• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
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  • 优先权
    • 专利摘要:
    The present invention relates to Pseudomonas sp. SY5 ( Pseudomonas sp. Strain SY5), and more specifically, Pseudomonas sp. Strain SY5, which is a new strain isolated from activated sludge in a sewage treatment plant, having excellent resolution of polychlorinated biphenyl (PCB) and useful for sewage treatment. ( Pseudomonas sp.strain SY5).
    公开号:KR19980044738A
    申请号:KR1019960062862
    申请日:1996-12-07
    公开日:1998-09-05
    发明作者:정선용；이재석
    申请人:강대승；주식회사 동성화학；
    IPC主号:
    专利说明:

    Pseudomonas sp.strain SY5
    The present invention relates to Pseudomonas sp. SY5 ( Pseudomonas sp. Strain SY5), and more specifically, Pseudomonas sp. Strain SY5, which is a new strain isolated from activated sludge in a sewage treatment plant, having excellent resolution of polychlorinated biphenyl (PCB) and useful for sewage treatment. ( Pseudomonas sp.strain SY5).
    PCB (polychlorinated biphenyl) is a compound in which 1 to 10 chlorine atoms are substituted in biphenyl (biphenyl) in which two benzene rings are bonded. In the past, PCB was used in large quantities in heat medium, insulating oil and gas oil. The PCB enters the sewage treatment plant as it is after use, causing environmental damage and ecosystem changes. It has been discontinued. However, in the world, 1 million tons of PCBs have been released into the natural world, and the amount of disposal disposal reaches 2 million tons, and in Korea, there are many PCBs to be treated at present, and their efficient disassembly method is urgently needed. Incineration of PCBs is legally prohibited because it causes secondary pollutants such as dioxine and dibenzofuran.
    In 1992, Frank from the University of California, U.S.A., reported that it is safe to disintegrate PCBs, rather than physically and chemically, to produce secondary toxic substances. As a result, researches on how to effectively decompose PCB by microorganisms have been actively conducted. However, due to the lack of good microorganisms that can withstand high concentrations of PCBs and the degradation of the degrading enzymes, there have been many difficulties in industrial approach.
    As far as is known, the strains of Corynebacterium sp. MB1 and Alcaligenes Eutrophus H850 are known to have excellent PCB resolution, but these strains are generally PCBs with high chlorine substitution. Its low resolution and only disassemble PCB of very limited structure.
    PCB, which is a problem in environmental pollution, is a case where most of the chlorine substituents of various structures are substituted. Therefore, the conventional PCB degradation bacteria are not suitable for application for environmental purification.
    The inventors of the present invention endeavored to isolate and identify excellent microorganisms capable of withstanding high concentration of PCB while having excellent PCB resolution. As a result, the present invention was completed by culturing the sample collected at the sewage terminal treatment plant under appropriate conditions, and selecting a microorganism having a degrading ability for PCB or a derivative thereof having a high chlorine substitution from the cultured microorganism.
    Accordingly, an object of the present invention is to provide a strain Pseudomonas sp. Strain SY5 ( Pseudomonas sp. Strain SY5) useful for sewage treatment with excellent PCB resolution.
    Figure 1 is an electron micrograph of Pseudomonas sp. Strain SY5 Pseudomonas strain SY5.
    Figure 2 is a gas chromatogram showing the PCB degradation activity of Pseudomonas sp. Strain SY5 ( Pseudomonas sp. Strain SY5),
    a) is the gas chromatogram of the standard sample without microorganism inoculation,
    b) is a gas chromatogram showing the degree of PCB degradation by Pseudomonas sp. Strain SY5.
    The present invention is characterized by Pseudomonas sp. Strain SY5.
    Referring to the present invention in more detail as follows.
    The present invention relates to a strain Pseudomonas sp. SY5 ( Pseudomonas sp. Strain SY5) excellent in PCB resolution, the isolation and identification of this strain is as follows.
