• Title/Summary/Keyword: meta-cleavage

Search Result 51, Processing Time 0.025 seconds

Cloning of pcb Genes in Pseudomonas sp.P20 Specifying Degradation of 4-Clorobiphenyl (4-Chlorobiphenyl을 분해하는 Pseudomonas sp. P20의 pcb 유전자군의 클로닝)

  • 남정현;김치경
    • Microbiology and Biotechnology Letters
    • /
    • v.22 no.4
    • /
    • pp.353-359
    • /
    • 1994
  • Pseudomonas sp. P20 was a bacterial isolate which has the ability to degrade 4-chlorobi- phenyl(4CB) to 4-chlorobenzoic acid via the process of meta-cleavage. The recombinant plasmid pCK1 was constructed by insetting the 14-kb EcoRI fragment of the chromosomal DNA containing the 4CB-degrading genes into the vector pBluescript SK(+). Subsequently, E. coli XL1-Blue was transformed with the hybrid plasmid producing the recombinant E. coli CK1. The recombinant cells degraded 4CB and 2,3-dihydroxybiphenyl(2,3-DHBP) by the pcbAB and pcbCD gene products, respectively. The pcbC gene was expressed most abundantly at the late exponential phase in E. coli CK1 as well as in Pseudomonas sp. P20, and the level of the pcbC gene product, 2,3-DHBP dioxygenase, expressed in E. coli CK1 was about two-times higher than in Pseudomonas sp. P20. The activities of 2,3-DHBP dioxygenase on catechol and 3-methylcatechol were about 26 to 31% of its activity on 2,3-DHBP, but the enzyme did not reveal any activities on 4-methylcatechol and 4-chlorocatechol.

  • PDF

Characterization of biphenyl biodegradation, and regulation of iphenyl catabolism in alcaligenes xylosoxydans

  • Lee, Na-Ri;On, Hwa-Young;Jeong, Min-Seong;Kim, Chi-Kyung;Park, Yong-Keun;Ka, Jong-Ok;Min, Kyung-Hee
    • Journal of Microbiology
    • /
    • v.35 no.2
    • /
    • pp.141-148
    • /
    • 1997
  • Alcaligenes xylosoxydans strain SMN3 capable of utilizing biphenyl grew not only on phenol, and benzoate, but also on salicylate. Catabolisms of biphenyl and salicylate appear to be interrelated since benzoate is a common metabolic intermediate of these compounds. Enzyme levels in the excatechol 2. 3-dioxygenas which is meta-cleavage enzyme of catechol, but did not induce catechol 1, 2-dioxygenase. All the oxidative enzymes of biphenyl and 2, 3,-dihydroxybiphenyl (23DHBP) were induced when the cells were grown on biphenyl and salicylate, respectively. Biphenyl and salicylate could be a good inducer in the oxidation of biphenyl and 2, 3-dihydroxybiphenyl. The two enzymes for the degradation of biphenyl and salicylate were induced after growth on either biphenyl or salicylate, suggesting the presence of a common regulatory element. However, benzoate could not induce the enzymes responsible for the oxidation of these compounds. Biphenyl and salicylate were good inducers for indigo formation due to the activity of biphenyl dioxygenase. These results suggested that indole oxidation is a property of bacterial dioxygenase that form cis-dihydrodiols from aromatic hydrocarbon including biphenyl.

