• Microbiology and Biotechnology Letters
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• v.36 no.4
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• pp.268-275
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• 2008

In order to develop the multi-functional rhizobacteria that can exert positive effect on the growth of plants growing in the coastal sand dune located along East Coast of Korea, rhizospheral bacteria of 11 different plants from this area were isolated 1,330 rhizobacteria. Among these, 23 strains were able to produce auxin and had spectrum of antagonism toward various phytopathogenic microbes. To know the mechanism of this antifungal activity, these 23 strains were subjected to further analyses; 19 strains of these produced siderophore as determined by color reaction on CAS-blue plate, 4 strains produced antifungal cellulase as judged by color change on CMC-Congo red plate, 17 strains were able to utilized insoluble phosphate salts, also determined by clear zone formation on PVK medium. Identification of the strain was assigned to all 23 strains by l6s rDNA sequence analysed, and all were identified to be in the genus of Bacillus and Pseudomonas. One strain of these, denoted Pseudomonas fluorescens IB4-14, showed ACC deaminase activity which is known to be involved in the resistance of environmental stress such as salt and drought. Also, P. fluorescens IB4-l4 showed the germination stimulation and roots growth promoting activity on the in vivo assay of Lysimachia mauritiana Lam. (spoonleaf yellow loosestrife).

• Korean Journal of Environmental Biology
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• v.25 no.3
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• pp.273-278
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• 2007

The cell numbers of heterotrophic bacteria inhabiting the surface and bottom sea water harvested from the 5 stations in the marine ranching ground of Tongyeong coastal waters in $2003{\sim}2007$ were examined, and species composition of the heterotrophic bacterial population and dominant species were analyzed as well. Sea water samples collected in summer season contained much higher number of heterotrophic bacteria than those harvested in winter, spring and autumn seasons due to the higher sea water temperature. However the cell number of heterotrophic bacteria did not show a significant dependence on the location of the sampling stations. The cell number of heterotrophic bacteria in the surface sea water harvested in October 2003 and in September 2004 was not discernibly different from that in the bottom sea water and sometimes the former was even fewer than the latter because of the typhoon and localized torrential downpour. The number of heterotrophic bacteria decreased every year. The main bacterial species were Pseudomonas fluorescens TY1, Pseudomonas stutzeri TY2, Acinetobacter lwoffii TY3, Sphingomonas paucimobilis TY4, Burkholderia mallei TY5, Pasteurella haemolytica TY6, Pasteurella multocida TY7, Comamonas acidovorans TY8, Actinobacillus ureae TY9 and Chryseobacterium indologenes TY10. P. fluorescens TY1 and A. lwoffii TY3 were found to be the dominant species.

• The Korean Journal of Mycology
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• v.25 no.1 s.80
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• pp.10-19
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• 1997

Fusarium wilt of radish (Raphanus sativus L.) is caused by the Fusarium oxysporum f. sp. raphani (FOR) which mainly attacks Raphanus spp. The pathogen is a soil-borne and forms chlamydospores in infected plant residues in soil. Infected pathogen colonizes the vascular tissue, leading to necrosis of the vascular tissue. Growth promoting beneficial organisms such as Pseudomonas fluorescens WCS374 (strain WCS374), P. putida RE10 (strain RE10) and Pseudomonas sp. EN415 (strain EN415) were used for microorganisms-mediated induction of systemic resistance in radish against Fusarium wilt. In this bioassy, the pathogens and bacteria were treated into soil separately or concurrently, and mixed the bacteria with the different level of combination. Significant suppression of the disease by bacterial treatments was generally observed in pot bioassy. The disease incidence of the control recorded 46.5% in the internal observation and 21.1% in the external observation, respectively. The disease incidence of P. putida RE10 recorded 12.2% in the internal observation and 7.8% in the external observation, respectively. However, the disease incidence of P. fluorescens WCS374 which was proved to be highly suppressive to Fusarium wilt indicated 45.6% in the internal observation and 27.8% in the external observation, respectively. The disease incidence of P. putida RE10 mixed with P. fluorescens WCS374 or Pseudomonas sp. EN415 was in the range of 10.0-22.1%. On the other hand, the disease incidence of P. putida RE10 mixed with Pseudomonas sp. EN415 was in the range of 7.8-20.2%. The colonization by FOR was observed in the range of $2.4-5.1{\times}10^3/g$ on the root surface and $0.7-1.3{\times}10^3/g$ in the soil, but the numbers were not statistically different. As compared with $3.8{\times}10^3/g$ root of the control, the colonization of infested ROR indicated $2.9{\times}10^3/g$ root in separate treatments of P. putida RE10, and less than $3.8{\times}10^3/g$ root of the control. Also, the colonization of FOR recorded $5.1{\times}10^3/g$ root in mixed treatments of 3 bacterial strains such as P. putida RE10, P. fluorescens WCS374 and Pseudomonas sp. EN415. The colonization of FOR in soil was less than that of FOR in root part. Based on soil or root part, the colonization of ROR didn't indicate a significant difference. The colonization of introduced 3 fluorescent pseudomonads was observed in the range of $2.3-4.0{\times}10^7/g$ in the root surface and $0.9-1.8{\times}10^7/g$ in soil, but the bacterial densities were significantly different. When growth promoting organisms were introduced into the soil, the population of Pseudomonas sp. in the root part treated with P. putida RE10 was similar in number to the control and recorded the low numerical value as compared with any other treatments. The population density of Pseudomonas sp. in the treatment of P. putida RE10 indicated significant differences in the root part, but didn't show significant differences in soil. The population densities of infested FOR and introduced bacteria on the root were high in contrast to those of soil. P. putida RE10 and Pseudomonas sp. EN415 used in this experiment appeared to induce the resistance of the host against Fusarium wilt.

