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Characterization of Acetobacter sp. Strain CV1 Isolated from a Fermented Vinegar (고산도 생성 초산균의 분리 및 발효특성)

  • Baek, Chang-ho;Baek, Seong-yeol;Lee, Se Hee;Kang, Ji-Eun;Choi, Han-Seok;Kim, Jae-Hyun;Yeo, Soo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.126-133
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    • 2015
  • Ten types of farm-made brewing vinegars were collected and four high acetic acid-producing strains (CV1, CV3, CV5, and CV6) were isolated. Among them strain CV1, exhibiting highly alcohol-resistant and acetic acid-producing properties, was selected and its taxonomic properties were investigated by phenotypic (particularly chemotaxonomic) characterization and phylogenetic inference based on 16S rRNA gene sequence analysis. On SM broth agar, cells of strain CV1 were gram-stainingnegative and formed pale white colonies with smooth to rough surfaces. Strain CV1 produced acetate from ethanol and was resistant to up to 8% (v/v) ethanol in LM broth. Strain CV1 had a G+C content of 61.0 mol%, contained meso-DAP as the cell wall amino acid, and possessed Q-10 as the major ubiquinone. A comparison of 16S rRNA gene sequences showed that strain CV1 was most closely related to Gluconacetobacter saccharivorans (≥99.0% identity). In liquid media, the optimum growth conditions for acetic acid production were 30℃ and pH >3.0 and strain CV1 produced 9.3% and 8.4% acetic acids from 10% and 9% alcohol concentrations, respectively.

Identification of a Protein Kinase using a FITC-labelled Synthetic Peptide in Streptomyces griseus IFO 13350 (형광 Peptide를 이용한 Streptomyces griseus IFO 13350의 인산화 단백질 동정)

  • 허진행;정용훈;김종희;신수경;현창구;홍순광
    • Microbiology and Biotechnology Letters
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    • v.30 no.3
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    • pp.235-240
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    • 2002
  • Streptomycetes is a group of Gram-positive soil bacteria that growas a branching vegetative mycelium leading to the formation of spores, and display a physiological differenti-ation related to the synthesis of many secondary metabolites including antibiotics. Their complex life cycle and multicellular differentiation require various levels of regulation and types of signal transduction systems including eukaryotic-type serine/threonine protein kinases and prokaryotic-type histidine/aspartic acid protein kinases. Akt kinase that was found in cells is a sorine/threonine kinase controlling signal pathway for multi-tude of important cellular events. The activation or inactivation of Akt kinase in the cell is one of the critical regulatory points to deliver cell proliferation, differentiation, survival or apoptosis signal. To find the regula-tory protein homologous to Akt in Streptomyces, the fluorescien-labeled synthetic peptide (FITC-TRRSR-TESIT) was designed from the consensus sequence of target proteins for Akt kinase. From the difference of the mobility between the nonphosphorylated and phosphorylated synthetic peptides on Agarose gel electro-phoresis, the Akt-phosphorylating activity was monitored. The cell-free extract prepared from Streptomyces griseus IFO 13350 and the Akt homologous protein was purified by ammonium sulfate fractionation and many steps of column chromatographies such as, DEAE-Sepharose, Mono Q, Resource Phenyl-Soporose and Gel permeation column chromatographies. As a result, the protein phosphorylating the fluorescien-labeled Akt substrate was identified and it's molecular weight was estimated as 39 kDa on SDS-PAGE.

Evaluation of Aluminum and Copper Biosorption in Two-Metal System using Algal Biosorbent