  • PDF

Regulation of Phenol Metabolism in Ralstonia eutropha JMP134

  • Kim Youngjun
    • Proceedings of the Microbiological Society of Korea Conference
    • /
    • 2002.10a
    • /
    • pp.27-30
    • /
    • 2002
  • Ralstonia eutrupha JMP134 is a well-known soil bacterium which can metabolite diverse aromatic compounds and xenobiotics, such as phenol, 2,4-dichlorophenoxy acetic acid (2, 4-D), and trichloroethylene (TCE), etc. Phenol is degraded through chromosomally encoded phenol degradation pathway. Phenol is first metabolized into catechol by a multicomponent phenol hydroxylase, which is further metabolized to TCA cycle intermediates via a meta-cleavage pathway. The nucleotide sequences of the genes for the phenol hydroxylase have previously been determined, and found to composed of eight genes phlKLMNOPRX in an operon structure. The phlR, whose gene product is a NtrC-like transcriptional activator, was found to be located at the internal region of the structural genes, which is not the case in most bacteria where the regulatory genes lie near the structural genes. In addition to this regulatory gene, we found other regulatory genes, the phlA and phlR2, downstream of the phlX. These genes were found to be overlapped and hence likely to be co-transcribed. The protein similarity analysis has revealed that the PhlA belongs to the GntR family, which are known to be negative regulators, whereas the PhlR2 shares high homology with the NtrC-type family of transcriptional activators like the PhlR. Disruption of the phlA by insertional mutation has led to the constitutive expression of the activity of phenol hydroxylase in JMP134, indicating that PhlA is a negative regulator. Possible regulatory mechanisms of phenol metabolism in R. eutropha JMP134 has been discussed.

  • PDF

Isolation and Characterization of a Phenol-Degrading Strain Acinetobacter sp.GEM2 (Phenol을 분해하는 Acinetobacter sp. GEM2의 분리 및 특성)

  • Lee, Chang-Ho;Oh, Hee-Mock;Kwon, Tae-Jong;Kwon, Gi-Seok;Lee, Sung-Gie;Suh, Hyun-Hyo;Yoon, Byung-Dae
    • Microbiology and Biotechnology Letters
    • /
    • v.22 no.6
    • /
    • pp.692-699
    • /
    • 1994
  • A bacterial strain which formed a distinct colony on agar plate containing phenol as a vapor phase and grew well in a liquid minimal medium was isolated and identified as Acinetobac- ter sp. GEM2. The optimal temperature and initial pH for the growth of Acinetobacter sp. GEM2 were 30$\circ$C and 7.0, respectively. Cell growth was inhibited by phenol at the concentration over 1500 ppm. Cell growth dramatically increased from 10 hours after cultivation and almost showed a stationary phase within 24 hours at which 95% of phenol was concomitantly degraded. Acinetobac- ter sp. GEM2 was capable of growing on aromatic compounds, such as benzoic acid, phenol, m- cresol, o-cresol, P-cresol, catechol, gentisic acid, and toluene, but did not grow on benzene, salicylic acid, p-toluic acid, and p-xylene. By the analysis of catechol dioxygenase, it seemed that catechol was degraded through both meta- and ortho-cleavage pathway. The growth-limiting log P value of Acinetobacter sp. GEM2 on organic solvents was 2.0.

  • PDF

Spectral and Mechanistic Investigation of Oxidative Decarboxylation of Phenylsulfinylacetic Acid by Cr(VI)

  • Subramaniam, Perumal;Selvi, Natesan Thamil;Devi, Soundarapandian Sugirtha
    • Journal of the Korean Chemical Society
    • /
    • v.58 no.1
    • /
    • pp.17-24
    • /
    • 2014
  • The oxidative decarboxylation of phenylsulfinylacetic acid (PSAA) by Cr(VI) in 20% acetonitrile -80% water (v/v) medium follows overall second order kinetics, first order each with respect to [PSAA] and [Cr(VI)] at constant [$H^+$] and ionic strength. The reaction is acid catalysed, the order with respect to [$H^+$] is unity and the active oxidizing species is found to be $HCrO_3^+$. The reaction mechanism involves the rate determining nucleophilic attack of sulfur atom of PSAA on chromium of $HCrO_3^+$ forming a sulfonium ion intermediate. The intermediate then undergoes ${\alpha}$,${\beta}$-cleavage leading to the liberation of $CO_2$. The product of the reaction is found to be methyl phenyl sulfone. The operation of substituent effect shows that PSAA containing electron-releasing groups in the meta- and para-positions accelerate the reaction rate while electron withdrawing groups retard the rate. An excellent correlation is found to exist between log $k_2$ and Hammett ${\sigma}$ constants with a negative value of reaction constant. The ${\rho}$ value decreases with increase in temperature evidencing the high reactivity and low selectivity in the case of substituted PSAAs.

