• Title/Summary/Keyword: biodegradation,

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Analysis of Enzymes of Stenotrophomonas maltophilia LK-24 Associated with Phenol Degradation (Stenotrophomonas maltophilia LK-24의 페놀분해 관련 효소)

  • Kim, Jeong-Dong;Kang, Kook-Hee
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.37-46
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    • 2004
  • The analysis of enzymes associated with metabolism of phenolics by Stenotrophomonas maltophilia LK-24 was conducted. To identify metabolites of phenol and phenol compound, we investigated enzymes of S. maltophilia LK-24 associated with degradation of phenolics. We found that phenol hydrolase, catechol-2.3-dioxygenase, 2-hydroxymuconic semialdehyde dehydrogenase, 2-hydroxymuconic semialdehyde hydroxylase and acetaldehyde dehydrogenase were activated. The results showed that phenolics were gone through the meta-pathway ring cleavage. The results will contribute greatly to understand metabolic pathways of phenol and it is possible to make some assessment of the feasibility of using S. maltophilia LK-24 for the treatments of phenolic-contaminated waste streams.

Direct membrane filtration of wastewater under very short hydraulic retention time

  • Yoon, Seong-Hoon
    • Advances in environmental research
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    • v.7 no.1
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    • pp.39-52
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    • 2018
  • Direct membrane filtration (DMF) of wastewater has many advantages over conventional biological wastewater treatment processes. DMF is not only compact, but potentially energy efficient due to the lack of biological aeration. It also produces more biosolids that can be used to produce methane gas through anaerobic digestion. Most of ammoniacal nitrogen in wastewater is preserved in effluent and is used as fertilizer when effluent is recycled for irrigation. In this study, a technical feasibility of DMF was explored. Organic and nitrogen removal efficiencies were compared between DMF and membrane bioreactor (MBR). Despite the extremely high F/V ratio, e.g., $14.4kg\;COD/m^3/d$, DMF provided very high COD removal efficiencies at ~93%. Soluble microbial products (SMP) and extracellular polymeric substances (EPS) were less in DMF sludge, but membrane fouling rate was far greater than in MBR. The diversity of microbial community in DMF appeared very narrow based on the morphological observation using optical microscope. On the contrary, highly diverse microbial community was observed in the MBR. Microorganisms tended to form jelly globs and attach on reactor wall in DMF. FT-IR study revealed that the biological globs were structurally supported by feather-like materials made of secondary amines. Confocal laser scanning microscopy (CLSM) study showed microorganisms mainly resided on the external surface of microbial globs rather than the internal spaces.

Isolation of a Pseudomonas sp. Capable of Utilizing 4-Nonylphenol in the Presence of Phenol

  • Chakraborty Joydeep;Dutta Tapan K.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1740-1746
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    • 2006
  • Enrichment techniques led to the isolation of a Pseudomonas sp. strain P2 from municipal waste-contaminated soil sample, which could utilize different isomers of a commercial mixture of 4-nonylphenol when grown in the presence of phenol. The isolate was identified as Pseudomonas sp., based on the morphological, nutritional, and biochemical characteristics and 16S rDNA sequence analysis. The ${\beta}$-ketoadipate pathway was found to be involved in the degradation of phenol by Pseudomonas sp. strain P2. Gas chromatography-mass spectrometric analysis of the culture media indicated degradation of various major isomers of 4-nonylphenol in the range of 29-50%. However, the selected ion monitoring mode of analysis of biodegraded products of 4-nonylphenol indicated the absence of any aromatic compounds other than those of the isomers of 4-nonylphenol. Moreover, Pseudomonas sp. strain P2 was incapable of utilizing various alkanes individually as sole carbon source, whereas the degradation of 4-nonylphenol was observed only when the test organism was induced with phenol, suggesting that the degradation of 4-nonylphenol was possibly initiated from the phenolic moiety of the molecule, but not from the alkyl side-chain.