  • Lee, Hak-Sung;Volesky, Bohumil
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • v.2 no.2
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    • pp.149-158
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    • 1998
  • Biomass of non-living brown seaweed Sargassun fluitans pretreated with NaOH is capable of taking up more than $10\%$ $(q_{max}$ : 3.85 mmol/g for Al and 1.48 mmol/g for Cu) of its dry weight in the Al and Cu at pH of 4.5. However, the maximum Al and Cu uptakes calculated from Langmuir isotherm were 1.58 mmol/g for Al and 1.35 mmol/g for Cu at pH 3.5. Equilibrium batch sorption study was performed using two-metal system containing Al and Cu. The mathematical model of the two-metal sorption system enabled quantitative estimation of one-metal biosorption inhibition due to the influence of a second metal. NaOH-treated S. fluitans contained 2.19 mmol $(43\;wt.\%)$ carboxyl groups per gram of biomass. A modified form of Langmuir, which assumes binding of Cu as $Cu^{2+}$ and Al as $Al(OH)_2^+,$ was used to model the experimental data. This result agrees with the one of mono-valent sorption for Al in single-metal system. The modified Langmuir model gives the following affinity correlated coefficients: 0.196 for Cu and 6.820 for Ah at pH 4.5, and 2.904 for Cu and 3.131 for Al at pH 3.5. The interference of Al in Cu biosorptive uptake was assessed by `cutting' the three dimensional uptake isotherm surfaces at constant second-metal final concentrations. Equimolar final equilibrium concentrations of Cu and Al of 1 mM at pH 4.5 give Cu and hi uptakes reduced by $82.5\%\;and\;5.4\%,$ respectively. However, these values at pH 3.5 were $55\%\;(Cu)\;and\;31\%$ (Al).

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Isolation and Characterization of Marine Bacterium Producing Arylsulfatase

  • BYUN , DAE-SEOK;KIM, DOO-SANG;J. SAMUEL GODBER,;NAM, SOO-WAN;OH, MYONG-JOO;SHIM, HANG-SUN;KIM, HYEUNG-RAK
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1134-1141
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    • 2004
  • A bacterial strain capable of hydrolyzing sulfate ester bonds in p-nitrophenyl sulfate and agar was isolated from the Southeast coast of Korea. The isolated strain (AS6330) is aerobic, Gram-negative, rod-shaped, and motile. Octadecanoic acid was the major cellular fatty acid in the isolate. An almost complete 16S rDNA sequence of the isolate was determined and the sequence similarity of the 16S rDNA with those of known Sphingomonas spp. was found to be at most $96.4\%$, implying that the isolate was a new Sphingomonas species. The organism was grown optimally at NaCl concentration of $1.5-3.5\%$. Optimum culture conditions were determined to be $30^{\circ}C$ and pH 7.0 for 48 h fermentation using a laboratory fermentor under constant culture conditions. Partially purified arylsulfatase through Q-Sepharose and phenyl­Sepharose chromatographies catalyzed hydrolysis of sulfate ester bonds in agar, and $97\%$ of sulfates in agar were removed after 4 h reaction at $45^{\circ}C$ and pH 7.0. The arylsulfatase from the isolated bacterium might be useful for the removal of sulfate groups in agar.

Sphingopyxis panaciterrae sp. nov., Isolated from Soil of Ginseng Field

  • Lee, Hae-Won;Ten, Irina L.;Jung, Hae-Min;Liu, Qing-Mei;Im, Wan-Taek;Lee, Sung-Taik
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1011-1015
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    • 2008
  • A Gram-negative, strictly aerobic, motile bacterial strain, designated Gsoil $124^T$, was isolated from a soil sample taken from a ginseng field in Pocheon Province (South Korea). The isolate contained Q-10 as the predominant lipoquinone, plus $C_{18:1}\;{\omega}7c$ and summed feature 4 ($C_{16:1}\;{\omega}6c$ and/or iso-$C_{15:0}$ 2-OH) as the major fatty acids. The G+C content of the genomic DNA was 68.1 mol%, and the major polar lipids consisted of sphingoglycolipid, phosphatidylglycerol, phosphatidylcholine, and phosphatidylethanolamine. A comparative 16S rRNA gene sequence analysis showed that strain Gsoil $124^T$ was most closely related to Sphingopyxis chilensis (98.7%), Sphingopyxis alaskensis (98.2%), Sphingopyxis witflariensis (98.2%), Sphingopyxis taejonensis (98.0%), and Sphingopyxis macrogoltabida (97.6%). However, the DNA-DNA relatedness between strain Gsoil $124^T$ and its phylogenetically closest neighbors was less than 22%. Thus, on the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil $124^T$ should be classified as representing a novel species in the genus Sphingopyxis, for which the name Sphingopyxis panaciterrae sp. nov. is proposed. The type strain is Gsoil $124^T$ (=KCTC $12580^T$=LMG $24003^T$).

