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Method for preparing half-fin anchovy antibacterial peptides

An antibacterial peptide and protein technology, which is applied in the field of preparation of the antibacterial peptide of yellow crucian carp protein, and achieves the effects of simple preparation method, good absorption and easy operation.

Inactive Publication Date: 2011-05-11
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The research on the preparation of antimicrobial peptides from food-source proteins is mainly focused on milk-source proteins at home and abroad, but there are few research reports on the direct enzymatic digestion of other food-source proteins to obtain peptides with antibacterial activity

Method used

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  • Method for preparing half-fin anchovy antibacterial peptides
  • Method for preparing half-fin anchovy antibacterial peptides
  • Method for preparing half-fin anchovy antibacterial peptides

Examples

Experimental program
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Effect test

Embodiment 1

[0020] 1) Remove the internal organs of the yellow crucian carp, wash and cut into small pieces, and use a tissue grinder to grind it evenly to make minced fish. Take 50 grams of minced fish and add 200 grams of deionized water;

[0021] 2) Use 6mol / L hydrochloric acid and 6mol / L sodium hydroxide to adjust the pH value of the minced fish and water mixture to 2.0 and keep it constant;

[0022] 3) Add 55,000 U of pepsin into the mixture, and perform enzymatic hydrolysis for 2.4 hours while maintaining the enzymatic hydrolysis temperature at 37°C;

[0023] 4) Heat up to 95-100°C to passivate and inactivate the enzyme, and the inactivation time is 10 minutes;

[0024] 5) Cool to below 4°C and perform centrifugation for 10 minutes at a speed of 7000r / min;

[0025] 6) Filtrate, and take the intermediate filtrate and store it frozen or freeze-dried at -20°C.

[0026] Trichloroacetic acid precipitation (TCA) combined with Kjeldahl method to determine the extraction rate of soluble p...

Embodiment 2

[0048] 1) Remove the internal organs of the yellow crucian carp, wash and cut into small pieces, and use a tissue grinder to grind it evenly to make minced fish. Take 50 grams of minced fish and add 150 grams of deionized water;

[0049] 2) Use 6mol / L hydrochloric acid and 6mol / L sodium hydroxide to adjust the pH value of the minced fish and water mixture to 2.0 and keep it constant;

[0050] 3) Add 65,000 U of pepsin into the mixture, and perform enzymatic hydrolysis for 3 hours while maintaining the enzymatic hydrolysis temperature at 35°C;

[0051] 4) Heating to 100° C. to passivate and inactivate the enzyme, and the inactivation time is 5 minutes;

[0052] 5) Cool to below 5°C and perform centrifugation for 15 minutes at a speed of 6000r / min;

[0053] 6) Filtrate, and take the intermediate filtrate and store it frozen or freeze-dried at -22°C.

Embodiment 3

[0055] 1) Remove the viscera of the yellow crucian carp, wash and cut into small pieces, and grind it evenly with a tissue grinder to make minced fish. Take 50 grams of minced fish and add 250 grams of deionized water;

[0056] 2) Use 6mol / L hydrochloric acid and 6mol / L sodium hydroxide to adjust the pH value of the minced fish and water mixture to 2.0 and keep it constant;

[0057] 3) Add 50,000 U of pepsin into the mixture, and perform enzymatic hydrolysis for 2 hours while maintaining the enzymatic hydrolysis temperature at 45°C;

[0058] 4) Heat up to 95° C. to passivate and inactivate the enzyme, and the inactivation time is 15 minutes;

[0059] 5) Cool to below 4°C and perform centrifugation for 5 minutes at a speed of 8000r / min;

[0060] 6) Filtrate, and take the intermediate filtrate and store it frozen or freeze-dried at -25°C.

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Abstract

The invention relates to a method for preparing half-fin anchovy antibacterial peptides, which comprises the following steps of: homogenating fish into fish paste, adding pepsin into the fish paste according to a ratio of 1,000-1,300U / g under the condition that the pH value is 1.5-2.5, raising the temperature to 35-45DEG C, performing enzymolysis for 2 to 3 hours, heating enzymolysis liquid at the temperature of between 95 and 100DEG C for 5 to 15 minutes for killing enzyme, cooling to the temperature of between 3 and 5DEG C, centrifuging at a rate of 6,000 to 8,000r / min for 5 to 15 minutes, filtering, taking intermediate filtrate, and performing frozen storage or freeze-dried storage at -20DEG C at least. The antibacterial peptides is prepared from a pure natural low-value marine product, namely half-fin anchovy; the half-fin anchovy antibacterial peptides are rich in amino acid necessary for human bodies, and have stable physical and chemical properties; the antibacterial peptides prepared from food-derived protein through enzymolysis have high safety, good absorptivity and good antibacterial effect, and can serve as a nutritional and functional additive in foods or is developed into drug resistant bacteria antibiotics. The preparation method is simple, economic and easy to operate, can be carried out in a common laboratory, and is suitable for industrial expanded production.

Description

technical field [0001] The invention relates to a method for preparing protein antibacterial peptides, in particular to a method for preparing protein antibacterial peptides of crucian carp. Background technique [0002] Today, with the abuse of antibiotics and the continuous emergence of drug-resistant strains, the research on antimicrobial peptides has attracted the attention of scholars at home and abroad. People are constantly looking for new antimicrobial peptides and exploring new mechanisms of action of antimicrobial peptides. Antibacterial peptide (ABP), also known as antimicrobial peptide (AMP), is widely distributed in bacteria, viruses, various animals and plants, and is an important part of biological natural immune defense system. Most of AMPs are a class of small molecule peptides with a molecular weight of less than 10kDa, cationic amphiphilic peptides with diversity in chain length, sequence and structure (Boman H G.Antibactericidal peptides: key components n...

Claims

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Application Information

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IPC IPC(8): C12P21/06
Inventor 宋茹韦荣编
Owner ZHEJIANG OCEAN UNIV
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