• BMB Reports
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• 제37권4호
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• pp.387-393
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• 2004

The L-asparaginase (E. C. 3. 5. 1. 1) enzyme was purified to homogeneity from Pseudomonas aeruginosa 50071 cells that were grown on solid-state fermentation. Different purification steps (including ammonium sulfate fractionation followed by separation on Sephadex G-100 gel filtration and CM-Sephadex C50) were applied to the crude culture filtrate to obtain a pure enzyme preparation. The enzyme was purified 106-fold and showed a final specific activity of 1900 IU/mg with a 43% yield. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme revealed it was one peptide chain with $M_r$ of 160 kDa. A Lineweaver-Burk analysis showed a $K_m$ value of 0.147 mM and $V_{max}$ of 35.7 IU. The enzyme showed maximum activity at pH 9 when incubated at $37^{\circ}C$ for 30 min. The amino acid composition of the purified enzyme was also determined.

• Journal of Veterinary Science
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• 제21권3호
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• pp.46.1-46.18
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• 2020

Background: High concentrations of particulate matter less than 2.5 ㎛ in diameter (PM2.5) in poultry houses is an important cause of respiratory disease in animals and humans. Pseudomonas aeruginosa is an opportunistic pathogen that can induce severe respiratory disease in animals under stress or with abnormal immune functions. When excessively high concentrations of PM2.5 in poultry houses damage the respiratory system and impair host immunity, secondary infections with P. aeruginosa can occur and produce a more intense inflammatory response, resulting in more severe lung injury. Objectives: In this study, we focused on the synergistic induction of inflammatory injury in the respiratory system and the related molecular mechanisms induced by PM2.5 and P. aeruginosa in poultry houses. Methods: High-throughput 16S rDNA sequence analysis was used for characterizing the bacterial diversity and relative abundance of the PM2.5 samples, and the effects of PM2.5 and P. aeruginosa stimulation on inflammation were detected by in vitro and in vivo. Results: Sequencing results indicated that the PM2.5 in poultry houses contained a high abundance of potentially pathogenic genera, such as Pseudomonas (2.94%). The lung tissues of mice had more significant pathological damage when co-stimulated by PM2.5 and P. aeruginosa, and it can increase the expression levels of interleukin (IL)-6, IL-8, and tumor necrosis factor-α through nuclear factor (NF)-κB pathway in vivo and in vitro. Conclusions: The results confirmed that poultry house PM2.5 in combination with P. aeruginosa could aggravate the inflammatory response and cause more severe respiratory system injuries through a process closely related to the activation of the NF-κB pathway.

• 대한임상검사과학회지
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• 제50권4호
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• pp.406-413
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• 2018

최근 P. aeruginosa의 carbapenem에 대한 내성은 전 세계적으로 증가하고 있는 실정이다. 특히 $metallo-{\beta}-lactamases$ (MBLs)는 carbapenem의 고도 내성에 관여하고 있는 것으로 보고되고 있다. 한편, Sequence type 235 (ST235)는 다제내성 클론으로써 국제적으로 보고되고 있으며, IMP-6와 VIM-2 유전자의 확산에도 관여하는 것으로 알려져 있다. 본 연구에서는 2008년 3월부터 2014년 6월까지, 대전지역의 3차 병원에서 분리된 carbapenem 내성 P. aeruginosa에서 MBL 유전자를 분석하고 이에 대한 역학관계를 조사하고자 하였다. 항균제 감수성 양상은 디스크 확산법으로 확인하였고, MBL 유전자의 분석을 위해 PCR과 염기서열분석을 수행하였다. 더불어, 역학 관계를 조사하기 위해 multilocus sequence typing (MLST)를 실시하였다. 110 균주의 carbapenem 내성 P. aeruginosa 중, 32균주(29.1%)가 MBL를 생성하였고, IMP-6 (29균주, 90.6%)가 주요하게 확인되었다. VIM-2는 3균주(9.4%)에서 확인되었으며, 모두 ST357로 확인되었다. IMP-6를 생성하는 P. aeruginosa는 모두 다제내성을 보였고, ST235로 확인되었다. ST235 (55균주, 50.0%)는 가장 높은 비율로 확인된 클론이며 7년 동안 지속적으로 확인되었다. 이러한 다제내성 ST235의 확산을 방지하기 위해, carbapenem의 과도한 사용을 제한하고, 지속적으로 모니터링하는 전략이 개발되어야 할 것으로 사료된다.

