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PEG-ACS/si-luxR compound and application thereof, and method for reducing content of biogenic amines in large yellow croakers during storage

A technology of PEG-ACS and compound, which is applied in food ingredients as antimicrobial preservation, chemical preservation of meat/fish, food science, etc. It can solve the problems of biogenic amine safety and reduce the quality of large yellow croaker

Active Publication Date: 2018-12-04
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, studies have found that during the storage of large yellow croaker, the main spoilage bacteria, Shewanella baltica, can degrade nitrogenous substances such as proteins and amino acids in large yellow croaker into biogenic amines, thereby reducing the quality of large yellow croakers and causing safety problems of biogenic amines

Method used

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  • PEG-ACS/si-luxR compound and application thereof, and method for reducing content of biogenic amines in large yellow croakers during storage
  • PEG-ACS/si-luxR compound and application thereof, and method for reducing content of biogenic amines in large yellow croakers during storage
  • PEG-ACS/si-luxR compound and application thereof, and method for reducing content of biogenic amines in large yellow croakers during storage

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Experimental program
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Effect test

Embodiment 1

[0040] Preparation of PEG-ACS / siRNA complex:

[0041] (1) The si-luxR sequence was designed according to the related sequence of the quorum-sensing receptor protein synthesis regulatory gene of the main amine-producing strain Shewanella baltica in storage of large yellow croaker (see Appendix). The si-luxR sequence is:

[0042] Sense strand: 5'-GUUCCGAAUUGUUGCGAUACC-3';

[0043] Antisense strand: 5'-UAUCGCAACAAUUCGAGACUU-3'.

[0044] (2) Preparation of si-luxR: T7 promoter template DNA is 5'-TAATACGACTCACTATA GGAGACAGG-3', take 3 μL siRNA molecule sense strand (100umol / L) and 3 μL T7 promoter primer template (100umol / L) and mix, 95 Pre-denaturation at ℃ for 4 min, ice bath for 20 min, then add 3 μL 10x Klenow buffer, 8 μL deoxynucleoside triphosphate (dNTP) (10 μmol / L), 4 μL deionized water, 3 μL Klenow enzyme (2U / μL), 30-40 °C Water bath for 20-40 minutes. Take 6 μL of reaction solution, add 4 μL of buffer, 7 μL of nucleoside triphosphate (rNTP) mixture (25 mmol / L), 4 μL ...

Embodiment 2

[0057] PEG-ACS / siRNA complex

[0058] Wherein the preparation of siRNA is the same as in Example 1.

[0059] (1) Preparation of ACS: Take 1g chitosan, dissolve it in the TEMED / HCl buffer solution of pH 4.5, then add coupling agent EDC 1.13g, NHS 0.685g in this solution, after stirring evenly, then add and Arginine with half the molar amount of the chitosan amino group was reacted at room temperature for 7 hours under magnetic stirring, and then desalted water dialysis and freeze-drying were performed to obtain arginine-modified chitosan.

[0060] (2) 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.

[0061] (3) Preparation of siRNA solution: 10D siRNA was dissolved in DEPC water to prepare an siRNA solution with a concentration of 17 μM, and stored at -...

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Abstract

The invention discloses a PEG-ACS / si-luxR compound and a method and application of the thereof to reduction of the content of biogenic amines of large yellow croakers during storage. The method mainlycomprises the following steps of (1) designing and preparing small disturbance ribonucleic acid(siRNA)si-luxR according to Baltic shewanella (Shewanella baltica OS678) group induction receptor protein regulatory gene luxR sequence; (2) making PEG-ACS / si-luxR particles by using PEG arginine modified chitosan as a carrier; and (3) making the obtained compound into a solution with a certain concentration, soaking the large yellow croakers in the solution for 3-5min, performing draining, and performing cooling so as to obtain finished products. The PEG-ACS / si-luxR compound and the method and application disclosed by the invention have the advantages that the method is simple, the content of the biogenic amines in the large yellow croakers during storage can be effectively and significantly reduced by 65%-80%, and due sense organ characteristics including textures, smell and the like of the large yellow croakers can be effectively maintained.

Description

technical field [0001] The invention belongs to the field of food quality and safety, and specifically relates to the preparation of a PEG-ACS / si-luxR complex and its application in the storage of large yellow croaker, aiming at reducing the content of biogenic amines in the storage. Background technique [0002] Biogenic amine is a low molecular weight nitrogen-containing organic base, including aliphatic, aromatic and heterocyclic structures, and belongs to polar or semi-polar compounds. Biogenic amines are mainly produced by the action of amino acid decarboxylase produced by bacteria on amino acids or the transamination of aldehydes and ketones by bacteria. When the human body ingests excess biogenic amines, it can cause headaches, nausea, hypotension, high blood pressure, migraines, skin allergies, and digestive problems such as food poisoning. Moreover, biogenic amines are precursors of nitrosamines, which have a certain relationship with carcinogenic and mutagenic act...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23B4/20
CPCA23B4/20A23V2002/00A23V2200/10
Inventor 傅玲琳王彦波吴哲铭王飞飞黎凡王翀周瑾茹
Owner ZHEJIANG GONGSHANG UNIVERSITY
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