• Title/Summary/Keyword: Pseudomonas putida

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Effect of Dissolved Oxygen Concentration on the Metabolism of Glucose in Pseudomonas putida BM014

  • Park, Won-Jae;Lee, Eun-Yeol;Park, Cha-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.109-111
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    • 1998
  • The effect of dissolved oxygen concentration on the metabolism of glucose in Pseudomonas putida BM014 was investigated. Glucose was completely converted to 2-ketogluconate via extracellular oxidative pathway and then taken up for cell growth under the condition of sufficient dissolved oxygen concentration. On the other hand, oxygen limitation below dissolved oxygen tension (DOT) value of 20% of air saturation caused the shift of glucose metabolism from the extracellular oxidative pathway to the intracellular phosphorylative pathway. Specific activities of hexokinase and gluconate kinase in intracellular phosphorylation pathway decreased as the DOT increased, while 2-ketogluconokinase activity in extracellular oxidative pathway increased under the same condition. This result can be usefully applied to microbial transformation of glucose to 2-ketogluconate, the synthetic precursor for iso-vitamine C, with almost 100% yield via extracellular oxidation by simple DOT control.

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Characterization of the Biogenic Manganese Oxides Produced by Pseudomonas putida strain MnB1

  • Jiang, Shaofeng;Kim, Do-Gun;Kim, Jeong-Hyun;Ko, Seok-Oh
    • Environmental Engineering Research
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    • v.15 no.4
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    • pp.183-190
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    • 2010
  • Biogenic Mn oxides are expected to have great potential in the control of water pollution due to their high catalytic activity, although information on biological Mn oxidation is not currently sufficient. In this study, the growth of a Mn oxidizing microorganism, Pseudomonas putida MnB1, was examined, with the Mn oxides formed by this strain characterized. The growth of P. putida MnB1 was not significantly influenced by Mn(II), but showed a slightly decreased growth rate in the presence of Pb(II) and EE2, indicating their insignificant adsorption onto the cell surface. Mn oxides were formed by P. putida MnB1, but the liquid growth medium and resulting biogenic solids were poorly crystalline, nano-sized particles. Biogenic Mn oxidation by P. putida MnB1 followed Michaelis-Menten kinetics, with stoichiometric amounts of Mn oxides formed, which corresponded with the initial Mn(II) concentration. However, the formation of Mn oxides was inhibited at high initial Mn(II) concentration, suggesting mass transfer obstruction of Mn(II) due to the accumulation of Mn oxides on the extracellular layer. Mn oxidation by P. putida MnB1 was very sensitive to pH and temperature, showing sharp decreases in the Mn oxidation rates outside of the optimum ranges, i.e. pH 7.43-8.22 and around 20-$26^{\circ}C$.

Isolation and Detection of Genes Responsible for Pyoverdines Biosynthesis in Pseudomonas putida KNUK9

  • Hussein, Khalid A.;Joo, Jin Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.48 no.2
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    • pp.119-124
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    • 2015
  • Pyoverdines (PVDs) are organic compounds produced by the fluorescent Pseudomonads under iron starvation conditions. Among the isolated rhizosphere pseudomonads strains, P. putida KNUK9 showed the highest production of PVDs and its production reached to 62.81% siderophores units. DNA isolation, ligation, PCR amplification, and transformation using E. coli $DH5{\alpha}$ cells were carried out for preparing the strong pyoverdine producer strains. We detected seven genes playing the fundamental roles in the pyoverdine metabolism in Pseudomonads. According to data and analysis obtained from the study, we deduced that the strain P. putida KNUK9 contains the essential genes required for pyoverdine biosynthesis.

Characterization of a salicylate-degrading strain of pseudomonas putida (Salicylate를 생분해하는 pseudomonas putida의 특성)

  • 홍순우;하영칠;이계준;배경숙
    • Korean Journal of Microbiology
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    • v.24 no.1
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    • pp.51-56
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    • 1986
  • Eight strains of the bacteria capable of growing on salicylate as the sole carbon source were isolated form soil and river water. Three of these isolates were identified as Acinetobacter calcoaceticus (AcBl), Pseuomonas putida biotype B (PpB2), and P. putida biotype A (PpB3). Effects of temperature, pH and C source concentration on biodegradation of salicylate by PpB3 were wxamined. The optimum conditions were as follows; $30^{\circ}C$ for temperature, 7.0 for pH, and 10mM for C source concentration. Ultraviolet scanning spectrum of the salicylate was measured. The spectrum has two peaks at 225nm and 292nm. The spectra of the culture filtrates indicate that ring degradation of salicylate is accomplished.

