• Title/Summary/Keyword: Sequence of 16S rDNA.

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The Importance of Weathered Crude Oil as a Source of Hydrocarbonoclastic Microorganisms in Contaminated Seawater

  • Sheppard, Petra J.;Simons, Keryn L.;Kadali, Krishna K.;Patil, Sayali S.;Ball, Andrew S.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1185-1192
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    • 2012
  • This study investigated the hydrocarbonoclastic microbial community present on weathered crude oil and their ability to degrade weathered oil in seawater obtained from the Gulf St. Vincent (SA, Australia). Examination of the native seawater communities capable of utilizing hydrocarbon as the sole carbon source identified a maximum recovery of just $6.6{\times}10^1\;CFU/ml$, with these values dramatically increased in the weathered oil, reaching $4.1{\times}10^4\;CFU/ml$. The weathered oil (dominated by > $C_{30}$ fractions; $750,000{\pm}150,000mg/l$) was subject to an 8 week laboratory-based degradation microcosm study. By day 56, the natural inoculums degraded the soluble hydrocarbons (initial concentrations $3,400{\pm}700mg/l$ and $1,700{\pm}340mg/l$ for the control and seawater, respectively) to below detectable levels, and biodegradation of the residual oil reached 62% ($254,000{\pm}40,000mg/l$) and 66% ($285,000{\pm}45,000mg/l$) in the control and seawater sources, respectively. In addition, the residual oil gas chromatogram profiles changed with the presence of short and intermediate hydrocarbon chains. 16S rDNA DGGE sequence analysis revealed species affiliated with the genera Roseobacter, Alteromonas, Yeosuana aromativorans, and Pseudomonas, renowned oil-degrading organisms previously thought to be associated with the environment where the oil contaminated rather than also being present in the contaminating oil. This study highlights the importance of microbiological techniques for isolation and characterisation, coupled with molecular techniques for identification, in understanding the role and function of native oil communities.

Isolation and Identification of Stenotrophomonas maltophilia BW-13 Active Against Rhizoctonia solani Causing Crisphead Lettuce Bottom Rot (Rhizoctonia solani에 의한 결구상추 밑둥썩음병 방제균주 Stenotrophomonas maltophilia BW-13의 분리 및 동정)

  • Kim Han-Woo;Park Jong-Young;Kim Hyun-Ju;Lee Kwang-Youll;Lee Jin-Woo;Choi Woobong;Lee Seon-Woo;Moon Byung-Ju
    • Research in Plant Disease
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    • v.11 no.2
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    • pp.152-157
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    • 2005
  • In a course of searching for biofungicide to control crisphead lettuce bottom rot caused by Rhizoctonia solani, we have isolated an antagonistic bacterium from lettuce rhisophere soil. A total of 702 bacterial isolates were isolated and tested for in vitro growth inhibition of R. solani. Seven strains appeared to have strong antagonistic effect against R. solani in in vitro growth inhibition assay. In the pot experiments, a strain BW-13 showed the most potent disease control effect on the both lettuce seedlings and adults plants. Therefore, the BW-13 was selected as a biocotrol candidate against crisphead lettuce bottom rot. Based on its morphology, physiological characteristics, and 165 rRNA gene analysis, the BW-13 was finally identified as Stenotrophomonas maltophilia. This study indicated that S. maltophilia BW-13 could be used as a biocontrol agent to control crisphead lettuce bottom rot.

Cloning and Characterization of a Gene for Fibrinolytic Enzyme from Bacillus subtilis BB-1 Isolated from Black Bean Chung-kuk (흑두로 제조한 청국에서 분리된 Bacillus subtillus BB-1으로 부터 혈전용해효소 유전자 크로닝 및 특성규명)

