• Journal of Microbiology and Biotechnology
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• 제13권2호
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• pp.243-250
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• 2003

Eight numerically dominant 4-(2,4-dichlorophenoxy) butyric acid (2,4-DB)-degrading bacteria and three pairs of bacteria showing syntrophic metabolism of 2,4-DB were isolated from soils, and their phylogenetic and phenotypic characteristics were investigated. The isolates were able to utilize 2,4-DB as a sole source of carbon and energy, and their 2.4-DB degradative enzymes were induced by the presence of 2.4-DB. Analysis of 16S rDNA sequences indicated that the isolates were related to members of the genera, Variovorax, Sphingomonas, Bradyrhizobium, and Pseudomonas. The chromosomal DNA patterns of the isolates obtained by polymerase-chain-reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences were distinct from each other. Four of the isolates had plasmids, but only one strain, DB 1, rad a transmissible 2,4-D degradative plasmid. When analyzed with PCR using primers targeted to the tfdA, B, and C genes, only strains DB2 and DB9a produced DNA bands of the expected sizes with the tfdA and C primers, respectively. All of the isolates were able to degrade 2,4-D as well as 2,4-DB, suggesting that the degradation pathways of these compounds were closely related to each other, but respiratory activities of many isolates adapted to 2,4-DB metabolism were quite low with 2,4-D.

• Applied Biological Chemistry
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• 제39권5호
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• pp.403-408
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• 1996

토양에서 세균군집의 DNA를 추출하여 이사디 분해세균의 밀도와 군집변화를 tdfA 유전자와 Spa Probe를 이용하여 조사하였다. 이사디 분해균주인 Pseudomonas cepacia/pJP4을 토양에 여러 가지 밀도로 접종한 후 추출된 토양세균군집의 DNA를 Southern blot에서 분석한 결과, 본 실험에 사용된 DNA probe method에 의해 이 세균을 $10^5\;cells/g$ soil 수준까지 검출할 수 있는 것으로 나타났다. 이사디를 가해준 microcosm 토양에서 추출된 세균군집의 DNA를 분석한 실험에서는 Pseudemonas pickettii와 Sphingomonas Paucimobilis가 우점종으로 검출되었고, 사용된 두 가지의 DNA probes는 토양의 이사디 분해미생물에 대해 매우 높은 특이성을 가지고 있는 것으로 나타났다. 밭에 이사디를 장기 적으로 가해준 후 추출된 토양세균군집의 DNA를 분석 한 실험에서는 이사디를 최소한 10 ppm 이상 가해주어야 토양의 이사디 분해세균을 DNA probe method에 의해 검출할 수 있었고, tfdA 유전자는 실제의 밭토양에서도 높은 특이성을 나타냈으나 Spa probe는 일부의 토착세균에 비특이적으로 반응하는 것으로 나타났다. 토양에서 추출된 세균군집의 DNA를 분석하는 DNA probe method는 Southern blot과 함께 사용되었을 때 토양에 존재하는 이사디 분해미생물을 실험실 배지에 배양하지 않고 검출할 수 있었고,이 미생물들의 밀도, 군집변화, 유전적 변화 등을 효과적으로 분석할 수 있는 것으로 나타났다.

• Journal of Microbiology
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• 제34권4호
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• pp.320-326
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• 1996

To analyze the effects of host versus plasmid on survival of 2, 4-degrading bacteria in environmental samples, strains Pseudomonas cepacia/pJP4, Alcaligenes JMP228/pJP4, P. cepacia/p712, and Alcaligenes JMP228/p712 were separately inoculated into samples of field soil, paddy soil, lake water, and river water, and then the changes of their populations were measured. The strains used contained a 2, 4-D degradative plasmid, either pJP4 conferring fast-growing property to the host or p712 conferring slow-growing property, and were resistant to antibiotics such that the inoculated strains could be enumerated against the indigenous microbial populations. In sterile environmental samples, these strains were stably maintained at the levels used for inoculation, except in sterile paddy soil where Alcaligenes JMP228 strains died drapidly. In natural soil samples for four strains declined steadily with time, but in naturla water samples their polulations fell rapidly at the early phase and then remained almost constant. When the environmentla samples were treated with 2, 4-D, P. cepacia/pJP4 and P. cepacia/p712 maintained significant numbers, while Alcaligenes JMP228/pJP4 and Alcaligenes JMP228/p712 declined significantly in most of the samples. The results indicated that the survivability of genetically modified microorganisms could vary depending on the environments and that their abundance in the environments under s2, 4-D selection was markedly influenced by the nature of the 2, 4-D degradative plasmid as well as type of the host strain.

