• Title/Summary/Keyword: Soil DNA

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Sinomonas terrae sp. nov., Isolated from an Agricultural Soil

  • Hyosun Lee;Ji Yeon Han;Dong-Uk Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.909-914
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    • 2023
  • While searching for the bacteria which are responsible for degradation of pesticide in soybean field soil, a novel bacterial strain, designated 5-5T, was isolated. The cells of the strain were Gram-staining-positive, aerobic and non-motile rods. Growth occurred at 10-42℃ (optimum, 30℃), pH 5.5-9.0 (optimum, pH 7.0-7.5), and 0-2% (w/v) NaCl (optimum, 1%). The predominant fatty acids were C15:0 anteiso, C17:0 anteiso, and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). The predominant menaquinone was MK-9 (H2). Diphosphatidylglycerol, glycolipids, phosphatidylinositol, and phosphatidylglycerol were the major polar lipids. Phylogenetic analysis of 16S rRNA gene sequences indicated that strain 5-5T is a member of the genus Sinomonas and its closest relative is Sinomonas humi MUSC 117T, sharing a genetic similarity of 98.4%. The draft genome of strain 5-5T was 4,727,205 bp long with an N50 contig of 4,464,284 bp. Genomic DNA G+C content of strain 5-5T was68.0 mol%. The average nucleotide identity (ANI) values between strain 5-5T and its closest strains S. humi MUSC 117T and S. susongensis A31T were 87.0, and 84.3 % respectively. In silico DNA-DNA hybridization values between strain 5-5T and its closest strains S. humi MUSC 117T and S. susongensis A31T were 32.5% and 27.9% respectively. Based on the ANI and in silico DNA-DNA hybridization analyses, the 5-5T strain was considered as novel species belonging to the genus Sinomonas. On the basis of the results from phenotypic, genotypic and chemotaxonomic analyses, strain 5-5T represents a novel speciesof the genus Sinomonas, for which the name Sinomonas terrae sp. nov. is proposed. The type strain is 5-5T (=KCTC 49650T =NBRC 115790T).

Analysis of Microbial Community Structure in Soil and Crop Root System II. Analysis of soil microbial community structure in different soil Environmental conditions by MIDI and DNA analyses (토양과 작물근계의 미생물군집 구조 해석 II. MIDI 및 DNA 분석에 의한 토양환경별 미생물 군집 해석)

  • Ryu, Jin-Chang;Kwon, Soon-Wo;Kim, Jong-Shik;Suh, Jang-Sun;Jung, Beung-Gan;Choi, Sun-Shik
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.2
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    • pp.118-126
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    • 2002
  • To evaluate the correlations of microbial populations with soil healthiness and crop production and establish the criteria for microbial population of soil types. We analyzed the microbial community structure of 13 soils which were different in physical and chemical properties and cultivation methods. According to the analysis of microbial population suing the dilution plate method, the large differences of the microbial population structures among soil types were shown: aerobic bacteria $2-27{\times}10^6$, fluorescent Pseudomonas $1-1,364{\times}10^5$, Gram negative bacteria $1-126{\times}10^4$, and mesophilic Bacillus $1-110{\times}10^5$. The density of Gram negative bacteria was highest on red pepper cultivating soils (sample no. 4 and 6) of Umsung and Gesan, Chungbuk, and the density of the fluorescent Pseudomonas was highest on greenhouse soil (sample no. 7) of Jinju, Kyungnam. The crop productivity of three soils was high as compared with those of other soils. It was supposed that the density of fluorescent Pseudomonas and mesophilic Bacillus were correlated with the incresed crop production. By MIDI analysis, 579 strains isolated from 13 soils composed of a variety of microbes including 102 isolates of Agrobacterium, 112 isolates of Bacillus, 32 isolates of Pseudomonas, 44 isolates of Kocuria, and 34 isolates of Pseudomonas. Among the 624 isolates of Gram negative bacteria, Pseudomonas including P. putida and p. fluorescens occupied the highest density (51%), and Stenotrophomonas maltophilia and Burkholderia cepacia also appeared at high density. From RAPD analysis, the fluorescent Pseudomonas strains isolated from 13 soil types showed a high level of strain diversities and were grouped into 2 - 14 patterns according to soil types. Many of unknown bacteria were recovered from the paddy soil, and needed to be further characterized on the molecular basis.