Identification of Yarrowia lipolytica Y103 and Its Degradability of Phenol and 4-Chlorophenol

  • Lee, Jeong-Soon;Kang, Eun-Jeong;Kim, Min-Ok;Lee, Dong-Hun;Bae, Kyung-Sook;Kim, Chi-Kyung
    • Journal of Microbiology and Biotechnology
    • /
    • v.11 no.1
    • /
    • pp.112-117
    • /
    • 2001
  • A nonconventional yeast strain Y103 capable of degrading several aromatic hydrocarbons was isolated from the wastewater of the Yocheon industrial complex. The strain Y103 was identified as Yarrowia lipolytica on the basis of its unique dimorphic and biochemical characteristics as determined by a Biolog test. Y. lipolytica Y103 was found to degrade phenol and 4-chlorophenol to produce catechol. The catechol then will be further degraded to produce 2-hydroxymuconic semialdehyde via meta-cleavage. These results indicate that strain Y103 degrades 4-chlorophenol, phenol, and catechol through a consecutive reaction to produce 2-hydroxymuconic semialdehyde. The most active degradation of phenol by Y. lipolytica Y103 occurred with a 0.5 mM phenl concentration in an MM2 medium at $30^{\circ}C$ and pH 7.0.

  • PDF

Structural Layers and History of Folding in the Western Part of the Baegunsan Syncline, Samcheog Coalfield, South Korea (백운산향사(白雲山向斜) 서단부(西端部)의 구조계층(構造階層)과 습곡작용(褶曲作用)의 과정(過程))

  • RHI, Jae-Young;PARK, Bong-Soon
    • Economic and Environmental Geology
    • /
    • v.15 no.1
    • /
    • pp.41-47
    • /
    • 1982
  • Typical stratigraphic sequences of Paleozoic form the Baegunsan Syncline (equal to Hambaeg Syncline) and Taebaegsan Group is the basement in the southern part of Kangweon Province. Deformation pattern depends on their stratigraphic site and their internal properties. In the biotite schists and meta-sandstones of Pre-Cambrian sequences, flow folds and ptygmatic folds are developed, and high strain deformation is pervasive. Deformational patterns of Cambro-Ordovician are variable because of their various formations. Fracturing is pervasive in the Jangsan Quartzite and Daegi and Maggol Limestone. Welldeveloped slaty cleavage and angular folds of kink and chevron types are the prevailing structures of pelitic rocks. The most characteristic feature of limestone alternated with argillite is the "Compositional cleavage (Author's proposal)" known as the "Worm-eatern" structures. It was known that this structures have a sedimentary origin. But their preferred orienation of long axis of erosional lime holes, originated from tectonic deformation of folding. And this structures have the same character as the axial plane cleavages. Fracturing and long wave concentric folds are dominant characters of the Upper Paleozic sequences. In this area, two folds are superimposed. Field studying and analysis of structures show that the cross-folds of NS-direction are pervasive and were deformed by the EW-directional Baegunsan Syncline. The cross-folding is the major and penetrative deformation and prior to Baegunsan Syncline.

  • PDF

Isolation of a Phenol-degrading Bacterial Strain and Biological Treatment of Wastewater Containing Phenols (Phenol 분해균주의 분리 및 페놀함유 폐수의 생물학적 처리)