• Korean Journal of Environmental Agriculture
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• v.12 no.2
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• pp.144-152
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• 1993

A bacterium which degrades efficiently synthetic detergents was isolated from the polluted waters, activated sludge of wastewater treatment plants or polluted soil. This bacterium showed considerably higher growth rate in the agar plate containing $2,000{\mu}g/ml$ of synthetic detergents than any other isolated strains, was identified as a Pseudomonas fluorescens or strains similar to it. The strain was named as a Pseudomonas fluorescens S1. Optimum pH and temperature for the growth of the Pseudomonas fluorescens S1 were pH 7.0 and $30^{\circ}C$, respectively. The strain was resistant to streptomycin and gentamycin, but sensitive to kanamycin. The strain was greatly resistant to zinc chloride, lead nitrate and copper sulfate, but unable to grow in the presence of relatively low concentrations of mercury chloride and silver nitrate. This strain utilized benzene, catechol, cyclohexane and xylene as a sole carbon source. The strain was well grown in the medium containing ABS 10,000${\mu}g$/ml. Degradation of ABS was 55% and 60% at 20${\mu}g$/ml and 100${\mu}g$/ml of ABS, respectively. Benzene ring was degraded 45% in 100${\mu}g$/ml of ABS. During the incubation of the strain in the medium containing ABS 100${\mu}g$/ml and COD 10,000${\mu}g$/ml for 4 days, degradation of ABS and COD were reduced to 40${\mu}g$/ml and 3,200${\mu}g$/ml, respectively. Total amino acid content of the Pseudomonas fluorescens S1 grown with 1,000${\mu}g$/ml of ABS was 115mg/g cell, whereas its content was decreased in the bacterium grown without synthetic detergent by 9.4%.

• Korean Journal of Microbiology
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• v.13 no.4
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• pp.143-156
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• 1975

A rotting bacterium was isolated from decayed root of ginseng (Panax ginseng Meyer), cultured purely, and its pathogenicity was confirmed by reinoculation test. The strain causing ginseng root rot was identified as Pseudomonas fluoresens biotype II. The strain was somewhat different from P.marginalis and P.talaasii, considering the number of flagella, pathotype and ability of indole production. The strain did not exhibit pathogenicity to other plants tested, such as red kidney bean(Phasolus vulgaris L.), soy bean (Glycine max Merr.), cucumber (Cucumis sativus L.) lettuce (Lactuca sativa L.) and cowpea bean (Vigna sinensis Savi.).

• The Plant Pathology Journal
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• v.26 no.2
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• pp.189-193
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• 2010

The in vitro antibacterial properties of two kinds of chitosan solutions and their effect in protection of broccoli from bacterial head rot disease were evaluated. Results showed that the two kinds of chitosan solution at different concentrations exhibited strong antibacterial activity against Pseudomonas fluorescens. However, the antibacterial activity of chitosan A solution increased with the increase of chitosan concentration up to 0.10 mg/ml while the antibacterial activity of chitosan B solution increased with the increase of chitosan concentration up to 0.05 mg/ml. In addition, the antibacterial activity of chitosan A and chitosan B solution of 0.10 mg/ml increased with the incubation time within 12 h and 24 h, respectively. The disease incidence and the lesion diameter of broccoli inoculated with P. fluorescens were significantly reduced when plants were either pretreated or post-treated with six different combinations of chitosan solutions. Overall, the results indicated that the two kinds of chitosan solutions had a potential in controlling bacterial head rot of broccoli.