Characterization of Naphthalene-Degrading Pseudomonas Species Isolated from Pollutant-Contaminated Sites: Oxidative Stress During their Growth on Naphthalene

  • Kang, Yoon-Suk;Kim, Young-Jun;Jeon, Che-Ok;Park, Woo-Jun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1819-1825
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    • 2006
  • Four naphthalene-degrading bacteria (Pseudomonas sp. strains O1, W1, As1, and G1) were isolated feom pollutant-contaminated sites. Examination of their substrate utilization and analyses of key naphthalene-catabolic regulatory genes revealed that the pathway and regulation of naphthalene-degradation in all four strains resemble those of NAH7 from P. putida G7. Superoxide anion production, superoxide dismutase activity, and catalase activity during their growth on naphthalene-amended medium increased significantly, compared with those with glucose-amended medium. Addition of ascorbate, an antioxidant, or ferrous iron ($Fe^{2+}$) increased the growth rates of all tested microorganisms on naphthalene. Northern blot and HPLC analyses showed that both nahA gene expression and naphthalene degradation increased under those conditions. Our data suggest that naphthalene degradation can impose severe oxidative stress, and defenses against oxidative stress would play an important role in the metabolism of naphthalene.

Isolation and Characterization of Stenotrophomonas maltophilia Strains Capable of Degrading Aniline. (아닐린 분해 세균인 Stenotrophomonas maltophilia의 분리 및 특성)

  • 김현주;김진철;김흥태;최경자;최도일;김홍기;조광연
    • Microbiology and Biotechnology Letters
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    • v.28 no.4
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    • pp.202-208
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    • 2000
  • Four bacteria capable of using aniline as a sole source of carbon and energy we4e isolated from river waters. Among them, two strains were identified as Stenotrophomonas maltophilia based on their physiological and biochemical characteristics and 16SrRNA gene sequence and the others as delftia acidovorans. The four strains were able to grow on the mineral salt media containing aniline at concentrations up to 6,000 $\mu\textrm{g}$/ml. Since aniline degradation by S. maltophilia has not been reported so far, the two strains A-s and 51-4 were selected for further studies. They completely utilized aniline in a mineral salt medium containing 300 $\mu\textrm{g}$/ml of aniline as a sole carbon and energy source within 24 hours. Optimum pH and temperature for aniline degradation and cell growth of both strains were 7.0 and $35^{\circ}C$, respectively. In addition, they effectively degraded aniline is waste, underground and river waters containing 300 $\mu\textrm{g}$/ml of aniline. This is the first report of aniline degradation by S. maltophilia strains.

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Purification and Characterization of 2,3-Dihydroxybiphenyl 1,2-Dioxygenase from Comamonas sp. SMN4

  • Lee, Na-Ri;Lee, Jang-Mi;Min, Kyung-Hee;Kwon, Dae-Young
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.487-494
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    • 2003
  • 2,3-Dihydroxybiphenyl 1,2-dioxygenase (23DBDO), an enzyme of the biphenyl biodegradation pathway encoded by the bphC gene of Comnmonas sp. SMN4, was expressed and purified using column chromatographies. SDS-PAGE of purified 23DBDO showed a single band with a molecular mass of 32 kDa, which was consistent with the data from the gel filtration chromatography (GFC). The purified enzyme exhibited a maximum 23DBDO activity at pH 9.0 and was stable at pH 8.0. The enzyme showed maximum activity at $40^{\circ}C$ and maintained activity at $30^{\circ}C$ for 24 h. Kinetic parameters represented by Michaelis-Menten constants such as $K_m\;and\;V_{max}$ values for various substrates were determined by Lineweaver-Burk plots: The purified enzyme 23DBDO from Comamonas sp. SMN4 had the highest catalytic activity for 2,3-dihydroxybiphenyl and 3-methylcatechol, and had very poor activity with catechol and 4-methylcatechol.

Benzene Biodegradation Using the Polyurethane Biofilter Immobilized with Stenotrophomonas maltophilia T3-c

  • Kwon, Heock-Hoi;Lee, Eun-Young;Cho, Kyung-Suk;Ryu, Hee-Wook
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.70-76
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    • 2003
  • The benzene removal characteritics of the polyurethane (PU) biofilter immobilized with S. maltophilia T3-c, that could efficiently degrade benzene, was investigated. Maximum capacity to eliminate benzene was maintained at $100-110g{\cdot}m^-3{\cdot}h^-1$ when space velocity (SV) ranged from 100 to $300 h^-1$ -1/, however, it decreased sharply to $55 g{\cdot}m^-3{\cdot}h^-1^$ as SV increased to $400 h^-1$. The critical elimination capacities that guaranteed $90\%$ removal of inlet loading of the PU biofilter were determined to be 70,30, and $15 g{\cdot}m^-3{\cdot}h^-1$ at SV 100,200, and $300 h^-1$, respectively. Based on the result of a kinetic analysis of the PU biofilter, maximum benzene elimination velocity ($V_m$) was $125 g{\cdot}m^-3^\;of\;PU{\cdot}h^-1$ and saturation constant ($K_m$) was $0.22 g{\cdot}m^-3^$ of benzene ($65{\mu}{\cdot}I^-1$). This study suggests that the biofilter utilizing S. maltophilia T3-c and polyurethane is a very promising technology for effectively degrading benzene.