  • Lee, Hyun Don;Lee, Myoung Eun;Kim, Hyung Gab;Suh, Hyun-Hyo
    • Journal of Life Science
    • /
    • v.23 no.10
    • /
    • pp.1273-1279
    • /
    • 2013
  • Aromatic hydrocarbons, such as phenol, have been detected frequently in wastewater, soil, and groundwater because of the extensive use of oil products. Bacterial strains (56 isolates) that degraded phenol were isolated from soil and industrial wastewater contaminated with hydrocarbons. GN13, which showed the best cell growth and phenol degradation, was selected for further analysis. The GN13 isolate was identified as Neisseria sp. based on the results of morphological, physiological, and biochemical taxonomic analyses and designated as Neisseria sp. GN13. The optimum temperature and pH for phenol removal of Neisseria sp. GN13 was $32^{\circ}C$ and 7.0, respectively. The highest cell growth occurred after cultivation for 30 hours in a jar fermentor using optimized medium containing 1,000 mg/l of phenol as the sole carbon source. Phenol was not detected after 27 hours of cultivation. Based on the analysis of catechol dioxygenase, it seemed that catechol was degraded through the meta- and ortho-cleavage pathway. Analysis of the biodegradation of phenol by Neisseria sp. GN13 in artificial wastewater containing phenol showed that the removal rate of phenol was 97% during incubation of 30 hours. The removal rate of total organic carbon (TOC) by Neisseria sp. GN13 and activated sludge was 83% and 78%, respectively. The COD removal rate by Neisseria sp. GN13 from petrochemical wastewater was about 1.3 times higher than that of a control containing only activated sludge.

Kinetic Property and Phylogenie Relationship of 2-Hydroxy-muconic Semialdehyde Dehydrogenase Encoded in tomC Gene of Burkholderia cepacia G4

  • Reddy, Alavala-Matta;Min, Kyung-Rak;Lee, Kyoung;Lim, Jai-Yun;Kim, Chi-Kyung;Kim, Young-Soo
    • Archives of Pharmacal Research
    • /
    • v.27 no.5
    • /
    • pp.570-575
    • /
    • 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.

Improvement of 4-chlorobiphenyl degradation bya recombinant strain, pseudomonas sp. DJ12-C

  • Kim, Ji-Young;Kim, Young-Chang;You, Lim-Jai;Lee, Ki-Sung;Ok, Ka-Jong;Hee, Min-Kyung;Kim, Chi-Kyung
    • Journal of Microbiology
    • /
    • v.35 no.1
    • /
    • pp.53-60
    • /
    • 1997
  • Pseudomonas sp. P20 and Pseudomonas sp. DJ-12 isolated from the polluted environment are capable of degrading biphenyl and 4-chlorobiphenyl (4CB) to produce benzoic acid and 4-chlorobenzoic acid (4CBA) respectively, by pcbABCD-encoded enzymes. 4CBA can be further degraded by Pseudomonas sp. DJ-12, but not by Pseudomonas sp P20. However, the meta-cleavage activities of 2, 3-dihydroxybiphenyl (2, 3-DHBP) and 4-chloro-2, 3-DHBP dioxygenases (2, 3-DHBD) encoded by pcbC in Pseudomonas sp. P20 were stronger than Pseudomonas sp. DJ-12. In this study, the pcbC gene encoding 2, 3-DHBD was cloned from the genomic DNA of Pseudomonas sp. P20 by using pKT230. A hybrid plasmid pKK1 was constructed and E. coli KK1 transformant was selected by transforming the pKK1 hybrid plasmid carrying pcbC into E. coli XL1-Blue. By transferring the pKK1 plasmide of E. coli KK1 into Pseudomonas sp. DJ-12 by conjugation, a recombinant strain Pseudomonas sp. P20, Pseudomonas sp. DJ-12, and the recombinant cell assay methods. Pseudomonas sp. DJ12-C readily degraded 4CB and 2, 3-DHBP to produce 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoic acid (HOPDA), and the resulting 4CBA and benzoic acid were continuously catabolized. Pseudomonas sp. DJ12-C degraded 1 mM 4CB completely after incubation for 20 h, but Pseudomonas sp. P20 and Pseudomonas sp. DJ-12 showed only 90% and Pseudomonas sp. DJ-12 had, but its degradation activity to 2, 3-DHBP, 3-methylcatechol, and catechol was improved.

  • PDF