• 미생물학회지
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• 제22권1호
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• pp.29-34
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• 1984

서울 근교에서 채취한 河川水 및 土壤標品으로부터 238菌株의 탄화수소자화세균을 分離하였다. 分離된 菌株중 68菌株는 Pseudomonas屬으로 同定되었고, 11菌株는 Alcaligenes屬으로, 10菌株는 Acinetobacter屬으로 同定되었다. 68菌株의 Pseudomona중, 35菌株는 P. aeruginosa, 5菌株는 P. fluorescence, 10菌株는 P. putida, 그리고 2菌株는 P. mendocina로 각각 同定되었다.

• Journal of Microbiology and Biotechnology
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• 제8권4호
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• pp.416-421
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• 1998

A carboxyl group modifier, l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was used to study the interactions between three components of toluene dioxygenase (TDO) from Pseudomonas putida FI. $Ferredoxin_{TOL}$ activity was increased by the treatment with EDC; however, the activity was rapidly declined in the prolonged incubation. In covalent cross-linking experiments with EDC, $Ferredoxin_{TOL}$ made a one-to-one complex with $Reductase_{TOL}$ or the large subunit of $ISP_{TOL}$. These results provide evidence for the involvement of electrostatic interactions in the TDO electron transfer system.

• 미생물학회지
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• 제26권3호
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• pp.167-172
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• 1988

A genetic map of the IncP-1 group plasmid pKU10 has been prepared through the construction of recombinant plasmids containing various fragments of pKU10. Phenotypic analysis of these derivatives has identified the location of genes encoding resistance to ampicillin, tetracyclin, and chloramphenicol. The region involved in conferring resistance to ampicillin was located around two PstI sites that are 1.0Kb apart. The tetracyclin resistance gene was mapped on the region of HindIII E fragment and a part of HindIII D fragment, and the determinant for chloramphenicol resistance gene was localized on HindIII D fragment.

• Journal of Microbiology and Biotechnology
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• 제16권7호
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• pp.1032-1040
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• 2006

Some Pseudomonas species can grow on styrene as a sole carbon and energy source. From the new isolate Pseudomonas putida SN1, the genes for styrene catabolism were cloned and sequenced. They were composed of four structural genes for styrene monooxygenase (styA and styB), styrene oxide isomerase (styC), and phenylacetaldehyde dehydrogenase (styD), along with two genes for the regulatory system (styS and styR). All the genes showed high DNA sequence (91% to 99%) and amino acid sequence (94% to 100%) similarities with the corresponding genes of the previously reported styrene-degrading Pseudomonas strains. A recombinant Escherichia coli to contain the styrene monooxygenase from the SN1 was constructed under the control of the T7 promoter for the production of enantiopure (S)-styrene oxide, which is an important chiral building block in organic synthesis. The recombinant E. coli could convert styrene into an enantiopure (S)-styrene oxide (ee >99%) when induced by IPTG The maximum activity was observed as 140 U/g cell, when induced with 1 mM IPTG at $15^{\circ}C$.

• 대한임상검사과학회지
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• 제39권3호
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• pp.167-177
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• 2007

From the total 121,294 clinical materials submitted to the Department of Laboratory Medicine of "C" hospital from December 1, 2004 to November 30, 2006, 3,408 Pseudomonas spp. were isolated. The isolation frequencies of Pseudomonas spp. were as follows, P. aeruginosa 95.5%, P. putida 2.5%, P. fluorescens 0.8%, along with low frequencies of P. luteola, P. alcaligenes, P. stutzeri, P. oryzihabitants, P. mendocina and unidentified Pseudomonas species. The isolation rates of Pseudomonas spp. according to season and sex were evenly distributed. The isolated frequency of Pseudomonas spp. in male was two times higher than that of in female showing significantly more male patients in surgical areas and more female patients in internal areas (p<0.001). In monthly analysis, Pseudomonas spp. were the most frequently isolated in July (10.4%), but lowest in February (5.6%). Half of Pseudomonas spp. were isolated from sputum (48.2%). In the susceptibility analysis of Pseudomonas spp. by VITEK II AST cards, the Pseudomonas spp showing higher susceptibility against antimicrobial agents were piperacillin/tazobactam (82.7%) in P. aeruginosa; amikacin (84.7%), colistin (83.3%) in P. putida; and amikacin (96.3%), cefepime (87.5%), ceftazidime (87.5%) ciprofloxacin (92.3%), colistin (88.5%) gentamicin (96.2%), isepamicin (96,1%), meropenem (92.3%), netilmicin (96.0%), piperacillin/ tazobactam (95.4%) and tobramycin (92.6%) in P. fluorescens.

