• Korean Journal of Microbiology
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• v.42 no.3
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• pp.223-229
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• 2006

To develop multifunctional microbial inoculant, an insluble phosphate-solubilizing bacterium with antifungal activity was isolated from plant rhizospheric soil. On the basis of its morphological, cultural and physiological characteristics and Biolog analysis, this bacterium was identified as Pseudomonas fluorescens RAF15. P. fluorescens RAF15 showed antifungal activities against phytopathogenic fungi Botrytis cinerea and Rhizoctonia solani. The optimal medium composition and cultural conditions for the solubilization of insoluble phosphate by P. fluorescens RAF15 were 1.5% of glucose, 0.005% of urea, 0.3% $MgCl_2{\cdot}6H_2\;0.01%\;of\;MgSO_4{\cdot}7H_2O\;0.01%,\;of\;CaCl_2{\cdot}2H_2O$, and 0.05% of NaCl along with initial pH 7.0 at $30^{\circ}C$. The soluble phosphate production under optimum condition was 863 mg/L after 5 days of cultivation. The solubilization of insoluble phosphates was associated with a drop in the pH of the culture medium. P. fluorescens RAF15 showed resistance against different environmental stresses like $10-35^{\circ}C$ temperature, 1-4% salt concentration and pH 2-11 range. The strain produced soluble phosphate to the culture broth with the concentrations of 971-1121 mg/L against $CaHPO_4$, 791-908 mg/L against $Ca_3(PO_4){_2}$, and 844 mg/L against hydroxyapatite, respectively. However, the strain produced soluble phosphate to the culture broth with the concentrations of 15 mg/L against $FePO_4$, and 5 mg/L against $AlPO_4$, respectively.

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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• pp.415-421
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• 2003

A bacterium, Pseudomonas fluorescens 2112, that is antagonistic against a red-pepper blight-causing fungus, Phytophthora capsici, was isolated from the local soil of Gyongju, Korea. This strain formed an orange-colored clear halo zone on chrome azurol S (CAS) blue agar, suggesting the production of a siderophore in addition to an antifungal antibiotic. The optimal culture conditions for siderophore production by P. fluorescens 2112 were 30-h cultivation at $25^{\circ}C$ and pH 6.5 in King's B medium. The presence of $20{\mu}g/ml\;of\;Fe^3+$ ion or EDDHA promoted the production of siderophore in King's B medium. The siderophore was purified from culture broth by CM-Sephadex C-25 and Sephadex G-25 column chromatographies. The UV spectra of the purified siderophore was the same as that of pyoverdins or pseudobactins. The molecular mass was 1,958 Da determined by FAB-rlass spectrometer, and the amino acid composition analysis showed that the purified siderophore consisted of glycine/threonine/serine/glutamic acid/alanine/lysine with the molar ratio of 3:2:1:1:1:1, DL-Threo-${\beta}$-hydroxyaspartic acid and $N^{\delta}$-hydroxyornithine, two of the essential constituents of pyoverdin, were also found. The purified siderophore pyoverdin showed strong in vitro and in vivo antagonistic activities against phytophthora blight-causing P. capsici. Especially in an in vivo pot test, the siderophore protected red-pepper Capsicum annum L. very well from the attack of P. capsici. These results indicated that the purified siderophore of P. fluorescens 2112 played a critical role in the biocontrol of the red-pepper blight disease, equivalent to treatment by P.fluorescens 2112 cells.

• Journal of agriculture & life science
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• v.45 no.3
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• pp.89-96
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• 2011

The objective of this study is to evaluate reduction efficiency of various air purifiers on airborne microorganism causing air infected animal disease according to sample collection method. Collection efficiencies of MS2 virus and Pseudomonas fluorescens by biosampler was significantly higher than those by button sampler (p<0.05). Regardless of types of air purifier and sample collection method, temporal reduction efficiencies of MS2 virus and P. fluorescens compared to initial background concentration were >50% and >45% on 5 minutes, >70% and >50% on 15 minutes, >80% and >70% on 30 minutes and >90% and >75% on 60 minutes after operating air purifier, respectively. The air purifier of ionizer type showed the highest reduction efficiency on MS2 virus followed by air purifier of electronic precipitation, water filter and dry filter while the reduction efficiency of air purifier on P. fluorescens was highest in the electronic precipitation type followed by ionizer type, dry filter type and water filter type (p<0.05). Based on the results obtained from this study, temporal reduction efficiency of air purifier on MS2 virus was relatively higher than P. fluorescens.