• Journal of Microbiology and Biotechnology
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• 제12권4호
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• pp.583-591
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• 2002

Biodegradation of fat, oil, and grease (FOGs) plays an Important role in wastewater management and water pollution control. However, many industrial food-processing and food restaurants generate FOG-containing waste waters for which there Is no acceptable technology for their pretreatment. To solve these problems, this study evaluated the feasibility of effective FOG-degrading microorganisms on the biodegradation of olive oil and FOG-containing wastewater. Twenty-two strains capable of degrading FOGs were isolated from five FOG-contaminated sites for the evaluation of their FOG degradation capabilities. Among twenty-two strains tested, the lipase-producing Pseudomonas sp. strain D2D3 was selected for actual FOG wastewater treatment. Its biodegradability was performed at 3$0^{\circ}C$ and pH 8. The extent of FOG removal efficiency was varied for each FOG tested, being the highest for olive oil and animal fat (94.5% and 94.4%), and the lowest for safflower oil (62%). The addition of organic nitrogen sources such as yeast extract, soytone, and peptone enhanced the removal efficiency of FOGs, but the addition of the inorganic nitrogen nutrients such as $NH_4$Cl and $(NH_4)_2SO_4$ did not increase. The $KH_2PO_4$ sources in 0.25% to 0.5% concentrations showed more than 90% degradability. As a result, the main pathway for the oxidation of fatty acids results in the removal of two carbon atoms as acetyl-CoA with each reaction sequence: $\beta$-oxidation. Its lipase activity showed 38.5 U/g DCW using the optimal media after 9 h. Real wastewater and FOGs were used for determining the removal efficiency by using Pseudomonas sp. strain D2D3 bioadditive. The degradation by Pseudomonas sp. strain D2D3 was 41% higher than that of the naturally occurring bacteria. This result indicated that the use of isolated Pseudomonas sp. strain D2D3 in a bioaugmentating grease trap or other processes might possibly be sufficient to acclimate biological processes for degrading FOGs.

• 한국토양비료학회지
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• 제33권1호
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• pp.47-51
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• 2000

화합물을 분해하는 우수균주 개발의 기초연구로서, 대전 근교 지역의 논과 밭에서 채취한 토양 표품으로부터 100균주의 세균을 분리하였고, 그 중 2,4,5-T를 단일 탄소원으로 하는 고체 최소 배지에서 잘 자라는 균주 19균주를 선별하였다. 이들 균주를 등정한 결과 Pseudomonas속이 11균주 Acinetobacter속이 4균주, Alcaligenes속이 1균주이고 3균주는 미동정되었다. Pseudomonas속으로 밝혀진 MU19와 MU92는 네가지 염소계 화합물들(2,4-D, 2,4,5-T, MCPA 그리고 3CB)을 모두 분해하는 것으로 나타났다. 최소 액체배지에서 배양한 경우 Acinetobacter로 동정된 MU38균주가 접종 48시간 후에 가장 높은 분해도를 나타내었고, MUl9, MU57, MU73과 MU92는 그 다음으로 높은 분해도를 나타냈다. 실험결과 선별된 19균주 중 Acinetobacter sp. MU38 그리고 Pseudomonas sp. MU19과 MU92는 염소계 방향쪽 화합물에 대한 넓은 분해능을 갖고 있으며, 특히 2,4,5-T에 대한 높은 분해도를 나타내는 것으로 조사되었다.

• 한국환경과학회지
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• 제21권2호
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• pp.253-261
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• 2012

We isolated and characterized novel duck feather-degrading bacteria producing keratinase. Twelve strains were isolated from soil and faces at poultry farm, and decayed feathers. They were identified as Bacillus methylotrophicus, Pseudomonas geniculata, Pseudomonas hibiscicola, Exiquobacterium profundum, Bacillus pumilus, Bacillus amyloliquefaciens, Chryseobacterium indologenes, Bacillus thuringiensis, Thermomonas koreensis, respectively, by phenotypic characters and 16S rRNA gene analysis. Generally, the level of keratinase production was not proportional to feather degradation rate. The highest keratinolytic activity was observed in the culture inoculated with Chryseobacterium indologenes D27. Although all strains did not degrade human hair, strains tested effectively degraded chicken feather(53.8-91.4%), wool(40.4-93.0%) and human nail (51.0-82.9%). These results suggest that strains isolated could be not only used to improve the nutritional value of recalcitrant feather waste but also is a potential candidate for biotechnological processes of keratin hydrolysis.

• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.376-383
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• 2005

Alcaligenes sp. JMP228 carrying 2,4­dichlorophenoxyacetic acid (2,4-D) degradative plasmid pJP4 was inoculated into natural soil, and transfer of the plasmid pJP4 to indigenous soil bacteria was investigated with and without 2,4-D amendment. Plasmid pJP4 transfer was enhanced in the soils treated with 2,4-D, compared to the soils not amended with 2,4-D. Several different transconjugants were isolated from the soils treated with 2,4-D, while no indigenous transconjugants were obtained from the unamended soils. Inoculation of the soils with both the donor Alcaligenes sp. JMP228/pJP4 and a recipient Burkholderia cepacia DBO 1 produced less diverse transconjugants than the soils inoculated with the donor alone. Repetitive extragenic palindromic-polymerase chain reaction (REP-PCR) analysis of the transconjugants exhibited seven distinct genomic DNA fingerprints. Analysis of 16S rDNA sequences indicated that the transconjugants were related to members of the genera Burkholderia and Pandoraea. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that inoculation of the donor caused clear changes in the bacterial community structure of the 2,4-D­amended soils. The new 16S rRNA gene bands in the DGGE profile corresponded with the 16S rRNA genes of 2,4-D­degrading transconjugants isolated from the soil. The results indicate that introduction of the 2,4-D degradative plasmid as Alcaligenes sp. JMP228/pJP4 has a substantial impact on the bacterial community structure in the 2,4-D-amended soil.

