• 제목/요약/키워드: Soil DNA

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한국 선충 포식성곰팡이 분류검색표 및 Arthrobotrys flagrans 와 A. superba의 재기재 (Key to the Korean Nematode-Trapping Fungi with Additional Descriptions of Arthrobotrys flagrans and A. superba)

  • 서종민;강헌일;권기윤;박남숙;배창환;최인수
    • 한국균학회지
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    • 제47권4호
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    • pp.291-301
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    • 2019
  • 선충 포식성곰팡이는 선충 포획 기관을 만들어 선충 포획 후 양분을 섭취하는 곰팡이다. 이들 중 Arthrobotrys flagrans와A. superba에 대한 특성들 중 보고될 때 생략된 것이 있어 두 종을 토양으로 부터 분리하고 순수배양하여 추가적으로 조사하였다. 선충 종류별 포식력, 포식기관의 형태·크기, 분생포자의 형태·크기, 분생포자병의 형태, 후막포자 등을 조사하였으며, ITS rDNA 염기서열의 분자계통학적 분석을 토대로 종 동정을 실시하였다. 또한 1981년 처음으로 선충 포식성곰팡이가 국내에서 보고된 이래로 현재까지 총 21종이 발견되었으나 이들에 대한 분류 체계와 주요 식별 형질이 없는 상황이었기에 제공하였다. 이를 바탕으로 아직 연구가 많이 진행되지 않은 국내 선충 포식성곰팡이 연구에 기초자료가 될 수 있기를 기대한다.

Isolation, Screening, and Identification of Actinomycetes with Antifungal and Enzyme Activity Assays against Colletotrichum dematium of Sarcandra glabra

  • Song, Lisha;Jiang, Ni;Wei, Shugen;Lan, Zuzai;Pan, Limei
    • Mycobiology
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    • 제48권1호
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    • pp.37-43
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    • 2020
  • A serious leaf disease caused by Colletotrichum dematium was found during the cultivation of Sarcandra glabra in Jingxi, Rong'an, and Donglan Counties in Guangxi Province, which inflicted huge losses to plant productivity. Biological control gradually became an effective control method for plant pathogens. Many studies showed that the application of actinomycetes in biological control has been effective. Therefore, it may be of great significance to study the application of actinomycetes on controlling the diseases caused by S. glabra. Strains of antifungal actinomycetes capable of inhibiting C. dematium were identified, isolated and screened from healthy plants tissues and the rhizospheres in soils containing S. glabra. In this study, 15 actinomycetes strains were isolated and among these, strains JT-2F, DT-3F, and JJ-3F, appeared to show antagonistic effects against anthracnose of S. glabra. The strains JT-2F and DT-3F were isolated from soil, while JJ-3F was isolated from plant stems. The antagonism rate of strain JT-2F was 86.75%, which was the highest value among the three strains. Additionally, the JT-2F strain also had the strongest antagonistic activity when the antagonistic activities were tested against seven plant pathogens. Strain JT-2F is able to produce proteases and cellulase to degrade the protein and cellulose components of cell walls of C. dematium, respectively. This results in mycelia damage which leads to inhibition of the growth of C. dematium. Strain JT-2F was identified as Streptomyces tsukiyonensis based on morphological traits and 16S rDNA sequence analysis.

