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peg-acs/si-luxr complex and its application and method for reducing biogenic amine content in large yellow croaker in storage

A compound, the technology of large yellow croaker, which is applied in the fields of botanical equipment and methods, biochemical equipment and methods, food ingredients as antimicrobial preservation, etc., can solve the problems of reducing the quality of large yellow croaker, biogenic amine safety, etc. Compatibility, quality improvement effect

Active Publication Date: 2022-04-08
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 complex and its application and method for reducing biogenic amine content in large yellow croaker in storage
  • peg-acs/si-luxr complex and its application and method for reducing biogenic amine content in large yellow croaker in storage
  • peg-acs/si-luxr complex and its application and method for reducing biogenic amine content in large yellow croaker in storage

Examples

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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'-TAATACGACTCACTATAGGAGACAGG-3', take 3 μL siRNA sense strand (100umol / L) and 3μL T7 promoter primer template (100umol / L) and mix, 95℃ Pre-denaturation for 4 minutes, ice bath for 20 minutes, 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 20-40min. Take 6 μL of reaction solution, add 4 μL of buffer, 7 μL of nucleoside triphosphate (rNTP) mixture (25 mmol / L), 4 μL of deionized ...

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 its method and application for reducing the content of biogenic amines in large yellow croaker storage. It mainly includes the following steps: (1) designing and preparing the small interfering ribonucleic acid (siRNA) si-luxR according to the quorum sensing receptor protein regulatory gene luxR sequence of Shewanella baltica OS678; (2) after PEGylation Arginine-modified chitosan was used as a carrier to make PEG‑ACS / si‑luxR particles. (3) The obtained compound is made into a solution with a certain concentration, soaked in large yellow croaker for 3min-5min, drained and cooled to make a finished product. The invention has the advantages of simple method, can effectively and significantly reduce the content of biogenic amine in the storage of large yellow croaker by 65%-80%, and can effectively maintain the proper texture, smell and other sensory characteristics of the large yellow croaker.

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 Patents(China)
IPC IPC(8): C12N15/00A23B4/20
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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