阅读说明：本技术 泥蚶血红蛋白抗菌肽在食品防腐保鲜中的应用 (Application of tegillarca granosa hemoglobin antibacterial peptide in food preservation and freshness keeping ) 是由 杨燊 董玉婷 李健 谢婷玉 章跃陵 刘光明 于 2020-03-10 设计创作，主要内容包括：本发明提出了泥蚶血红蛋白抗菌肽在食品防腐保鲜中的应用,所述泥蚶血红蛋白抗菌肽的氨基酸序列如SEQ ID NO:1所示。该抗菌肽对单增李斯特菌具有很强的抗菌作用,可吸附在单增李斯特菌细胞膜表面并对细胞膜进行破坏导致单增李斯特菌死亡,可改变单增李斯特菌细胞膜的通透性并抑制细胞膜生成。从而为寻找新的食品防腐剂提供实验依据,促进我国食品行业的健康、持续发展。(The invention provides an application of tegillarca granosa hemoglobin antibacterial peptide in food preservation and fresh keeping, wherein the amino acid sequence of the tegillarca hemoglobin antibacterial peptide is shown as SEQ ID NO. 1. The antibacterial peptide has strong antibacterial effect on Listeria monocytogenes, can be adsorbed on the surface of the cell membrane of Listeria monocytogenes and can destroy the cell membrane to cause the death of Listeria monocytogenes, can change the permeability of the cell membrane of Listeria monocytogenes and inhibit the generation of the cell membrane. Thereby providing experimental basis for searching new food preservative and promoting the healthy and sustainable development of food industry in China.)

1. An application of tegillarca granosa hemoglobin antibacterial peptide in food preservation and fresh keeping is characterized in that the amino acid sequence of the tegillarca granosa hemoglobin antibacterial peptide is shown as SEQ ID NO. 1.

2. The application of the tegillarca granosa hemoglobin antimicrobial peptide of claim 1 in food preservation and freshness preservation, which is characterized in that the application is the inhibition of the growth of Listeria monocytogenes.

3. The application of the tegillarca granosa hemoglobin antimicrobial peptide of claim 1 in food preservation and fresh keeping, wherein the application is that the tegillarca granosa hemoglobin antimicrobial peptide is adsorbed on the surface of a cell membrane of Listeria monocytogenes and damages the cell membrane to cause the death of Listeria monocytogenes.

4. The application of the tegillarca granosa hemoglobin antimicrobial peptide of claim 1 in food preservation and fresh-keeping, which is characterized in that the application is changing the permeability of Listeria monocytogenes cell membranes and inhibiting the generation of the cell membranes.

5. The application of the tegillarca granosa hemoglobin antibacterial peptide of claim 1 in food preservation and fresh-keeping is characterized in that the tegillarca granosa hemoglobin antibacterial peptide is directly sprayed on the surface of solid food in the form of solution or is mixed in liquid food.

6. An antiseptic preservative is characterized by comprising tegillarca granosa hemoglobin antibacterial peptide and auxiliary materials acceptable in bromatology, wherein the amino acid sequence of the tegillarca granosa hemoglobin antibacterial peptide is shown as SEQ ID NO. 1.

Technical Field

The invention relates to the technical field of food microorganisms, in particular to application of tegillarca granosa hemoglobin antibacterial peptide in food preservation and freshness preservation.

Background

Listeria monocytogenes is a food-borne pathogen, often detected in food, which is one of the four major food-borne pathogens defined by the world health organization. The bacteria can enter human bodies and livestock bodies through polluted food, and symptoms such as septicemia, meningitis, mononucleosis and the like are caused. In addition, the food-borne pathogenic bacteria not only have influence on human health, but also cause huge economic loss in the food processing and storage processes.

Tegillarca granosa (Tegillarca granosa) is a bivalve mollusk distributed in a wide range of regions from the east coast of south africa to china, southeast asia and australia. Unlike most molluscs that use hemocyanin for oxygen transport, blood clam uses hemoglobin for oxygen transport. The hemoglobin of the blood clam not only can be used as an oxygen carrier, but also plays a certain role in immune reaction. The antibacterial effect of the tegillarca granosa hemoglobin antibacterial peptide obtained by separation by an enzymolysis method in the related technology is not ideal, and whether the tegillarca granosa hemoglobin and polypeptide thereof have antibacterial activity to Listeria monocytogenes has not been reported.