Reports of Drawida (Oligochaeta: Moniligastridae) from far East Asia

  • Blakemore, Robert J.;Lee, Seunghan;Seo, Hong-Yul
    • Journal of Species Research
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    • 제3권2호
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    • pp.127-166
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    • 2014
  • Moniligastrids are an important yet often ignored earthworm group commonly found in cultivated soils, especially paddy, in the tropical East. Seven new taxa are: Drawida koreana austri, D. koreana nanjiro, D. koreana shindo, D. odaesan, D. jeombongsan, D. companio and D. csuzdii Blakemore spp. or sub-spp. nov. from Korea. Drawida csuzdii is the first new species from North Korea since Lumbricidae Eisenia koreana (Zicsi, 1972). Historical East Asian moniligastrids are reviewed chronologically and Drawida barwelli (Beddard, 1886), D. japonica (Michaelsen, 1892) and D. siemsseni Michaelsen, 1910 are compared on their museum types. These three taxa were thought similar and related to D. nepalensis Michaelsen, 1907 and its possible synonym D. burchardi Michaelsen, 1903 (priority!) and both of these to prior D. uniqua (Bourne, 1887). Indian Drawida calebi Gates, 1945 is compared to new material of D. japonica from Japan, and D. willsi Michaelsen, 1907 to the new sub-species of D. koreana Kobayashi, 1938 from Korea. Where available, mtDNA COI gene barcodes are provided to help objective determinations and a phylogram is provided with outgroup Ocnerodrilidae Eukerria saltensis (Beddard, 1895) itself found in rice paddy/irrigation. The challenge now is comparison of all early taxa in their various homelands in order to assess the genetic variability and taxonomic boundaries acceptable, especially for unpigmented D. barwelli and also for pink/grey D. japonica and blue/grey D. koreana. A checklist of moniligastrids is appended showing 22 species from China (including Hainan and Taiwan), 21 from Korea, nine from Japan and the Drawida ghilarovi Gates, 1969 species-complex from far eastern Russian (Siberia). Recent Drawida dandongensis Zhang & Sun, 2014 from Sino-Korean border is misdescribed and cannot be meaningfully compared to any other Drawidas.

유기용매내성 세균 Bacillus sp. BCNU 5005의 유용성에 대한 검증 (Evaluation of the Potential of Organic Solvent Tolerant Bacillus sp. BCNU 5005)

  • 최혜정;황민정;정영기;주우홍
    • 생명과학회지
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    • 제21권5호
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    • pp.700-705
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    • 2011
  • 농화배양법을 이용하여 울산공단 일대의 폐수 및 토양에서 유기용매 내성 Bacillus sp. BCNU 5005를 분리하였다. 16S 리보좀DNA 염기서열 분석결과 BCNU 5005 균주는 B. subtilis와 98% 상동성을 가진 것으로 나타났으며 계통학적으로도 B. subtilis임이 확인되었다. 일반적으로 대부분의 세균과 그들의 효소는 고농도 유기용매하에서 불활성화되거나 파괴된다. 그러나. Bacillus sp. BCNU 5005의 lipase 활성은 chloroform, ethylbenzene 그리고 decane을 제외한 다양한 종류의 유기용매(25%, v/v)에서 매우 안정함을 보였다. 게다가 BCNU 5005는 유기용매를 분해하는 능력을 가진 것으로 확인하였다. 유기용매 내성 Bacillus sp. BCNU 5005는 생물전환과 생물복구산업을 위한 새로운 잠재적인 자원으로서 이용될 수 있다.

Isolation and Characterization of Ethanol-Producing Schizosaccharomyces pombe CHFY0201

  • Choi, Gi-Wook;Um, Hyun-Ju;Kim, Mi-Na;Kim, Yule;Kang, Hyun-Woo;Chung, Bong-Woo;Kim, Yang-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제20권4호
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    • pp.828-834
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    • 2010
  • An ethanol-producing yeast strain, CHFY0201, was isolated from soil in South Korea using an enrichment technique in a yeast peptone dextrose medium supplemented with 5% (w/v) ethanol at $30^{\circ}C$. The phenotypic and physiological characteristics, as well as molecular phylogenetic analysis based on the D1/D2 domains of the large subunit (26S) rDNA gene and the internally transcribed spacer (ITS) 1+2 regions, suggested that the CHFY0201 was a novel strain of Schizosaccharomyces pombe. During shaking flask cultivation, the highest ethanol productivity and theoretical yield of S. pombe CHFY0201 in YPD media containing 9.5% total sugars were $0.59{\pm}0.01$ g/l/h and $88.4{\pm}0.91%$, respectively. Simultaneous saccharification and fermentation for ethanol production was carried out using liquefied cassava (Manihot esculenta) powder in a 5-l lab-scale jar fermenter at $32^{\circ}C$ for 66 h with an agitation speed of 120 rpm. Under these conditions, S. pombe CHFY0201 yielded a final ethanol concentration of $72.1{\pm}0.27$ g/l and a theoretical yield of $82.7{\pm}1.52%$ at a maximum ethanol productivity of $1.16{\pm}0.07$ g/l/h. These results suggest that S. pombe CHFY0201 is a potential producer for industrial bioethanol production.