• 한국미생물·생명공학회지
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• 제39권2호
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• pp.161-167
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• 2011

본 연구실에서 확보한 diesel 분해 고효율 균주 Pseudomonas putida KDi 19, kerosene 분해 고효율 균주 P. aeruginosa K14, gasoline 분해 고효율 균주 P. putida G8, BTEX 분해 고효율 균주 P. putida BJ10, P. putida E41의 5개의 고효율 균주를 컬럼 및 반응기에 적용하여 TPH의 생물학적 분해 실험에 적용하였다. 영양염류 및 산소 농도, 균농도 등 최적의 환경인자 도출을 통해 최적의 생물학적 처리 효율을 TPH의 경우, MSM 및 activator I을 주입하여 25일 동안 76.3%의 제거 효율과 제거속도상수 K=0.711를 나타냈으며, diesel의 경우 40일 동안 99.2%의 제거 효율을 보였다. 또한, TPH 오염 토양의 lab-scale bioremediation 실험에서 고효율 균주를 적용한 결과 45일 운전 기간 동안 7,209.9 $mg{\cdot}kg^{-1}$을 825.6 $mg{\cdot}kg^{-1}$까지 88.5% 제거하였다. 본 연구에서 도출된 TPH로 오염된 토양의 bioremediation을 위한 고효율 균주 확보와 최적의 환경 인자 도출은 현재 부족한 생물학적 처리 연구와 물리적 화학적 처리의 문제를 해소하기 위한 기초적 실험 자료로서 기여할 것으로 사료된다.

• BMB Reports
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• 제30권2호
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• pp.132-137
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• 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.

• 한국동물위생학회지
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• 제44권3호
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• pp.133-140
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• 2021

This study was aimed to investigate the prevalence and antimicrobial resistance of Pseudomonas spp. isolated from bovine mastitis milk samples. A total of 50 (4.9%) Pseudomonas spp. was isolated from 1,023 samples, those collected between 2018 and 2021, derived from 110 dairy farms. The prevalence of the identified species of Pseudomonas isolates was as follows; P. aeruginosa (70.0%), P. fluorescens (14.0%), P. putida (10.0%), P. fragi (4.0%), and P. chlororaphis (2.0%). Most of somatic cell counts in the quarter milk carrying Pseudomonas spp. were less than 3,000,000 cell/ml (90.0%). The isolates of Pseudomonas spp. showed high susceptibility to cefepime (98.0%), ciprofloxacin (98.0%), ceftazidime (96.0%), and colistin (96.0%). The rate of antibiotic resistance in the isolates was highest to ceftiofur (92.0%), followed by the resistance rate to chloramphenicol (86.0%) and trimethoprim/sulphamethoxazole (80.0%). In addition, there is a remarkable difference in antimicrobial resistance pattern among Pseudomonas species. P. aeruginosa and P. putida showed a similar resistance pattern, whereas P. fluorescens showed exceptionally lower resistance to trimethoprim/sulphamethoxazole and chloramphenicol than that of the other species. This study showed that prevalence of Pseudomonas spp. other than P. aeruginosa were 30.0% in bovine mastitis milk, and the occurrence rate of antibiotic resistance were similar or higher level, compared with the previous reports on the mastitisderived Pseudomonas spp. isolated in Korea.

• Mycobiology
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• 제33권3호
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• pp.131-136
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• 2005

Infection structures were observed at the penetration sites on the leaves of cucumber plants inoculated with Colletotrichum orbiculare using a fluorescence microscope. The cucumber plants were previously drenched with suspension of bacterial strains Pseudomonas putida or Micrococcus luteus. The plants pre-inoculated with both bacterial strains were resistant against anthracnose after inoculation with C. orbiculare. To investigate the resistance mechanism by both bacterial strains, the surface of infected leaves was observed at the different time after challenge inoculation. At 3 days after inoculation there were no differences in the germination and appressorium formation of conidia of C. orbiculare as well as in the callose formation of the plants between both bacteria pre-inoculated and non-treated. At 5 days, the germination and appressorium formation of the fungal conidia were, however, significantly decreased on the leaves of plants pre-inoculated with M. luteus at the concentration with $1.0{\times}10^7\;cfu/ml$. Furthermore, callose formation of plants cells at the penetration sites was apparently increased. In contrast, there were no defense reactions of the plants at the concentration with $1.0{\times}10^6\;cfu/ml$ of M. luteus. Similarly, inoculation P. putida caused no plant resistance at the low concentration, whereas increase of callose formation was observed at the higher concentration. The results of this study suggest that the resistant mechanisms might be differently expressed by the concentration of pre-treatment with bacterial suspension.

• Journal of Microbiology and Biotechnology
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• 제24권1호
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• pp.59-69
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• 2014

One of the major challenges in metabolic engineering for enhanced synthesis of value-added chemicals is to design and develop new strains that can be translated into well-controlled fermentation processes using bioreactors. The aim of this study was to assess the influence of various fed-batch strategies in the performance of metabolically engineered Pseudomonas putida strains, ${\Delta}gcd$ and ${\Delta}gcd-pgl$, for improving production of medium-chain-length polyhydroxyalkanoates (mcl-PHAs) using glucose as the only carbon source. First we developed a fed-batch process that comprised an initial phase of biomass accumulation based on an exponential feeding carbon-limited strategy. For the mcl-PHA accumulation stage, three induction techniques were tested under nitrogen limitation. The substrate-pulse feeding was more efficient than the constant-feeding approach to promote the accumulation of the desirable product. Nonetheless, the most efficient approach for maximum PHA synthesis was the application of a dissolved-oxygen-stat feeding strategy (DO-stat), where P. putida ${\Delta}gcd$ mutant strain showed a final PHA content and specific PHA productivity of 67% and $0.83g{\cdot}l^{-1}{\cdot}h^{-1}$, respectively. To our knowledge, this mcl-PHA titer is the highest value that has been ever reported using glucose as the sole carbon and energy source. Our results also highlighted the effect of different fed-batch strategies upon the extent of realization of the intended metabolic modification of the mutant strains.