  • Lee Young-Hoon;Lee Sung-Ho;Jeon Ju-Mi;Kim Hong-Chul;Cho Yong-Un;Park Ki-Hoon;Choi Young-Ju;Gal Sang-Wan
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.513-521
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    • 2005
  • A bacterium producing five fibrinolytic isozymes was isolated from black bean chung kuk. The bacterium was identified as Bacillus subtilis BB-1 by 16s rDNA sequence homology search. A gene out of five fibrinolytic genes in the Bacillus subtilis BB-1 was cloned by shot-gun method. A Cla I DNA fragment of B. subtilis BB-1 chromosome was cloned in to pBluescript II SK(-) and showed the fibrinolytic activity to bacterial cells. The Cla I DNA fragment was sequenced and the sequences did not show homology with gene for protease or fibrinolytic enzyme genes in other organisms. The Cla I DNA fragment was reduced to 2,142 bp by activity-guided PCR cloning method. The optimum pH and temperature of the enzyme were 5.0 and $35^{\circ}C$, respectively. Substrate specificity of the fibrinolytic enzyme was detected in skim milk, casein, gelatin and blood agar plates. The activity of the enzyme was not detected with these substrates. Taken together, this enzyme is a new fibrinolytic enzyme and may be used to prevent thrombosis and arteriosclerosis.

Characterization of Cellulase and Xylanase from Bacillus subtilis NC1 Isolated from Environmental Soil and Determination of Its Genes (Bacillus subtilis NC1 유래 cellulase와 xylanase의 특성 규명 및 효소 유전자의 규명)

  • Park, Chang-Su;Kang, Dae-Ook;Choi, Nack-Shick
    • Journal of Life Science
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    • v.22 no.7
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    • pp.912-919
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    • 2012
  • A Bacillus sp. strain producing celluase and xylanase was isolated from environmental soil with LB agar plate containing carboxymethylcellulose (CM-cellulose) and beechwood xylan stained with trypan blue as substrates, respectively. Based on the 16S rRNA gene sequence and API 50 CHL test, the strain was identified as B. subtilis and named B. subtilis NC1. The cellulase and xylanase from B. subtilis NC1 exhibited the highest activities for CM-cellulose and beechwood xylan as substrate, respectively, and both enzymes showed the maximum activity at pH 5.0 and $50^{\circ}C$. We cloned and sequenced the genes for cellulase and xylanase from genomic DNA of the B. subtilis NC1 by the shot-gun cloning method. The cloned cellulase and xylanase genes consisted of a 1,500 bp open reading frame (ORF) encoding a 499 amino acid protein with a calculated molecular mass of 55,251 Da and a 1,269 bp ORF encoding a 422 amino acid protein with a calculated molecular mass of 47,423 Da, respectively. The deduced amino acid sequences from the genes of cellulase and xylanase showed high identity with glycosyl hydrolases family (GH) 5 and 30, respectively.

The Use of the Pathogen-specific Bacteriophage BCP8-2 to Develop a Rice Straw-derived Bacillus cereus-free Starter Culture (단일 박테리오파지를 이용한 볏짚 유래 Bacillus cereus free 스타터 컬쳐의 개발)

  • Bandara, Nadeeka;Chung, Seo-Jin;Jeong, Do-Youn;Kim, Kwang-Pyo
    • Korean Journal of Food Science and Technology
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    • v.46 no.1
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    • pp.115-120
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    • 2014
  • The purpose of this study was to develop a rice straw-derived Bacillus cereus (B. cereus)-free starter culture for traditional soybean fermented products using a B. cereus-specific bacteriophage, BCP8-2. To determine the optimal medium that supports the growth of rice straw-derived microorganisms and BCP8-2 activity, 5 different culture media were tested. The 5% ground bean (GB) medium was selected for further study. No B. cereus was detected in the BCP8-2-treated rice straw in GB medium, whereas B. cereus at a level of $10^7$ CFU/mL was recovered in the no-phage control. The total bacterial count reached approximately $10^9$ CFU/mL regardless of phage addition. When the 16S rRNA sequence-based microbial community was monitored using denaturing gradient gel electrophoresis (DGGE) and pyrosequencing, a similar microbial community was observed in the phage-treated and control samples. In conclusion, we demonstrate that phage can be used to prepare a rice straw-derived B. cereus-free starter culture with minimal effect on natural microflora.

