• Title/Summary/Keyword: Soil DNA

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Studies on the Isolation and Identification of Xylanase and Mannanase Producing Aspergillus niger (Xylanase와 Mannanase를 생산하는 Aspergillus niger의 분리와 동정에 관한 연구)

  • Kim, Byoung-Suk;Cho, Jin-Kook;Song, Jin-Ook;Lee, Hak-Kyo;Hwang, Seong-Gu
    • Journal of Animal Science and Technology
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    • v.51 no.5
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    • pp.427-432
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    • 2009
  • This study was undertaken to screen a high xylanase and mannanase producing microbes. In the first experiment, screening was undertaken against 50 samples of microorganisms having xylanase and mannanase activities from soil and fallen leaves. The screening process has focused on picking out fungi having high xylanase and mannanase activities under the solid-state fermentation. The xylanase and mannanase activities of 6 screened microbes were 0.9~1.6 unit/mL and 0.2~0.4 unit/mL, respectively, under the submerged fermentation condition. However, under the solid-state fermentation, xylanase and mannanase activities were 103.7~220.0 unit/g and 20.1~40.3 unit/g, respectively. Finally one microbe (E-3) was selected and its xylanase and mannanase activities were 197.3 unit/g and 39.9 unit/g, respectively. The morphological and molecular biological classification of E-3 showed 99% homology with the Aspergillus niger.

Physiological properties and transformation of alkaline-tolerant bacteria (알카리내성 세균의 생리적 특성 및 형질전환)

  • 유주현;정용준;정건섭;오두환
    • Microbiology and Biotechnology Letters
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    • v.14 no.3
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    • pp.239-244
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    • 1986
  • To develop the potential use as new host strain for gene cloning, alkaline-tolerant isolates from soil were examined for amylase activity, protease activity, antimicrobial activity and transformability by using plasmid pUB 110. Of these strains, one was selected and identified as Bacillus sp. YA-14. in the enzymatic properties of Bacillus sp. YA-14 the optimal conditions for the reaction of amylase and protease were at pH 0.8 and pH 7.5 respectively. The antimicrobial activity of Bacillus sp. YA-14 was also found. For the transformation, Bacillus sp. YA-14 was cultured to late logarithmic growth phase ai 37$^{\circ}C$ in modified SPI medium (pH 8.0) containing 0.4% MgSO$_4$. The presence of pUB 110 plasmid DNA in transformants was confirmed by electrophoresis and stably maintained in the new host.

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Selection and Antagonistic Mechanism of Pseudomonas fluorescens 4059 Against Phytophthora Blight Disease (고추역병과 시들음병을 방제하는 토착길항세균 Pseudomonas fluorescens 4059의 선발과 길항기작)

  • Jeong, Hui-Gyeong;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.32 no.4
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    • pp.312-316
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    • 2004
  • In oder to select the powerful rhizophere-dorminatable biocontrol agent, we had isolated an indigenous antagonistic bacterium which produced antibiotic and siderophore from a disease suppressive local field soil of Gyungsan, Korea. And we could select the Pseudomosp. 4059 which can strongly antagonize against Fusarium oxysporum and Phytophthora capsici by two kinds of antifungal mechanism that can be caused by the antibiotic of Phenazin, a siderophore and a auxin like subThe selected strain was identified as Pseudomonas fluorescens (biotype A) 4059 by biochemical tests, API $\textregistered$ test, MicroLog TM system and 16S rDNA analysis. The selected antagonistic microorganism, Pseudomosp. 4059 had an antifungal mechanism of antifungal antibiotic and sidrophore. And we were confirmed the antagonistic activity of P fluorescens 4059 with in vitro antifungal test against Phytophthora capsici and in vivo by red-pepper.

Diversities of Arbuscular Mycorrhizal Fungi in Cultivated Field Soils of Korean Ginseng (금산의 인삼 재배 토양에 분포하는 수지상균근균의 다양성)

  • Kil, Yi-Jong;Eo, Ju-Kyeong;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.40 no.1
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    • pp.1-6
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    • 2012
  • In this study, soil samples were collected from cultivated fields of 1-5 year old Korean ginseng in Geumsan, Korea. Spores of arbuscular mycorrhizal fungi were extracted from soils and identified using morphological characteristics and 18s rDNA sequences of the spores. Total 10 species of AMF were identified: Acaulospora longula, Archaeospora trappei, Glomus caledonium, Glomus etunicatum, Glomus intraradices, Glomus mosseae, Glomus sp., Paraglomus occultum, Paraglomus brasilianum, and Scutellospora heterogama. Relative abundance of spores of A. trappei were increased with increase of cultivation period of the ginseng. However, relative abundance of other species of AMF and Shannon diversity (H') of AMF were significantly decreased with the increase of cultivation periods of the ginseng.

