• Journal of Microbiology and Biotechnology
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• v.13 no.5
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• pp.700-705
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• 2003

Pseudomonas sp. S-47 is capable of degrading benzoate and 4-chlorobenzoate as well as catechol and 4-chlorocatechol via the meta-cleavage pathway. The three enzymes of 2-oxopenta-4-enoate hydratase (OEH), acetaldehyde dehydrogenase (acylating) (ADA), and 2-oxo-4-hydroxypentonate aldolase (HOA) encoded by xylJQK genes are responsible for the three steps after the meta-cleavage of catechol. The nucleotide sequence of the xylJQK genes located in the chromosomal DNA was cloned and analyzed. GC content of xylJ, xylQ, and xylK was 65% and consisted of 786, 924, and 1,041 nucleotides, respectively. The deduced amino acid sequences of xylJ, xylQ, and xylK genes from Pseudomonas sp. S-47 showed 93%, 99%, and 99% identity, compared with those of nahT, nahH, and nahI in Pseudomonas stutzeri An10. However, there were only about 53% to 85% identity with xylJQK of Pseudomonas putida mt-2, dmpEFG of P. putida CF600, aphEFG of Comamonas testosteroni TA441, and ipbEGF of P. putida RE204. On the other hand, the xylLTEGF genes located upstream of xylJQK in the strain S-47 showed high homology with those of TOL plasmid from Pseudomonas putida mt-2. These findings suggested that the xylLTEGFIJQK of Pseudomonas sp. S-47 responsible for complete degradation of benzoate and then catechol via the meta-pathway were phylogenetically recombinated from the genes of Pseudomonas putida mt-2 and Pseudomonas stutzeri An10.

• Journal of Species Research
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• v.6 no.1
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• pp.15-24
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• 2017

During the extensive survey of the prokaryotic species diversity in Korea, bacterial strains belonging to Betaproteobacteria and Epsilonproteobacteria were isolated from various sources including freshwater, sediment, soil and fish. A total of 23 isolates were obtained, among which 22 strains were assigned to the class Betaproteobacteria and one strain to the class Epsilonproteobacteria. The 22 betaproteobacterial strains were further assigned to Comamonadaceae (11 strains), Burkholderiaceae (6 strains), Oxalobacteraceae (2 strains), Neisseriaceae (1 strain) and unclassified family groups (2 strains). For the strains of Burkholderiaceae, 3 strains were identified as 3 species of Burkholderia, and 2 strains were as 2 species of Cupriavidus. For the strains of Comamonadaceae, 4 strains were identified as 2 species of the genus Hydrogenophaga, 2 strains as 2 species of Acidovorax, 2 strains as 2 species of Limnohabitans, and each of the remaining strains as single species of Comamonas, Curvibacter and Rhodoferax, respectively. For the strains of Oxalobacteraceae, 1 strain was identified as a species of Undibacterium, and the other strain as a species of Herbaspirillum. The strain belonging to Neisseriaceae was identified as a species of Iodobacter. The remaining strains of Betaproteobacteria were identified as species of Sphaerotilus and Methylibium respectively (family unassigned). The epsilonproteobacterial strain was identified as a species of Arcobacter of the family Camplyobacteraceae. The detailed description of each unrecorded species is provided.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.8 no.1
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• pp.67-75
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• 1998

The objectives of this study were both to evaluate the level and correlations of hazardous agents and to suggest measures to control industrial hygiene problems caused by using water-soluble metalworking f1uids(MWF). Geometric mean of formaldehyde(0.039 ppm) was higher than criteria of NIOSH(0.016ppm). Formaldehyde, originally existed in the biocide, is released and used to kill microbes in soluble MWF. Microbe concentrations were above $10^4No./mL$ in 14 MWF tanks among 20 tanks surveyed. Nitrosamines that is formed by reaction of nitrosating group and amines was detected to $18.4-47.1{\mu}g/m^3$. Formaldehyde concentration was low when microbes were abundant(r=-0.67, p=0.011), and high when open tank area was wide(r=0.75. p=0.012). The significant relationship between pH and microbes(r=-0.76. p=0.003) was also observed. The predominant bacteria species in MWF were Pseudomonas spp., Bacillus spp., Comamonas testosteroni, Acinetobacter haemolyticus, Bordertella bronchiseptica in order. Therefore, hazardous agents emitted by using water-soluble MWF seems to be correlated microbial growth. In order to minimize worker's exposure to several hazardous agents by an water-soluble MWF and to increase productivity, microbial growth must be controlled to the lowest level as possible. Administrative control as well as engineering control must comprehensively be applied to control microbe's growth in water-soluble MWF.