Labrenzia callyspongiae sp. nov., Isolated from Marine Sponge Callyspongia elegans in Jeju Island

  • Park, So Hyun;Kim, Ji Young;Heo, Moon Soo
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1969-1974
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    • 2019
  • A Gram-staining-negative, aerobic, light brown pigment bacterium, designated strain CE80T was isolated from marine sponge Callyspongia elegans in Jeju Island, Republic of Korea. Strain CE80T grew optimally at 25℃, in the range of pH 5.0-11.0 (optimum 7.0-8.0), and with 1.0-5.0% NaCl (optimum 1-3% (w/v)). Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain CE80T belonged to the genus Labrenzia and was closely related to L. suaedae YC6927T (98.3%), L. alexandrii DFL-11T (96.6%), L. aggregata IAM 12614T (96.6%) L. marina mano18T (96.5%) and L. alba CECT 5094T (96.2%). The major fatty acids of strain CE80T were C18:1 ω7c, and summed feature. The polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamin, one unidentified aminolipid, one phospholipid and four unidentified lipids. The DNA G+C content of strain CE80T was 55.9 mol%. The major respiratory quinone was Q-10. DNA-DNA relatedness between strain CE80T and L. suaedae YC6927T was 56.1±2.8%. On the basis of physiological and biochemical characterization and phylogenetic and chemotaxonomic analysis, strain CE80T represents a novel species of the Labrenzia, for which the name Labrenzia callyspongiae sp. nov., is proposed. The type strain is CE80T (=KCTC 42849T =JCM 31309T).

Caulobacter ginsengisoli sp. nov., a Novel Stalked Bacterium Isolated from Ginseng Cultivating Soil

  • Liu, Qing-Mei;Ten, Leonid N.;Im, Wan-Taek;Lee, Sung-Taik;Yoon, Min-Ho
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.15-20
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    • 2010
  • A Gram negative, aerobic, nonspore-forming, straight or curved rod-shaped bacterium, designated Gsoil $317^T$, was isolated from soil of a ginseng field in Pocheon Province (South Korea) and was characterized using a polyphasic approach. Cells were dimorphic, with stalk (or prostheca) and nonmotile or nonstalked and motile, by means of a single polar flagellum. Comparative analysis of 16S rRNA gene sequences revealed that strain Gsoil $317^T$ was most closely related to Caulobacter mirabilis LMG $24261^T$ (97.2%), Caulobacter fusiformis ATCC $15257^T$ (97.1 %), Caulobacter segnis LMG $17158^T$ (97.0%), Caulobacter vibrioides DSM $9893^T$ (96.8%), and Caulobacter henricii ATCC $15253^T$ (96.7%). The sequence similarities to any other recognized species within Alphaproteobacteria were less than 96.0%. The detection of Q-10 as the major respiratory quinone and a fatty acid profile with summed feature 7 ($C_{18:1}\;{\omega}7c$ and/or $C_{18:1}\;{\omega}9t$ and/or $C_{18:1}\;{\omega}12t;$ 56.6%) and $C_{16:0}$ (15.9%) as the major fatty acids supported the affiliation of strain Gsoil $317^T$ to the genus Caulobacter. The G+C content of the genomic DNA was 65.5 mol%. DNA-DNA hybridization experiments showed that the DNA-DNA relatedness values between strain Gsoil $317^T$ and its closest phylogenetic neighbors were below 11%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil $317^T$ should be classified as representing a novel species in the genus Caulobacter, for which the name Caulobacter ginsengisoli sp. novo is proposed. The type strain is Gsoil $317^T$ (=KCTC $12788^T=DSM\;18695^T$).