Purification and Characterization of Cell Wall Hydrolase from Alkalophilic Bacillus mutanolyticus YU5215

  • 옥승호;남승우;김진만;유윤정;배동훈
    • Journal of Microbiology and Biotechnology
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    • 제14권6호
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    • pp.1142-1149
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    • 2004
  • Streptococcus mutans has the capacity of inducing dental caries. Thus, to develop a novel way of preventing dental caries, a cell wall hydrolase-producing strain was isolated and its characteristics were investigated. Among 200 alkalophilic strains isolated from soil, 8 strains exhibited lytic activities against Streptococcus mutans. However, strain YU5215 with the highest cell wall hydrolase activity was selected for further study. Strain YU5215 was identified as a novel strain of Bacillus based on analyzing its 16S rDNA sequence and Bergey's Manual of Systematic Bacteriology, and thus designated as Bacillus mutanolyticus YU5215. The optimal conditions for the production of the cell wall hydrolase from Bacillus mutanolyticus YU5215 consisted of glucose ($0.8\%$), yeast extract ($1.2\%$), polypeptone ($0.5\%$), $K_{2}HPO_{4}\;(0.1\%$), $MgSO_{4}{\cdot}7H_{2}O$ ($0.02\%$), and $Na_{2}CO_{3}\;(1.0\%$) at pH 10.0. Bacillus mutanolyticus YU5215 was cultured at 30^{circ}C for 72 h to produce the cell wall hydrolase, which was then purified by acetone precipitation and CM-agarose column chromatography. The molecular weight of the lytic enzyme was determined as 22,700 Da by SDS-PAGE. When the cell wall peptidoglycan of Streptococcus mutans was digested with the lytic enzyme, no increase in the reducing sugars was observed, while the free amino acids increased, indicating that the lytic enzyme had an endopeptidase-like property. The amino terminus of the cell wall peptidoglycan digested by the lytic enzyme was determined as a glutamic acid, while the lytic site of the lytic enzyme in the Streptococcus mutans peptidoglycan was identified as the peptide linkage of L-Ala and D-Glu.

Gene Cloning of Streptomyces Phospholipase D P821 Suitable for Synthesis of Phosphatidylserine

  • Moon Min-Woo;Lee Jung-Kee;Oh Tae-Kwang;Shin Chul-Soo;Kim Hyung-Kwoun
    • Journal of Microbiology and Biotechnology
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    • 제16권3호
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    • pp.408-413
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    • 2006
  • A strain, P821, with phospholipase D activity was isolated from soil and identified as a Streptomyces species. The phospholipase D enzyme was purified from a culture broth of the isolated strain using ammonium sulfate precipitation and DEAE-Sepharose, phenyl-Sepharose, and Superose 12 HR column chromatographies. The purified enzyme exhibited an optimum temperature and pH of $55^{\circ}C$ and 6.0, respectively, in the hydrolysis of phosphatidylcholine and remained stable up to $60^{\circ}C$ within a pH range of 3.5-8.0. The enzyme also catalyzed a transphosphatidylation reaction to produce phosphatidylserine with phosphatidylcholine and serine substrates. The optimum conditions for the transphosphatidylation were $30^{\circ}C$ and pH 5.0, indicating quite different optimum conditions for the hydrolysis and transphosphatidylation reactions. The gene encoding the enzyme was cloned by Southern hybridization and colony hybridization using a DNA probe designed from the conserved regions of other known phospholipase D enzymes. The resulting amino acid sequence was most similar to that of the PLD enzyme from Streptomyces halstedii (89.5%). Therefore, the enzyme was confirmed to be a phospholipase D with potential use in the production of phosphatidylserine.

An Antifungal Antibiotic Purified from Bacillus megaterium KL39, a Biocontrol Agent of Red-Pepper Phytophthora-Blight Disease

