bimetallic catalyst supported on $Al_2O_3$ for methanation was prepared using deposition-precipitation method. For the comparison, Ni, Co monometallic catalyst was prepared using the same method. The prepared catalysts were characterized by TEM, XRD and TPR and applied to methanation reaction. The catalysts prepared using deposition-precipitation method showed the high metal dispersion. The activity of Ni-Co bimetallic catalyst was higher than that of Ni, Co monometallic catalyst. TPR measurements indicated that Ni-Co bimetallic catalyst had more active hydrogen species than Ni, Co monometallic catalyst due to the synergetic effect in the presence of Ni and Co.

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Assessment of Monitored Natural Attenuation as Remediation Approach for a BTEX Contaminated Site in Uiwang City (의왕시내 BTEX 오염 부지에서의 자연 정화법 이용 적합성 고찰)

  • 이민효;윤정기;박종환;이문순;강진규;이석영
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 1999.04a
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    • pp.149-156
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    • 1999
  • In the United States (U.S.), the monitored natural attenuation (MNA) approach has been used as an alternative remedial option for organic and inorganic compounds retained in soil and dissolved in groundwater. The U.S. Environmental Protection Agency (EPA) defines the MNA as“in-situ naturally-occurring processes include biodegradation, diffusion, dilution, sorption, volatilization, and/or chemical and biochemical stabilization of contaminants and reduce contaminant toxicity, mobility or volume to the levels that are protective of human health and the environment”. The Department of Soil Environment. National Institute Environmental Research (NIER) is in the process for demonstrating the MNA approach as a potential remedial option for the BTEX contaminated site in Uiwang City. The project is charactering the research site in terms of the nature and extend of contamination, biological degradation rate, and geochemical and hydrological properties. The microbial-degradation rate and effectiveness of nutrient and redox supplements will be determined through laboratory batch and column tests. The geochemical process will be monitored for determining the concentration changes of chemical species involved in the electron transfer processes that include methanogenesis, sulfate and iron reduction, denitrification, and aerobic respiration. Through field works, critical soil and hydrogeologic parameters will be acquired to simulate the effects of dispersion, advection, sorption, and biodegradation on the fate and transport of the dissolved-phase BTEX plume using Bioplume III model. The objectives of this multi-years research project are (1) to evaluate the MNA approach using the BTEX contaminated site in Uiwang City, (2) to establish a standard protocol for future application of the approach, (3) to investigate applicability of the passive approach as a secondary treatment remedy after active treatments. In this presentation, the overall picture and philosophy behind the MNA approach will be reviewed. Detailed discussions of the site characterization/monitoring plans and risk-based decision-making processes for the demonstration site will be included.

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Biological and Molecular Characterization of a Korean Isolate of Cucurbit aphidborne yellows virus Infecting Cucumis Species in Korea

  • Choi, Seung-Kook;Yoon, Ju-Yeon;Choi, Gug-Seoun
    • The Plant Pathology Journal
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    • v.31 no.4
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    • pp.371-378
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    • 2015
  • Surveys of yellowing viruses in plastic tunnels and in open field crops of melon (Cucumis melo cultivar catalupo), oriental melon (C. melo cultivar oriental melon), and cucumber (C. sativus) were carried out in two melon-growing areas in 2014, Korea. Severe yellowing symptoms on older leaves of melon and chlorotic spots on younger leaves of melon were observed in the plastic tunnels. The symptoms were widespread and included initial chlorotic lesions followed by yellowing of whole leaves and thickening of older leaves. RT-PCR analysis using total RNA extracted from diseased leaves did not show any synthesized products for four cucurbit-infecting viruses; Beet pseudo-yellows virus, Cucumber mosaic virus, Cucurbit yellows stunting disorder virus, and Melon necrotic spot virus. Virus identification using RT-PCR showed Cucurbit aphid-borne yellows Virus (CABYV) was largely distributed in melon, oriental melon and cucumber. This result was verified by aphid (Aphis gossypii) transmission of CABYV. The complete coat protein (CP) gene amplified from melon was cloned and sequenced. The CP gene nucleotide and the deduced amino acid sequence comparisons as well as phylogenetic tree analysis of CABYV CPs showed that the CABYV isolates were undivided into subgroups. Although the low incidence of CABYV in infections to cucurbit crops in this survey, CABYV may become an important treat for cucurbit crops in many different regions in Korea, suggesting that CABYV should be taken into account in disease control of cucurbit crops in Korea.

Characterization of the Rosellinia necatrix Transcriptome and Genes Related to Pathogenesis by Single-Molecule mRNA Sequencing

  • Kim, Hyeongmin;Lee, Seung Jae;Jo, Ick-Hyun;Lee, Jinsu;Bae, Wonsil;Kim, Hyemin;Won, Kyungho;Hyun, Tae Kyung;Ryu, Hojin
    • The Plant Pathology Journal
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    • v.33 no.4
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    • pp.362-369
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    • 2017
  • White root rot disease, caused by the pathogen Rosellinia necatrix, is one of the world's most devastating plant fungal diseases and affects several commercially important species of fruit trees and crops. Recent global outbreaks of R. necatrix and advances in molecular techniques have both increased interest in this pathogen. However, the lack of information regarding the genomic structure and transcriptome of R. necatrix has been a barrier to the progress of functional genomic research and the control of this harmful pathogen. Here, we identified 10,616 novel full-length transcripts from the filamentous hyphal tissue of R. necatrix (KACC 40445 strain) using PacBio single-molecule sequencing technology. After annotation of the unigene sets, we selected 14 cell cycle-related genes, which are likely either positively or negatively involved in hyphal growth by cell cycle control. The expression of the selected genes was further compared between two strains that displayed different growth rates on nutritional media. Furthermore, we predicted pathogen-related effector genes and cell wall-degrading enzymes from the annotated gene sets. These results provide the most comprehensive transcriptomal resources for R. necatrix, and could facilitate functional genomics and further analyses of this important phytopathogen.

Chemosystematic and Phenotypic Characterization of Gram-positive Bacteria from Coastal Seawater, Korea (한국 근해에서 분리한 그람양성 세균의 화학 분류학적 및 표현형적 특성)

  • 전정훈;박진숙
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.167-172
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    • 2000
  • Twenty-five halotolerant gram-positlve bacteria were isolated from the coastal seawater 01 Cheju Island and Incheon J&yakdo Chemosystematic and phenotypic characteristics were used to iuvestigate the taxonomic position of these bacteria. According to their chemosystematic characteristics, the twenty-tive isolates were divided into 4 groups. Group 1 bacteria possesed 40.1 to 49.9 inol% m DNA G+C content, menaquinone-7 as a major quinone, and meso-Alpm as a diamino acid of peptidoglycan. Group 1 tam were identified as Bacilluspumilis, Bacillus lichenifbrrizis, Bacillus megaterium, Bncill~rsubtilis. Group 2 bacteria possessed 63.9 to 66.4 mol% and MK-8. They were all in the genus Arth~obaclm-. Group 3 bacteria possessed 31.0 to 37.6 mol% and MK-7. They were identified as Staphylococcus haeniol.viicvs, Siaph~~lococc~is sapropl~j~ticns, and Siaphylococcus intermediru.. Group 4 bacterium possessed 72.0mol% and MK-8 and was identified as ~Lficrococcus ltdtezm. Bacillus species accounted for 68% of h e total isolates.

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