• Journal of Environmental Science International
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• v.23 no.1
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• pp.129-142
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• 2014

To treat chromaticity contained in effluents of dyeing wastewater efficiently, potent dye-degrading microorganisms were isolated from influent water, aeration- tank sludge, recycle water and settling-tank sludge located in leather and dyeing treatment plant. Six potent strains were finally isolated and identified as Comamonas testosteroni, Methylobacteriaceae bacterium, Stenotrophomonas sp., Kluyveromyces fragilis, Ascomycetes sp. and Basidiomycetes sp. When Basidiomycetes sp. was inoculated into ME medium containing basal mixed-dyes, 93% of color was removed after 8 days incubation. In the same experiment, the 1:1 mixed culture of Basidiomycetes sp. and photosynthetic bacterium exhibited 88% of color removal; however, it showed better color removal for single-color dyes. The aeration-tank and settling-tank samples revealed higher color removal (95-96%) for black dyes. The settling-tank sample also revealed higher color removal on basal mixed-dyes, which resulted in 90% color removal after 6-h incubation. From the above results, it is expected to achieve a higher color removal using the mixed microorganisms that were isolated from aeration-tank and settling-tank samples.

• Journal of Species Research
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• v.7 no.2
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• pp.97-103
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• 2018

In 2016, as a subset study to discover indigenous prokaryotic species in Korea, a total of 15 bacterial strains were isolated and assigned to the class Betaproteobacteria. From the high 16S rRNA gene sequence similarity (>98.8%) and formation of a robust phylogenetic clade with the closest species, it was determined that each strain belonged to each independent and predefined bacterial species. There is no official report that these 15 species have been described in Korea; therefore, 1 strain of the Aquitalea, 5 strains of the Paraburkholderia, 2 strains of the Comamonas, 1 strain of the Cupriavidus, 1 strain of the Diaphorobacter, 2 strains of the Hydrogenophaga, 1 strain of the Iodobacter, 1 strain of the Massilia and 1 strain of the Rhodoferax within the Betaproteobacteria are described for unreported bacterial species in Korea. Gram reaction, colony and cell morphology, basic biochemical characteristics, and isolation sources are also described in the species description section.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.560-570
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• 2007

The effective and accurate assessment of the total microbial community diversity is one of the primary challenges in modem microbial ecology, especially for the detection and characterization of unculturable populations and populations with a low abundance. Accordingly, this study was undertaken to investigate the diversity of the microbial community during the biodegradation of cis- and trans-dichloroethenes in soil and wastewater enrichment cultures. Community profiling using PCR targeting the l6S rRNA gene and denaturing gradient gel electrophoresis (PCR-DGGE) revealed an alteration in the bacterial community profiles with time. Exposure to cis- and trans-dichloroethenes led to the disappearance of certain genospecies that were initially observed in the untreated samples. A cluster analysis of the bacterial DGGE community profiles at various sampling times during the degradation process indicated that the community profile became stable after day 10 of the enrichment. DNA sequencing and phylogenetic analysis of selected DGGE bands revealed that the genera Acinetobacter, Pseudomonas, Bacillus, Comamonas, and Arthrobacter, plus several other important uncultured bacterial phylotypes, dominated the enrichment cultures. Thus, the identified dominant phylotypes may play an important role in the degradation of cis- and trans-dichloroethenes.

• Archives of Pharmacal Research
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• v.27 no.5
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• pp.570-575
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• 2004

2-Hydroxymuconic semialdehyde (2-HMS) dehydrogenase catalyzes the conversion of 2-HMS to 4-oxalocrotonate, which is a step in the meta cleavage pathway of aromatic hydrocarbons in bacteria. A tomC gene that encodes 2-HMS dehydrogenase of Burkholderia cepacia G4, a soil bacterium that can grow on toluene, cresol, phenol, or benzene, was overexpressed into E. coli HB 101, and its gene product was characterized in this study. 2-HMS dehydrogenase from B. cepacia G4 has a high catalytic efficiency in terms of V$_{max}$K$_{max}$ towards 2-hydroxy-5-methyl-muconic semialdehyde followed by 2-HMS but has a very low efficiency for 5-chloro-2-hydroxymuconic semialdehyde. However, the enzyme did not utilize 2-hydroxy-6-oxo-hepta 2,4-dienoic acid and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid as substrates. The molecular weight of 2-HMS dehydrogenase from B. cepacia G4 was predicted to be 52 kDa containing 485 amino acid residues from the nucleotide sequence of the tomC gene, and it exhibited the highest identity of 78% with the amino acid sequence of 2-HMS dehydrogenase that is encoded in the aphC gene of Comamonas testosteroni TA441. 2-HMS dehydrogenase from B. cepacia G4 showed a significant phylogenetic relationship not only with other 2-HMS dehydrogenases, but also with different dehydrogenases from evolutionarily distant organisms.sms.

