• Title/Summary/Keyword: Microbial isolation

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Isolation and Characterization of Plant Growth Promoting Bacteria Pseudomonas sp. SH-26 from Peat Soil (이탄 토양으로부터 식물생육촉진세균 Pseudomonas sp. SH-26의 분리 및 특성)

  • Ho-Young Shin;Da-Son Kim;Chang-Ho Lee;Dong-Soek Lee;Song-Ih Han
    • Journal of the Korean Applied Science and Technology
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    • v.41 no.2
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    • pp.199-207
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    • 2024
  • We conducted to investigate both plant growth-promoting and plant disease-controlling activities of bacterial strains isolated from soil. Among the 48 isolated strains, SH-23, SH-26, SH-29, and SH-33 were identified as excellent strains for the production of β-glucosidase, cellulase, amylase, and protease. These 4 strains exhibited antifungal activity against plant pathogenic fungi (Botrytis cinerea, Rhizoctonia solani, Fusarium oxysporum, Colletotrichum acutatum). Strain SH-26, which exhibited excellent organic matter decomposition and antifungal activity against plant pathogenic fungi, was selected as the final superior strain. Upon determining the 16S rRNA gene sequence of the selected SH-26 strain, it exhibited 100% similarity with Pseudomonas knackmussii HG322950 B13T, Pseudomonas citronellolis BCZY01000096 NBRC 103043T, and Pseudomonas delhiensis jgi.1118306 RLD-1T. Furthermore, it was confirmed that the Pseudomonas sp. SH-26 exhibited siderophore production, nitrogen fixation ability, and the production of Indole-3-acetic acid.

Isolation and characterization analysis of the halophilic archaea isolated from solar saltern, Gomso (곰소 염전에서 분리한 호염성 고세균의 특성 분석)

  • Koh, Hyeon-Woo;Kim, So-Jeong;Rhee, Sung-Keun;Park, Soo-Je
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.427-434
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    • 2015
  • Most of halophilic archaea are found in the various hypersaline environments including solar saltern, salt lake with very high salt concentration. The present study is about isolation and characterization of halphilic archaea from Gomso solar saltern known as a representative high salt environment in Korea. In order to isolate the halophilic archaea, we prepared and used high salt medium. Finally, total 7 strains obtained were tentatively identified based on comparative similarity analysis for 16S rRNA gene sequence and physiological traits. All halophilic archaea belonged to Haloruburm, Halogeometriucm, Halobacterium, and Haloarcula genera. These isolates were all Gram-staining negative, and growth was not observed using nitrate as an alternative electron acceptor under anaerobic conditions. In addition, all isolates required about 12-30% (w/v, NaCl) salt. This case study might provide basic information on microbial isolation technologies and related research in halophilic microorganisms from domestic halophilic environments, and contribute to obtaining useful indigenous halophilic archaea in a variety of extreme environmental conditions.

Analysis of Prokaryote Communities in Korean Traditional Fermented Food, Jeotgal, Using Culture-Dependent Method and Isolation of a Novel Strain (배양 분리법을 통한 젓갈 내 원핵 세균 군집 분석 및 신규 미생물의 분리)

  • Kim, Min-Soo;Park, Eun-Jin;Jung, Mi-Ja;Roh, Seong-Woon;Bae, Jin-Woo
    • Korean Journal of Microbiology
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    • v.45 no.1
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    • pp.26-31
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    • 2009
  • This study was aimed at the analysis of prokaryote communities in Korean traditional fermented food, jeotgal, and isolation of a novel strain from jeotgal by using culture-dependent and molecular biological approaches. Seventeen kinds of jeotgal were selected on the basis of its origins and sources. The samples were inoculated on 12 kinds of media. 308 isolates were selected randomly by morphological features, and its 16S rRNA gene sequences was amplified by PCR technique with bacteria and archaea specific primers (8F, 21F, and 1492R). The 16S rRNA gene sequences were compared with those in EzTaxon and GenBank databases. DNA-DNA hybridization was performed to identify a novel strain. As a result, the majority of the isolates were lactic acid bacteria (Leuconostoc, Weisella, Lactococcus, Lactobacillus, Carnobacterium, Marinilactibacillus), Bacillus, Pseudomonas, Micrococcus, Brevibacterium, Microbacterium and Kocuria in 17 kinds of jeotgal. The strains belonging to Salinicoccus, Halomonas, Cobetia, Lentibacillus, Paracoccus, and Psychrobacter were isolated as minor ones. Fourteen novel species were identified based on phylogenetic analysis.

