• Title/Summary/Keyword: bacterial sp

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Production and Characterization of Cyclodextrin Glucanotransferase fronm Bacillus sp. JK-43 Isolated from Kimchi (김치 분리균인 Bacillus sp. JK-43이 생산하는 Cyclodextrin Glucanotransferase의 생산 및 특성)

  • Jun, Hong-Ki;Bae, Kyung-Mi;Kim, Young-Hee;Baik, Hyung-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.1
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    • pp.41-48
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    • 2000
  • A bacterial strain, designated as JK-43, producing extracellular cyclodextrin glucanotransferase (CGTase)[EC 2.4.1.19] was isolated from kimchi. The CGTase from isolated strain JK-43 showed the transglucosylation activity from soluble starch to L-ascorbic acid(AA) compared to those obtained from other strains. A main product formed by this reaction was identified as $2-O-{\alpha}-glucopyranosyl$ L-ascorbic acid(AA-2G) by testing its susceptibility to ${\alpha}-glucosidase$ hydrolysis, the HPLC profiles, and through the elementary analysis. the ${\beta}-CD,\;{\gamma}-CD$, potato starch and corn starch were identified to be suitable glucosyl donor for transglucosylation reaction on AA by CGTase. Acceptor specificity on AA-2G production was examined by use of AA, Iso-AA and AA-2P. Transglucosylation was observed toward AA-2P as well as AA and Iso-AA. The microorganism isolated from kimchi was identified as a strain of Bacillus sp. JK-43 based on the morphological, cultural, biochemical characteristics and partial 16SrDNA sequence analysis. The maximal CGTase production was observed in a medium containing 1.0% soluble starch, 1.0% yeast extract, 1.0% $Na_2CO_3\;0.1%\;K_2HPO_4,\;and\;0.02%\;MgSO_4{\cdot}7H_2O$ with initial pH 7.0. The strain was cultured at $37^{\circ}C$ for 26 hrs with reciprocal shaking.

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Specific Detection of Xanthomonas oryzae pv. oryzicola in Infected Rice Plant by Use of PCR Assay Targeting a Membrane Fusion Protein Gene

  • Kang, Man-Jung;Shim, Jae-Kyung;Cho, Min-Seok;Seol, Young-Joo;Hahn, Jang-Ho;Hwang, Duk-Ju;Park, Dong-Suk
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1492-1495
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    • 2008
  • Successful control of Xanthomonas oryzae pv. oryzicola, the causal agent of bacterial leaf streak, requires a specific and reliable diagnostic tool. A pathovar-specific PCR assay was developed for the rapid and accurate detection ofthe plant pathogenic bacterium Xanthomonas oryzae pv. oryzicola in diseased plant. Based on differences in a membrane fusion protein gene of Xanthomonas oryzae pv. oryzicola and other microorganisms, which was generated from NCBI (http://www.ncbi.nlm.nih.gov/) and CMR (http://cmr.tigr.org/) BLAST searches, one pair of pathovar-specific primers, XOCMF/XOCMR, was synthesized. Primers XOCMF and XOCMR from a membrane fusion protein gene were used to amplity a 488-bp DNA fragment. The PCR product was only produced from 4 isolates of Xanthomonas oryzae pv. oryzicola among 37 isolates of other pathovars and species of Xanthomonas, Pectobacterium, Pseudomonas, Burkholderia, Escherichia coli, and Fusarium oxysporum f.sp. dianthi. The results suggested that the assay detected the pathogen more rapidly and accurately than standard isolation methods.

Rapid Detection and Isolation of Known and Putative $\alpha-L-Arabinofuranosidase$ Genes Using Degenerate PCR Primers

  • Park, Jung-Mi;Han, Nam-Soo;Kim, Tae-Jip
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.481-489
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    • 2007
  • [ $\alpha$ ]-L-Arabinofuranosidases (AFases; EC 3.2.1.55) are exo-type enzymes, which hydrolyze terminal nonreducing arabinose residues from various polysaccharides such as arabinan and arabinoxylan. Genome-wide BLAST search showed that various bacterial strains possess the putative AFase genes with well-conserved motif sequences at the nucleotide and amino acid sequence levels. In this study, two sets of degenerate PCR primers were designed and tested to detect putative AFase genes, based on their three highly conserved amino acid blocks (PGGNFV, GNEMDG; and DEWNVW). Among 20 Bacillus-associated species, 13 species were revealed to have putative AFase genes in their genome and they share over 67% of amino acid identities with each other. Based on the partial sequence obtained from an isolate, an AFase from Geobacillus sp. was cloned and expressed in E. coli. Enzymatic characterization has verified that the resulting enzyme corresponds to a typical AFase. Accordingly, degenerate PCR primers developed in this work can be used for fast, easy, and specific detection and isolation of putative AFase genes from bacterial cells.

