• Research in Plant Disease
• /
• v.10 no.4
• /
• pp.310-312
• /
• 2004

Pseudomonas syringae pv. actinidiae is the causative agent of bacterial canker in kiwifruit. A nested PCR detection method that uses primers designed from the cfl gene, involved in production of the phytotoxin coronatine, was applied on soil samples. These primers yielded 665 and 310-bp fragments in consecutive PCR amplification step with DNA from soil inoculated with Korean strain of P. syringae pv. actinidiae. This system was applied to survey soil samples from a kiwifruit orchard destroyed by bacterial canker. A specific 310-bp PCR product was obtained from all six samples of soil tested.

• Korean Journal of Microbiology
• /
• v.24 no.3
• /
• pp.270-275
• /
• 1986

The localization of carbon monoxide dehydrogenases (CO-DHs) in Pseudomonas carvoxydovorans and Acinetobacter sp.1 was examined by comparison of the distribution of CO-oxidizing activity between soluble and particulate fractions obtained after disruption of CO-grown cells by sonic oscillation and of spheroplasts by osmotic shock. When the cells were broken by sonic oscillation, most of the CO-DH activity was recovered from soluble fractions. However, disryption by osmotic lysis of spheroplasts revealed that the enzyme activity is present in the cell membrane. The results indicated the CO-DHs in these cells are loosely attached to the cytoplasmic membrane.

• Korean Journal of Microbiology
• /
• v.34 no.3
• /
• pp.115-119
• /
• 1998

We cloned the 4.8 kb BglII fragment containing genes downstream pHENX7 from Pseudomonas sp. strain DJ77. The restriction map of the resultant clone, recombinant plasmid pYCS500, was determined. Sequencing analysis of the 465 bp HindIII-ClaI fragment revealed an open reading frame of 282 bp that was then designated phnM. The deduced polypeptide is 93 amino acid residues long with a $M_r$ of 10,008. The PhnM has 37.3-53.9% identity with plant-type ferredoxin proteins such as NahT, XylT, DmpQ, AtdS, PhlG, PhhQ and TbuW and contains the motif similar to well-conserved functional domains of those proteins.

• BMB Reports
• /
• v.30 no.2
• /
• pp.132-137
• /
• 1997

In order to compare molecular structure, malonate decarboxylases from Acinetobacter calcoaceticus, Pseudomonas fluorescens, and Pseudomonas putida aerobically grown on malonate, were purified by the method employing streptomycin sulfate treatment, chromatography with PBE 94 and ${\omega}-aminohexyl$ agarose. Molecular masses were estimated to be 185, 200, and 200 kDa, respectively. All malonate decarboxylases were multimeric enzymes consisting of four different subunits, $2{\alpha},\;1{\beta},\;1{\gamma},\;and\;1{\delta}$. The molecular masses of the Pseudomonas enzyme subunits were $65({\alpha})$, $33({\beta})$, $30({\gamma})$, and $11kDa({\delta})$; which are very similar to those, $65({\alpha})$, $32({\beta})$, $25({\gamma})$, and $11kDa({\delta})$ of Acinetobacter enzyme. The ${\delta}-subunit$ of the active form of the enzymes was acetylated. The acetyl group may form a thioester bond with the thiol group of the prosthetic group covalently linked to the enzyme. It suggests that such molecular organization is common in all malonate decarboxylases.

• Microbiology and Biotechnology Letters
• /
• v.29 no.3
• /
• pp.134-141
• /
• 2001

A bacterial strain KP-364 producing extracellular keratinolytic protease was isolated from the soil of the poultry fac-tory. It was identified as Pseudomonas sp. based on its morphological and physiological characteristics, The optimal culture conditions for the production of keratinolytic protease by Pseudomonas sp. KP-364 were investigated. The composition of optimal medium for the keratinolytic protease was 2.0% glucose, 0.5% soybean meal. 0.5% $NaNO_3$ and 0.2% KCI Optimal initial pH for production of Keratinolytic protease production were 6.5 and $37^{\circ}C$ respec- tively. The keratinolytic protease production reached a maximum of 1,270 U/ml/hr after 48 hours cultivation under the optimal culture conditions.

• The Korean Journal of Food And Nutrition
• /
• v.15 no.4
• /
• pp.300-306
• /
• 2002

141 methanol extracts from 125 plant species which populate in Korea were screened for antimicrobial activity against various food-borne pathogens and food spoilage microorganisms. Those plants were selected from 3 different plant groups: traditional herbs, edible plants and flowers. The methanol extracts were tested by using the disk diffusion assay against five bacteria: Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Enterobacter aerogenes, Escherichia coli. From the evaluation of the inhibition zone diameter of microbial growth, the most significant antimicrobial activity against Bacillus subtilis, Staphylococcus auresus, Pseudomonas aeruginosa, Enterobacter aerogenes, Escherichia coli was observed from the extract of Schizandra chinensis (Turcz.) Baill., Rheum officinale Baill., Schizandra chinensis (Turcz.) Baill., Koelreuteria paniculata Lax and Crataegus pinnatifida Bunge, respectively. The extract from many plants - Koelreuteria paniculata Lax, Chaenomeles sinensis Koehne, Scutellaria bacicalensis Georgi, Castanea crenata Sieb. et Zucc., Rosa centifolia L., Allium fistulosum L. var. giganteum Makino, Crataegus pinnatifida Bunge, Schizandra chinensis (Turcz.) Baill., Lonicera japonica - showed antimicrobial activity all four tested bacteria.