Acinetobacter antiviralis sp. nov., from Tobacco Plant Roots

  • Lee, Jung-Sook;Lee, Keun-Chul;Kim, Kwang-Kyu;Hwang, In-Cheon;Jang, Cheol;Kim, Nam-Gyu;Yeo, Woon-Hyung;Kim, Beom-Seok;Yu, Yong-Man;Ahn, Jong-Seog
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.250-256
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    • 2009
  • Acinetobacter strain $KNF2022^T$ was isolated from tobacco plant roots during the screening of antiviral substances having inhibitory effects on Tobacco mosaic virus (TMV) and examined by phenotypic, chemotaxonomic, and genetic characterization. It was a nonmotile, Gram-negative bacterium. This strain contained Q-9 as the main respiratory quinone. The major cellular fatty acids of the isolate were 16:0, 18:1 w9c, and 16:1 w7c/15 iso 2OH. The DNA base composition was 44 mol%. Phylogenetic analysis based on the 16S rRNA sequence revealed that the isolate formed an evolutionary lineage distinct from other Acinetobacter species. Based on the evaluation of morphologic, physiologic, and chemotaxonomic characteristics, DNA-DNA hybridization values, and 16S rRNA sequence comparison, we propose the new species Acinetobacter antiviralis sp. nov., the type strain of which is $KNF2022^T$ (=KCTC $0699BP^T$).

Purification and Characterization of 2,4-Dichlorophenol Oxidizing Peroxidase from Streptomyces sp. AD001

  • Jeon, Jeong-Ho;Yun-Jon Han;Tae-Gu kang;Eung-Soo Kim;Soon-Kwang Hong;Byeong-Chul Jeong
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.972-978
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    • 2002
  • Streptomyces sp. AD001 is a Gram-positive soil actinomycetes secreting an uncharacterized 2,4-dichlorophenol (DCP) oxidizing enzyme, whose activity is similar to the previously known Actinomycetes lignin-peroxidase (ALiP). This extracellular peroxidase was purified from Streptomyces sp. AD001 as a single protein band on an SDS-PACE by ammonium sulfate fractionation, Q-sepharose, concanavalin A, and Bio-Gel HTP column chromatographies. The molecular mass of the purified peroxidase was determined by SDS-PAGE to be 45.2 kDa, and 49.7 kDa with MALDI-TOF-MS, respectively. The highest level of peroxidase activity was observed at pH 7.5 and $30^{\circ}C$. The amino terminal sequence of the purified peroxidase (G-E-P-E-E-G-N-V-D-G-T-L) showed no significant homologies to my known proteins, suggesting that Streptomyces sp. AD001 may secrete a novel kind of bacterial peroxidase Initial rate kinetic data of the 2,4-DCP oxidation were best modeled with a random-binding bireactant system.

Acinetobacter marinus sp. novo and Acinetobacter seohaensis sp. nov., Isolated from Sea Water of the Yellow Sea in Korea

  • Yoon, Jung-Hoon;Kim, In-Gi;Oh, Tae-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1743-1750
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    • 2007
  • Two Gram-negative, nonmotile, coccobacilli, SW-$3^T$ and SW-$100^T$, were isolated from sea water of the Yellow Sea in Korea. Strains SW-$3^T$ and SW-$100^T$ contained ubiquinone-9 (Q-9) as the predominant respiratory lipoquinone and $C_{18:1}\;{\omega}9c$ and $C_{16:0}$ as the major fatty acids. The DNA G+C contents of strains SW-$3^T$ and SW- $100^T$ were 44.1 mol% and 41.9 mol%, respectively. A neighbor-joining tree based on l6S rRNA gene sequences showed that the two isolates fell within the evolutionary radiation enclosed by the genus Acinetobacter. Strains SW-$3^T$ and SW-$100^T$ exhibited a l6S rRNA gene similarity value of 95.7% and a mean DNA-DNA relatedness level of 9.2%. Strain SW-$3^T$ exhibited l6S rRNA gene sequence similarity levels of 93.5-96.9% to the validly described Acinetobacter species and fifteen Acinetobacter genomic species. Strain SW-$100^T$ exhibited l6S rRNA gene sequence similarity levels of less than 97.0% to the other Acinetobacter species except Acinetobacter towneri DSM $14962^T$ (98.0% similarity). Strains SW-$3^T$ and SW-$100^T$ exhibited mean levels of DNA-DNA relatedness of 7.3-l6.7% to the type strains of some phylogenetically related Acinetobacter species. On the basis of phenotypic, phylogenetic, and genetic data, strains SW-$3^T$ and SW-$100^T$ were classified in the genus Acinetobacter as two distinct novel species, for which the names Acinetobacter marinus sp. novo (type strain SW-$3^T$=KCTC $12259^T$=DSM $16312^T$) and Acinetobacter seohaensis sp. novo (type strain SW-$100^T$=KCTC $12260^T$=DSM $16313^T$) are proposed, respectively.