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Preparing methods and applications of microbacterium paraoxidans and broad-spectrum polychlorinated biphenylase preparation thereof

The technology of a Microbacillus oxidans and a culture method is applied in the field of preparation of the Microbacillus paraoxidans and its broad-spectrum polychlorinated biphenylase preparation, which can solve the problems such as the inability of polychlorinated biphenyls to meet the requirements, achieve environmental protection, large-scale operation, and convenience. The effect of large-scale operation and simple preparation process

Active Publication Date: 2019-05-17
ECOLOGY INST SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the treatment speed of the above-mentioned strains and the types of PCBs still cannot meet the needs of actual treatment. The rapid repair of PCBs compound pollution under aerobic conditions has become a current research hotspot.

Method used

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  • Preparing methods and applications of microbacterium paraoxidans and broad-spectrum polychlorinated biphenylase preparation thereof
  • Preparing methods and applications of microbacterium paraoxidans and broad-spectrum polychlorinated biphenylase preparation thereof
  • Preparing methods and applications of microbacterium paraoxidans and broad-spectrum polychlorinated biphenylase preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Take the leachate from POPs-contaminated soil and dilute it to a concentration of 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 5 concentration gradients. Spread the diluted bacterial suspension on the solid medium containing biphenyl, and culture at 30°C for 1-3 days. Pick out fast-growing bacterial colonies with typical shapes, and then separate and purify them by streaking three times on plates, then pick out single colonies in inorganic salt liquid medium, culture them at 30°C and 150 rpm for 3 days, and take 1.5 mL of the culture Add 0.5mL glycerin, mix well and store in -80°C refrigerator for a long time.

[0050] The solid medium used to coat the bacterial suspension is LB solid medium, and the components are as follows:

[0051] Peptone 10g, yeast extract 5g, sodium chloride 10g, agar 20g, water to 1L, pH natural.

[0052] The components of the inorganic salt liquid medium for single colony cultivation are as follows:

[0053] Potassium dihydrogen phosphate 0.5g, di...

Embodiment 2

[0080] Utilize the method for preparing broad-spectrum polychlorinated biphenyl degrading enzyme preparation by the microbacterium paraoxydans (Microbacterium paraoxydans) ECO-2 described in embodiment 1, the steps are as follows:

[0081] (1) Take Microbacterium paraoxydans (Microbacterium paraoxydans) ECO-2 and streak it on LB solid medium, and activate it upside down at 35°C for 2 days to obtain the activated strain;

[0082] (2) Inoculate the activated bacterial strain obtained in step (1) into LB liquid medium, and culture it on a shaking table for 2 days at 35°C with a rotation speed of 200 rpm to obtain a seed solution;

[0083] (3) Get the seed liquid that step (2) makes, transfer in the inorganic salt culture medium that contains biphenyl 0.5g / L by volume percentage 10%, under the condition of 35 ℃, dissolved oxygen 70%, expand culture 2 days, the bacteria solution was prepared;

[0084] (4) Take the bacterium solution prepared in step (3), centrifuge it for 10 minut...

experiment example

[0090] After pentachlorobiphenyl PCB114, PCB101, and trichlorobiphenyl PCB28 with a concentration of 25mg / L were mixed with broad-spectrum polychlorinated biphenyl enzyme preparation and PBS buffer solution in a ratio of 1:5:19 (volume ratio), at 30 ℃ and pH 7.0 for 10h, 13h, and 10h respectively, adding 10mL of n-hexane for extraction three times, taking the extract, and detecting the degradation rate of the above-mentioned PCBs by GC-MS.

[0091] After testing, it was found that the broad-spectrum polychlorinated biphenyl degrading enzyme preparation obtained in Example 2, under aerobic conditions, the degradation rate of p-trichlorobiphenyl PCB28 can reach 90% within 10 hours, and the degradation rate of p-dioxin-like pentachlorobiphenyl The degradation rate of PCB114 can reach 57.1% within 10 hours, and the degradation rate of PCB101 can reach 100% within 13 hours.

[0092] In Comparative Example 2, only the degradation rate of the dioxin-like pentachlorobiphenyl PCB114 ca...

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Abstract

The invention relates to preparing methods and applications of microbacterium paraoxidans and a broad-spectrum polychlorinated biphenylase preparation thereof. The microbacterium paraoxydans ECO-2 ispreserved in China General Microbiological Culture Collection Center on June 01, 2018 and has the bacterial species preservation number of CGMCC No.15836. An intracellular complex enzyme obtained by induction culture of the microbacterium paraoxidans ECO-2 with a single substrate of biphenyl can degrade PCB28, PCB101 and / or PCB114 and other low-chloro and high-chloro PCBs, and the enzyme preparation has wide degradation spectrum, has significant difference from a fact that a product obtained from conventional known single induction culture can only degrade one component, and has wide application prospects.

Description

technical field [0001] The invention relates to a preparation method and application of a strain of Microbacterium paraoxidans and a broad-spectrum polychlorinated diphenylase preparation, belonging to the technical field of microorganisms. Background technique [0002] Persistent organic pollutants polychlorinated biphenyls (PCBs) are highly toxic, bioaccumulative, long-distance migration, and have a long half-life, seriously polluting soil, aquatic ecosystems, and drinking water sources, causing teratogenic and carcinogenic effects. Both the Taiwan oil disease incident and the Japanese rice bran oil incident caused by PCBs pollution caused heavy loss of life and property. Due to thermal and chemical stability, flame retardancy, insulation and oxidation resistance, PCBs are widely used in chemical industry, electric power, electronics and machinery industries. At present, there are a large amount of PCBs and their pollutants in my country, and there are a lot of electronic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/00B09C1/10C12R1/01
CPCB09C1/10C12N1/20C12N9/00C12N1/04A62D3/02C12R2001/01C12N1/205Y02A50/20
Inventor 季蕾张强傅晓文王加宁陈贯虹宋繁永李天元郭书海
Owner ECOLOGY INST SHANDONG ACAD OF SCI
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