阅读说明：本技术 一种生物膜抑制剂及其制备方法和应用 (Biofilm inhibitor and preparation method and application thereof ) 是由 李睿 卢洁元 周敏 毕旺来 于 2020-07-23 设计创作，主要内容包括：本发明公开一种生物膜抑制剂及其制备方法和应用,所述生物膜抑制剂包括茶多酚、柠檬酸、Na<Sub>2</Sub>EDTA和无菌水,其中,所述茶多酚、柠檬酸和Na<Sub>2</Sub>EDTA的质量比为(0.1～3)：(25～60)：(0.1～2.5)。本发明提供的生物膜抑制剂包括茶多酚、柠檬酸和Na<Sub>2</Sub>EDTA,对食品的优势腐败菌生长具有显著的抑制效果,能够抑制食源性致病菌的生长,减少细菌生物膜的形成,可应用于食品保鲜处理,也可以用于清除食品加工设备的生物膜。(The invention discloses a biomembrane inhibitor and a preparation method and application thereof, wherein the biomembrane inhibitor comprises tea polyphenol, citric acid and Na 2 EDTA and sterile water, wherein the tea polyphenol, citric acid and Na 2 The mass ratio of the EDTA is (0.1-3): (25-60): (0.1-2.5). The biomembrane inhibitor provided by the invention comprises tea polyphenol, citric acid and Na 2 EDTA has obvious effect of inhibiting the growth of dominant putrefying bacteria in food, can inhibit the growth of food-borne pathogenic bacteria and reduce the formation of bacterial biomembrane, can be applied to food preservation treatment and can also be used for removing foodBiofilm of product processing equipment.)

1. A biofilm inhibitor comprises tea polyphenols, citric acid and Na₂EDTA and sterile water, wherein the tea polyphenol, citric acid and Na₂The mass ratio of the EDTA is (0.1-3): (25-60): (0.1-2.5).

2. The biofilm inhibitor of claim 1, wherein said tea polyphenol, citric acid and Na₂The mass ratio of the EDTA is (0.5-2): (40-55): (0.2 to 1).

3. The biofilm inhibitor of claim 2, wherein said tea polyphenol, citric acid and Na₂The mass ratio of EDTA is 1:50: 0.5.

4. A method for preparing the biofilm inhibitor according to any one of claims 1 to 3, comprising the steps of:

mixing citric acid and Na₂Sequentially adding EDTA and tea polyphenol into sterile water, stirring to completely dissolve, and making into biomembrane inhibitor.

5. A method for keeping food fresh is characterized by comprising the following steps:

spraying a biofilm inhibitor on the surface of food to be subjected to fresh-keeping treatment, and then packaging and cold-chain storing the food to be subjected to fresh-keeping treatment; wherein the biofilm inhibitor is as defined in any one of claims 1 to 3.

6. The method for keeping food fresh as claimed in claim 5, wherein the food to be treated for freshness is chilled meat or crawfish.

7. The method for keeping food fresh as claimed in claim 5, wherein the step of spraying the biofilm inhibitor on the surface of the food to be treated for fresh keeping, and then packaging and cold-chain storing the food to be treated for fresh keeping comprises the steps of:

tea polyphenol, citric acid and Na in the biomembrane inhibitor₂The total concentration of EDTA used was 0.1g/L, 5g/L and 0.05g/L, respectively.

8. An antibacterial cleaning method for food processing equipment is characterized by comprising the following steps:

spraying a biofilm inhibitor on cleaned food processing equipment; wherein the biofilm inhibitor is as defined in any one of claims 1 to 3.

9. The bacteriostatic cleaning method of food processing equipment according to claim 8, wherein the step of spraying the biofilm inhibitor on the cleaned food processing equipment comprises the following steps:

tea polyphenol, citric acid and Na in the biomembrane inhibitor₂The total concentration of EDTA used was 0.1g/L, 5g/L and 0.05g/L, respectively.

Technical Field

The invention relates to the technical field of food processing, in particular to a biofilm inhibitor and a preparation method and application thereof.

