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Active nano-particles for degrading organophosphorus, as well as preparation method and applications of active nano-particles

A technology for degrading activity and nanoparticles, applied in biochemical equipment and methods, chemical instruments and methods, and methods based on microorganisms, etc., can solve the problems of cumbersome operation and high cost, and achieve high recycling rate, low production cost, Good effect of repeated use

Active Publication Date: 2018-09-28
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The immobilization of conventional enzymes requires the preparation of free enzyme proteins and immobilized carriers separately, which is cumbersome and relatively expensive

Method used

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  • Active nano-particles for degrading organophosphorus, as well as preparation method and applications of active nano-particles
  • Active nano-particles for degrading organophosphorus, as well as preparation method and applications of active nano-particles
  • Active nano-particles for degrading organophosphorus, as well as preparation method and applications of active nano-particles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1.1 Extraction of Cupriavidus necator genomic DNA

[0039] (1) Cultivate 5 mL of actinomycete culture, take 1 mL of culture and centrifuge at 12000 rpm for 2 min;

[0040] (2) Add 567 μL TE buffer to the precipitate, and resuspend it by pipetting repeatedly. Add 30μL 20mg / mL lysozyme, and bathe in water at 37℃ for 1h;

[0041] (3) Add 300 μL of 10% SDS and 30 μL of 20 mg / mL proteinase K, mix well, and bathe in water at 37°C for 1 hour;

[0042] (4) Add 100 μL 5M NaCl, mix thoroughly, then add 80 μL CTAB / NaCl solution, mix well, and bathe in 65°C water for 10 minutes;

[0043] (5) Add equal volumes of phenol / chloroform respectively, and extract once with chloroform solution;

[0044] (6) Add 0.6 times the volume of isopropanol, mix gently, centrifuge at 12,000 rpm for 5 min, discard the supernatant, wash twice with 70% ethanol, and dry in an ultra-clean typhoon;

[0045] (7) Add 50 μL TE buffer and dissolve the DNA overnight at 4°C.

[0046] 1.2 Amplification of the...

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Abstract

The invention discloses active nano-particles for degrading organophosphorus, as well as a preparation method and applications of the active nano-particles. According to the technical scheme, by constructing engineering bacteria expressing beta-keto-thiolase, acetoacetyl CoA reductase, fusion protein of organophosphorus acid anhydride hydrolase and polyhydroxyalkanoate synthetase, and fusion protein of organophosphorus acid anhydride hydrolase and polyhydroxyalkanoate binding protein, the obtained engineered organophosphorus acid anhydride hydrolase is fixed to the surfaces of the polyhydroxyalkanoate nano-particles in a combined manner through two methods including covalence and non-covalence, the nano-particles with the activity of degrading the organophosphorus is efficiently produced at one step through a fermentation method, and the high stability and recycling effect are achieved. Compared with the traditional enzyme immobilization method, the method provided by the invention hasthe advantages that the zymoprotein and the carrier for immobilization do not need to be respectively prepared, the production of the immobilization particles can be completed through one step, and the production cost is low. The active nano-particles have wide applications in the aspects of the purification for pesticide residue of fruits and vegetables and the treatment for the environmental pollutants.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an organophosphorus degrading active nanoparticle, its production engineering strain, its fermentation preparation method and its application. Background technique [0002] Human production activities discharge a large amount of organic pollutants, causing environmental pollution and threatening ecological security and human health. Organophosphorus compounds are a class of esters or thiol derivatives of phosphonic acid or phosphoramidate, which exert their toxicity by covalently binding to serine, the active center of acetylcholinesterase. Since it was first synthesized in 1937, it has been widely used in human warfare and production activities. G-type and V-type organophosphorus nerve agents were important chemical weapons during World War II. In modern industrial and agricultural fields, organophosphates are widely used as pesticides, flame retardants, petroleum addit...

Claims

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Application Information

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IPC IPC(8): C12N1/21A23L5/20C12R1/19
CPCA23L5/27C07K14/00C07K2319/00C12N9/001C12N9/14
Inventor 龙丽娟杨键李茹
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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