Two-year Quaternary Isolation of Gram-positive Bacilli Using MALDI-TOF MS in Positive Blood Culture of a University Hospital (일개 대학병원의 혈액배양에서 MALDI-TOF MS를 이용한 Gram-positive Bacilli의 2년간 분기별 분리율)

  • Choi, Jin-Un;Yu, Young-Bin;Kim, Sang-Ha;Won, Seungho;Kim, Young-Kwon
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.4
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    • pp.414-421
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    • 2018
  • In this study, Gram-positive bacilli (GPB) were identified by MALDI-TOF MS and analyzed according to the quaternary and microbial strains in the blood culture medium over a two year period at a university hospital. The results were as follows. The overall positive rate of blood culture was 9.97%. In 713 isolated GPB, 410 strains (57.5%) were identified using a microflex MALDI Biotyper. The positive rate of GPB among the blood culture positive bacteria was 8.2%, and the quarterly isolation rate was 9.8% in the third quarter of 2015, 8.7% in the second quarter of 2016, 8.1% in the third quarter of 2016, 8.1% in the first quarter of 2015, 7.9% in the first quarter of 2015, 7.9% in the second quarter of 2015, 6.8% in the first quarter of 2016, and 6.7% in the fourth quarter of 2015. The isolates were Corynebacterium striatum 89 (12.4%), Bacillus cereus 60 (8.4%), Bacillus subtilis 30 (4.2%), Paenibacillus urinalis 29 (4.1%), and Listeria monocytogenes 25 (3.5%). The results of 16S rRNA sequencing of 43 isolates (86.0%) were consistent with those of the other 50 isolates. Five out of the seven unmatched weeks were not identified by MALDI-TOF MS.

Isolation and Identification of Antagonistic Microorganisms for Biological Control to Major Diseases of Apple Tree(Malus domestica Borkh) (사과 주요 병해 방제를 위한 길항미생물 분리 및 동정)

  • 박흥섭;조정일
    • Korean Journal of Organic Agriculture
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    • v.5 no.1
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    • pp.137-147
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    • 1996
  • For the purpose of acquiring microbial agents that can be utilized to biologically control the major airborne diseases to apple trees, such as canker(Botryosphaeria dothidea), bitter rot(Glomerella cingulata), alternaria leaf spot(Alternaria mali), root rot(rosellinia necatrix), canker(Valsa ceratosperma) and gray mold rot(Botrytis cinerea), the effective microorgaisms were isolated, tested for antagonistic activity to the pathogens causing major diseases to apple trees and identifed. Screening of more than 5,000 species of microorganisms collected in nature for them antagonistic action to the pathogens causing 5 major diseases to apple trees resulted in selection of effective species. Out of the 11 species, one species designated as CAP134 demonstrated outstanding activity. The bacterial strain, CAP134 exerted antagonistic efficiency of 57% on an isolated strain and 40% on a donated strain of Botryosphaeria dothidea., 52% on an isolated strain and 46% on a purchased strain of Alternaria mali, 60% on Valsa ceratosperma 25% on Glomerella cingulata, and 64% Rosellinia necatrix. The CAP134 was identified as a bacterial strain to Bacillus subtilis ATCC 6633 based on morephology, culture conditions, and physio-biochemical characteristics.

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Isolation and Identification of Pathogenic Bacteria from Salads of Fast Food Restaurants (유통 샐러드중의 병원성 미생물 오염 실태조사)

  • Park Yong-Bae;Kang Jeong-Bok;Kim Jung-Beom;Kim Jong-Chan
    • Journal of environmental and Sanitary engineering
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    • v.20 no.1 s.55
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    • pp.23-31
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    • 2005
  • On the purpose of epidemiological survey relate to food poisoning, a total of 114 samples of different salads collected from fast food Restaurants in Gyeonggi-do were for the presence of pathogenic microorganisms. Microbial assessment of salads revealed that TPC($1.1\times10\;\~\;8.4\times10^{5}\;CFU/g$) and coliforms($0\~5.4\times10^{4}\;CFU/g$) exceeded the standards by Solberg et al. ($TPC:10^{5}\;CFU/g,\;coliforms:10^{2}\;CFU/g$). Two pathogenic bacteria were isolated from salad samples, and identified by biochemical methods, including API identification systems. Isolates from PALCAM agar and MYP agar media were in 98.6, $99.8\%$ agreements with Listeria monocytogenes, Bacillus cereus at the species level, respectively. All 7 strains of Bacillus cereus isolates produced enterotoxin as revealed with CRET-RPLA.