Background

The biofilm is a special matrix structure formed by bacteria under adverse living conditions, is a membrane-shaped compound formed by bacteria adhered to a solid or liquid surface in a growth process, and mainly consists of the bacteria and substances secreted by the bacteria, such as protein, polysaccharide, nucleic acid and the like. The formation process of the biological film comprises the following stages: reversible processes, irreversible processes, microcolony formation stages, biofilm maturation stages, and propagation stages. Once the bacterial biofilm is formed, the bacterial biofilm is difficult to remove, and antibiotics, heat stress, disinfectants and the like are difficult to penetrate through a biofilm barrier and act on bacteria in the biofilm, so that the stress resistance of the bacteria in the biofilm is greatly enhanced.

In the food field, bacterial biofilms can be dropped and doped into food after being formed on pipeline instruments in the food processing production process, bacteria continuously adhere to the surface of the food and form biofilms, and food poisoning can be caused once a human body eats the bacteria by mistake. Reports have shown that 80% of food-borne diseases are caused by biofilm formation by food-borne pathogenic bacteria. Secondly, the sterilization measures can be disabled after the microorganisms in the food form a biological film, the shelf life of the food is greatly shortened, and for example, meat such as cold fresh meat, crayfish and the like is rich and is easy to deteriorate under the action of microorganism propagation. In the prior art, an inhibitor for food-borne biomembranes is lacked, and particularly, the inhibitor for the biomembranes has high-efficiency and safe antibacterial effects on foods such as chilled meat, crayfish and the like which are easy to generate putrefying bacteria biomembranes.

Disclosure of Invention

The invention mainly aims to provide a biofilm inhibitor, a preparation method and application thereof, and aims to provide an inhibitor capable of safely and efficiently inhibiting food dominant spoilage bacteria from growing to form a biofilm.

In order to achieve the purpose, the invention provides a biofilm inhibitor which comprises tea polyphenol, citric acid and Na₂EDTA and sterile water, wherein the tea polyphenol, citric acid and Na₂The mass ratio of the EDTA is (0.1-3): (25-60): (0.1-2.5).

Optionally, the tea polyphenols, citric acid and Na₂The mass ratio of the EDTA is (0.5-2): (40-55): (0.2 to 1).

Optionally, the tea polyphenols, citric acid and Na₂The mass ratio of EDTA is 1:50: 0.5.

Further, the invention also provides a preparation method of the biofilm inhibitor, which comprises the following steps:

mixing citric acid and Na₂Sequentially adding EDTA and tea polyphenol into sterile water, stirring to completely dissolve, and making into biomembrane inhibitor.

In addition, the invention also provides a food fresh-keeping method, which comprises the following steps:

spraying a biofilm inhibitor on the surface of food to be subjected to fresh-keeping treatment, and then packaging and cold-chain storing the food to be subjected to fresh-keeping treatment; wherein the biofilm inhibitor is the biofilm inhibitor as described above.

Optionally, the food to be freshness-treated is chilled meat or crawfish.

Optionally, the steps of spraying the biofilm inhibitor on the surface of the food to be subjected to the fresh-keeping treatment, and then packaging and cold-chain storing the food to be subjected to the fresh-keeping treatment comprise:

tea polyphenol, citric acid and Na in the biomembrane inhibitor₂The total concentration of EDTA used was 0.1g/L, 5g/L and 0.05g/L, respectively.

In addition, the invention also provides a bacteriostatic cleaning method of food processing equipment, which comprises the following steps:

spraying a biofilm inhibitor on cleaned food processing equipment; wherein the biofilm inhibitor is the biofilm inhibitor as described above.

Optionally, the step of spraying the biofilm inhibitor on cleaned food processing equipment comprises:

tea polyphenol, citric acid and Na in the biomembrane inhibitor₂The total concentration of EDTA used was 0.1g/L, 5g/L and 0.05g/L, respectively.