A Green Fluorescent Protein-based Whole-Cell Bioreporter for the Detection of Phenylacetic Acid

  • Kim, Ju-Hyun;Jeon, Che-Ok;Park, Woo-Jun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.10
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    • pp.1727-1732
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    • 2007
  • Phenylacetic acid (PAA) is produced by many bacteria as an antifungal agent and also appears to be an environmentally toxic chemical. The object of this study was to detect PAA using Pseudomonas putida harboring a reporter plasmid that has a PAA-inducible promoter fused to a green fluorescent protein (GFP) gene. Pseudomonas putida KT2440 was used to construct a green fluorescent protein-based reporter fusion using the paaA promoter region to detect the presence of PAA. The reporter strain exhibited a high level of gfp expression in minimal medium containing PAA; however, the level of GFP expression diminished when glucose was added to the medium, whereas other carbon sources, such as succinate and pyruvate, showed no catabolic repression. Interestingly, overexpression of a paaF gene encoding PAA-CoA ligase minimized catabolic repression. The reporter strain could also successfully detect PAA produced by other PAA-producing bacteria. This GFP-based bioreporter provides a useful tool for detecting bacteria producing PAA.

Genome Analysis of Naphthalene-Degrading Pseudomonas sp. AS1 Harboring the Megaplasmid pAS1

  • Kim, Jisun;Park, Woojun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.330-337
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    • 2018
  • Polycyclic aromatic hydrocarbons (PAHs), including naphthalene, are widely distributed in nature. Naphthalene has been regarded as a model PAH compound for investigating the mechanisms of bacterial PAH biodegradation. Pseudomonas sp. AS1 isolated from an arseniccontaminated site is capable of growing on various aromatic compounds such as naphthalene, salicylate, and catechol, but not on gentisate. The genome of strain AS1 consists of a 6,126,864 bp circular chromosome and the 81,841 bp circular plasmid pAS1. Pseudomonas sp. AS1 has multiple dioxygenases and related enzymes involved in the degradation of aromatic compounds, which might contribute to the metabolic versatility of this isolate. The pAS1 plasmid exhibits extremely high similarity in size and sequences to the well-known naphthalene-degrading plasmid pDTG1 in Pseudomonas putida strain NCIB 9816-4. Two gene clusters involved in the naphthalene degradation pathway were identified on pAS1. The expression of several nah genes on the plasmid was upregulated by more than 2-fold when naphthalene was used as a sole carbon source. Strains have been isolated at different times and places with different characteristics, but similar genes involved in the degradation of aromatic compounds have been identified on their plasmids, which suggests that the transmissibility of the plasmids might play an important role in the adaptation of the microorganisms to mineralize the compounds.

Biodegradation of Feather Waste Keratin by the Keratin-Degrading Strain Bacillus subtilis 8

  • He, Zhoufeng;Sun, Rong;Tang, Zizhong;Bu, Tongliang;Wu, Qi;Li, Chenlei;Chen, Hui
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.314-322
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    • 2018
  • Bacillus subtilis 8 is highly efficient at degrading feather keratin. We observed integrated feather degradation over the course of 48 h in basic culture medium while studying the entire process with scanning electron microscopy. Large amounts of ammonia, sulfite, and $\text\tiny{L}$-cysteic acid were detected in the fermented liquid. In addition, four enzymes (gamma-glutamyltranspeptidase, peptidase T, serine protease, and cystathionine gamma-synthase) were identified that play an important role in this degradation pathway, all of which were verified with molecular cloning and prokaryotic expression. To the best of our knowledge, this report is the first to demonstrate that cystathionine gamma-synthase secreted by B. subtilis 8 is involved in the decomposition of feather keratin. This study provides new data characterizing the molecular mechanism of feather degradation by bacteria, as well as potential guidance for future industrial utilization of waste keratin.