• Journal of Microbiology and Biotechnology
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• v.25 no.6
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• pp.887-892
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• 2015

Uricase is an important microbial enzyme that can be used in the clinical treatment of gout, hyperuricemia, and tumor lysis syndrome. A total of 127 clinical isolates of Pseudomonas aeruginosa were tested for uricase production. A Pseudomonas strain named Ps43 showed the highest level of native uricase enzyme expression. The open reading frame of the uricase enzyme was amplified from Ps43 and cloned into the expression vector pRSET-B. Uricase was expressed using E. coli BL21 (DE3). The ORF was sequenced and assigned GenBank Accession No. KJ718888. The nucleotide sequence analysis was identical to the coding sequence of uricase gene puuDof P. aeruginosa PAO1. We report the successful expression of P. aeruginosa uricase in Escherichia coli. E. coli showed an induced protein with a molecular mass of about 58 kDa that was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. We also established efficient protein purification using the Ni-Sepharose column with activity of the purified enzyme of 2.16 IU and a 2-fold increase in the specific activity of the pure enzyme compared with the crude enzyme.

• Biomedical Science Letters
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• v.18 no.1
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• pp.16-21
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• 2012

Although there are many potential cytotoxic molecules released from bacteria, the role of these molecules on the apoptosis of various cancer cells is not well understood. Pseudomonas aeruginosa (P. aeruginosa) is a Gram-negative, aerobic and rod-shaped bacterium, and has a number of virulence factors. To understand the cytotoxic effect of bacterial extracts on the colorectal cancer cell line, HCT 116 cells, we examined alteration of the cell viability, proliferation, cell cycle and apoptosis of HCT 116 cells after treatment with extract of P. aeruginosa (PaE). These cytotoxicity of PaE occurred in a time- and a dose-dependent manners. In addition, PaE arrested the cell cycle of HCT 116 cell in a time-dependent manner. PaE inhibited the protein levels of Bcl-2 and induced the release of cytochrome c from mitochondria of HCT 116 cells. The decrease of procaspase-3 was induced by the treatment of PaE. These results indicate that PaE has a cytotoxicity in HCT 116 cells via the induction of apoptosis associated with mitochondrial pathway. Therefore, PaE may used as the potential target for the treatment of colorectal cancer.

• Journal of Microbiology and Biotechnology
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• v.25 no.7
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• pp.1154-1162
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• 2015

The emergence of carbapenem resistance among Pseudomonas aeruginosa is an increasing problem in many parts of the world. In particular, metallo-$\beta$-lactamases (MBLs) and AmpC $\beta$lactamases are responsible for high-level resistance to carbapenem and cephalosporin. We studied the diversity and frequency of $\beta$-lactamases and characterized chromosomal AmpC $\beta$lactamase from carbapenem-resistant P. aeruginosa isolates. Sixty-one carbapenem-resistant P. aeruginosa isolates were collected from patients in a tertiary hospital in Daejeon, Korea, from January 2011 to June 2014. Minimum inhibitory concentrations (MICs) of four antimicrobial agents were determined using the agar-dilution method. Polymerase chain reaction and sequencing were used to identify the various $\beta$-lactamase genes, class 1 integrons, and chromosomally encoded and plasmid-mediated ampC genes. In addition, the epidemiological relationship was investigated by multilocus sequence typing. Among 61 carbapenem-resistant P. aeruginosa isolates, 25 isolates (41.0%) were MBL producers. Additionally, 30 isolates producing PDC (Pseudomonas-derived cephalosporinase)-2 were highly resistant to ceftazidime (MIC₅₀ = $256{\mu}g/ml$) and cefepime (MIC₅₀ = $256{\mu}g/ml$). Of all the PDC variants, 25 isolates harboring MBL genes showed high levels of cephalosporin and carbapenem resistance, whereas 36 isolates that did not harbor MBL genes revealed relatively low-level resistance (ceftazidime, p < 0.001; cefepime, p < 0.001; imipenem, p = 0.003; meropenem, p < 0.001). The coexistence of MBLs and AmpC $\beta$-lactamases suggests that these may be important contributing factors for cephalosporin and carbapenem resistance. Therefore, efficient detection and intervention to control drug resistance are necessary to prevent the emergence of P. aeruginosa possessing this combination of $\beta$-lactamases.