• Culinary science and hospitality research
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• v.9 no.4
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• pp.113-122
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• 2003

This study was conducted to examine the antagonism of useful microbes against soybean sprout rotting pathogens and their effect on the growth of soybean sprouts. The antagonism against soybean sprout rotting pathogens and the effect on the growth of soybean sprouts were examined by using P. areofacience 14H-3, P. fluorescens R1-12 and B. cereus Yell, bacteria were shown to inhibit mycellial growth of Rhizotonia solani strongly. The results of this study are summarized as follows. P. areofacience 14H-3 and B. cereus Yell were highly antagonistic against Rizoctonia solani, while they were especially highly antagonistic against bacterial diseases. The effect of inhibiting the proliferation of soybean sprout rotting pathogens was also examined by adding the culture solution for antagonistic bacteria to the PDA. Both P. areofacience 14H-3 and P. fluorescens Rl-12 showed the inhibition rate of 78.8%, while B. cereus Yell did 52.9%. The fresh weight and length of soybean sprouts were measured after raising them with added antagonistic microbes and culture medium. Soybean sprouts treated with B. cereus Yell showed increased higher, compared with those not treated with it. Soybean sprouts were also raised in the culture solution with antagonistic bacteria to examine the growth of soybean sprouts. Soybean sprouts treated with the culture solution of 200 times showed better growth than those not treated with it. Analyze proximate composition in soybean sprout showed that moisture, ash, total sugar did not appear difference, but in case of crude protein B. cereus Yell(8.9%) increased about 2 times than control(3.6%), but occasion of crude fat and crude fiber were P. areofacience 14H-3, P. fluorescens Rl-12 increased about each 2 times than control. In occasion of vitamin, bacterial antagonist(9.4∼10.8mg%) was more higher than control(9.9mg%).

• Applied Biological Chemistry
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• v.38 no.5
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• pp.371-375
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• 1995

The antagonistic rhizobacteria Pseudomonas(P.) fluorescens against F. oxysporum and R. solani were isolated and selected, and then, their biological and physiological characteristics were investigated. The posibility and optimum condition of the electroporation of antagonistic rhizobacteria with Ps70, one of the selected one, and plasmid pSV2-neo was studied. Its optimum condition was found with HGEB which contains 1 mM (pH 7.0) hepes and 10% glycerol at setting of 200 resistance, 25 ${\mu}F$ capacitance, and 2.5 kV applied voltage. In addition, the transformation efficiency obtained with pSV2-neo was compared to other plasmids with different sizes. The applied voltage, the buffer composition and the parallel resistor (time constant) were shown to have the greatest effect on transformation efficiency in electroporation. And the rest of the selected rhizobacteria were also successfully transformed with pSV2-neo by electroporation.

• Journal of Microbiology and Biotechnology
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• v.28 no.7
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• pp.1133-1140
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• 2018

Pseudomonas fluorescens KLR101 was found to be capable of producing polyhydroxyalkanoate (PHA) using various sugars and fatty acids with carbon numbers ranging from 2 to 6. The PHA granules consisted mainly of a poly(3-hydroxybutyrate) homopolymer and/or poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer. Genomic DNA of P. fluorescens was fractionated and cloned into a lambda library, in which a 5.8-kb fragment that hybridized to a heterologous phaC probe from Ralstonia eutropha was identified. In vivo expression in Klebsiella aerogenes KC2671 (pUMS), restriction mapping, Southern hybridization experiments, and sequencing data revealed that PHA biosynthesis by P. fluorescens relied upon a polypeptide encoded by a 1,683-bp non-operonal ORF, which was preceded by a possible -24/-12 promoter and highly similar to DNA sequences of a gene encoding PHA synthase in the genus Pseudomonas. In vivo expression of the putative PHA synthase gene ($phaC_{Pf}$) in a recombinant Escherichia coli strain was investigated by using glucose and decanoate as substrates. E. coli (${phaC_{Pf}}^+$, pUMS) grown in medium containing glucose accumulated PHA granules consisting mainly of 3-hydroxybutyrate, whereas only a trace amount of 3-hydroxydecanoate was detected from an E. coli fadR mutant (${phaC_{Pf}}^+$) grown in medium containing decanoate. In vitro enzymatic assessment experiments showed that 3-hydroxybutyryl-CoA was efficiently used as a substrate of purified $PhaC_{Pf}$, suggesting that the putative PHA synthase of P. fluorescens utilizes mainly short-chain-length PHA precursors as a substrate.