Comparison of Characteristics between Insecticidal and Noninsecticidal Basillus thuringiensis Strains belonging to Serotype H8a8b

  • Roh, Jong-Yul;Je, Yeon-Ho;Park, Hyun-Woo;Chang, Jin-Hee;Jin, Byung-Rae;Lee, Dae-Weon;Ziwen Yang;Kang, Seok-Kwon
    • Journal of Sericultural and Entomological Science
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    • v.40 no.2
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    • pp.126-130
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    • 1998
  • A noninsecticidal strain, Bacillus thuringiensis NTB-88, isolated from Korean soil, had a typical bipyramidal parasporal inclusion and its serotype is identical to B. thuringiensis subspmorrisoni (H8a8b). To elucidate differences between insecticidal and noninsecticidal strains, we compared strain NTB-88 to other toxic B. thuringiensis subsp. morrisoni strains (HD-12 and PG-14). Restriction endonucleases digested plasmid DNA patterns showed that strain NTB-88 was different from lepidopteran-toxic strain, HD-12, but it was similar to dipteran-toxic strain, PG-14. The gene type of strain NTB-88 was different from those of other insecticidal strains, Furthermore, the NH2-terminal amino acid sequence of crystal protein of strain NTB-88 had no relation to those of the previously known $\delta$-endotoxins in other toxic strains as well as HD-12 and PG-14 strains. Therefore, the noninsecticidal crystal protein in strain NTB-88 is novel and its property is different from insecticidal ones.

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Enhanced Biosynthesis of ${\alpha}$-tocopherol in Transgenic Soybean by Introducing ${\gamma}$-TMT gene

  • Kim Young-Jin;Seo Hong-Yul;Park Tae-Il;Baek So-Hyeon;Shin Woon-Chul;Kim Hyun-Soon;Kim Jung-Gon;Choi Yong-Eui;Yun Song-Joong
    • Journal of Plant Biotechnology
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    • v.7 no.3
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    • pp.203-209
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    • 2005
  • This study was conducted to improve tocopherol (vitamin E) composition in soybean (Glycine max) by introducing a gamma-tocopherol methyl transferase (${\gamma}$-TMT) gene via Agrobacterium tumefaciens-mediated transformation. Immature cotyledon explants were cocultivated with Agrobacterium tumefaciens. Putative transgenic embryos were selected from immature cotyledons on MS medium supplemented with 40 mg/L 2,4-D containing 100 mg/L kanamycin, 500 mg/L carbenicillin and 250 mg/L cefotaxime. Plantlets were developed from somatic embryos, and then transferred to soil. Nineteen regenerated plantlets obtained on the selection medium from 1,460 cotyledons. However, only 9 plantlets were confirmed as transformed plants. Integration of the transgene into the soybean genomic DNA was confirmed by PCR and Southern blot analysis. HPLC analysis showed that the content of ${\alpha}$-tocopherol in transgenic soybean seeds (AT-1) was approximately 4-fold higher than that of non-transgenic plants. Conclusively, we obtained the transgenic soybean having increased ${\alpha}$-tocopherol content by the overexpression of ${\gamma}$-TMT transgene.

Isolation and Identification of Streptomyces sp. Producing Anti-vancomycin Resistant Staphylococcus aureus Substance (반코마이신 내성 Staphylococcus aureus 억제 물질 생산 Streptomyces sp.의 분리 및 동정)

  • Oh Se-Teak;Lee Jun-Jae;Lee Ji-Youn;Kim Jin-Kyu;Yang Si-Yong;Kim Yang-Soo;Song Min-Dong
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.90-95
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    • 2005
  • An Actinomycetes producing an anti-VRSA (vancomycin-resistant Staphylococcus aureus) substance was isolated from soil. The cultural, morphological, physiological and phylogenetic analyses of an isolated strain were investigated for identification. Cultural characteristics based on ISP (International Streptomyces Project) were as follows: white aerial mycelium, yellow reverse side, and good growth on various medium. Also, the isolate did not produce the soluble pigment. Morphological characteristics were showed cylindrical spore chain and smooth spore surface by SEM (Scanning Electron Microscope). Physiological characteristics were showed LL-type by DAP isomer analysis and detected glycine, glutamic acid and alanine. A phylogenetic analysis of the 16S rDNA provided a clue that the isolated strain was actually a member of the genus Streptomyces, because the determined sequence exhibited a higher homology with Streptomyces echinatus. The isolate was identified to be a genus of Streptomyces sp.. The optimal culture conditions for the maximum production of anti-VRSA substance by Streptomyces sp. were attained in a culture medium composed of $2.0\%$ (w/v) glucose, and $0.4\%$ (w/v) yeast extract. The anti-VRSA substance was highly produced after 5 days of culture. Optimal pH and temperature conditions for the production of anti-VRSA substance were pH 7.0 and $28^{\circ}C$, respectively.