• Korean Journal of Microbiology
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• v.50 no.4
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• pp.281-284
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• 2014

Immune defense responses against Pseudomonas aeruginosa infection play an important role in maintaining homeostasis in the human body. Previously, we reported that expression of the bradykinin receptor (BR) is induced in response to P. aeruginosa infection. However, the factors responsible for the induction was uncertain. Here, we found that the type III secretion system (T3SS) is responsible for the induction of BR expression, and nucleoside diphosphate kinase (Ndk), as a novel T3SS effector, mediates the upregulation. The Ndk-mediated expression of BR was not induced by fliC mutant treatment, indicating the involvement of flagellin, one of the well-known pathogen-associated molecular patterns (PAMPs). Taken together, this study demonstrated that Ndk cooperates with flagella in the development of defense responses against P. aeruginosa infection.

• Microbiology and Biotechnology Letters
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• v.31 no.2
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• pp.171-176
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• 2003

Productivity of biosurfactant (rhamnolipid) by Pseudomonas aeuginosa F722 was investigated in the several culture conditions and culture composition. Biosurfactant production by P. aeuginosa F722 was amounted to 0.78 g/l as the result of the nitrogen sources and carbon sources without investing of optimum conditions. As for that one was investigated, biosurfactant production by P. aeruginosa F722 was amounted to 1.66 g/l. Biosurfactant production increased twofold because the composition of a modified C-medium was investigated efficiently. $NE_4$Cl or $NaNO_2$ inorganic nitrogens and yeast extract or trypton organic nitrogens were effective, but others inorganic nitrogens and organic nitrogens tested were not efficient far biosurfactant production by P. aeruginosa F722. The optimum concentration of $NH_4$Cl; inorganic nitrogen and yeast extract; organic nitrogen were 0.05% and 0.1%, respectively. In various carbon sources, others with the exception of hydrophobic property substrate (n-alkane) and hydrophilic property substrate (glucose, glycol) were not found to be effective fur biosurfactant production, and 3.0% was better in yield than other concentration of glucose. This yielded C-to-N ratios between 17 and 20. In our experiment, the highest biosurfactant production by P. aeruginosa F722 were observed in 5 days cultivation, containing glucose 3.0%, $NH_4$Cl 0.05%, and yeast extract 0.1% and C-to-N ratio was 20. Optimal pH and temperature for biosurfactant production were 7.0 and $35^{\circ}C$, respectively. Under the optimal culture conditions with glucose, biosurfactant production was amounted to 1.66 g/l. Velocity of biosurfactant production and strain growth increased after nitrogen depletion. The average surface tension of 30 mN/m after the 3 days of incubation under optimal culture condition was measured by ring tensionmeter.

• Korean Journal of Microbiology
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• v.48 no.3
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• pp.200-206
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• 2012

A bacterial strain capable of synthesizing polyhydroxyalkanoates (PHAs) with an unusual pattern of monomer units was isolated from activated sludge using the enrichment culture technique. The organism, identified as Pseudomonas aeruginosa P-5, produced polyesters consisting of 3-hydroxyvalerate and medium-chain-length (MCL) 3-hydroxyalkanoate monomer units when $C_{-odd}$ alkanoic acids such as nonanoic acid and heptanoic acid were fed as the sole carbon source. Solvent fractionation experiments using chloroform and hexane revealed that the 3-hydroxyalkanoate monomer units in these polyesters were copolymerized. The molar concentration of 3-hydroxyvalerate in the polyesters produced were significantly elevated up to 26 mol% by adding 1.0 g/L valeric acid as the cosubstrate. These copolyesters were sticky with low degrees of crystallinity. The PHA synthase genes were cloned, and the deduced amino acid sequences were determined. P. aeruginosa P-5 possessed genes encoding MCL-PHA synthases (PhaC1 and PhaC2) but lacked the short-chain-length PHA synthase gene, suggesting that the MCL-PHA synthases from P. aeruginosa P-5 are uniquely active for polymerizing (R)-3-hydroxyvaleryl-CoA as well as MCL (R)-3-hydroxyacyl-CoAs.