    -Isolation of mycobacteria-
    Samples are collected from the sewage treatment plant and placed in a test tube containing 10 ml of inorganic medium, and cultured in a liquid state at 30 ° C. for 1 week. At this time, as a carbon source, a PCB derivative having a basic skeleton, such as PCB or biphenyl, is used at a concentration of 1%. After one week, the test tubes in which the bacteria are grown are screened, inoculated 1% in the test tube containing the same medium, and incubated for another week. Repeat this twice, and then plated in an organic solid medium and allowed to incubate in solid state at 30-37 ° C. for 3 days, and then acetone containing 2,3-dihydroxybiphenyl on the resulting colonies. Spray the solution to select PCB digestion microorganisms.
    -Identification of mycobacteria-
    The isolated strain was identified (Bergey's Manual of systematic Bacteriology 9ed.). The items tested for identification and the results are shown in Table 1 below.
    According to the results of Table 1, the strain isolated in the present invention is found to belong to Pseudomonas sp. This strain is named Pseudomonas sp.strain SY5.
    Pseudomonas spp SY5 (Pseudomonas sp. Strain SY5) was commissioned on October 9, 1996 in the Center for Genetic Resources, Korea Institute of Science and Technology R & D Biotechnology Institute, it was given an accession number KCTC 8764P.
    The present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto.
    Example 1 Culture of Pseudomonas Strain SY5
    Pseudomonas sp SY5 (Pseudomonas sp. Strain SY5) is capable of liquid culture and solid culture.
    1) Liquid culture in LB medium
    8 ml of LB liquid medium (tryptone 10g, yeast extract 5g, NaCl 10g, ion exchanged water 1000ml) was sterilized and cooled, and then inoculated with Pseudomonas sp. Strain SY5 (30-37 ℃) The shaker was incubated for one day in a shaker at 150 rpm.
    2) Liquid culture in C-medium
    C-medium ((NH 4 ) 2 SO 4 5g, K 2 HPO 4 2g, KH 2 PO 4 1g, MgSO 4 7H 2 O 0.2g, CaCl 2 10mg, FeSO 4 7H 2 O 10mg, NaCl 2g , Yeast extract 0.2g, trace element solution 2ml, ion-exchanged water 1000ml, pH 7.0; trace element solution: MoO 3 1.0mg, ZnSO 4 · 7H 2 O 7.0mg, CuSO 4 · 5H 2 O 0.5mg, H 3 1.0 mg of BO 3, 1.0 mg of MnSO 4 · 5H 2 O, 1.0 mg of CoCl 2 · 6H 2 O, 1.0 mg of NiSO 4 · 7H 2 O, 1000 mL of ion-exchanged water), which were dispensed into 8 mL of test tube and sterilized. After cooling, 0.1% of biphenyl or p-chlorobiphenyl was added and inoculated with Pseudomonas sp. Strain SY5, and then cultured in a shaker at 30-37 ° C. and 150 rpm for one day.
    3) Solid culture on LB plate
    LB agar plates (10 g of tryptone, 5 g of yeast extract, 10 g of NaCl, 15 g of agar, 1000 ml of ion-exchanged water) were prepared and sterilized, plated and cooled, and then Pseudomonas sp. Strain SY5 on the surface of the plate. Was plated and incubated in a 30-37 ° C. incubator.
    Example 2: Magnetization Test
    The magnetization ability of Pseudomonas sp. Strain SY5 of the present invention was tested using various aromatic compounds instead of PCB as a carbon source.
    In order to test the magnetizing ability, the Pseudomonas sp. Strain SY5 of the present invention was cultured in a C-media containing 0.1% of the carbon source shown in Table 2, and the magnetizing ability of the strain was examined. In other words, the strain was incubated in a medium containing each carbon source for 3 days, and the absorbance (wavelength 600 nm) was measured, and the growth state of the bacteria was compared with the turbidity of the medium as an index. The results are shown in Table 2 below.
