peg-acs/luxr-siRNA nanocomplex and its application and method for reducing biogenic amines in Litopenaeus vannamei

A PEG-ACS and nanocomposite technology, which is applied in the field of food quality and safety, can solve the problems of high operation requirements, lower food quality, and ineffective effects, and achieve wide application prospects, lower biogenic amine content, and delay the process of corruption. Effect

Active Publication Date: 2021-03-02
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have disadvantages such as high operation requirements, high cost, inconspicuous effects, and easy reduction of food quality, so it is difficult to apply and promote them in production.

Method used

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  • peg-acs/luxr-siRNA nanocomplex and its application and method for reducing biogenic amines in Litopenaeus vannamei
  • peg-acs/luxr-siRNA nanocomplex and its application and method for reducing biogenic amines in Litopenaeus vannamei
  • peg-acs/luxr-siRNA nanocomplex and its application and method for reducing biogenic amines in Litopenaeus vannamei

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Effect of PEG-ACS / luxR-siRNA nanocomplex in Litopenaeus vannamei

[0056] (1) Preparation of ACS: Weigh 1g of chitosan, dissolve it in TEMED / HCl buffer solution with pH 5.0, then add 1130mg of EDC and 685mg of NHS coupling agent to the solution, stir well, then add Arginine with 80% molar weight of the polysaccharide amino group was reacted at room temperature for 8 hours under magnetic stirring, and then desalted water dialysis and freeze-drying were performed to obtain arginine-modified chitosan.

[0057] (4) Preparation of PEG-ACS: Take 2.5 mL of 10 mg / mL arginine-modified chitosan, add 25 mg of PEG-SPA, and react at room temperature for 4 h. Use a dialysis bag with a molecular weight cut-off of 14,000 to remove unreacted PEG. PEGylated arginine-modified chitosan was obtained.

[0058] (5) Preparation of siRNA solution: 10D siRNA was dissolved in 125 μL DEPC water to prepare siRNA solution with a concentration of 20 μM, and stored at -20° C. for later us...

Embodiment 2

[0071] Example 2: Effect of PEG-ACS / luxR-siRNA nanocomplex in Litopenaeus vannamei

[0072] (1) The preparation of ACS: take by weighing 0.5g chitosan, be dissolved in the TEMED / HCl buffer solution of pH 4.5, then add the EDC of 567mg and 340mg NHS coupling agent in this solution, after stirring evenly, then add and Arginine with 50% molar weight of the chitosan amino group was reacted at room temperature for 6 hours under magnetic stirring, and then desalted water dialysis and freeze-drying were performed to obtain arginine-modified chitosan.

[0073] (4) Preparation of PEG-ACS: Take 2.5 mL of 10 mg / mL arginine-modified chitosan, add 25 mg of PEG-SPA, and react at room temperature for 4 h. Use a dialysis bag with a molecular weight cut-off of 14,000 to remove unreacted PEG. PEGylated arginine-modified chitosan was obtained.

[0074] (5) Preparation of siRNA solution: 10D siRNA was dissolved in DEPC water to prepare an siRNA solution with a concentration of 18 μM, and stored...

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Abstract

The invention discloses a PEG-ACS / luxR-siRNA nanocomposite, a method for reducing the biogenic amine content in Litopenaeus vannamei and an application thereof. It mainly includes the following steps: (1) designing and preparing small interfering ribonucleic acid (siRNA) according to the luxR sequence of the main amine-producing bacterium Shewanella putrefaciens amine-producing gene luxR in Litopenaeus vannamei; (2) using the PEGylated refined Amino acid-modified chitosan was used as a carrier to obtain a stable PEG-ACS / luxR-siRNA nanocomplex; (3) PEG-ACS / luxR-siRNA nanocomplex was added to Litopenaeus vannamei in a certain proportion. Experiments have proved that the method does not affect the sensory characteristics of Litopenaeus vannamei, and has a significant effect of inhibiting the content of biogenic amines, which can reduce the content of biogenic amines in Litopenaeus vannamei by more than 80%, and has important promotion significance for the control of biogenic amines in other aquatic products.

Description

technical field [0001] The invention belongs to the field of food quality and safety, and in particular relates to a preparation method of a PEG-ACS / luxR-siRNA nanocomposite and a strategy for reducing biogenic amines in Litopenaeus vannamei. Background technique [0002] Litopenaeus vannamei, also known as Penaeus vannamei, is native to South America and is one of the aquatic products with the highest aquaculture output in the world. Since it was introduced into my country in 1998, Penaeus vannamei has gradually become the first of the three major economic shrimps in my country. Litopenaeus vannamei tastes delicious and is rich in nutrition. However, its rich protein is converted into polypeptides and amino acids under the action of protease, and these small molecular substances can be easily further converted into biogenic amines, and can reach a fairly high concentration. The study found that Shewanella putrefaciens is the dominant spoilage bacterium in the storage proce...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 傅玲琳王彦波黎凡王飞飞吴哲铭王翀周瑾茹
Owner ZHEJIANG GONGSHANG UNIVERSITY
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