• 한국미생물·생명공학회지
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• 제34권2호
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• pp.109-114
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• 2006

본 연구는 부패 생마로부터 저온성 부패균을 분리, 동정하고 분리균의 생육특성과 다양한 효소의 부패관련성을 조사하여, 생마 저온 장기저장 및 생마 가공식품 개발을 위한 기초자료를 제시할 목적으로 수행되었다. 저온 장기저장중의 부패생마로부터 서로 다른 13종의 저온세균을 분리, 동정하였으며, 다양한 온도에서 생마 절편을 이용한 부패력 측정 결과 분리균주 중 YAM-10및 YAM-12균주가 저온부패에 직접적으로 관련됨을 확인하였으며, 이들은 각각 Pseudomonas cepacia 및 Pseudomonas rhodesiae 동정되었다. 이들 균주는 20$^{\circ}C$에서도 우수한 amylase, CMCase, xylanase 활성을 나타내었으며 , 특히 amylase 활성은 생마 부패에 중요한 역할을 하는 것으로 판단되었다. YAM-10 및 YAM-12균주는 양파절편을 부패시키지는 못하였으며, 4$\sim$12$^{\circ}C$의 온도에서도 생육가능하며, pH 5 이하 및 pH 10 이상에서는 생육이 급격히 억제되었다. 현재 생마의 저온 장기저장 및 부패억제를 위한 저온성 Pseudomonas sp.의 제어와 생마부패에 관련되는 효소들의 특성과 저해에 대한 연구 및 저장성, 관능성이 강화된 산장, 염장 등을 통한 생마 가공식품 개발이 진행 중이다.

• Archives of Pharmacal Research
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• 제23권2호
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• pp.187-195
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• 2000

Nucleotide sequence extending 2,3-dihydroxybiphenyl 1,2-dioxygenase gene (pcbC) and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase gene (pcbD) of Pseudomonas sp. DJ-12 was previously analyzed and the two genes were present in the order of pcbD-pcbC preceded by a promoter from Pseudomonas sp. DJ-12. In this study, a 3.8-kb nucleotide sequence located downstream of the pcbC gene was analyzed to have three open reading frames (ORFs) that are designated as orf1, pcbE and orf2 genes. All of the ORFs were preceded by each ribosome-binding sequence of 5-GGAXA-3 (X=G or A). However, no promoter-like sequence and transcription terminator sequence were found in the analyzed region, downstream of pcbC gene. Therefore, the gene cluster appeared to be present in the order of pcbD-pcbC-orf1-pcbE-orf2 as an operon, which is unique organization characterized so far in biphenyl- and PCB-degrading bacteria. The orf1 gene was composed of 1,224 base pairs which can encode a polypeptide of molecular weight 44,950 containing 405 amino acid residues. A deduced amino acid sequence of the orf1 gene product exhibited 21-33％ identity with those of indole dioxygenase and phenol hydroxylase components. The pcbE gene was composed of 783 base pairs encoding 2-hydroxypenta-2,4-dienoate hydratase involved in the 4-chlorobiphenyl catabolism. The orf2 gene was composed of 1,017 base pairs encoding a polypeptide of molecular weight 37,378 containing 338 amino acid residues. A deduced amino acid sequence of the orf2 gene product exhibited 31％ identity with that of a nitrilotriacetate monooxygenase component.

• 상하수도학회지
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• 제10권4호
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• pp.119-126
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• 1996

Influence factors and efficiency characteristics for treatment of wastewater containing phenol were studied with using Pseudomonas sp. B3. It took 130 hours to remove phenol, when only activated sludge of terminal disposal palnt of sewage was innoculated in batch culture, but it was required just 36 hours, when bacteria degrading phenol and activated sludge were simultaneously innoculated. If only phenol an carbon source was used, it necessary 36 hours for biodegradation of phenol, while glucose was added to medium, it took 73 hours. It was revealed as excellent effluent and SVI, when the F/M ratio, COD and phenol concentration were 53mg/l and 1.2mg/l, respectively, and optimum F/M ratio was revealed 0.31. The reactor were seriously shocked as reducing hydraulic retention time at constant phenol concentration more than increasing phenol concentration at constant hydraulic retention time, when volumetric loading rate was increased to $0.8kg\;phenol/m^3{\codt}d$ from $1.6kg\;phenol/m^3{\codt}d$. And also the effluent phenol concentration was 34mg/l after starting 12 hours of shocking and reactor was recovered as steady state after 65 hours of changing in the former test. Although the effluent phenol concentration was maximum value with 12mg/l after starting 20 hours of shocking and reactor was recovered as steady state after 54 hours of changing in the later test.