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Biodegradation of Aniline by Pseudomonas putida FW (Pseudomonas putida FW에 의한 Aniline의 생물학적 분해)

  • Park, Y.K.;Oh, J.S.;Ban, C.I.;Yoon, S.J.;Choi, M.S.
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.413-420
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    • 1988
  • The strain capable of growing on minimal medium containing aniline as a sole source of carbon was isolated from activated sludges and identified as Pseudomonas putida biotype A. The characterizations of the strain were determined. The optimum concentration for growth of the strain was 1-20 mM of aniline. No changes of pH were detected during cultivation. Some metabolic products of biodegradation of aniline were detected after cultivation of the strain on 10 mM aniline for 48 hours. The strain showed to be resistant to streptomycin, tetracycline, trimethoprim, and sulfanilamide. The strain was also capable of utilizing other aromatic compounds related to aniline as a sole source of carbon. One plasmid carried by this strain was detected. The properties of some of the mutant strains treated with mitomycin C were also discussed. The results suggest that separate, regulatory enzyme systems capable of degrading aniline may exist in plasmid DNA.

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Cloning and Sequence Analysis of the xyIL Gene Responsible for 4CBA-Dihydrodiol Dehydrogenase from Pseudomonas sp. S-47

  • Park, Dong-Woo;Kim, Youngsoo;Lee, Sang-Mahn;Ka, Jong-Ok;Kim, Chi-Kyung
    • Journal of Microbiology
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    • v.38 no.4
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    • pp.275-280
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    • 2000
  • Pseudomonas sp. S-47 is capable of catabolizing 4-chlorobenzoate (4CBA) as rarbon and energy sources under aerobic conditions via the mesa-cleavage pathway. 4CBA-dioxygenase and 4CBA-dihydrodiol dehydrogenase (4CBA-DD) catalyzed the degradation af 4CBA to produce 4-chlorocatechol in the pathway. In this study, the xylL gene encoding 4CBA-DD was cloned from the chromosomal DNA of Pseudomonas sp. S-47 and its nucleotide sequence was analyzed. The xylL gene was found to be composed of 777 nucleotide pairs and to encode a polypeptide of 28 kDa with 258 amino acid residues. The deduced amino acid sequence of the dehydrogenase (XylL) from strain S-47 exhibited 98% and 60% homologies with these of the corresponding enzymes, Pseudomonas putida mt-2 (XyIL) and Acinetobacter calcoaceticus (BenD), respectively. However, the amino arid sequences show 30% or less homology with those of Pseudomonas putida (BnzE), Pseudomonas putida Fl (TodD), Pseudomonas pseudoalcaligenes KF707 (BphB), and Pseudomonas sp. C18 (NahB). Therefore, the 4CBA-dihydrodiol dehdrogenase of strain S-47 belongs to the group I dehydrogenase involved in the degradation of mono-aryls with a carboxyl group.

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Degradation Characteristics of Methyl Ethyl Ketone and Methyl Isobuthyl Ketone by Pseudomonas putida KT-3. (Pseudomonas putida KT-3의 Methyl Ethyl Ketone 및 Methyl Isobuthyl Ketone 분해 특성)

  • 김민주;이태호;이경미;류희욱;조경숙
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.395-401
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    • 2002
  • Methyl ethyl ketone (MEK) and methyl isobutyl ketone (MIBK) have been widely used as solvents in various industries. Biodegradation of MEK and MIBK by Pseudomonas putida KT-3, which could utilize MEK or MIBK as a sole carbon source, was characterized, and the cosubstrate interaction in MEK/MIBK mixture was also studied. Within the range of initial MEK concentration (from 0.5 to 5.5 mM), an increased substrate concentration increased the specific degradation rate of MEK by P putida KT-3 (from 3.15 to 10.58 mmol/g DCW$\cdot$h), but the rate sightly increased at 11.0 mM of initial MEK concentation (11.28 mmol/g DCW$\cdot$h). The similar degradation rates of MIBK (4.69-4.92 mmol/g DCW$\cdot$h) were obtained at more than 3.0 mM of initial MIBK concentation. Kinetic analysis on the degradation of MEK/MIBK mixture by P. putida KT-3 showed that MEK or MIBK acted as a competitive inhibitor. Maximum degradation rate ($V_{max}$), saturation constant ($K_{m}$) and inhibition constant ($K_{1}$) were as follows: $V_{max,MEK}$=12.94 mmol/g DCW$\cdot$h; $K_{m,MEK}$=1.72 mmol/L; $K_{l,MEK}$=1.30 mmol/L; $V_{max,MIBK}$=5.00 mmol/g-DCW$\cdot$h; $K_{m,MIBK}$=0.42 mmol/L; $K_{l,MEK}$=0.77 mmol/L.

