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

Carboxylesterase and application thereof in degradation of pesticides malathion and carbaryl

A technology of carboxylesterase and malathion, applied in the field of bioengineering, can solve the problems of limited enzyme production ability of wild strains, high production cost, long fermentation cycle, etc., achieve efficient degradation of malathion, easy to obtain pure Effect of improving protein and degradation efficiency

Inactive Publication Date: 2013-03-13
YUNNAN NORMAL UNIV
View PDF0 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The use of microorganisms to degrade pesticide residues mostly relies on the enzymatic hydrolysis of intracellular enzymes. However, due to the limited enzyme production capacity of wild strains, long fermentation cycle, complicated purification, difficulty in mass production, and high production costs, its popularization and application are limited.

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
  • Carboxylesterase and application thereof in degradation of pesticides malathion and carbaryl
  • Carboxylesterase and application thereof in degradation of pesticides malathion and carbaryl
  • Carboxylesterase and application thereof in degradation of pesticides malathion and carbaryl

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Carboxylesterase Gene D-1CarE3 clone

[0047] Extract the genomic DNA of thermophilic Alicyclobacillus D-1: CTAB lysis method, the specific steps are: centrifuge the fresh bacterial liquid cultured in liquid for 12 hours, and add 800 μL solution Ⅰ (20 mM Tris, pH8.0, 2 mM Na 2 -EDTA, final concentration 20 mg / mL lysozyme), incubated at 37°C for 30 min, added 100 μL 10% SDS and mixed up and down, then added 10 μL 10 mg / mL proteinase K, incubated at 37°C for 30 min, added 150 μL μL 5 M NaCl and 150 μL 10% CTAB solution were mixed upside down, incubated at 65°C for 20 min, divided into 600 μL tubes, and then added 600 μL phenol / chloroform / isopropanol (volume ratio 25:24:1) For extraction, centrifuge at 12000 rpm for 10 min at 4 °C, take 300 μL of the supernatant and extract again in 300 μL chloroform / isopropanol (volume ratio 24:1) to remove impurities, and centrifuge at 12000 rpm for 10 min at 4 °C , then take the supernatant and add 2 times the volume of a...

Embodiment 2

[0054] Embodiment 2: Preparation of recombinant carboxylesterase D-1CarE3

[0055] pET -D-1CarE3 of Escherichia coli BL21(DE3) strain and pET-only - Empty plasmid for 28a(+) Escherichia coli BL21(DE3) strain was inoculated in LB (50 μg / mL Kan) medium with 0.1% inoculum, and shaken rapidly at 37 °C for 16 h. Then inoculate the activated bacterial solution into fresh LB (50 μg / mL Kan) culture solution with 1% inoculum, and culture with rapid shaking for about 2–3 h (OD 600 After reaching 0.6–1.0), add IPTG at a final concentration of 0.7 mM for induction, and continue shaking culture at 20 °C for about 20 h or at 26 °C for about 8 h. Centrifuge at 12000 rpm for 5 min to collect the cells. After suspending the cells with an appropriate amount of pH7.0 Tris-HCl buffer solution, the cells were disrupted by ultrasonic waves in a low-temperature water bath. The above intracellular concentrated crude enzyme solution was centrifuged at 13,000 rpm for 10 min, the supernat...

Embodiment 3

[0070] Application of Purified Recombinant Carboxylesterase D-1CarE3 in Degradation of Pesticides Malathion and Carbaryl

[0071] 1. The degradation effect of recombinant carboxylesterase D-1CarE3 on the pesticide malathion:

[0072] Take 1 mL of purified recombinant carboxylesterase D-1CarE3 diluted 100 times and add it to 1 / 15 mol / L, pH7.0 disodium hydrogen phosphate-dihydrogen phosphate containing 5 mg / L pesticide malathion In the potassium buffer solution, the total reaction system was 3 mL, and the reaction was shaken at 37 °C; in the control group, the enzyme solution was not added, and the enzyme solution was replaced by 1 mL of disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution with pH 7.0, every 20 min Take out 0.5 mL of the reaction solution and mix it with an equal volume of n-hexane, shake it, centrifuge at 12000 rpm for 2 min, add 0.2 g of anhydrous sodium sulfate to the upper layer of the extract to remove water, centrifuge at 12000 rpm fo...

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

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention relates to the application of carboxylesterase D-1CarE3, gene D-1CarE3 and carboxylesterase D-1CarE3 in the degradation of pesticides malathion and carbaryl. The amino acid sequence of carboxylesterase D-1CarE3 is as shown in SEQ ID NO.1, the amino acid sequence of the encoding gene D-1CarE3 of the carboxylesterase is as shown in SEQ ID NO.2, and the carboxylesterase comprises recombinant vector of carboxylesterase D-1CarE3 and the recombination strain of the carboxylesterase D-1CarE3. The invention further provides the application of the carboxylesterase D-1CarE3 in the degradation of pesticides malathion and carbaryl. The carboxylesterase has the advantages that the cultivation period is short, the pure protein is easy to obtain, and the malathion can be degraded at high efficiency, and compared with the reported carboxylesterase from insects, the carboxylesterase is higher in efficiency for depredating the pesticide malathion and carbaryl. The invention has an important significance in the aspect of the treatment of pesticide pollution.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a carboxylesterase D-1CarE3 and its application in the degradation of pesticides malathion and carbaryl. Background technique [0002] Carboxylesterases (Carboxylesterases, E C3.1.1.1) are a class of non-specific esterases that can catalyze the hydrolysis of carboxylic acid esters to generate carboxylic acids and alcohols. They belong to the α / β hydrolase family of lipases. Their amino acid sequence Contains Ser-Asp-His triplet structure, Ser, Asp, His constitute the catalytic active center of carboxylate lipase (Nardini, M, et al., Current Opinion in Structural Biology . 1999, 9(6):732-737.). [0003] Carboxylate lipase widely exists in animals, plants and microorganisms (Melloney J., et al., J of Molecular Catalysis B : Enzymatic . 2005, 32:261-270.), wherein microbial-derived carboxylesterase is an important industrial enzyme, which has important application va...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/18C12N15/55C12N15/63C12N1/15C12N1/19C12N1/21A62D3/02A62D101/04A62D101/28A62D101/26
Inventor 黄遵锡谢振荣丁俊美周峻沛李俊俊唐湘华杨云娟许波
Owner YUNNAN NORMAL UNIV
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