Application of Tegillarca granosa hemoglobin antimicrobial peptide in food preservation and fresh-keeping

A hemoglobin and food antiseptic technology, applied in the field of food microorganisms, can solve the problems of unsatisfactory antibacterial effect and achieve good growth effect

Active Publication Date: 2020-07-14
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the related art, the antibacterial effect of clam hemoglobin antimicrobial peptides isolated by enzymatic hydrolysis is not satisfactory, and whether the clam hemoglobin and its polypeptides have antibacterial activity against Listeria monocytogenes has not been reported.

Method used

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  • Application of Tegillarca granosa hemoglobin antimicrobial peptide in food preservation and fresh-keeping
  • Application of Tegillarca granosa hemoglobin antimicrobial peptide in food preservation and fresh-keeping
  • Application of Tegillarca granosa hemoglobin antimicrobial peptide in food preservation and fresh-keeping

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Screening of clam hemoglobin-derived antimicrobial peptides

[0029] The hemoglobin sequence of cockles consists of 152 amino acids:

[0030] Because the cell membrane of bacteria is negatively charged, negatively charged amino acids are not conducive to the antibacterial activity of peptides, and there are more negatively charged regions in the hemoglobin sequence of mud cockles, only three regions (sequence number 22-39, sequence number 59-81, sequence number 92-109) are less negatively charged. TWEMVSGKKKNGVVLMIK has a net positive charge of +3.

[0031] Lysamino (K) has the function of stabilizing the secondary structure of the polypeptide, and the secondary structure can ensure the realization of the antibacterial effect of the polypeptide. In the hemoglobin sequence of the clam, only the fragment with sequence number 22-39 has 4 lysines (K), while the two fragments with sequence numbers 59-81 and 92-109 have 1 and 3 lysines (K) respectively. 2.

[00...

Embodiment 2

[0034] Embodiment 2 Minimum Inhibitory Concentration (MIC) Determination

[0035] Cultivate Listeria monocytogenes at 37°C for 12h to logarithmic growth phase, dilute to 10 in 0.01M pH 7.2 phosphate buffer 6 CFU / mL. The antimicrobial peptide TGH1 of Example 1 was dissolved in phosphate buffer, and mixed with Listeria monocytogenes in equal volume at 37° C. for 2 hours. The minimum inhibitory concentration (MIC) is the lowest concentration of an antimicrobial peptide at which no bacterial growth is visible from the microtiter plate after overnight incubation at 37°C. The results showed that the minimum inhibitory concentration (MIC) of the antimicrobial peptide TGH1 to Listeria monocytogenes was 15.6 μg / mL, as figure 1 shown. The antimicrobial peptide TGH1 has an MIC greater than 1000 μg / mL to Escherichia coli, greater than 1000 μg / mL to Streptococcus B, and greater than 1000 μg / mL to Staphylococcus aureus. The antimicrobial peptide TGH1 has a very strong inhibitory effect ...

Embodiment 3

[0036] Example 3 Antimicrobial Peptide TGH1 Time and Lethal Relationship Analysis

[0037] The time-killing kinetics of antimicrobial peptides was assessed by plate counting. Adjust the cultured Listeria monocytogenes to approximately 10 6 CFU / mL concentration, exposed to different concentrations of antimicrobial peptide TGH1, then incubated at 37°C for 5h, and observed the number of residual bacteria at 0, 1, 2, 3, 4, and 5 hours, respectively. The indicator strain cultured in phosphate buffer (0.01M, pH 7.2) was used as negative control.

[0038] combine figure 2 , where, a: sterile water; b: 0.5mg / mL ampicillin; c: 1.0mg / mL antimicrobial peptide TGH1; d: 0.5mg / mL antimicrobial peptide TGH1; e: 0.25mg / mL antimicrobial peptide TGH1; f: 0.125 mg / mL antimicrobial peptide TGH1; g: 62.5 μg / mL antimicrobial peptide TGH1; h: 31.25 μg / mL antimicrobial peptide TGH1; i: 15.6 μg / mL antimicrobial peptide TGH1. It can also be seen from the time-kill curve that the number of bacteria...

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Abstract

The invention provides application of Tegillarca granosa hemoglobin antimicrobial peptide in food preservation and fresh-keeping, An amino acid sequence of the Tegillarca granosa hemoglobin antimicrobial peptide is shown as SEQ ID NO:1. The Tegillarca granosa hemoglobin antimicrobial peptide has a high antibacterial effect on Listeria monocytogenes, can be adsorbed on the cell membrane surface ofListeria monocytogenes to destroy the cell membrane to cause the death of Listeria monocytogenes, the permeability of the cell membrane of Listeria monocytogenes can be changed, and the cell membraneformation is inhibited. Experimental basis is provided for finding new food preservatives, and the healthy and sustainable development of national food industry are promoted.

Description

technical field [0001] The invention relates to the technical field of food microorganisms, in particular to the application of a clam hemoglobin antibacterial peptide in food preservation and preservation. Background technique [0002] Listeria monocytogenes is a foodborne pathogen that is often detected in food and is one of the four major foodborne pathogens defined by the World Health Organization. The bacteria can enter the human and animal body through contaminated food, causing symptoms such as sepsis, meningitis and mononucleosis. In addition, food-borne pathogens not only affect human health, but also cause huge economic losses during food processing and storage. [0003] The mud cockle (Tegillarca granosa) is a bivalve mollusk that is distributed over a wide area from the east coast of South Africa to China, Southeast Asia and Australia. Unlike most molluscs, which use hemocyanin for oxygen transport, cockles use hemoglobin to transport oxygen. The hemoglobin of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23L3/3526
CPCA23L3/3526
Inventor 杨燊董玉婷李健谢婷玉章跃陵刘光明
Owner JIMEI UNIV
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