  • JUNG HEE KYOUNG;KIM SANG-DAL
    • Journal of Microbiology and Biotechnology
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    • 제15권5호
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    • pp.1001-1010
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    • 2005
  • Bacillus megaterium KL39, an antibiotic-producing plant growth promoting rhizobacterium (PGPR), was selected from soil. The antifungal antibiotic, denoted KL39, was purified from culture filtrate by column chromatography using Dion HP-20, Silica gel, Sephadex LH-20, and prep-HPLC. Thin layer chromatography, employing the solvent system of ethanol:ammonia:water=8:1:1, showed the $R_{f}$. value of 0.32. The antibiotic KL39 showed a negative reaction with ninhydrin solution, positive with iodine vapor, and also positive with Ehrlich reagent. It was soluble in methanol, ethanol, butanol, and acetonitrile, but insoluble in chloroform, toluene, hexane, ethyl ether, or acetone. Its UV spectrum had the maximum absorption at 208 nm. Amino acid composition, FAB-mass, $^{1}H-NMR,\;^{13}C-NMR$, and atomic analyses showed that the antibiotic KL39 (MW=1,071) has a structure very similar to iturin E. The antibiotic KL39 has a broad antifungal spectrum against a variety of plant pathogenic fungi including Rhizoctonia solani, Pyricularia oryzae, Monilinia froeticola, Botrytis cinenea, Altenaria kikuchiana, Fusarium oxysporum, and F. solani. An MIC value of $10\;{\mu}g/ml$ was determined for Phytophthora capsici. Macromolecular incorporation studies with P. capsici using radioactive [$^{3}H-adenine$] as the precursor, indicated that the antibiotic KL39 strongly inhibits the DNA biosynthesis of the fungal cell. Microscopic observation of the antifungal action showed abnormal hyphal swelling of P. capsici. The purified antibiotic KL39 was very effective for the biocontrol of in vivo Phytophthora-blight disease of pepper.

Isolation and Biological Properties of Novel Cell Cycle Inhibitor, HY558, Isolated from Penicillium minioluteum F558

  • Lee, Chul-Hoon;Lim, Hae-Young;Kim, Min-Kyoung;Cho, Youl-Hee;Oh, Deok-Kun;Kim, Chang-Jin;Lim, Yoon-Gho
    • Journal of Microbiology and Biotechnology
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    • 제12권3호
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    • pp.470-475
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    • 2002
  • In the course of screening for a novel cell cycle inhibitor, a potent Cdk 1 inhibitor, HY558, was found from the culture broth of Penicillium minioluteum F558 isolated from a soil sample. The molecular ion of HY558 was identified at m/z 329 (MH+) with a molecular formula of $C_20H_44ON_2$. HY558 exhibited selective antiproliferative effects on various human cancer cell lines. Its $IC_50$ values were estimated to be 0.29 mM on HepG2, 0.30 mM on HeLa, 0.30 mM on HL6O, 0.33 mM on HT-29, and 0.25 mM on AGS cells. Interestingly, Hy558 demonstrated no antiproliferative effect with normal lymphocytes used as the control, and a low level of inhibition on the proliferation of A549 cancer cells. A flow cytometric analysis of HepG2 cells revealed an appreciable arrest of cells at the G1 and G2/M phases of the cell cycle following treatment with Hy558. furthermore, DNA fragmentation due to apoptosis was observed in HeLa cells treated with 0.46 mM of HY558.

Purification and Biochemical Characterization of a Novel Fibrinolytic Enzyme from Streptomyces sp. P3

  • Cheng, Guangyan;He, Liying;Sun, Zhibin;Cui, Zhongli;Du, Yingxiang;Kong, Yi
    • Journal of Microbiology and Biotechnology
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    • 제25권9호
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    • pp.1449-1459
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    • 2015
  • A novel proteolytic enzyme with fibrinolytic activity, FSP3, was purified from the recently isolated Streptomyces sp. P3, which is a novel bacterial strain isolated from soil. FSP3 was purified to electrophoretic homogeneity by ammonium sulfate precipitation, anion exchange, and gel filtration. FSP3 is considered to be a single peptide chain with a molecular mass of 44 kDa. The maximum activity of the enzyme was observed at 50℃ and pH 6.5, and the enzyme was stable between pH 6 and 8 and below 40℃. In a fibrin plate assay, FSP3 showed more potent fibrinolytic activity than urokinase, which is a clinical thrombolytic agent acting as a plasminogen activitor. The activity was strongly inhibited by the serine protease inhibitor PMSF, indicating that it is a serine protease. Additionally, metal ions showed different effects on the activity. It was significantly suppressed by Mg2+ and Ca2+ and completely inhibited by Cu2+, but slightly enhanced by Fe2+. According to LC-MS/MS results, its partial amino acid sequences are significantly dissimilar from those of previously reported fibrinolytic enzymes. The sequence of a DNA fragment encoding FSP3 contained an open reading frame of 1287 base pairs encoding 428 amino acids. FSP3 is a bifunctional enzyme in nature. It hydrolyzes the fibrin directly and activates plasminogen, which may reduce the occurrence of side effects. These results suggest that FSP3 is a novel serine protease with potential applications in thrombolytic therapy.