Phylogenetic Diversity and Antibacterial Activity in Bacterium from Balloon Fish (Diodon holocanthus) of Jeju Island (제주 연안의 가시복(Diodon holoanthus)에서 분리된 세균의 다양성 및 항균활성 효과)

  • Moon, Chae-Yun;Ko, Jun-Cheol;Kim, Min-Seon;Heo, Moon-Soo
    • Microbiology and Biotechnology Letters
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    • v.48 no.1
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    • pp.57-63
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    • 2020
  • Over the past 20 years, global warming has transformed the marine ecosystem of the Jeju Island into a subtropical zone making it conducive to the production of tropical fishes. Recently, the balloon fish (Diodon holoanthus) has been found off the coast of the Jeju Island. In this study, we analyzed the diversity of its intestinal microorganisms as a representative for the surrounding environment. In addition, the isolates were evaluated for their antibacterial activity. A total of 161 strains of various species were identified and isolated using 16S ribosomal RNA gene sequence analysis. They were separated into three groups, of which Phylum Proteobacteria was found to be the most dominant with 91% sequence similarity. This includes the class γ-proteobacteria that is made up of twelve genera and twenty-four hundred species. The second group comprised strains of the genus Vibrio, made up of 35% Photobacteria, 32% Shewanella, and 6% Psychrobacter. It was also determined that 4% of the isolates were Acinetobacter, 3% were Enterovibrio, while Moraxella_g2 accounted for 1% of the total isolates. Class α-proteobactera includes five genera and five species; Brevundimonas, Allorhizobium, Pseudoceanicola and Erythrobcter, each accounting for 1% of the total isolates. The Firmicute strains belonged to six genera and ten species. 5% of the strains were Terribacillus, while Paenibacillus, Salinicoccus, Staphylococcus and Streptococcus accounted for 1% each of the total isolates. Actinobacteria accounted for the final phylum with strains belonging to three genera and ten species with Janibacter, Micrococcus and Isoptericola each accounting for 1% of the total isolates.

Plant Growth-Promoting Potential of Endophytic Bacteria Isolated from Roots of Coastal Sand Dune Plants

  • Shin, Dong-Sung;Park, Myung-Soo;Jung, Se-Ra;Lee, Myoung-Sook;Lee, Kang-Hyun;Bae, Kyung-Sook;Kim, Seung-Bum
    • Journal of Microbiology and Biotechnology
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    • v.17 no.8
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    • pp.1361-1368
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    • 2007
  • Endophytic bacteria associated with the roots of coastal sand dune plants were isolated, taxonomically characterized, and tested for their plant growth-promoting activities. Ninety-one endophytic bacterial isolates were collected and assigned to 17 different genera of 6 major bacterial phyla based on partial 16S rDNA sequence analyses. Gammaproteobacteria represented the majority of the isolates (65.9%), and members of Pseudomonas constituted 49.5% of the total isolates. When testing for antagonism towards plant pathogenic fungi, 25 strains were antagonistic towards Rhizoctonia solani, 57 strains were antagonistic towards Pythium ultimum, 53 strains were antagonistic towards Fusarium oxysporum, and 41 strains were antagonistic towards Botrytis cinerea. Seven strains were shown to produce indole acetic acid (IAA), 33 to produce siderophores, 23 to produce protease, 37 to produce pectinase, and 38 to produce chitinase. The broadest spectra of activities were observed among the Pseudomonas strains, indicating outstanding plant growth-promoting potential. The isolates from C. kobomugi and M. sibirica also exhibited good plant growth-promoting potential. The correlations among individual plant growth-promoting activities were examined using phi coefficients, and the resulting data indicated that the production of protease, pectinase, chitinase, and siderophores was highly related.

Isolation and Characterization of Three Kinds of Lipopeptides Produced by Bacillus subtilis JKK238 from Jeot-Kal of Korean Traditional Fermented Fishes (한국 전통젓갈에서 분리한 Bacillus subtilis JKK238 균주 유래 세 종류 Lipopeptide의 분리 및 특성)

