Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Detoxication engineering bacteria, and preparation method and application thereof

A technology of engineering bacteria and detoxifying esters, applied in the directions of botanical equipment and methods, biochemical equipment and methods, and applications, can solve the problems of foreign trade export losses of agricultural products, excessive pesticide residues, and restricted export scale, and achieves solutions to pollution problems, The effect of broadening the degradation spectrum and eliminating losses

Inactive Publication Date: 2011-01-05
INST OF ZOOLOGY CHINESE ACAD OF SCI
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the lack of standardized management of the use of pesticides in my country, pesticide residues in agricultural products generally exceed the standard, resulting in heavy losses in the export of agricultural products in my country. Traditional export products, such as fruits, tea and Chinese herbal medicines, have repeatedly returned in large quantities, and the export scale has been severely restricted.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Detoxication engineering bacteria, and preparation method and application thereof
  • Detoxication engineering bacteria, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1. Construction of engineering bacteria

[0040] 1. Construction of recombinant plasmid (pETDuet-E4-mpd)

[0041] 1. Preparation of hydrolase gene

[0042] The DNA shown in sequence 4 is artificially synthesized, and nucleotides 11 to 901 from the 5'end of sequence 4 are the hydrolase (mpd) gene, the upstream of which is the BamHI site, and the downstream is the HindIII site.

[0043] 2. Preparation of detoxification esterase gene

[0044] The DNA shown in sequence 2 is artificially synthesized, and nucleotides 8 to 1666 from the 5'end of sequence 2 are the detoxification esterase (E4) gene, the upstream of which is the BglII site, and the downstream is the XhoI site.

[0045] 3. Construction of recombinant plasmid (pETDuet-E4-mpd)

[0046] ① Use restriction enzymes BamHI and HindIII to cut the hydrolase gene of step 1.

[0047] ②The plasmid pETDuet-1 was digested with restriction enzymes BamHI and HindIII.

[0048] ③ Connect the digested product of step ① and step ② to obtai...

Embodiment 2

[0082] Example 2. Degradation of carbaryl and malathion by crude enzyme solution of Pet30-mpd strain

[0083] 1. Degradation of carbaryl by the crude enzyme solution of Pet30-mpd

[0084] The degradation of carbaryl was determined according to the method of Hayatsu and Nagata (1993), as follows:

[0085] Buffer: 10mM KH 2 PO 4 / KOH buffer; pH7.5.

[0086] Color developer (DBLS): Dissolve 4.5 mg of Fast Blue B salt in 4.5 ml of water, and then add 10.5 ml of 5% SDS solution.

[0087] Reaction system (20ml): Add Carbaryl and the crude enzyme solution of Pet30-mpd strain prepared in Example 1 into the buffer. The reaction system contains 130μM Carbaryl and 5ml crude enzyme solution (107U ml -1 ).

[0088] Control system (20ml): Use deionized water instead of crude enzyme solution, and the other reaction systems are the same.

[0089] The reaction temperature was 37°C, and 2 ml was sampled every 30 minutes. 0.5ml of freshly prepared DBLS solution was immediately added to the sample, and the ...

Embodiment 3

[0098] Example 3 Detoxification experiment of chlorpyrifos gallisepticum poisoning

[0099] Adult aircraft hens (Gallus gallus) (12 months old, chicken weighing about 1.5 kg) were purchased from the experimental chicken farm of Beijing Agricultural University. The chickens were re-purchased and fed separately, and the experiment was started after at least one week of acclimatization. Free access to food and water. During the experiment, the temperature of the chicken house was controlled at about 20℃, and the light was about 10 hours per day.

[0100] Divide the experimental chickens into seven groups, 10 in each group:

[0101] Test group: Simultaneous administration of chlorpyrifos (packed in empty capsules, at a dose of 170 mg / kg, orally) and Pet30-mpd (engineered bacteria) (0.5g wet bacteria into empty capsules, orally);

[0102] Experimental group two: first administer chlorpyrifos (filled in empty capsules, the dose is 170mg / kg, orally), 40 minutes later, give Pet30-mpd (engine...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses detoxication engineering bacteria, and a preparation method and application thereof. The invention provides a recombinant plasmid which is obtained by introducing coded genes of detoxication esterase and coded genes of hydrolase into an enzyme cutting site in plasmid pETDuet-1. The engineering bacteria of the invention can be obtained by introducing the recombinant plasmidinto host bacteria. The engineering bacteria comprise the coded genes of the detoxication esterase and the hydrolase and can realize inducible expression of the detoxication esterase and the hydrolase. Crude enzyme solution obtained by fermentation of the engineering bacteria can also be protected. The engineering bacteria and the crude enzyme solution thereof can simultaneously degrade organic acid esters such as malathion, parathion, high-efficiency cypermethrin, acetofenate and the like and organophosphorus pesticides, and can be used for detoxication of organophosphorus poisoned warm-blooded animals. The invention provides a new way for biological pollution treatment of toxic and harmful compounds such as pesticides remained in water sources, soil and the like by using natural resources of eukaryote and prokaryote.

Description

Technical field [0001] The invention relates to a detoxification engineering bacteria and its preparation method and application. Background technique [0002] Modern agricultural production has played a huge role in satisfying people’s desire for high food yield. It was once regarded as a panacea for third world countries to get rid of poverty and hunger. However, in the face of the decline in the quality of agricultural and sideline products and the reduction of biodiversity in modern agriculture, With the increasing rampant crop diseases and insect pests, the waste of agricultural resources, and the degradation of the agricultural environment, in the face of the unabated food crisis in Africa, people are rejoicing, and gradually a kind of confusion and doubt have arisen, and they have begun to re-examine modern agriculture. Pros and cons, re-understand the connotation of agricultural resources and environment, and rethink the direction of agricultural research and decision-mak...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/63C12N15/55C12N9/16C12N9/14C12N1/21A62D3/02A61K38/46A61P39/02C12R1/19A62D101/04
Inventor 乔传令杨超兰文升
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com