454 Pyrosequencing Analysis of Bacterial Diversity Revealed by a Comparative Study of Soils from Mining Subsidence and Reclamation Areas

  • Li, Yuanyuan;Chen, Longqian;Wen, Hongyu;Zhou, Tianjian;Zhang, Ting;Gao, Xiali
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.313-323
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    • 2014
  • Significant alteration in the microbial community can occur across reclamation areas suffering subsidence from mining. A reclamation site undergoing fertilization practices and an adjacent coal-excavated subsidence site (sites A and B, respectively) were examined to characterize the bacterial diversity using 454 high-throughput 16S rDNA sequencing. The dominant taxonomic groups in both the sites were Proteobacteria, Acidobacteria, Bacteroidetes, Betaproteobacteria, Actinobacteria, Gammaproteobacteria, Alphaproteobacteria, Deltaproteobacteria, Chloroflexi, and Firmicutes. However, the bacterial communities' abundance, diversity, and composition differed significantly between the sites. Site A presented higher bacterial diversity and more complex community structures than site B. The majority of sequences related to Proteobacteria, Gemmatimonadetes, Chloroflexi, Nitrospirae, Firmicutes, Betaproteobacteria, Deltaproteobacteria, and Anaerolineae were from site A; whereas those related to Actinobacteria, Planctomycetes, Bacteroidetes, Verrucomicrobia, Gammaproteobacteria, Nitriliruptoria, Alphaproteobacteria, and Phycisphaerae originated from site B. The distribution of some bacterial groups and subgroups in the two sites correlated with soil properties and vegetation due to reclamation practice. Site A exhibited enriched bacterial community, soil organic matter (SOM), and total nitrogen (TN), suggesting the presence of relatively diverse microorganisms. SOM and TN were important factors shaping the underlying microbial communities. Furthermore, the specific plant functional group (legumes) was also an important factor influencing soil microbial community composition. Thus, the effectiveness of 454 pyrosequencing in analyzing soil bacterial diversity was validated and an association between land ecological system restoration, mostly mediated by microbial communities, and an improvement in soil properties in coal-mining reclamation areas was suggested.

Molecular Cloning and Expression of Bacillus stearothermophilus $\beta$-D-Xylosidase Gene in E. coli (Bacillus stearothermophilus $\beta$-D-Xylosidase 유전자의 크로닝 및 Escherichia coli에서의 발현)

  • 오세욱;박성수;최용진;박영인
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.136-142
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    • 1992
  • Bacillus stearothemophilus isolated from soil was identified to express multiple extracellular xylanases. Two HindIII restriction fragments of 5.4 and 6.4 kb from B. stearothermophilus genomic DNA were cloned into pBR322 to obtain recombinant plasmids pMG0l and pMG02, respectively, which enabled E. coli HBlOl cells to produce $\beta$-D-xylosidase activity. By subcloning into pUC18 and Southern blotting, the loci of the $\beta$-D-xyiosidase genes were elucidated to be on non-homologous DNA fragments of 2.2 kb from pMGOl(pMG1) and 1.0 kb from pMG02(pMG2), respectively. The two enzymes produced in E. coli cleaved xylobiose, xylotriose, xylotetrose and xylotetrose to produce xylose as a major end product. The gene on pMG1, distinct from that on pMG2 was observed to encode a bifunctional protein that displayed both P-D-xylosidase (EC.3.2.1.37) and a-L-arabinofuranosidase activities (EC.3.2.1.55).