In the technical scheme provided by the invention, the biofilm inhibitor comprises tea polyphenol, citric acid and Na₂EDTA has remarkable inhibitory effect on the growth of dominant putrefying bacteria in foodCan inhibit the growth of food-borne pathogenic bacteria and reduce the formation of bacterial biofilms; wherein the tea polyphenol, citric acid and Na₂EDTA is a common and safe additive in food, and tea polyphenol has good antibacterial, antioxidant and anti-biofilm effects; citric acid is a common acid-base regulator, antioxidant and flavor enhancer in food, and has certain bacteriostatic action; na (Na)₂EDTA is commonly used as a chelating agent, preservative and antioxidant in food additives, in this case complexing metal ions, Na₂EDTA and citric acid act synergistically with tea polyphenol to enhance the antibacterial and anti-biofilm effects of tea polyphenol.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other related drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a comparison graph of the inhibition effect of the biofilm inhibitor provided by the invention on common bacillus biofilms;

FIG. 2 is a comparison graph of the effect of the biofilm inhibitor provided by the invention on the inhibition of the biofilm of the dominant spoilage bacteria of crayfish;

FIG. 3 is a comparison graph of the effect of the biofilm inhibitor provided by the invention on the biofilm of the dominant spoilage bacteria of meat.

The implementation, functional features and advantages of the objects of the present invention will be further explained with reference to the accompanying drawings.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially. In addition, the meaning of "and/or" appearing throughout includes three juxtapositions, exemplified by "A and/or B" including either A or B or both A and B. In addition, technical solutions between various embodiments may be combined with each other, but must be realized by a person skilled in the art, and when the technical solutions are contradictory or cannot be realized, such a combination should not be considered to exist, and is not within the protection scope of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The bacterial biofilm has a barrier function, so that bacteria in the biofilm have strong stress resistance and have higher tolerance to antibiotics, disinfectants and preservatives. In the food field, bacterial biofilms can be dropped and doped into food after being formed on pipeline instruments in the food processing production process, bacteria continuously adhere to the surface of the food and form biofilms, and food poisoning can be caused once a human body eats the bacteria by mistake. Reports have shown that 80% of food-borne diseases are caused by biofilm formation by food-borne pathogenic bacteria. Secondly, the sterilization measures can be disabled after the microorganisms in the food form a biological film, the shelf life of the food is greatly shortened, and for example, meat such as cold fresh meat, crayfish and the like is rich and is easy to deteriorate under the action of microorganism propagation. The chilled meat mainly contains predominant putrefying bacteria such as lactic acid bacteria, pseudomonas aeruginosa, staphylococcus aureus, escherichia coli and the like in the storage process, the crayfish mainly contains predominant putrefying bacteria such as enterobacter cloacae, bacillus cereus, bacillus thuringiensis, hydrophilus, metacoccus caseii and the like, and the bacteria can grow in a mixing manner and form a biological membrane in the food refrigeration process, so that the food is accelerated to be putrescent. Different foods have different dominant spoilage bacteria types and different film-producing characteristics, and different antibacterial and anti-biofilm strategies are adopted, so that the prior art is lack of an inhibitor for food-borne biofilms, particularly a biofilm inhibitor which has high-efficiency and safe antibacterial effects on foods such as chilled meat, crayfish and the like which are easy to produce spoilage bacteria biofilms.

To solveThe invention provides a biomembrane inhibitor, which solves the problem that the prior art is lack of an inhibitor capable of efficiently and safely inhibiting food-borne biomembranes, and is prepared by matching tea polyphenol with citric acid and Na₂EDTA, which is effective in inhibiting the growth of food-borne pathogenic bacteria. In an embodiment of the biofilm inhibitor provided by the present invention, the biofilm inhibitor comprises tea polyphenol, citric acid, Na₂EDTA and sterile water, wherein the tea polyphenol, citric acid and Na₂The mass ratio of the EDTA is (0.1-3): (25-60): (0.1-2.5). The sterile water can be plain boiled water or sterile water subjected to sterilization treatment by a chemical method, a physical filtration method and the like, the addition amount of the sterile water is not limited, and the biofilm inhibitor can be prepared into a high-concentration concentrated solution or a low-concentration diluent which can be directly used according to actual use requirements.