The Characteristics of Tetrachloroethylene (PCE) Degradation by Pseudomonas putida BJ10 (Pseudomonas putida BJ10의 Tetrachloroethylene (PCE) 분해 특성)

  • Choi, Myung-Hoon;Kim, Jai-Soo;Lee, Sang-Seob
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.311-316
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    • 2008
  • In this study, biological PCE degradation by using a BTEX degrading bacterium, named BJ10, under aerobic conditions in the presence of toluene was examined. According to morphological, physiological characteristics, 16S rDNA sequencing and fatty acid analysis, BJ10 was classified as Pseudomonas putida. As a result of biological PCE degradation at low PCE concentrations (5 mg/L), PCE removal efficiency by P. putida BJ10 was 52.8% for 10 days, and PCE removal rate was 5.9 nmol/hr (toluene concentration 50 mg/L, initial cell density 1.0 g (wet weight)/L, temperature 30, pH 7 and DO $3.0{\sim}4.2\;mg/L$. At high PCE concentration (100 mg/L), PCE removal efficiency by P. putida BJ10 was 20.3% for 10 days, and PCE removal rate was 46.0 nmol/hr under the same conditions. The effects of various toluene concentration (5, 25, 50, 100, 200 mg/L) on PCE degradation were examined under the same incubation conditions. The highest PCE removal efficiency of PCE was 57.0% in the initial PCE concentration of 10 mg/L in the presence of 200 mg/L toluene for 10 days. Furthermore, the additional injection of 5.5 mg/L PCE (total 7.6 mg/L) made 63.0% degradation for 8 days in the presence of 50 mg/L toluene under the same conditions. Its removal rate was 13.5 nmol/hr, which was better than the initial removal rate (8.1 nmol/hr).

Application of the genetically engineered strains of pseudomonas degrading various persistent aromatic hydrocarbons to wastewater (다양한 난분해성 방향족 탄화수소를 분해하는 pseudomonas 개발균주의 폐수에의 적용)

  • 송재욱;박경량;박용근;이영록
    • Korean Journal of Microbiology
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    • v.28 no.3
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    • pp.243-248
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    • 1990
  • Genetically engineered strains of Pseudomonas, which could degrade the various aromatic hydrocarbons, were acclimated to the synthetic wastewater with increasing substrate concentration. All of the tested strains except KUD101 showed 70-90% of the COD removal efficiencies. Acclimated strains, Pseudomonas putida KUD106, KUD107 and KUD108 were inoculated into the ordinary activated sludge and these sludges were used in the dyeing or alkylbenzene wastewater treatment system. In the case of the wastewater containing alkylbenzene compounds, COD removal efficiency was 12-14% higher than that of the ordinary activated sludge, while it was not effective for dyeing wastewater. On the other hand, the floc formation of the activated sludge inoculated with genetically engineered strains was more rapid in both wastewaters tested than that of the activated sludge by the ordinary natural strains.

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Degradation of Toluene and Acetic Acid Using Cell-Free Enzyme System from Single Cell-Strain (Single cell-strain부터 유래된 무세포 효소 시스템을 이용한 톨루엔 및 아세트산 분해)

  • Jang, Jae Hyun;Kim, Yeji;Roh, Tae Yong;Park, Joong Kon
    • Korean Chemical Engineering Research
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    • v.54 no.5
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    • pp.665-670
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    • 2016
  • This study deals with the possible degradation of toluene and acetic acid when subjected to cell-free enzyme system from the toluene degrading bacteria Pseudomonas putida and acetic acid degrading bacteria Cupriavidus necator. P. putida produces toluene dioxygenase only under the existence of toluene in culture medium and toluene is degraded to cis-toluene dihydrodiol by this enzyme. C. necator produces acetyl coenzyme A synthetase-1 and converts acetic acid to acetyl CoA in order to synthesize ATP to need for growth or PHA which is biodegradable polymer. In case of toluene degradation, the experiment was conducted before and after production of toluene dioxygenase as this enzyme, produced by P. putida, is an inducible enzyme. Toluene was detected using gas chromatography (GC). Similar amount of toluene was found in control group and before production of toluene dioxygenase (experimental group 1). However, reduction in toluene was detected after the production of toluene dioxygenase (experimental group 2). Acetic acid was detected through application of gas chromatography-mass spectrometer (GC-MS). The results showed the acetic acid peak was not detected in the experimental group to apply cell-free enzyme system. These results show that the cell-free enzyme system obtained from P. putida and C. necator retained the ability to degrade toluene and acetic acid. However, P. putida needs to produce the inducible enzyme before preparation of the cell-free enzyme system.