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Transconjugation for Molecular Genetic Study of Streptomyces platensis Producing Transglutaminase (Transglutaminase를 생산하는 Streptomyces platensis의 분자생물학적인 연구를 위한 접합 전달법 확립)

  • Bae, Se-Joung;Jo, Yang-Ho;Choi, Sun-Uk
    • Journal of Life Science
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    • v.20 no.1
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    • pp.97-102
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    • 2010
  • Streptomyces platensis YK-2, newly isolated from forest soil, produces transglutaminase (TGase), which catalyses an acyl transfer reaction between the primary grade amine and protein or $\gamma$-carboxyamide group of peptide bound glutamine residues. For a molecular genetic study of S. platensis, an effective transformation method was established by using a conjugal transfer of DNA from Escherichia coli to spores of actinomycetes. The highest transconjugation frequency of S. platensis was obtained on an MS medium containing 50 mM $MgCl_2$, using $5{\times}10^7\;E$. coli as a DNA donor and $1{\times}10^8$ spores without heat treatment as a host. We also identified that S. platensis contains a single attB site within an ORF encoding a pirin-homolog, and that its attB site sequence shows high homology to that of S. logisporoflavus. In addition, it was confirmed by phenotypic analyses of exconjugants that the introduction of heterologous DNA into the attB site of the S. platensis chromosome does not affect its morphological differentiation and TGase production.

Physiological and Phylogenetic Analysis of Burkholderia sp. HY1 Capable of Aniline Degradation

  • Kahng, Hyung-Yeel;Jerome J. Kukor;Oh, Kye-Heon
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.643-650
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    • 2000
  • A new aniline-utilizing microorganism, strain HY1 obtained from an orchard soil, was characterized by using the BIOLOG system, an analysis of the total cellular fatty acids, and a 16S rDNA sequence. Strain HY1 was identified as a Burkholderia species, and was designated Burkholderia sp. HY1. GC and HPLC analyses revealed that Burkholderia sp. HY1 was able to degrade aniline to produce catechol, which was subsequently converted to cis,cis-muconic acid through an ortho-ring fission pathway under aerobic conditions. Strain HY1 exhibited a drastic reduction in the rate of aniline degradation when glucose was added to the aniline media. However, the addition of peptone or nitrate to the aniline media dramatically accelerated the rate of aniline degradation. A fatty acid analysis showed that strain HY1 was able to produce lipids 16:0 2OH, and 11 methyl 18:1 ${\omega}7c$ approximately 3.7-, 2.2-, and 6-fold more, respectively, when grown on aniline media than when grown on TSA. An analysison the alignment of a 1,435 bp fragment. A phylogenetic analysis of the 16S rDNA sequence based on a 1,420 bp multi-alignment sowed of the 16s rDNA sequence revealed that strain HY1 was very closely related to Burkholderia graminis with 95% similarity based that strain HY1 was placed among three major clonal types of $\beta$-Proteobacteria, including Burkholderia graminis, Burkholderia phenazinium, and Burkholderia glathei. The sequence GAT(C or G)${\b{G}}$, which is highly conserved in several locations in the 16S rDNA gene among the major clonal type strains of $\beta$-Proteobacteria, was frequently replaced with GAT(C or G)${\b{A}}$ in the 16S rDNA sequence from strain HY1.

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Isolation and Characterization of Thermophilic Microorganism Producing Starch-hydrolyze Enzyme (한국 토양으로부터 전분가수분해효소를 생산하는 고온성 균주의 선별과 동정)

  • Choi, Wonseok;Bai, Dong-Hoon
    • Food Engineering Progress
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    • v.14 no.1
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    • pp.7-13
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    • 2010
  • A thermophilic microorganism, which is able to hydrolyze starch, was isolated from soil and compost in Korea. It was Gram-positive, rod-shaped, catalase positive, nonmotile, glucose and mannitol fermentative, xylose oxidative, and spore forming microorganism. It also has an ability to hydrolyze casein and gelatin. The color of colony was yellowish white. The sequence of 16S rDNA of strain 2719 showed 99.5% sequence homology with the sequence of 16S rDNA of Bacillus thermoglucosidasius. On the basis of biochemical and physiological properties and phylogenetic analysis, the isolated strain was named as Bacillus thermoglucosidasius 2719.

