• Journal of Microbiology and Biotechnology
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• 제13권5호
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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.

• 한국산업보건학회지
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• 제8권1호
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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.

• 한국환경과학회지
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• 제23권1호
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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.

• Archives of Pharmacal Research
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• 제27권5호
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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 Microbiology
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• 제42권2호
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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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• 제11권5호
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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.

• Fisheries and Aquatic Sciences
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• 제21권2호
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• pp.6.1-6.10
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• 2018

The intention of this study was to identify the bacterial pathogens infecting Oreochromis niloticus (Nile tilapia) and Clarias gariepinus (African catfish), and to establish the antibiotic susceptibility of fish bacteria in Uganda. A total of 288 fish samples from 40 fish farms (ponds, cages, and tanks) and 8 wild water sites were aseptically collected and bacteria isolated from the head kidney, liver, brain and spleen. The isolates were identified by their morphological characteristics, conventional biochemical tests and Analytical Profile Index test kits. Antibiotic susceptibility of selected bacteria was determined by the Kirby-Bauer disc diffusion method. The following well-known fish pathogens were identified at a farm prevalence of; Aeromonas hydrophila (43.8%), Aeromonas sobria (20.8%), Edwardsiella tarda (8.3%), Flavobacterium spp. (4.2%) and Streptococcus spp. (6.3%). Other bacteria with varying significance as fish pathogens were also identified including Plesiomonas shigelloides (25.0%), Chryseobacterium indoligenes (12.5%), Pseudomonas fluorescens (10.4%), Pseudomonas aeruginosa (4.2%), Pseudomonas stutzeri (2.1%), Vibrio cholerae (10.4%), Proteus spp. (6.3%), Citrobacter spp. (4.2%), Klebsiella spp. (4.2%) Serratia marcescens (4.2%), Burkholderia cepacia (2.1%), Comamonas testosteroni (8.3%) and Ralstonia picketti (2.1%). Aeromonas spp., Edwardsiella tarda and Streptococcus spp. were commonly isolated from diseased fish. Aeromonas spp. (n = 82) and Plesiomonas shigelloides (n = 73) were evaluated for antibiotic susceptibility. All isolates tested were susceptible to at-least ten (10) of the fourteen antibiotics evaluated. High levels of resistance were however expressed by all isolates to penicillin, oxacillin and ampicillin. This observed resistance is most probably intrinsic to those bacteria, suggesting minimal levels of acquired antibiotic resistance in fish bacteria from the study area. To our knowledge, this is the first study to establish the occurrence of several bacteria species infecting fish; and to determine antibiotic susceptibility of fish bacteria in Uganda. The current study provides baseline information for future reference and fish disease management in the country.

• 한국버섯학회지
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• 제19권2호
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• pp.103-108
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• 2021

본 시험은 재배과정 중 배지에서 분리한 미생물의 특성을 조사하여 이들 미생물과 버섯균의 생육과 연관관계를 밝히고자 하였다. 짚, 계분 등 배지 재료에서 분리한 미생물의 양송이 균주에 대한 억제 정도는 국내 육성 양송이 균주가 외국에서 수입한 양송이 균주 보다 생육 억제 정도가 낮은 경향을 보였다. 양송이 배지 발효 단계별 분리 미생물의 양송이 품종간 균사 생육 억제 정도는 국내 육성 품종인 새도가 다른 품종보다는 생육이 좋았다. 그리고 발효과정이 진행됨에 따라 버섯균에 대한 억제 정도는 약해지는 경향을 보였다. 양송이 배지에서 분리한 미생물을 합성배지(CDA)에 도말한 뒤 버섯균사를 접종하여 균사의 생육을 조사한 결과와 분리균을 도말하고 항온기에서 2일간 배양한 후 배지를 완전히 뒤집어 배지의 뒷면 중앙에 버섯 균사를 접종하여 균사 생육을 조사한 결과를 비교 분석한 결과 양송이균의 생육 정도가 비슷한 경향을 보이는 것으로 보아 양송이 배지에서 분리한 미생물은 분비성 물질을 통해서 버섯균의 생육과 증식에 영향을 미친다는 것을 알 수 있었다. 양송이 균의 생육을 촉진하는 미생물을 분리한 결과 21균주가 선발되었고 대부분 4차 뒤집기 이후에서 많이 선발되었다. 대표적인 균주로는 Alcaligenes faecalis B-4-28, Comamonas testosteroni B-4-31, Acinetobacter soli B-4-40 등이 있다.