  • Yoon Sang-Hong;Kim Jung-Bong;Lim Yoong-Ho;Hong Seong-Ryeul;Song Jae-Kyeung;Kim Sam-Sun;Kwon Soon-Wo;Park In-Cheol;Kim Soo-Jin;Yeo Yun-Soo;Koo Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.295-301
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    • 2005
  • About seven hundred bacterial strains were collected from Jeot-Kal, a Korean traditional fermented fishes, in various Korean districts. One of the strains designated JKK238 has its ability to antagonize in vitro the growth of a wide variety of plant pathogenic fungi responsible for diseases of economical importance. The JKK238 strain was isolated from Oh-Jeot, a kind of fermented shrimps, of Kangkyeung in Korea, and was identified as Bacillus subtilis based on its physiological characteristics, fatty acids compositions of cellular wall, and 16S rDNA sequence analysis. We isolated simply antimicrobial lipopeptides (AMLP) by $25\%$ ammonium sulfate precipitation of 3 days-old tryptic soy broth cultures of the JKK238 strain. Further analysis of AMLP revealed that B. subtilis JKK238 produces a wide variety of antifungal lipopeptide isomers from the iturin, fengycin and surfactin families simultaneously. Above results indicate that the JKK238 strain can be added to the limited number B. subtilis strains reported to co-produce the three kinds of lipopeptide families.

Phylogenetic Analysis on Wild Cordyceps Collected from Miryang Region of South Korea (밀양근교에서 채집한 야생 동충하초 계통의 PCR 산물에 근거한 계통 유전학적 연구)

  • Park, Hyeancheal;Lee, Sangmong;Park, Namsook
    • Korean Journal of Plant Resources
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    • v.34 no.1
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    • pp.1-16
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    • 2021
  • The phylogenetic relationships among thirty-two strains (P1~P32; including Cordyceps sp., Paecilomyces sp., Beauveria sp., Aranthomyces sp., Isaria sp. and Himenostilbe sp.) in Miryang region located in the southern part of Korea, were investigated based on internal transcribed spacer (ITS) sequences of ribosomal DNA. A fragment of ITS region was amplified by polymerase chain reaction (PCR) using the specific primer pairs ITS1 and ITS4. After obtained same size of PCR products from various strains, we cloned them into a pGEM-T easy vector to determine their sequences. BLAST analyses of the nucleotide sequence ITS1, 5.8S and ITS2 gene fragments revealed the identity and their phylogenetic relationship. Among 32 strains isolated from Miryang region, Cordyceps militaris was shared 100% sequences with Genbank (AY49191, EU825999, AY491992), while some species are not shared perfectly with reported sequences. For example, strain P17 (P. tenuipes in Ulju-gun Gaji Mountain) has some differences among the other strains of P. tenuipes (Miryang-si Jocheon-eup, Miryang-si Gaji Mountain) and those of gene bank. We conclude that ITS analyses with strains in the suburbs of Miryang in this study can be effectively used as a tool for classification, evaluation and collection of the natural eco-type genetic resources.

Comparison of Harboring the Resistance Gene and Disc Diffusion Susceptibility Test Result in Staphylococcus pseudintermedius from the Bacterial Dermatitis (세균성 피부염 개에서 분리된 Staphylococcus pseudintermedius에서 항생제 감수성 검사와 내성 유전자 획득의 비교)

  • Jang, Hye-Jin;Son, Hyoung-Won;Kang, Hyo-Min;Han, Jae-Ik;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.32 no.2
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    • pp.158-161
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    • 2015
  • Bacterial dermatitis is common disease that is necessary to treat with antibiotics. In recent, antibiotic-resistant bacteria is being increased in worldwide. The purpose of the present study was to evaluate the prevalence of resistant genes in Staphylococcus (S.) pseudintermedius isolated from dogs, and to compare the resistant gene profile with the result of antibiotic disc diffusion test. A total of seven S. pseudintermedius was included in the study. Bacterial identification was performed by 16S ribosomal RNA gene sequence analysis. S. pseudintermedius isolates had more than one antibiotic resistant gene (mecA, blaZ and aac(6')/aph(2"). While all isolates were PCR positive to blaZ gene, only two isolates were resistant to amoxicillin/clavulanate. Among five isolates harboring gentamicin resistance, one isolate was negative to aac(6')/aph(2")-targeted PCR. Taken together, the results suggest that resistant gene-targeted PCR and disc diffusion test are complementary to detect antibiotic resistance.