Disclosure of Invention

In order to solve one of the problems, the invention provides an application of tegillarca granosa hemoglobin antibacterial peptide in food preservation and fresh keeping. The antibacterial peptide has strong antibacterial effect on Listeria monocytogenes.

Therefore, in the first aspect of the invention, the invention provides the application of the tegillarca granosa hemoglobin antibacterial peptide in food preservation and freshness preservation, and the amino acid sequence of the tegillarca granosa hemoglobin antibacterial peptide is shown as SEQ ID NO. 1.

According to the embodiment of the invention, the molecular weight of the tegillarca granosa hemoglobin antibacterial peptide is 2049Da, the damage degree of the antibacterial cell membrane is observed by using a transmission electron microscope, whether the antibacterial peptide can be adsorbed on the surface of bacteria is observed by using a laser confocal microscope, the influence of TGH1 on the permeability rate and the film forming property of the Listeria monocytogenes cell membrane is verified, the contents of α -helix, β -fold and other secondary structures of the antibacterial peptide are measured by using circular dichroism, the secondary structure of the antibacterial peptide is preliminarily predicted, and the influence on the human safety of the antibacterial peptide is finally evaluated.

In addition, according to the embodiment of the present invention, the above application may further include at least one of the following additional technical features:

according to an embodiment of the invention, the use is for inhibiting the growth of listeria monocytogenes.

According to an embodiment of the present invention, the application is to adsorb on the surface of the cell membrane of the listeria monocytogenes and destroy the cell membrane, which results in the death of the listeria monocytogenes.

According to an embodiment of the invention, the use is for altering the permeability of a listeria monocytogenes cell membrane and inhibiting cell membrane production.

According to the embodiment of the invention, the tegillarca granosa hemoglobin antibacterial peptide is directly sprayed on the surface of solid food in the form of solution or is mixed in liquid food.

In a second aspect of the invention, the invention provides an antiseptic preservative, which comprises tegillarca granosa hemoglobin antibacterial peptide and an auxiliary material acceptable in bromatology, wherein the amino acid sequence of the tegillarca hemoglobin antibacterial peptide is shown in SEQ ID NO. 1.

The preservative and the fresh-keeping agent can better inhibit the growth of Listeria monocytogenes in food.

Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.

Drawings

FIG. 1 is a graph of TGH1 determination of the Minimum Inhibitory Concentration (MIC) for several common pathogenic bacteria according to an embodiment of the present invention;

FIG. 2 is a graph of the kill time of TGH1 versus Listeria monocytogenes according to an embodiment of the present invention;

FIG. 3 is a transmission electron microscope observation of Listeria monocytogenes according to an embodiment of the present invention;

FIG. 4 is a confocal microscope observation of TGH1 treated Listeria monocytogenes cells according to an embodiment of the present invention;

FIG. 5 shows the inner membrane permeability change of Listeria monocytogenes after TGH1 treatment according to an embodiment of the present invention;

FIG. 6 shows the inhibition of Listeria monocytogenes biofilm formation by TGH1 according to an embodiment of the present invention;

FIG. 7 is a secondary structure analysis of TGH1 according to an embodiment of the present invention;

FIG. 8 is a structural view of a predictive model of TGH1 according to an embodiment of the invention;

FIG. 9 shows the effect of TGH1 on the viability of normal human hepatocytes (L O2 cells) according to an embodiment of the present invention.

Detailed Description

The technical solution of the present invention is illustrated by specific examples below. It is to be understood that one or more method steps mentioned in the present invention do not exclude the presence of other method steps before or after the combination step or that other method steps may be inserted between the explicitly mentioned steps; it should also be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Moreover, unless otherwise indicated, the numbering of the various method steps is merely a convenient tool for identifying the various method steps, and is not intended to limit the order in which the method steps are arranged or the scope of the invention in which the invention may be practiced, and changes or modifications in the relative relationship may be made without substantially changing the technical content.

In order to better understand the above technical solutions, exemplary embodiments of the present invention are described in more detail below. While exemplary embodiments of the invention have been shown, it should be understood that the invention may be embodied in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

The following reagents and bacteria were purchased from the market.

The invention will now be described with reference to specific examples, which are intended to be illustrative only and not to be limiting in any way.