• The Plant Pathology Journal
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• v.21 no.3
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• pp.262-269
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• 2005

An antiviral producing bacterial strain was isolated from a ginseng rhizosphere in Kangwon province of Republic of Korea. In order to identify the bacterial strain, microbiological, physiological and biochemical tests were performed, along with RAPD, 16S rRNA, 16S-23S rRNA ITS (intergenic spacer region) and DNA-DNA hybridization analyses. The bacterium was found to be a strain of Pseudomonas fluorescens, which was designated as Gpf01. The strain was grown in Muller-Hinton (MH) broth, and the culture supernatant obtained was filtered through a $0.45{\mu}l$ filter. It was further boiled at $100^{\circ}C$ and tested in two experiments for its ability to control a yellow strain of Cucumber mosaic virus (CMV-Y). In the first experiment, boiled culture filtrate (RCF) was treated on one half of the leaves of Chenopodium amaranticolor followed by CMV- Y inoculation on both halves. In the second experiment, BCF was treated on the lower leaves of Nicotiana tobacum var. Xanthi-nc, with the CMV-Y mechanically inoculated onto the upper untreated leaves. In the first experiment, BCF treatment was able to considerably reduce the number of viral lesion, and in the second experiment, plants treated with BCF showed no visible viral symptoms compared to the Muller-Hinton (MH) media treated controls 15 days post inoculation (dpi), and remained symptomless throughout the study period. Thus, Gpf01, identified as P. fluorescence, was able to produce an antiviral component in the culture filtrate, which was found to be heat stable, non-phytotoxic and effective in local as well as systemic hosts of CMV.

• BMB Reports
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• v.29 no.4
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• pp.277-285
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• 1996

In Pseudomonas fluorescens grown on malonate as sole carbon source, acetyl-CoA synthetase was induced, suggesting that malonate is metabolized through acetate and then acetyl-CoA. Acetyl-CoA synthetase was purified 18.6-fold in 4 steps to apparent homogeneity. The native molecular mass of the enzyme estimated by a native acrylamide gel electrophoresis was 130 kDa. The enzyme was composed of two identical subunits with a molecular mass of 67 kDa. Optimum pH was 70. The acetyl-CoA synthetase showed typical Michaelis-Menten kinetics for the substrates, acetate, ATP and CoA, whose $K_m$ values were calculated to be 33.4, 74.8, and 40.7 mM respectively. Propionate. butyrate and pentanoate were also used as substrates by the enzyme, but the rate of the formation of the CoA derivatives was decreased in the order of the increase in carbon number. The enzyme was inhibited by the group-specific reagents diethylpyro-carbonate, 2,3-butanedione, pyridoxal-5'-phosphate and N-bromosuccinimide. In the presence of substrates the inactivation rate of the enzyme, by all of the group-specific reagents mentioned above decreased, indicating the presence of catalytically essential histidine, arginine, lysine and tryptophan residues at or near the active site. Preincubation of the enzyme with ATP, $Mg^{2+}$ resulted in the increase of its susceptibility to diethylpyrocarbonate, suggesting that ATP, $Mg^{2+}$ may induce a conformational change in the active site exposing the essential histidine residue to diethylpyrocarbonate. The enzyme was acetylated in the presence of acetyl-CoA, indicating that this is one of acyl-enzyme.

• Journal of Life Science
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• v.30 no.5
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• pp.420-427
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• 2020

A repebody, an artificial non-immunoglobulin protein scaffold, is expected to be a solution in the search for faster, cheaper, and customizable antibodies. However, the production of medical repebodies remains difficult due to their low yield and the complex purification processes required. The Pseudomonas fluorescens ABC transporter system has been suggested as an efficient and cost-effective method for repebody production, but the total yield is low because of the secreted protein's positive charge; thus, a repebody with a high isoelectric point needs to be changed into a more negatively charged protein for better secretion. To achieve this, we first attached oligo-aspartic acids to the N- and C-terminals of the repebody, but secretion efficiency was not enhanced significantly. Subsequently, we devised an alternative method for improved secretion efficiency by engineering fifteen positively charged amino acids to aspartic acid in the non-antigen binding sites of the repebody to give a high net negative charge. As a result, secretion efficiency was greatly enhanced from 21.2% (wildtype) to 58.5% (negatively supercharged). The negatively supercharged repebody was succussfully produced extracellularly by ABC transporter secretion system in P. fluorescens.

• Korean Journal of Food Science and Technology
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• v.40 no.5
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• pp.562-567
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• 2008

The primary objective of this study was to select a probiotic starter for cheonggukjang using 60 strains isolated from rice straw. Among isolated strains, only 8 strains including strain B-59 evidenced proteolytic, amylolytic and soybean activity. These 8 strains were all gram-positive, spore-forming rods. The B-59 strain evidenced antimicrobial activity against Staphylococcus aureus, Listeria monocytogenes, Pseudomonas fluorescens, and Vibrio parahaemolyticus. The antimicrobial activity of isolated B-59 was verified by its ability to inhibit the growth of S. aureus, L. monocytogenes, P. fluorescens, and V. parahaemolyticus. The selected B-59 strain was indentified as Bacillus licheniformis, as shown by a result of 99.0% homology upon API kit analysis. The selected B-59 strain also displayed viability in pH 2.5 artificial gastric juice, artificial bile acid, NaCl (2, 4, 8, 16, 32%), and ethanol (4, 8, 16, 32%). The antioxidative activity of the strain B-59 culture was assessed via a DPPH free radical scavenging activity assay. The activity increased with an increasing in the fermentation time of strain B-59 for 20 hours.