Cloning and Expression of $\beta$-Xylosidase Gene from Alkali-tolerant Bacillus sp. YA-14 in Escherichia coli (알카리 내성 Bacillus sp. YA-14의 $\beta$-Xylosidase 유전자의 Cloning 및 대장균에의 발현)

  • 박덕철;김진만;정용준;공인수;배동훈;유주현
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.574-579
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    • 1989
  • Chromosomal DNA fragments of Bacillus sp. YA-14, isolated from soil as a potent $\beta$-xylosidase producing bacterium, were ligated to a vector plasmid pBR322 and used to transfer Escherichia coli HB101 cells. The recombinant plasmid pYXL22 was found to enable the transformants to produce $\beta$-xylosidase. pYXL22 was found to contain the 7.0 kb HindIII DNA fragment originated from the Bacillus sp. YA-14 chromosomal DNA by Southern hybridization. The optimum temperature for the reaction of $\beta$-xylosidase produced by E. coli HB101 (pYXL22) was appeared at 3$0^{\circ}C$. The enzyme was maintained stably up to 4$0^{\circ}C$ when stored 1hr at 4$0^{\circ}C$. The $\beta$-xylosidase was repressed completely by 0.4% (w/v) glucose concentration in E. coli HB101 (pYXL22). The optimum concentration of xylose for the $\beta$-xylosidase production in Bacillus sp. YA-14 was 0.2% (w/v).

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Characterization and identification of heterotrophic nitrifying bacteria isolated from composting soil (퇴비로부터 분리한 종속영양 질화세균의 동정 및 특징에 관한 연구)

  • Kim, Young-Jun;Lee, Gun-Young;Kim, Jin-Yong
    • Journal of the Korea Organic Resources Recycling Association
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    • v.13 no.2
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    • pp.121-127
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    • 2005
  • A heterotrophic nitrifying bacterium was isolated from the compost and analyzed for its characteristics. This bacterium was found to be a Gram positive rod, catalase positive, and motile. Nitrite production was detected on the ammonium acetate medium through the violet color formation. BBL test showed that this strain has high homology with Bacillus strains. Phylogenetic analysis using 16S rDNA revealed that the bacterium has 94% of similarity with Mycobacterium smegmatis strain.

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Bioproduction and Anticancer Activity of Biosurfactant Produced by the Dematiaceous Fungus Exophiala dermatitidis SK80

  • Chiewpattanakul, Paramaporn;Phonnok, Sirinet;Durand, Alain;Marie, Emmanuelle;Thanomsub, Benjamas Wongsatayanon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1664-1671
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    • 2010
  • A new biosurfactant producer was isolated from palm-oil-contaminated soil and later identified through morphology and DNA sequencing as the yeast-like fungus Exophiala dermatitidis. Biosurfactant production was catalyzed by vegetable oil, supplemented with a basal medium. The culture conditions that provided the biosurfactant with the highest surface activity were found to be 5% palm oil with 0.08% $NH_4NO_3$, at a pH of 5.3, with shaking at 200 rpm, and a temperature of $30^{\circ}C$ for a 14-day period of incubation. The biosurfactant was purified, in accordance with surfactant properties, by solvent fractionation using silica gel column chromatography. The chemical structure of the strongest surface-active compound was elucidated through the use of NMR and mass spectroscopy, and noted to be monoolein, which then went on to demonstrate antiproliferative activity against cervical cancer (HeLa) and leukemia (U937) cell lines in a dose-dependent manner. Interestingly, no cytotoxicity was observed with normal cells even when high concentrations were used. Cell and DNA morphological changes, in both cancer cell lines, were observed to be cell shrinkage, membrane blebbling, and DNA fragmentation.