PCR-based Specific Detection of Ralstonia solanacearum by Amplification of Cytochrome c1 Signal Peptide Sequences

  • Kang, Man-Jung;Lee, Mi-Hee;Shim, Jae-Kyung;Seo, Sang-Tae;Shrestha, Rosemary;Cho, Min-Seok;Hahn, Jang-Ho;Park, Dong-Suk
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1765-1771
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    • 2007
  • A polymerase chain reaction (PCR)-based method was developed to detect the DNA of Ralstonia solanacearum, the causal agent of bacterial wilt in various crop plants. One pair of primers (RALSF and RALSR), designed using cytochrome c1 signal peptide sequences specific to R. solanacearum, produced a PCR product of 932 bp from 13 isolates of R. solanacearum from several countries. The primer specificity was then tested using DNA from 21 isolates of Ralstonia, Pseudomonas, Burkholderia, Xanthomonas, and Fusarium oxysporum f. sp. dianthi. The specificity of the cytochrome c1 signal peptide sequences in R. solanacearum was further confirmed by a DNA-dot blot analysis. Moreover, the primer pair was able to detect the pathogen in artificially inoculated soil and tomato plants. Therefore, the present results indicate that the primer pair can be effectively used for the detection of R. solanacearum in soil and host plants.

Random amplified polymorphic DNA analysis of bacterial pathogens using universal rice primers

  • Monoldorova, Sezim;Kim, Jinsol;Kim, Joon Hee;Jeon, Bo-Young
    • Korean Journal of Veterinary Service
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    • v.40 no.1
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    • pp.1-6
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    • 2017
  • Molecular typing of pathogenic microorganisms is important for epidemiological investigation of infectious disease outbreaks. In this study, we applied Universal Rice Primers (URP) that were originated from repetitive sequences in rice chromosomal DNA to random amplified polymorphic DNA (RAPD) analysis of pathogenic bacteria such as Escherichia coli, Listeria monocytogenes, and Salmonella sp. Of the twelve URP primers examined to date, seven primers (URP-2, -3, -4, -5, -6, -8, and -9) generated reproducible and polymorphic PCR products ranging from 1 to 13 bands. One of them, URP-6 was very effective in differentiating seven E. coli serotypes, seven L. monocytogenes clinical isolates, and eight Salmonella subspecies (ssp.) serovars. The results thus indicate that RAPD analysis using URP primers might be useful in typing bacterial pathogens including E. coli, L. monocytogenes, and Salmonella strains.

Biosequestration, Transformation, and Volatilization of Mercury by Lysinibacillus fusiformis Isolated from Industrial Effluent

  • Gupta, Saurabh;Goyal, Richa;Nirwan, Jashan;Cameotra, Swaranjit Singh;Tejoprakash, Nagaraja
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.684-689
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    • 2012
  • In the present study, an efficient mercury-tolerant bacterial strain (RS-5) was isolated from heavy-metalcontaminated industrial effluent. Under shake flask conditions, 97% of the supplemented mercuric chloride was sequestered by the biomass of RS-5 grown in a tryptone soy broth. The sequestered mercuric ions were transformed inside the bacterial cells, as an XRD analysis of the biomass confirmed the formation of mercurous chloride, which is only feasible following the reaction of the elemental mercury and the residual mercuric chloride present within the cells. Besides the sequestration and intracellular transformation, a significant fraction of the mercury (63%) was also volatilized. The 16S rRNA gene sequence of RS-5 revealed its phylogenetic relationship with the family Bacillaceae, and a 98% homology with Lysinibacillus fusiformis, a Gram-positive bacterium with swollen sporangia. This is the first observation of the sequestration and volatilization of mercuric ions by Lysinibacillus sp.

Monitoring and Characterization of Bacterial Contamination in a High-Purity Water System Used for Semiconductor Manufacturing

  • Kim, In -Seop;Lee, Geon-Hyoung;Lee, Kye-Joon
    • Journal of Microbiology
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    • v.38 no.2
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    • pp.99-104
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    • 2000
  • Hydrogen peroxide has been used in cleaning the piping of an advanced high-purity water system that supplies ultra-high purity water (UHPW) for 16 megabyte DRAM semiconductor manufacturing. The level of hydrogen peroxide-resistant bacteria in UHPW water was monitored prior to and after disinfecting the piping with hydrogen peroxide. Most of the bacteria isolated after hydrogen peroxide disinfection were highly resistant to hydrogen peroxide. However, the percentage of resistant bacteria decreased with time. The hydrogen peroxide-resistant bacteria were identified as Micrococcus luteus, Bacillus cereus, Alcaligenes latus, Xanthomonas sp. and Flavobacterium indologenes. The susceptibility of the bacteria to hydrogen peroxide was tested as either planktonic cells or attached cells on glass. Attached bacteria as the biofilm on glass exhibited increased hydrogen peroxide resistnace, with the resistance increasing with respect to the age of the biofilm regrowth on piping after hydrogen peroxide treatment. In order to optimize the cleaning strategy for piping of the high-purity water system, the disinfecting effect of hydrogen preoxide and peracetic acid on the bacteria was evaluated. The combined use of hydrogen peroxide and peracetic acid was very effective in killing attached bacteria as well as planktonic bacteria.