Isolation of Myxobacteria Carrying Soraphen Biosynthetic Gene Clusters (Soraphen 생합성 유전자군을 갖는 점액세균의 분리)

  • Lee, Cha-Yul;Hyun, Hye-Sook;Cho, Kung-Yun
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.10-16
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    • 2009
  • Polymerase chain reactin (PCR) could be a simple way to screen new microbial strains producing useful secondary metabolites if their biosynthetic genes are known and candidate strains to be screened are available. In this study, we have screened two myxobacterial strains, KYC3047 and KYC3076, carrying genes appeared to be biosynthetic genes of soraphen A, a potent antifungal substance, out of 50 cellulose degrading myxobacteria using PCR. The two strains were identified as Sorangium cellulosum based on morphological, physiological, and molecular biological characteristics. Both of the strains produced substances having strong antifungal activities as expected against Candida albicans, a causative agent of candidiasis, and Colletotrichum acutatum, a causative agent of anthracnose on pepper.

New Yeast Cell-Based Assay System for Screening Histone Deacetylase 1 Complex Disruptor

  • Jeon, Kwon-Ho;Kim, Min-Jung;Kim, Seung-Young
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.286-291
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    • 2002
  • Histone deacetylase I (HDAC1) works as one of the components in a nucleosome remodeling (NuRD) complex that consists of several proteins, including metastasis-associated protein 1 (MTA1). Since the protein-protein interaction of HDAC1 and MTA1 would appear to be important for both the integrity and functionality of the HDAC1 complex, the interruption of the HDAC1 and MTA1 interaction may be an efficient way to regulate the biological function of the HDAC1 complex. Based on this idea, a yeast two-hybrid system was constructed with HDAC1 and MTA1 expressing vectors in the DNA binding and activation domains, respectively. To verify the efficiency of the assay system, 3,500 microbial metabolite libraries were tested using the paper disc method, and KB0699 was found to inhibit the HDAC1 and MTA1 interaction without any toxicity to the wild-type yeast. Furthermore, KB0699 blocked the interaction of HDAC1 and MTA1 in an in vitro GST pull down assay and induced morphological changes in B16/BL6 melanoma cells, indicating the interruption of the HDAC1 complex function. Accordingly, these results demonstrated that the yeast assay strain developed in this study could be a valuable tool for the isolation of a HDAC1 complex disruptor.

Isolation and Characterization of the Eicosapentaenoic Acid Biosynthesis Gene Cluster from Shewanella sp. BR-2

  • Lee, Su-Jin;Seo, Pil-Soo;Kim, Chul-Ho;Kwon, Oh-Suk;Hur, Byung-Ki;Seo, Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.881-887
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    • 2009
  • Forty-four eicosapentaenoic acid (EPA)-producing microbial strains were isolated from the intestines of marine fishes. Among them, one strain showing a maximum level of EPA (4.78% of total fatty acids) was identified as Shewanella sp. BR-2 on the basis of its 168 rRNA sequence. The EPA content reached a maximum level during the mid-exponential phase of cell growth, and gradually decreased with further growth of the cells. A cosmid DNA including the EPA biosynthesis gene cluster consisting of pfaA-E was isolated from a cosmid library of genomic DNA of Shewanella sp. BR-2, named pCosEPA-BR2. An E. coli clone harboring pCosEPA-BR2 produced EPA at a maximum level of 7.5% of total fatty acids, confirming the EPA biosynthesis activity of the cloned gene cluster.

Isolation of Leuconostoc and Weissella Species Inhibiting the Growth of Lactobacillus sakei from Kimchi (김치로부터 Lactobacillus sakei 생육저해 Leuconostoc 및 Weissella 속 균주의 분리)

  • Lee, Kwang-Hee;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.175-181
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    • 2011
  • Kimchi is a group of traditional fermented vegetable foods in Korea and known to be the product of a natural mixed-fermentation process carried out principally by lactic acid bacteria (LAB). According to microbial results based on conventional identification, Leuconostoc mesenteroides and Lactobacillus plantarum were considered to be responsible for the good taste and over-ripening of kimchi, respectively. However, with the application of phylogenetic identification, based on 16S ribosomal RNA gene similarities, a variety of Leuconostoc and Lactobacillus species not detected in the previous studies have been isolated, together with a species in the genus Weissella. Additionally, Lactobacillus sakei has been accepted as the most populous LAB in over-ripened kimchi. In this study, Leuconostoc and Weissella species inhibiting the growth of Lb. sakei were isolated from kimchi for future applications to do with kimchi fermentation. From 25 kimchi samples, 378 strains in the genera Leuconostoc and Weissella were isolated and 68 strains identified as Lc. mesenteroides, Lc. citreum, Lc. lactis, W. cibaria, W. confusa, and W. paramesenteroides exhibited growth inhibition against Lb. sakei. Most of the strains also had antagonistic activities against Lb. brevis, Lb. curvatus, Lb. paraplantarum, Lb. pentosus, and Lb. plantarum. Their antagonistic activities against Lb. sakei were more remarkable at lower temperatures of incubation.