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PEG-ACS/M-siRNA nano-composite, application thereof, and method for reducing histamine content of fish meal in storage process

A PEG-ACS and nanocomposite technology, applied in the field of feed industry, can solve the problems of high cytotoxicity and in vivo toxicity, low efficiency, poor targeting, etc., and achieve the effect of wide application prospects and delaying the process of corruption

Active Publication Date: 2018-11-23
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 nano-composite, application thereof, and method for reducing histamine content of fish meal in storage process
  • PEG-ACS/M-siRNA nano-composite, application thereof, and method for reducing histamine content of fish meal in storage process
  • PEG-ACS/M-siRNA nano-composite, application thereof, and method for reducing histamine content of fish meal in storage process

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] (1) Design 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 0.5 g of chitosan, dissolve it in a TEMED / HCl buffer solution with pH 5.0, then add 567 mg of EDC and 340 mg of NHS coupling agent to the solution, stir evenly, and then add the Arginine with 80% molar amount of polysaccharide amino group was reacted under magnetic stirring at room temperature for 8 hours, and then dialysis and freeze-drying of desalted water were carried out 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. The unreacted PEG was removed by dialysis using a dialysis bag with a molecular weight cut-off of 14,000. The PEGylated arginine-modified chitosan is obtained, which is freeze-dried for later use.

...

Embodiment 2

[0075] (1) Design 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: 0.8 g of chitosan was weighed and dissolved in TEMED / HCl buffer solution with pH 4.5, then 900 mg of EDC and 550 mg of NHS coupling agent were added to the solution, and after stirring evenly, Arginine in equimolar amount of polysaccharide amino group was reacted at room temperature for 7 hours under magnetic stirring, and then arginine-modified chitosan was obtained by dialysis of desalinated water and freeze-drying.

[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. The unreacted PEG was removed by dialysis using a dialysis bag with a molecular weight cut-off of 14,000. The PEGylated arginine-modified chitosan is obtained, which is freeze-dried for later use.

[008...

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Abstract

The invention discloses a PEG-ACS / M-siRNA nano-composite, application thereof, and a method for reducing the histamine content of fish meal in a storage process. The method comprises the steps that (1) according to the sequence of histidine decarboxylase genes MU9_RS17900 (Gene ID: 14672734) of morganella morganii subsp. morganii KT (CP004345) which is the main histamine-producing strain, small interfering ribonucleic acid (si RNA) is designed and prepared; (2) PEGylated arginine-modified chitosan is used as a carrier, and thus the stable PEG-ACS / M-siRNA nano-composite is obtained; and (3) thePEG-ACS / M-siRNA nano-composite is added into the fish meal in certain proportion. The method has a significant effect on restraining the histamine content of the fish meal in the storage process, thehistamine content of the fish meal can be reduced by 49%-53%, and therefore the method has important significance for control over biogenic amine in the fish meal in the feed industry.

Description

technical field [0001] The invention belongs to the field of feed industry, and in particular 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 hazardous substance histamine in the storage of fish meal, and improving the fish meal feeding safety. Background technique [0002] Fish meal is a powder that is made from whole fish or fish scraps through cooking, pressing, dehydration, defatting, drying, crushing and other processes. As a high-quality animal protein feed material, fish meal has a crude protein content of more than 55%, and is rich in a variety of 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 storage, the abundant histidine in fish meal will un...

Claims

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Application Information

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IPC IPC(8): A23L3/3562A23L3/3463A23K10/22
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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