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Post-Infectional Biochemical Changes in Mulberry Due to Xanthomonas campestris pv. mori Induced Bacterial Leaf Spot

  • Maji, M.D.;Sengupta, T.;Das, C.;Urs, S.Raje
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.2
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    • pp.255-259
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    • 2004
  • Post-infectional biochemical changes due to Xanthomonas campestris pv. mori (Xcm) infection in five elite mulberry varieties viz., $S_1$, $S_{1635}$, $V_1$, RF $S_{175}$ and JRH was studied under inoculated condition. It was revealed that total soluble sugar and protein content was significantly declined in all the varieties due to X. campestris infection. Total phenol content was at par prior to inoculation in all varieties, but it was significantly increased in $S_1$, RF $S_{175}$, $S_{1635}$ and JRH 7 days after inoculation. The correlation coefficient (r) between total soluble sugar and total phenol content was found positive (r = 0.825) and statistically significant. Similarly, correlation coefficient (r) between total soluble protein and phenol content was found positive (r = 0.897) and statistically significant. The present study indicates that X. campestris infected leaves are nutritionally inferior in quality and the duration of phenol production in a mulberry variety play decisive role on disease resistance.nce.

Characterization of Prophages in Leuconostoc Derived from Kimchi and Genomic Analysis of the Induced Prophage in Leuconostoc lactis

  • Kim, Song-Hee;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.32 no.3
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    • pp.333-340
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    • 2022
  • Leuconostoc has been used as a principal starter in natural kimchi fermentation, but limited research has been conducted on its phages. In this study, prophage distribution and characterization in kimchi-derived Leuconostoc strains were investigated, and phage induction was performed. Except for one strain, 16 Leuconostoc strains had at least one prophage region with questionable and incomplete regions, which comprised 0.5-6.0% of the bacterial genome. Based on major capsid protein analysis, ten intact prophages and an induced incomplete prophage of Leu. lactis CBA3626 belonged to the Siphoviridae family and were similar to Lc-Nu-like, sha1-like, phiMH1-like, and TPA_asm groups. Bacterial immunology genes, such as superinfection exclusion proteins and methylase, were found on several prophages. One prophage of Leu. lactis CBA3626 was induced using mitomycin C and was confirmed as belonging to the Siphoviridae family. Homology of the induced prophage with 21 reported prophages was not high (< 4%), and 47% identity was confirmed only with TPA_asm from Siphoviridae sp. isolate ct3pk4. Therefore, it is suggested that Leuconostoc from kimchi had diverse prophages with less than 6% genome proportion and some immunological genes. Interestingly, the induced prophage was very different from the reported prophages of other Leuconostoc species.

Direct and Indirect Reduction of Cr(VI) by Fermentative Fe(III)-Reducing Cellulomonas sp. Strain Cellu-2a

  • Khanal, Anamika;Hur, Hor-Gil;Fredrickson, James K.;Lee, Ji-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.31 no.11
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    • pp.1519-1525
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    • 2021
  • Hexavalent chromium (Cr(VI)) is recognized to be carcinogenic and toxic and registered as a contaminant in many drinking water regulations. It occurs naturally and is also produced by industrial processes. The reduction of Cr(VI) to Cr(III) has been a central topic for chromium remediation since Cr(III) is less toxic and less mobile. In this study, fermentative Fe(III)-reducing bacterial strains (Cellu-2a, Cellu-5a, and Cellu-5b) were isolated from a groundwater sample and were phylogenetically related to species of Cellulomonas by 16S rRNA gene analysis. One selected strain, Cellu-2a showed its capacity of reduction of both soluble iron (ferric citrate) and solid iron (hydrous ferric oxide, HFO), as well as aqueous Cr(VI). The strain Cellu-2a was able to reduce 15 μM Cr(VI) directly with glucose or sucrose as a sole carbon source under the anaerobic condition and indirectly with one of the substrates and HFO in the same incubations. The heterogeneous reduction of Cr(VI) by the surface-associated reduced iron from HFO by Cellu-2a likely assisted the Cr(VI) reduction. Fermentative features such as large-scale cell growth may impose advantages on the application of bacterial Cr(VI) reduction over anaerobic respiratory reduction.