• Journal of Microbiology
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• 제45권4호
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• pp.358-363
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• 2007

Multi-drug resistant Pseudomonas aeruginosa has been implicated in a variety of serious therapeutic problems in clinical environments. Among the 968 P. aeruginosa isolates obtained from two hospitals in Daegu, Korea, we acquired 17 isolates that were resistant to all available tested antimicrobial agents, with the exception of colistin (colistin-only sensitive). We characterized the antimicrobial susceptibilities, $metallo-{\beta}-lactamases$, and epidemiological relatedness among the colistin-only sensitive P. aeruginosa isolates. All colistin-only sensitive isolates were positive in the modified Hodge test and imipenem-EDTA synergy test, thereby indicating the production of $metallo-{\beta}-lactamases$. 11 isolates from the secondary hospital and six isolates from the tertiary teaching hospital harbored $bla_{VIM-2}$ and $bla_{IMP-1}$, respectively. The pulsed-field gel electrophoretic analysis of the SpeI-digested DNA from P. aeruginosa isolates indicated that two different clones of colistin-only sensitive P. aeruginosa originated from each hospital, and had spread within the hospital environment. Overall, colistin-only sensitive P. aeruginosa was detected in Korea for the first time, but no pan-drug resistant bacteria were identified. Nationwide surveillance is required in order to monitor the emergence of colistin-only sensitive or pan-drug resistant bacteria.

• 미생물학회지
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• 제52권4호
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• pp.455-462
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• 2016

Pseudomonas aeruginosa P-5 균주는 홀수개의 탄소수를 갖는 지방산으로부터 3-hydroxyvalerate (3HV)와 medium-chain-length (MCL) 3-hydroxyalkanoates (3HAs) 단위체로 구성된 popolyhydroxyalkanoate (PHA) 공중합체를 생산하는 특이한 성질을 갖고 있다. 이 균주가 갖고 있는 2개의 MCL-PHA synthases ($PhaC1_{P-5}$와 $PhaC2_{P-5}$)의 탄소길이에 따른 기질특이성을 비교하기 위하여 각각의 유전자를 PHA 생합성능이 결여된 돌연변이주 Pseudomonas putida GPp104에 도입하고 발현시킨 결과, $PhaC2_{P-5}$는 3HV와 MCL 3HAs로 이루어진 공중합체를 생산하지만 $PhaC1_{P-5}$는 단지 MCL 3HAs로 구성된 공중합체를 생산하였다. 이는 $PhaC2_{P-5}$가 $PhaC1_{P-5}$과는 달리 보다 짧은 탄소길이의 3-hydroxyvaleryl Co-A를 기질로 인지하여 합성반응에 이용할 수 있음을 보여주는 것이다. 또한 $PhaC2_{P-5}$의 효소활성 및 기질특이성의 변화를 유도하기 위하여 위치지정 돌연변이생성을 수행하고 P. putida GPp104과 다른 PHA 생합성능 결여 돌연변이주인 Ralstonia eutropha $PHB^-4$에서 발현시킨 결과, $PhaC2_{P-5}$ 내 두 개 아미노산의 치환(Ser326Thr과 Gln482Lys)이 공중합체의 3HV 함량을 크게 증진시키는 효과를 보였다. 두 개 아미노산이 모두 치환된 $PhaC2_{P-5}$ 유전자($phaC2_{P-5}QKST$)를 갖는 P. putida GPp104를 nonanoic acid가 탄소원으로 함유된 배지에서 배양하였을 때, 모균주에 비해 공중합체 함량과 공중합체 내 3HV 함량이 각각 2.5배 및 3.5배 증가하였다. 따라서 $phaC2_{P-5}QKST$를 포함하는 재조합 균주는 개량된 물성의 신규PHAs 생산에 유용할 것으로 기대된다.

• KSBB Journal
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• 제7권2호
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• pp.118-125
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• 1992

1.에멀젼형 화장품시료의 오염정도는 P. aeruginosa의 증식을 촉진하는 fatty acids, waxs, oils, steroids 등의 유기물에 의존하였다. 2. 오염의 결과, 물성의 변화로서 화장품시료의 pH가 40일 경과시 7.6에서 6.0으로 변화되었다. P. aeruginosa의 영양원으로서는 상대적으로 낮은 굴절률을 갖는 물질들이 소모되어 화장품시료(물:오일=70:30)의 굴절률이 1.4430에서 1.4530으로 변화 되었다. 3. 오염이 진행되는 동안 화장품시료의 相의 안정서이 파괴되었으며, 약간의 변색, 변취와 함께 creaming 및 응집현상이 나타났다. 4. P. aeruginosa의 최적증식조건인 pH7.0, 온도$20^{\circ}C$에서, 물과 오일의 부피비에 의한 균증식은 70:30, 80:20, 30:70, 90:10, 50:50의 순서로 되었다. 5. 방부살균제를 첨가하여 challenge test를 한 결과, P. aeruginosa의 증식이 억제되었으며 40일 경과시 균수는 방부살균제를 첨가하지 않은 경우 $10^8$개/ml에서 p-hydroxy benzoic acid propyl ester + phosphoric acid buffer solution첨가로 $5{\times}10^3$ 개/ml로 감소되었다. 6. 시험균주인 P. aeruginosa에 대한 항균력은 p-hydroxy benzoic acid propyl ester + phosphoric acid buffer solution>p-hydroxy benzoic acid buthyl ester + acetic acid buffer solution>p-hydroxy benzoic acid methyl ester + phosphoric acid buffer solution>p-hydroxy benzoic acid methyl ester + p-hydroxy benzoic acid propyl ester>p-hydroxy benzoic acid buthyl ester + potassium chloride sodium hydroxide buffer solution>p-hydroxy benzoic acid buthyl ester + p-hydroxy benzoic acid propyl ester>p-hydroxy benzoic acid methyl ester + acetic acid vbuffer solution>acetic acid buffer solution _ potassium chloride sodium hydroxide buffer solution의 순으로 우수하였으며 pH, 농도, 균수등은 양호한 수준을 유지하였다.

