High-efficiency strain for degrading polycyclic aromatic hydrocarbon and construction method and application thereof

A polycyclic aromatic hydrocarbon and construction method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of ineffective degradation, etc., and achieve the effect of simple operation, simple equipment requirements and high adaptability

Active Publication Date: 2011-01-19
广州市佳境水处理技术工程有限公司
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Problems solved by technology

[0004] Microbial degradation of PAHs has become an important means of remediation of polluted environments at home and abroad. It has the advantages of various treatment forms, low cost, and small impact on the environment. The research is mainly to screen efficient degrading strains or build efficient bacterial groups. Efficient degrading strains are mainly based on low molecular weight PAHs (LM

Method used

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  • High-efficiency strain for degrading polycyclic aromatic hydrocarbon and construction method and application thereof
  • High-efficiency strain for degrading polycyclic aromatic hydrocarbon and construction method and application thereof
  • High-efficiency strain for degrading polycyclic aromatic hydrocarbon and construction method and application thereof

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Embodiment 1

[0034] In the invention, the protoplasts of Sphingomonas GY2B and Pseudomonas GP3A are firstly prepared and then fused. The following is the specific construction method:

[0035] A. Hypertonic buffer SMM: Add 0.02mol / l maleic acid and 0.02mol / l MgCl to a solution of 0.5mol / l sucrose 2 ·6H 2 O, adjust the pH to 6.5, and sterilize at 121°C for 20 minutes.

[0036] B. Induction solution: Prepared with hypertonic buffer SMM containing polyethylene glycol, CaCl 2 and dimethyl sulfoxide, polyethylene glycol 40% w / v, CaCl 2 10mmol / L, dimethyl sulfoxide 15% v / v, sterilized by 0.45μm membrane filtration, stored at room temperature.

[0037] C. Lysozyme: purchased from BBI company, enzyme activity > 20,000U / mg, weighed 20mg of lysozyme, dissolved in SMM hypertonic fluid, the final concentration was 5mg / ml, sterilized by 0.45μm membrane filtration, at -20 Store at ℃ for later use

[0038] D. Double-antibody interlayer regeneration medium is composed of upper and lower layers of me...

Embodiment 2

[0048] Application of Gram-negative bacteria GY2B and GP3A fusion construction engineering bacteria technology

[0049] As shown in Table 1, F14 can grow on the medium containing 80 μg / ml ceftazidime and 80 μg / ml piperacillin sodium at the same time, but each of the two parental strains cannot. F14 has wider adaptability to environmental conditions than the parent strain. F14 was inoculated in shake flasks with an initial concentration of 100mg / L, and the degradation rate at different temperatures was detected at a speed of 150r / min. The degradation rate of F14 can reach 100.0% at 30°C, 82.9% at 20°C, and 99.7% at 36°C. However, the degradation rate of GY2B is about 50% at 20°C and 35°C. The F14 was inoculated in shake flasks with different initial concentrations, the rotation speed was 150r / min, the temperature was 30°C, and the degradation rate was detected. GY2B degrades phenanthrene with an initial concentration of 230mg / L, and the degradation rate is about 70% at 48h, a...

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Abstract

The invention discloses a high-efficiency strain for degrading polycyclic aromatic hydrocarbon and a construction method and application thereof. Microbial parents provided by the invention are two gram negative bacteria, namely GY2B (sphingomonas) and GP3A (Pesudomonas); protoplasts of the two strains are fused; and genetic engineering special bacteria are constructed by recombining genes of the parents in the same cell. The high-efficiency strain combines the advantages of high degradability and high adaptability of two parent strains and is favorable for processing the polycyclic aromatic hydrocarbon PAHs.

Description

technical field [0001] The present invention relates to a special-effect bacterial strain and its construction method, especially a biological construction and application technology for efficient degradation of polycyclic aromatic hydrocarbons (PAHs) new microbial strains, more specifically two Gram strains widely used in the environment A special strain and its construction method for the fusion of protoplasts of Bacteria-negative bacteria. Background technique [0002] Polycyclic aromatic hydrocarbons (PAHs) refer to a class of unique persistent organic pollutants composed of two or more benzene rings in a linear, angular or clustered form. PAHs contain π The conjugated system formed by bonds, due to the strong mobility of electrons, makes the entire molecular system relatively stable. It is precisely because of this relatively strong stability that substances such as polycyclic aromatic hydrocarbons can widely exist in the environment. Some of these PAHs can produce va...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/03A62D3/02C12R1/38A62D101/20
Inventor 党志卢静郭楚玲杨琛易筱筠卢桂宁
Owner 广州市佳境水处理技术工程有限公司
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