peg-acs/m-siRNA nanocomposite and its application and method for reducing histamine content in fishmeal storage

A PEG-ACS, nanocomposite technology, applied in the field of feed industry, can solve the problems of high cytotoxicity and in vivo toxicity, low efficiency, weak targeting, etc. Effect

Active Publication Date: 2022-02-08
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, both liposome and polyethyleneimine have high cytotoxicity and in vivo toxicity, while chitosan has the disadvantages of low efficiency and poor targeting when used as a carrier.

Method used

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  • peg-acs/m-siRNA nanocomposite and its application and method for reducing histamine content in fishmeal storage
  • peg-acs/m-siRNA nanocomposite and its application and method for reducing histamine content in fishmeal storage
  • peg-acs/m-siRNA nanocomposite and its application and method for reducing histamine content in fishmeal storage

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] (1) Design the M-siRNA sequence.

[0058] Sense strand: 5'-GGAUCGUAUUUCUGUUGAAAU-3',

[0059] Antisense strand: 5'-UUCAACAGAAAUACGAUCCAC-3'.

[0060] (2) Preparation of M-siRNA.

[0061] (3) Preparation of ACS: Weigh 0.5g chitosan, dissolve it in TEMED / HCl buffer solution with pH 5.0, then add 567mg EDC and 340mg 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.

[0062] (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. The obtained PEGylated arginine-modified chitosan was freeze-dried for future use.

[0063] (5) Preparation of siRNA solution: siRNA w...

Embodiment 2

[0075] (1) Design the M-siRNA sequence.

[0076] Sense strand: 5'-GGAUCGUAUUUCUGUUGAAAU-3',

[0077] Antisense strand: 5'-UUCAACAGAAAUACGAUCCAC-3'.

[0078] (2) Preparation of M-siRNA.

[0079] (3) Preparation of ACS: Weigh 0.8g chitosan, dissolve it in TEMED / HCl buffer solution with pH 4.5, then add 900mg EDC and 550mg NHS coupling agent to the solution, stir well, then add Arginine with an equimolar amount of polysaccharide amino groups 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.

[0080] (4) Preparation of PEG-ACS: Take 3 mL of 8 mg / mL arginine-modified chitosan, add 24 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. The obtained PEGylated arginine-modified chitosan was freeze-dried for future use.

[0081] (5) Preparation of siRNA solution: siRNA was ...

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Abstract

The invention discloses a PEG-ACS / M-siRNA nanocomposite and its application and a method for reducing the histamine content in the fishmeal storage process: (1) according to the main histamine-producing bacterium Morganella morganii in the fishmeal subsp.morganii KT) (CP004345) histidine decarboxylase gene MU9_RS17900 (Gene ID: 14672734) sequence design and preparation of small interfering ribonucleic acid (siRNA); (2) chitosan modified with arginine after PEGylation as a carrier to obtain a stable PEG-ACS / M-siRNA nanocomplex; (3) adding the PEG-ACS / M-siRNA nanocomplex to fish meal in a certain proportion. The method has a significant inhibitory effect on the histamine content in the fishmeal storage process, can reduce the histamine content in the fishmeal by 49% to 53%, and has great significance for the control of biogenic amines in the fishmeal in the feed industry.

Description

technical field [0001] The invention belongs to the field of feed industry, and specifically relates to a preparation method of a PEG-ACS / M-siRNA nanocomposite and an application method in the storage of fish meal for feeding, aiming at reducing the content of the harmful histamine in the storage of fish meal and improving the fish meal feeding safety. Background technique [0002] Fishmeal is a powder made from whole fish or fish leftovers, which are processed through cooking, pressing, dehydration, degreasing, drying, crushing and other processes. As a high-quality animal protein feed raw material, fish meal has a crude protein content of more than 55%, and is rich in various essential amino acids, essential fatty acids, minerals, vitamins, etc. In addition, fish meal also contains a large number of "unknown growth factors" that can promote animal growth, which plays an important role in feed production. However, during the storage process, the abundant histidine in fish...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/00
CPCA23K10/22A23L3/3463A23L3/3562A23V2002/00A23V2250/511C07H21/02C12N15/52C12Y401/01022C08B37/003C08L5/08B82Y30/00C12N2310/351
Inventor 王彦波傅玲琳黎凡刘忠甫梁君波
Owner ZHEJIANG GONGSHANG UNIVERSITY
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