    According to the results of Table 2, all of the aromatic compounds used showed degradability. When the benzene ring cleaved when the PCB was decomposed, chlorobenzene acid was produced, and most of the conventional PCB decomposition bacteria did not decompose these chlorobenzene acids. However, Pseudomonas sp. Strain SY5 of the present invention Pseudomonas sp. Strain SY5 can decompose 3-chlorobenzoic acid and 4-chlorobenzoic acid. Therefore, the Pseudomonas sp SY5 (Pseudomonas sp. Strain SY5) is a useful strain for complete PCB decomposition treatment.
    Example 3: Substrate Specificity Tests for Meta-based Enzymes
    Pseudomonas sp. Strain SY5 ( Pseudomonas sp. Strain SY5) is incubated in 8 ml LB-medium (tryptone 10g, yeast extract 5g, NaCl 10g, ion-exchanged water 1000ml) for 1 day. 1 ml of the culture is placed in an eppendorf tube, centrifuged at 7000 rpm for 5 minutes, the supernatant is discarded, and saline is added and suspended. Centrifuge again at 7000 rpm for 5 minutes, discard the supernatant, suspend in saline solution. 10 μl of the suspension was added to a 1M acetone solution in which catechol, 3-methylcatechol, 4-methylcatechol, and 2,3-dihydroxy biphenyl were dissolved in acetone, followed by mixing for 3 minutes at room temperature. Leave and observe the color change of the solution. If the color of the solution turns yellow, this indicates that the benzene ring of each substrate is meta ring cleavage towards the meta. The results are shown in Table 3 below.
    According to the results of Table 3, the meta-pyrolytic enzyme of this strain showed a strong enzymatic reaction against the remaining three substrates other than 2,3-dihydroxybiphenyl. Therefore, it can be seen that meta-pyrolytic enzymes, which are central enzymes involved in PCB degradation of the strain, have various substrate specificities.
    Example 4: PCB degradation activity rate comparison
    Pseudomonas sp. Strain SY5 ( Pseudomonas sp. Strain SY5) was suspended in phosphate buffer solution at pH 7 and compared to a known strain ( Alcaligenes Eutrophus H850) and PCB degradation activity. While shaking culture. After incubation, 10 µl of an acetone solution containing 20 mM 2,3-dihydroxybiphenyl was added, and the color of the solution turned yellow. As a result, it was found that the PCB degradation strain was alive. The strain was placed in 50 ml of inorganic medium, and 10 ppm of PCB sample (Aroclor 1242) was injected and incubated for 2 days, and then PCB degradation rate was measured.
    Analysis of the degradation activity of Pseudomonas sp. Strain SY5 ( Pseudomonas sp. Strain SY5) among PCB-degrading microorganisms on PCB (Aroclor 1242) is shown by gas chromatograph (GC / ECD). Compared to Alcaligenes Eutrophus H850 (Applied and Environmental Microbiology, Volume 53, 1094 ~ 1102, 1987), the strain shows excellent degradability.
    In addition, the quantitative analysis results of various compounds contained in the PCB (Aroclor 1242) are shown in Tables 4a, 4b, and 4c.

    Therefore, Pseudomonas sp. Strain SY5, a novel strain according to the present invention, is very useful as a microorganism for PCB degradation because of its excellent PCB resolution.
    权利要求:
    Claims (2)
    [1" claim-type="Currently amended] Pseudomonas sp SY5 (Pseudomonas sp. Strain SY5) .
    [2" claim-type="Currently amended] A method for treating sewage containing polychlorinated biphenyl (PCB) using Pseudomonas sp. Strain SY5.
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    同族专利:
    公开号 | 公开日
    KR100187883B1|1999-06-01|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    法律状态:
    1996-12-07|Application filed by 강대승, 주식회사 동성화학
    1996-12-07|Priority to KR1019960062862A
    1998-09-05|Publication of KR19980044738A
    1999-06-01|Application granted
    1999-06-01|Publication of KR100187883B1
    优先权:
    申请号 | 申请日 | 专利标题
    KR1019960062862A|KR100187883B1|1996-12-07|1996-12-07|A pseudomonas sp. strain sy5|
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