• Journal of Environmental Science International
• /
• v.11 no.1
• /
• pp.33-40
• /
• 2002

An antimicrobial substance-producing microorganism was isolated from soil samples. Based of the taxonomic characteristics of its morphological, cultural, physiological properties and 16s rRNA sequence alignment, this microorganism was identified as Pseudomonas aeruginosa, and we named Pseudomonas aeruginosa EL-KM. The optimal culture condition for production of antimicrobial substance was 1% mannitol, 0.4% yeast extract, 0.5% Nacl, 0.2% $K_2SO_4$, 100$\mu$M $MgSO_4$.$7H_2O$, 10$\mu$M $CaCl_2$.$2H_2O$, 1$\mu$M $FeSO_4$.$7H_2O$, 1$\mu$M $MnSO_4$.$4-5H_2O$, initial pH 7 and 200 rpm at 3$0^{\circ}C$. The purification of the antimicrbial substance was performed by silica gel column chromatographys, and fraction with TLC $R_f$ 0.77 value represented good antimicrobial activity. The crude antimicrbial substance was stable within a pH range of 3-10 and temperature range of 4$^{\circ}C$-121$^{\circ}C$ autoclaving. This crude antibacterial substance acted as bacteriolytic agent against Vibrio cholerae non-Ol ATCC 25872, and also exhibited excellent properties, when the substance was demonstrated against many other gram-positive, gram-negative bacteria, yeast and fungi.

• Journal of Environmental Science International
• /
• v.10 no.5
• /
• pp.359-364
• /
• 2001

Pseudomanas sp. EL-04J was previously isolated from phenol-acclimated activated sludge. This bacterium was capable of degrading phenol and cometabolizing trichloroethylene (TCE). After precultivation in the mineral salts medium containing phenol as a sole carbon source, Pseudomonas EL-04J degraded 90% of TCE $25 \mu\textrm{M}$ within 20 hours. Thus, phenol-induced Pseudomonas sp. EL-04J cells can bdegrade TCE. Followsing a transient lag period, Pseudomonas sp. EL-04J cells degraded TCE at concentrations of at least $250 \mu\textrm{M}$ with no apparent retardation in rate, but the transformance capacity of such cells was limited and depended on the cell concentration. The degradation rate of TCE followed the Michaelis-Menten kinetic model. The maximum degradation ratio ($V_{max}$) and saturation constant ($K_{m}$) were $7nmo {\ell}/min{\cdot}mg$ cell protein and $11 \mu\textrm{M}$, respectively. Cometabolism of TCE by phenol fed experiment was evaluated in $50m {\ell}$ serum vial that contained $10m {\ell}$ of meneral sals medium supplemented with $10 \mu\textrm{M}$ TCE degradation was inhibited in the initial period of 1 mM phenol addition, but after that time Pseudomonas sp. EL-04J cells degraded TCE and showed cell growth.

• Journal of Soil and Groundwater Environment
• /
• v.21 no.5
• /
• pp.16-24
• /
• 2016

A new strain of Pseudomonas sp. was isolated from mercury (Hg)-contaminated sites in Taiwan. This bacterium removed more than 80% of Hg present in the culture medium at 12 h incubation and was chosen for further analysis of the molecular mechanisms of Hg tolerance/removal abilities in this Pseudomonas sp. We used RNA-seq, one of the next-generation sequencing methods, to investigate the transcriptomic responses of the Pseudomonas sp. exposed to 60 mg/L of Hg²⁺. We de novo assembled 4,963 contigs, of which 10,533 up-regulated genes and 5,451 down-regulated genes were found to be regulated by Hg. The 40 genes most altered in expression levels were associated with tolerance to Hg stress and metabolism. Functional analysis showed that some Hg-tolerant genes were related to the mer operon, sulfate uptake and assimilation, the enzymatic antioxidant system, the HSP gene family, chaperones, and metal transporters. The transcriptome were analyzed further with Gene Ontology (GO) and Cluster of Orthologous Groups (COGs) of proteins and showed diverse biological functions and metabolic pathways under Hg stress.

• Journal of Applied Biological Chemistry
• /
• v.58 no.4
• /
• pp.349-354
• /
• 2015

Brown blotch disease in cultivated mushrooms is caused by Pseudomonas tolaasii, which secretes a lipodepsipeptide, tolaasin. Tolaasin is a pore-forming toxin in the cell membranes, thus destroying the fruiting body structure of mushroom. In this study, we isolated pathogenic bacteria from mushrooms that had symptoms of brown blotch disease. In order to identify these bacteria, their 16S rRNA genes were sequenced and analyzed. Pathogenic bacteria identified as Pseudomonas species were thirty five and classified into five subgroups: P1 to P5. Each subgroup showed different metabolic profile measured by API 20NE kit. Fifty percent of the bacteria were identified as P. tolaasii (P1 subgroup). All five subgroups caused the formation of brown blotches on mushroom tissues and the optimum temperature was 25oC, indicating that they may be able to secrete causal factors, such as tolaasin and similar peptide toxins. These results show that there are at least five different pathogenic Pseudomonas species as blotch-causing bacteria and, therefore, strains from the P2 to P5 subgroups should be also considered and studied as pathogens in order to improve the quality and yield of mushroom production.