• Journal of Environmental Science International
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• v.6 no.3
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• pp.249-258
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• 1997

Mercury- and cadmium-resistant bacteria were Isolated from an Industrial complex wastewater of Taejon area. All of them were motile, gram negative rods. In the results of physicochemical test and VITEK card test. HM1 was Identified with Achetobacter cd- coaceucus, CM3 was Identified 65 Commonas acidovorns, HM2, HM3, CMI , and CM4 were Pseudomonas sp., but HM4 and CM2 were unidenteed. They were tested for subceptlbility to 14 heavy metals. Mercury-resistant bacteria(HM1, HM2, HM3, and HM4) were sensitive to low concentration(100~400ppm) of $Cd^{2+}$, $Co^{2+}$. $Zn^{2+}$, and Ni2+, while cadmium-resistant bacteria(CM1, CM2, CM3, and CM4) showed resistance up to the high concentration(600~ 1, 200ppm) of these metal loons. As a result of resistance spectrum test of mercury-resistant bacteria, HM1 was broad-spectrum strain, HM2. HM3, and HM4 were narrow-spectrum stratas. Transmission electron microscopic examination of cell wall of HM1 culture grown with and without 100ppm of $HgCl_2$ showed remarkably merphological abnormalities. In the result of atomic absorption spectrometric analysis of cadmium-resistant bacteria grown at 200ppm of $CdCl_2$ for 6h, all of them accumulated cadrnium(14ppm~57ppm) In cell. In cadinium-resistant bacteria, CM1, CM2, and CM4 were spared from the Inhibitory effect of $Cd^{2+}$ by the addition of $Mn^{2+}$, CM4 were also spared from the Inhibitory effect of $Cd^{2+}$ by the addition of $Mn^{2+}$ as well as $Zn^{2+}$.

• Journal of Microbiology
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• v.42 no.2
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• pp.152-155
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• 2004

Pseudomonas sp. K82 has been reported to be an aniline-assimilating soil bacterium. However, this strain can use not only aniline as a sole carbon and energy source, but can also utilize benzoate, p-hydroxybenzoate, and aniline analogues. The strain accomplishes this metabolic diversity by using dif-ferent aerobic pathways. Pseudomonas sp. K82, when cultured in p-hydroxybenzoate, showed extradiol cleavage activity of protocatechuate. In accordance with those findings, our study attempted the puri-fication of protocatechuate 4,5-dioxygenase (PCD 4,5). However the purified PCD 4,5 was found to be very unstable during purification. After Q-sepharose chromatography was performed, the crude enzyme activity was augmented by a factor of approximately 4.7. From the Q-sepharose fraction which exhibited PCD 4,5 activity, two subunits of PCD4,5 (${\alpha}$ subunit and ${\beta}$ subunit) were identified using the N-terminal amino acid sequences of 15 amino acid residues. These subunits were found to have more than 90％ sequence homology with PmdA and PmdB of Comamonas testosteroni. The molecular weight of the native enzyme was estimated to be approximately 54 kDa, suggesting that PCD4,5 exists as a het-erodimer (${\alpha}$$_1$${\beta}$$_1$). PCD 4,5 exhibits stringent substrate specificity for protocatechuate and its optimal activity occurs at pH 9 and 15 $^{\circ}C$. PCR amplification of these two subunits of PCD4,5 revealed that the ${\alpha}$ subunit and ${\beta}$ subunit occurred in tandem. Our results suggest that Pseudomonas sp. K82 induced PCD 4,5 for the purpose of p-hydroxybenzoate degradation.

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.763-771
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• 2001

Biphenyl dioxygenase (BPDO), which catalyzes the first step in the bacterial degradation of biphenyl and polychlorinated biphenyls, was characterized in Pseudomonas sp. B4. The bphA locus containing the four structural genes encoding BPDO were cloned and sequenced. A regulatory gene as well as a putative regulatory sequence were identified upstream of this locus. A transposase-like gene was found within a 1-kb region further upstream, thereby suggesting that the bphA locus may be carried on a transposable element. The three components of the BPDO enzyme have been separately overexpressed and purified from E. coli. The ferredoxin and terminal dioxygenase components showed biochemical properties comparable to those of two previously characterized BPDOs, whereas the ferredoxin reductase exhibited an unusually high lability. The substrate selectivity of BPDO was examined in vivo using resting cell assays performed with mixtures of selected polychlorinated biphenyls. The results indicated that para-substituted congeners were the preferred substrates. In vitro studies were carried out on a BPDO complex where the reductase from strain B4 we replaced by the more stable isoform from Comamonas testosteroni B-356. The BPDO enzyme had a specific activity of $0.26{\pm}0.02 {\mu}mol {min^-1}{mg^-1}\;of\;ISP_{BPH}$ with biphenyl as the substrate. The 2,3-, 4,4'-, and 2,4,4'-chlorobiphenyls were converted to single dihydrodiols, while 2,4'-dichlorobiphenyl gave rise to two dihydrodiols. The current data also indicated that 2,4,4'-trichlorobiphenyl was a better substrate than the 4,4'-dichlorinated congener.