• Archives of Plastic Surgery
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• v.36 no.4
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• pp.372-379
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• 2009

Purpose: The purpose of this study was to compare the effect on antibacterial activities and wound healing effect of silver containing dressings with which of Betadine against Staphylococcus aureus and Pseudomonas aeruginosa. Methods: One full thickness skin defects in rats(n=72) were developed on the back and were given rise to infection with S. aureus(n=36) and P. aeruginosa(n=36). The 72 mice were divided into 6 groups : Acticoat$^{(R)}$, Aquacel$^{(R)}$-Ag, Medifoam silver$^{(R)}$, Polymen silver$^{(R)}$, Ilvadon$^{(R)}$ and Betadine(control group) dressing groups. Five silver containing dressings and Betadine were assesed on infected wound. Measurement of wound size change, bacterial colonies count and histologic findings was applied. Antibacterial activity was analyzed with bacterial restricted zone in Mueller Hinton agar. Results: On S. aureus, wound size was more decreased in all treated groups as compared with betadine, however Ilvadon$^{(R)}$ was less decreased on P. aeruginosa. In histologic findings, experimental group showed more effective findings than others on S. aureus, however on P. aeruginosa, which was showed similar. Acticoat$^{(R)}$ was best effective in wound healing against S. aureus and P. aeruginosa. The bacterial colonies count was increased in all treated groups except Acticoat$^{(R)}$ as compared with the control group on S. aureus, which was decreased in Acticoat$^{(R)}$ and Ilvadon$^{(R)}$ group on P. aeruginosa. There were not statistical differences. The restricted zone was shown in Mueller - Hinton agar of all groups except Medifoam silver$^{(R)}$ group on S. aureus, which was shown of all groups on P. aeruginosa. There were statistical differences. Conclusion: This study suggests that silver containing dressings may have not better potential than Betadine in assisting management of wounds at risk of infection on S. aureus and P. aeruginosa. However, which have better antibacterial activity on S. aureus and P. aeruginosa.

• Journal of Microbiology and Biotechnology
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• v.15 no.1
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• pp.86-90
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• 2005

Strain M-18 was isolated from the rhizosphere soil of sweet melon, using 1-aminocyclopropane-1-carboxylate (ACC) as a sole nitrogen source. Its phenotypic characteristics, metabolic tests, and 16S rDNA sequence were analyzed. The antibiotics secreted by strain M-18 were determined to be phenazine 1-carboxylic acid and pyoluteorin. These data showed that strain M-18 was a new fluorescent Pseudomonas strain that produced both phenazine 1-carboxylic acid and pyoluteorin, some features being similar to Pseudomonas aeruginosa and Pseudomonas fluorescens. Therefore, the strain M-18 appears to be the first pseudomonad described to date that is capable of producing both phenazine 1-carboxylic acid and pyoluteorin.

• Journal of Veterinary Clinics
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• v.36 no.5
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• pp.245-247
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• 2019

Taylorella equigenitalis (T. equigenitalis), Klebsiella pneumoniae (K. pneumoniae), and Pseudomonas aeruginosa (P. aeruginosa) are sexually transmittable bacteria known to cause venereal diseases (VD) in horses. T. equigenitalis causes contagious equine metritis (CEM), which is a considerable concern for equine breeding industry. K. pneumoniae and P. aeruginosa may cause endometritis and infertility in susceptible mares. The purpose of this study was to investigate the prevalence of these bacteria among breeding Thoroughbreds in South Korea. External genital swabs were collected from 178 breeding Thoroughbreds, including 11 stallions and 167 mares. The samples were tested using a commercial multiplex real-time PCR kit. T. equigenitalis, P. aeruginosa, and K. pneumoniae were present in 5.6%, 7.3%, and 5.6% of tested Thoroughbreds, respectively. The results highlight the need for regular testing of South Korean Thoroughbreds, particularly those used for breeding, for these bacteria. The regular pre-breeding test for these bacteria will prevent health complications for the horse and financial losses for the owner as a result of VD.

• Journal of Life Science
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• v.20 no.8
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• pp.1214-1220
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• 2010

Imipenem-resistant bacteria were isolated from clinical specimens taken from hospitalized patients in Suncheon, Korea. Fifty-four isolates were phylogenetically analyzed based on 16S rRNA gene and gyrB gene sequence comparisons. Isolates were affiliated with Pseudomonas aeruginosa (30 strains; 55.6%), Acinetobacter baumannii (21; 38.9%), Enterobacter hormaechei (2) and Pseudomonas putida (2). Twenty-two isolates produced metallo-$\beta$-lactamase (MBL); 12 Acinetobacter baumannii strains, 7 Pseudomonas aeruginosa strains, 2 P. putida strains and 1 Enterobacter hormaechei strain. Antibiotic susceptibility of the isolates was determined using the disc diffusion method and Vitek system. Strains producing metallo-$\beta$-lactamase (type IMP & VIM) were more resistant to antibiotics ceftazidime, aztreonam, amikacin and gentamicin than to strains producing OXA and SHV type of $\beta$-lactamase.