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Application of natural antibacterial peptide As-CATH4 in food preservation and fresh-keeping

A technology of food preservation and food preservatives, which is applied in the field of food chemistry and can solve problems such as degradation, narrow antibacterial spectrum, and poor stability of antimicrobial peptides

Active Publication Date: 2020-07-07
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently reported antimicrobial peptide biological preservatives have two disadvantages: first, they have a narrow antibacterial spectrum and can only kill some food spoilage bacteria, such as Nisin, which is only effective against Gram-positive spoilage bacteria
Secondly, most antimicrobial peptides have poor stability and are easily degraded by enzymes in food. They cannot be used as preservatives alone for a long time.

Method used

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  • Application of natural antibacterial peptide As-CATH4 in food preservation and fresh-keeping
  • Application of natural antibacterial peptide As-CATH4 in food preservation and fresh-keeping
  • Application of natural antibacterial peptide As-CATH4 in food preservation and fresh-keeping

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Preparation of As-CATH4

[0025] (1) The chemical synthesis method of As-CATH4: the complete sequence of As-CATH4 was synthesized by an automatic peptide synthesizer (433A, Applied Biosystems), and desalted and purified by HPLC reverse-phase column chromatography.

[0026] (2) The molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF).

[0027] (3) The purity of the purified As-CATH4 is identified by high performance liquid chromatography (HPLC), the molecular weight is determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF), the isoelectric point is determined by isoelectric focusing electrophoresis, and automatic amino acid sequencing is used Determination of amino acid sequence structure.

[0028] The natural antibacterial peptide As-CATH4 is composed of 38 amino acids, with a molecular weight of 4402.5Da and an isoelectric point of 12.78. The amino a...

Embodiment 2

[0031] Detection of antibacterial activity of As-CATH4 against common spoilage bacteria:

[0032] Determined by the minimum inhibitory concentration method. Select fish spoilage strains for MIC determination experiment. The test strains were inoculated into MH liquid medium (OXOID, UK), cultured with shaking at 37°C to the logarithmic growth phase, and then diluted to 2×10 with fresh MH liquid medium to the logarithmic growth phase. 5 CFU / ml for use.

[0033] Add 100 μl of LB liquid medium in advance to each well of a sterile 96-well plate, and then add 100 μl of As-CATH4 sample solution diluted to a certain concentration with MH liquid medium to the first well and filtered through a 0.22 μm filter membrane, After mixing evenly, take 100 μl and add it to the second well, serially dilute (see Table 1), suck out 100 μl from the ninth well and discard, and the tenth well is a control tube.

[0034] Table 1 Dilution method

[0035]

[0036] The 96-well plate was cultured at...

Embodiment 3

[0041] Determination of As-CATH4 sterilization speed:

[0042] Select four strains of spoilage bacteria (Acinetobacter Jones, Shewanella algae, Proteus mirabilis, and Bacillus subtilis) and culture them with MH liquid medium (OXOID, UK) at 37°C for 12 hours, and then culture them with fresh MH liquid base diluted to 10 6 CFU / ml bacterial suspension. The As-CATH4 sample dissolved in sterilized deionized water was added to the bacterial suspension so that the final concentration was 5×MIC. Place the bacterial solution added with the As-CATH4 sample in a 37°C incubator for shaking culture, take 10 μl of the bacterial solution to dilute 1000 times at 0, 15, 30, 60, 90, 120 and 180 minutes, and then take 50 μl of the diluted bacterial solution Spread it on the MH solid medium plate, and count the colonies after culturing overnight in a 37°C incubator. The experiment used butylparaben as a positive control and sterile deionized water as a negative control.

[0043] The results a...

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Abstract

The invention discloses application of a natural antibacterial peptide As-CATH4 in food preservation and fresh-keeping. The natural antibacterial peptide As-CATH4 in the present invention has a broad-spectrum and high-efficiency antibacterial effect on common food spoilage bacteria, has a rapid sterilization effect, and has a good synergistic effect with a traditional preservative butyl p-hydroxybenzoate. The combination of As-CATH4 and butyl p-hydroxybenzoate significantly inhibits the growth of spoilage bacteria, increases sensory evaluation score, and prolongs food storage time. In the storage experiment of salmon meat, cooked meat sauce and freshly squeezed fruit juice, As-CATH4 significantly inhibits the growth of spoilage bacteria, increases sensory evaluation score, and prolongs food storage time. In the storage experiment of orange juice, the combination of As-CATH4 and butyl p-hydroxybenzoate significantly inhibits the growth of spoilage bacteria, prolongs food storage time, and shows a good synergistic effect. In addition, As-CATH4 can be completely degraded by trypsin and has no toxic side effects on human body. As a new type of food preservative, As-CATH4 has wide application prospects.

Description

technical field [0001] The invention relates to the application of a natural antibacterial peptide As-CATH4 in food antiseptic and preservation, and belongs to the technical field of food chemistry. Background technique [0002] In the food industry, spoilage microorganisms are a major hazard in the process of food production and preservation, causing unnecessary waste and huge economic losses. Spoilage microorganisms can cause changes in the physical and chemical properties of food, thereby causing the food to lose its original quality, nutritional value and organizational characteristics, and ultimately lead to non-compliance with food hygiene standards. Bacteria, mold and yeast can cause food spoilage, of which bacteria and mold are the most common. In order to inhibit the growth of spoilage microorganisms, the food industry has adopted a variety of preservation methods, including chemical and physical preservation techniques. However, food spoilage remains a significan...

Claims

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Application Information

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IPC IPC(8): A23B4/20A23L2/44A23L3/3463A23L3/3517
CPCA23B4/20A23L2/44A23L3/3463A23L3/3517Y02A50/30
Inventor 王义鹏张芬于海宁章铭辉陈燕
Owner SUZHOU UNIV
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