• Journal of Microbiology and Biotechnology
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• 제34권5호
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• pp.1040-1050
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• 2024

To isolate and analyze bacteria with Verticillium wilt-resistant properties from the fermentation residue of kitchen wastes, as well as explore their potential for new applications of the residue. A total of six bacterial strains exhibiting Verticillium wilt-resistant capabilities were isolated from the biogas residue of kitchen waste fermentation. Using a polyphasic approach, strain ZL6, which displayed the highest antagonistic activity against cotton Verticillium wilt, was identified as belonging to the Pseudomonas aeruginosa. Bioassay results demonstrated that this strain possessed robust antagonistic abilities, effectively inhibiting V. dahliae spore germination and mycelial growth. Furthermore, P. aeruginosa ZL6 exhibited high temperature resistance (42℃), nitrogen fixation, and phosphorus removal activities. Pot experiments revealed that P. aeruginosa ZL6 fermentation broth treatment achieved a 47.72% biological control effect compared to the control group. Through activity tracking and protein mass spectrometry identification, a neutral metalloproteinase (Nml) was hypothesized as the main virulence factor. The mutant strain ZL6ߡNml exhibited a significant reduction in its ability to inhibit cotton Verticillium wilt compared to the strain P. aeruginosa ZL6. While the inhibitory activities could be partially restored by a complementation of nml gene in the mutant strain ZL6CMߡNml. This research provides a theoretical foundation for the future development and application of biogas residue as biocontrol agents against Verticillium wilt and as biological preservatives for agricultural products. Additionally, this study presents a novel approach for mitigating the substantial amount of biogas residue generated from kitchen waste fermentation.

• Pediatric Infection and Vaccine
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• 제13권2호
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• pp.180-185
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• 2006

침습적 Pseudomonas 감염은 대개 면역 저하 환자들에서 발생하며, 이는 높은 사망률과 관련이 있다. 그러나 드물게는 면역력이 정상인 숙주에서도 발생할 수 있다. 첫 번째 증례는 괴저성 농창과 Pseudomonas 패혈증이 발생한 5개월 여아이며, 두번째 증례는 9개월 남아에서 지역사회 감염으로 발생한 폐렴과 Pseudomonas 패혈증이다. 두 환아 모두에서 녹농균이 배양검사로 증명되어 감수성 있는 항생제를 사용하였으며, T 림프구, B 림프구, 보체, 식세포 등 면역력에 대한 검사는 모두 정상이었다. 저자들은 이전에 건강했던 영아들에서 발생한 침습성 녹농균 감염증 2례를 경험하였기에 이를 보고하는 바이다.

• 한국미생물·생명공학회지
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• 제21권2호
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• pp.132-138
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• 1993

The s-type pyocin was purified from the lysate of the mitomycin C-induced Pseudomonas aeruginosa 90-2-2205 cells by the other of ammonium sulfate precipitation, DEAE-Sephadex A-50 chromatography and Sephadex G-200 gel filtration. The purity was confirmed by the polyacry-lamide gel electrophoresis. The molecular weight of the purified pyocin was estimated 180, 000 by gel filtration. The pyocin was analyzed to be a complex of some polypeptides by the SDS-PAGE. The pyocin was stable by heat treatment and at pH 6-7.5 by adding 10-3% gelatin and 0.2M NaCl to the 10mM Tris-HCl buffer (pH7.5). Its killing action against the sensitive cells was assumably a single hit process.

• BMB Reports
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• 제47권4호
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• pp.203-208
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• 2014

To gain insights into the effect of MexB gene under the short interfering RNA (siRNA), we synthesized 21 bp siRNA duplexes against the MexB gene. RT-PCR was performed to determine whether the siRNA inhibited the expression of MexB mRNA. Changes in antibiotic susceptibility in response to siRNA were measured by the E-test method. The efficacy of siRNAs was determined in a murine model of chronic P. aeruginosa lung infection. MexB-siRNAs inhibited both mRNA expression and the activity of P. aeruginosa in vitro. In vivo, siRNA was effective in reducing the bacterial load in the model of chronic lung infection and the P. aeruginosa-induced pathological changes. MexB-siRNA treatment enhanced the production of inflammatory cytokines in the early infection stage (P < 0.05). Our results suggest that targeting of MexB with siRNA appears to be a novel strategy for treating P. aeruginosa infections.