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Preparation method of pulullan-animal esterase composite nanofiber

A technology of composite nanofibers and pullulan, applied in fiber treatment, hydrolytic enzymes, immobilized on/in organic carriers, etc., can solve the problems of difficult re-collection, decreased catalytic activity of enzymes, and insensitivity to pesticide responses, etc. problems, to achieve the effect of simple equipment and preparation process, good effect, and sensitive response to pesticides

Pending Publication Date: 2021-09-07
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the use of animal esterase is difficult to re-collect, and it is easily affected by environmental factors (light, heat, oxygen, pH, etc.) or chemical reactions, resulting in a decrease in the catalytic activity of the enzyme, which limits its scope of application in practical applications.
At present, the method for immobilizing animal esterase to detect pesticide residues mainly contains, using 96 microwell plates of polyvinyl chloride and polystyrene material as carrier physical adsorption method to immobilize chicken liver esterase (Tu etc., Anhui Agricultural Science, 39 (24 ): 14534-14536) and the micro-electromechanical processing method based on NCM9924 microcalorimetry chip (Yu Jinsong et al., Physical and Chemical Inspection (Chemical Volume). 2014, 50(11): 1338-1341), but there are still poor stability and poor response to pesticides. sensitive issues

Method used

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  • Preparation method of pulullan-animal esterase composite nanofiber
  • Preparation method of pulullan-animal esterase composite nanofiber
  • Preparation method of pulullan-animal esterase composite nanofiber

Examples

Experimental program
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Effect test

Embodiment 1

[0040] The preparation method of pullulan polysaccharide-animal esterase composite nanofiber comprises the steps:

[0041] (1) Extraction and purification of chicken liver esterase: crush the chicken liver and mix it with tromethamine-acetic acid buffer solution at a ratio of 30:100, add phenylmethylsulfonyl fluoride and 2,6-di-tert-butyl P-cresol was maintained at a concentration of 115 μmol / L and 45 μmol / L respectively. After the homogenate was centrifuged, 30 mL of the supernatant was taken, and ammonium sulfate was added to reach a saturation of 70%. The ratio of 1:10 was dialyzed in 0.1mol / L phosphate-buffered saline solution for 6 hours, and the supernatant was obtained after static centrifugation, which was the purified chicken liver esterase solution.

[0042] (2) Preparation of pullulan-chicken liver esterase electrospinning solution: according to the mass concentration of 0.15g / mL, the pullulan with a molecular weight of 150g / mol was dispersed in a phosphate buffered...

Embodiment 2

[0046] The preparation method of pullulan polysaccharide-porcine liver esterase composite nanofiber comprises the following steps:

[0047] (1) Extraction and purification of pig liver esterase: crush the pig liver and mix it with tromethamine-acetic acid buffer solution at a ratio of 20:200, add phenylmethylsulfonyl fluoride and 2,6-di-tert-butyl Keep the concentration of p-cresol at 100 μmol / L and 30 μmol / L respectively. After the homogenate is centrifuged, take 50 mL of the supernatant, add ammonium sulfate to 70% saturation, let it stand and centrifuge, take the precipitate and freeze-dry it, grind it into powder and use it as The ratio of 0.5:25 was dialyzed in 0.1mol / L phosphate buffered saline solution for 12 hours, and the supernatant was obtained after static centrifugation, which was the purified pig liver esterase solution.

[0048] (2) Preparation of pullulan-porcine liver esterase electrospinning solution: according to the mass concentration of 0.25g / mL, pullulan ...

Embodiment 3

[0052] The preparation method of pullulan polysaccharide-foie gras esterase composite nanofiber comprises the steps:

[0053] (1) Extraction and purification of foie gras esterase: crush the foie liver and mix it with tromethamine-acetic acid buffer solution at a ratio of 30:150, add phenylmethylsulfonyl fluoride and 2,6-di-tert-butyl P-cresol was maintained at a concentration of 130 μmol / L and 60 μmol / L respectively. After the homogenate was centrifuged, 40 mL of the supernatant was taken, and ammonium sulfate was added to reach a saturation of 70%. The ratio of 1.5:20 was dialyzed in 0.1mol / L phosphate-buffered saline solution for 8 hours, and the supernatant was obtained after static centrifugation, which was the purified foie gras esterase solution.

[0054] (2) Preparation of pullulan-foie gras esterase electrospinning solution: according to the mass concentration of 0.20g / mL, the pullulan with a molecular weight of 100g / mol was dispersed in a phosphate buffered saline so...

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Abstract

The invention discloses a preparation method of pulullan-animal esterase composite nanofiber. The method comprises the following steps of: extracting and purifying animal liver to prepare an animal esterase solution, then uniformly mixing the animal esterase solution with a pulullan solution to obtain a pulullan-animal esterase electrostatic spinning solution, and finally performing electrostatic spinning to prepare the pulullan-animal esterase composite nanofiber. The average diameter of the pulullan-animal esterase composite nanofiber is 70.39-108.79nm, animal esterase is uniformly distributed on the surface of the nanofiber, and the lower limit of detection on pesticide carbaryl is 0.81-1.10mg / L. The preparation method disclosed by the invention realizes immobilization of a nano material of the animal esterase, and has the advantages of simple process, low cost, environmental friendliness and the like.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzymes, and relates to a preparation method of pullulan-animal esterase composite nanofibers. Background technique [0002] The large-scale use of pesticides has a high probability of entering the ecological environment, which not only pollutes the soil and air, but also endangers fisheries and aquaculture, seriously affecting the sustainable development of agriculture. Pesticide residues in agricultural products can also cause multiple organ damage to humans and animals, and induce various diseases, such as cancer and endocrine system disorders. Therefore, pesticide residue detection has always been one of the research hotspots in agricultural product safety. [0003] According to the provisions of the national standard on the rapid detection methods of pesticide residues, the rapid detection card method and enzyme inhibition method have become the main methods for on-site rapid detection o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/10C12N9/16D01D5/00
CPCC12N11/10C12N9/16D01D5/0015
Inventor 王君袁亚明李松南
Owner YANGZHOU UNIV
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