• Korean Journal Plant Pathology
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• v.10 no.3
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• pp.151-156
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• 1994

Pseudomonas fluorescens produces the antibiotic, 2,4-diacetylphloroglucinol (Phl), which promotes plant growth by inhibiting bacteria and fungi. Cosmids (genomic library) were mobilized into Phl-nonproducing mutants through the triparental matings with pRK2013 as the helper plasmid at the frequency of 8.37$\times$10-4. Complemented mutants that showed antibiotic activity were selected among about 2,000 transconjugants. The complemented mutants were confirmed by acquired drug resistances (kanamycin and tetracycline). The antibiotic substances of wild type and complemented mutants showed the most excellent anti-bacterial activity. Inhibitory effects of complemented P. fluorescens against phytopathogenic fungi were equal to the parental strain. Complemented mutant and wild type of P. fluorescens were causal microbes of fungal morphological abnormalities. Complemented mutants in potato dextrose agar supplemented with bromothymol blue also showed restoration of glucose utilization as wild type. Plasmids of complemented mutants were isolated from transconjugant sand transformed into competent cells of E. coli DH5$\alpha$. The plamid DNA was reisolated from transformed E. coli DH5$\alpha$.

• Korean Journal of Food Science and Technology
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• v.23 no.4
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• pp.452-458
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• 1991

Psychrotrophs producing protease were isolated during ripening periods of Cheddar cheese and one of them containing the highest protease activity was identified as Pseudomonas fluorescens 65. The extracelluar proteolytic enzyme was partially purified from P. fluorescens 65 through the Sephadex G-100 gel filtration. The protease was eluted between 190 ml and 230 ml of elution volume of sodium phosphate buffer. The purified protease showed a single band in SDS-PAGE and its molecular weight was 47,000. The composition of amino acid for the protease was determined and the most abundant amino acids were glutamic acid (14.96%) and serine (13.86%). The optimum temperature and pH for the activity was $45{\sim}50^{\circ}C$ and 6.0, respectively.

• Journal of Environmental Science International
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• v.16 no.11
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• pp.1219-1223
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• 2007

The removal of nitrogen compounds from a wastewater is essential and it is often accomplished by bio-logical process. An aerobic nitrate-removing bacterium was isolated from a municipal sewage treatment plant and soil. On the basis of its morphological, cultural and physiological characteristics and 16S rRNA sequencing data, this strain was identified as Pseudomonas fluorescens, and named as P. fluorescens K4. The optimal conditions of the initial pH and temperature of media for its growth were $7.0{\sim}8.0$ and $30^{\circ}C$, respectively. P. fluorescens K4 was able to remove 99.9% of nitrate after 24 h in a culture. The strain could grow with a nitrate concentration up to 800 mg/l and was able to remove 99.9% of nitrate after 104 h of incubation. The optimal electron donor was sodium citrate for a nitrate removal. The strain K4 showed a capability of a complete nitrate removal when the initial C/N ratio was 1.0. An effect of the initial seed concentration was observed for a cell of 10% (v/v) for a nitrate removal. Especially P. fluorescens K4 could completely remove 200 mg/l ammonium for 3 days.

• Korean Journal of Microbiology
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• v.51 no.4
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• pp.448-452
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• 2015

The bacterial strain that was isolated from chinese cabbage rhizosphere, showed inhibition of yeast growth. This strain was identified as Pseudomonas fluorescens BB2 by API 20NE test and 16S rRNA gene sequence analysis. P. fluorescens BB2 strain produced antibiotics against yeast as a secondary metabolite effectively when the culture was carried out in YM medium with 3% glucose at $20^{\circ}C$. The protein antibiotic of BB2 strain which was concentrated by ammonium sulfate precipitation and n-butanol extraction inhibited the growth of yeast with the minimal inhibitory concentration of $10{\mu}g/ml$ against Candida albicans KCTC 7965, and the growth of yeast was completely inhibited at $80{\mu}g/ml$. The hydrophilic fraction of n-butanol extraction inhibited the growth of Bacillus cereus ATCC 21366, showed orange halo on chrome azurol S plate, which means the fraction contained iron chelating siderophore. The results of crystal violet uptake through the cell membrane showed that membrane permeability was increased about 9% than control, when the concentration of hydrophobic antibiotic against yeast C. albicans was $60{\mu}g/ml$. As a result, the antibiotic produced by P. fluorescens BB2 against yeast Candida is considered antimicrobial peptide, and this is the first report in the genus Pseudomonas.