Purification of Alkaline Restriction Endonuclease from Alkalophilic Bacillus sp. 8-13 (제한효소 생성능을 지닌 알칼리성 Bacillus sp. 8-13 균주로부터 알칼리성 제한효소의 정제)

  • Bae, Moo;Lee, Jee-Eun;Park, Kyoung-Sook;Lee, Kang-Man
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.289-294
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    • 1992
  • Twenty-four bacterial strains among alkalophillic bacteria isolated from soil samples were examined for the presence of type II restriction endonuclease in aerobic culture. One strain was found to contain specific enzyme to cleave lambda DNA. The characteristics of this microorganism is the ability to grow well in alkalophilic and high temperature condition, that is at pH 10.3 and $50^{\circ}C$. This strain was tentatively identified to Bacillus alkaloPhilus subsp. halodurans when morphological, physiological and biochemical characteristics were examined. The enzyme was purified from crude extract by streptomycin sulfate, ammonium sulfate precipitation, which was followed by DEAE-cellulose and phosphocellulose ion exchange column chromatography, and the subunit molecular weight was about, 32,000 daltons by polyacrylamide gel electrophoresis containing 0.1% SDS.

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Ramlibacter ginsenosidimutans sp. nov., with Ginsenoside-Converting Activity

  • Wang, Liang;An, Dong-Shan;Kim, Song-Gun;Jin, Feng-Xie;Kim, Sun-Chang;Lee, Sung-Taik;Im, Wan-Taek
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.311-315
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    • 2012
  • A novel ${\beta}$-proteobacterium, designated BXN5-$27^T$, was isolated from soil of a ginseng field of Baekdu Mountain in China, and was characterized using a polyphasic approach. The strain was Gram-staining-negative, aerobic, motile, non-spore-forming, and rod shaped. Strain BXN5-$27^T$ exhibited ${\beta}$-glucosidase activity that was responsible for its ability to transform ginsenoside $Rb_1$ (one of the dominant active components of ginseng) to compound Rd. Phylogenetic analysis based on 16S rRNA gene sequences showed that this strain belonged to the family Comamonadaceae; it was most closely related to Ramlibacter henchirensis $TMB834^T$ and Ramlibacter tataouinensis$TTB310^T$ (96.4% and 96.3% similarity, respectively). The G+C content of the genomic DNA was 68.1%. The major menaquinone was Q-8. The major fatty acids were $C_{16:0}$, summed feature 4 (comprising $C_{16:1}$ ${\omega}7c$ and/or iso-$C_{15:0}$ 2OH), and $C_{17:0}$ cyclo. Genomic and chemotaxonomic data supported the affiliation of strain BXN5-$27^T$ to the genus Ramlibacter. However, physiological and biochemical tests differentiated it phenotypically from the other established species of Ramlibacter. Therefore, the isolate represents a novel species, for which the name Ramlibacter ginsenosidimutans sp. nov. is proposed, with the type strain being BXN5-$27^T$ (=DSM $23480^T$ = LMG $24525^T$ = KCTC $22276^T$).

Biodegradation of JP-8 by Rhodococcus fascians Isolated from Petroleum Contaminated Soil (유류 오염 토양에서 분리된 Rhodococcus fascians를 이용한 JP-8의 분해 특성)

  • Nam, Bo Hyun;Park, Bong Je;Yun, Hyun Shik
    • Korean Chemical Engineering Research
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    • v.46 no.4
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    • pp.819-823
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    • 2008
  • Microorganisms isolated from petroleum contaminated site were tested for their ability to grow on JP-8 by culturing them on the culture medium that contains JP-8 as a carbon source. The microorganism which grew on JP-8 containing minimal salt medium was separated and identified as Rhodococcus fascians. Changes in JP-8 biodegradation of R. fascians that was isolated from petroleum contaminated site was investigated with various inoculums sizes, JP-8 concentrations, medium pHs, and culture temperatures. The amount of JP-8 was analyzed by TPH using Gas Chromatography.