In the technical scheme provided by the invention, the biofilm inhibitor comprises tea polyphenol, citric acid and Na₂EDTA and sterile water, and tea polyphenols, citric acid and Na₂The mass ratio of the EDTA is (0.1-3): (25-60): (0.1-2.5), the biofilm inhibitor obtained in the proportion range has a remarkable inhibition effect on the growth of dominant putrefying bacteria of food, can inhibit the growth of food-borne pathogenic bacteria, and reduces the formation of bacterial biofilms; wherein the tea polyphenol, citric acid and Na₂EDTA is a common and safe additive in food, and tea polyphenol has good antibacterial, antioxidant and anti-biofilm effects; citric acid is a common acid-base regulator, antioxidant and flavor enhancer in food, and has certain bacteriostatic action; na (Na)₂EDTA is commonly used as a chelating agent, preservative and antioxidant in food additives, in this case complexing metal ions, Na₂EDTA and citric acid act synergistically with tea polyphenol to enhance the antibacterial and anti-biofilm effects of tea polyphenol. In addition, the concentration of substances related to the biofilm inhibitor provided by the invention is low and is lower than the national limit standard of food additives in GB2760-2014 national food additive use standard, the biofilm inhibitor is safe, reliable and low in cost, is easy to commercially obtain, and can be used as a food preservative and a food preservativeThe biomembrane scavenger has good specificity and strong pertinence, and is particularly suitable for the requirements of antibiosis, biological membrane resistance, color protection and fresh keeping of the chilled meat and the crayfish.

As a preferred embodiment of the present invention, in the biofilm inhibitor: the tea polyphenols, citric acid and Na₂The mass ratio of the EDTA is (0.5-2): (40-55): (0.2 to 1). Within the proportion range, the biofilm inhibitor has better inhibition effect on the growth of the food-borne pathogenic bacteria.

As another preferred embodiment provided by the present invention, in the biofilm inhibitor: the tea polyphenols, citric acid and Na₂The mass ratio of EDTA is 1:50: 0.5. The biofilm inhibitor obtained by the adding proportion has a more remarkable inhibiting effect on common bacilli, including escherichia coli ATCC25922, salmonella, proteus mirabilis, bacillus subtilis and the like, and has a very remarkable inhibiting effect on meat dominant putrefying bacteria and crayfish dominant putrefying bacteria.

Furthermore, the invention also provides a preparation method of the biofilm inhibitor, which is characterized in that the tea polyphenol, the citric acid and the Na are proportionally mixed₂EDTA is added into sterile water, and stirred until the tea polyphenol, citric acid and Na are mixed₂And completely dissolving the EDTA to obtain the biofilm inhibitor. Comparing the citric acid and Na in consideration of the difficulty of dissolution of tea polyphenol₂EDTA is slightly higher, therefore, as a preferred embodiment of the biofilm inhibitor provided by the invention, the preparation method of the biofilm inhibitor comprises the following steps: mixing citric acid and Na₂Sequentially adding EDTA and tea polyphenol into sterile water, stirring to completely dissolve, and making into biomembrane inhibitor. Specifically, the citric acid and Na are firstly mixed₂EDTA is added into sterile water according to a certain proportion, and is stirred until the EDTA is completely dissolved, and then tea polyphenol is added and is stirred until the EDTA is completely dissolved, so that the problem that the tea polyphenol is difficult to dissolve can be solved, and the preparation difficulty of the biomembrane inhibitor is reduced.

The biofilm inhibitor provided by the embodiment of the invention has a remarkable inhibiting effect on food-borne pathogenic bacteria, can be directly used in a food preservation treatment process, is sprayed on the surface of food to inhibit the biofilm and prolong the quality guarantee period of the product, can also be applied to processing equipment used in a food processing process to perform bacteriostatic cleaning on the food processing equipment, plays a role in preventing corrosion and resisting the biofilm, and avoids pollution caused by the food-borne bacteria and the biofilm in a food processing production environment. Therefore, based on the provided biofilm inhibitor, the invention also provides a food fresh-keeping method, and the food is subjected to fresh-keeping treatment by using the provided biofilm inhibitor. Specifically, in an embodiment of the method for preserving food provided by the present invention, the method for preserving food comprises the following steps: spraying a biofilm inhibitor on the surface of food to be subjected to fresh-keeping treatment, and then packaging and cold-chain storing the food to be subjected to fresh-keeping treatment; wherein the biofilm inhibitor is the biofilm inhibitor as described above. Furthermore, the food to be subjected to fresh-keeping treatment is preferably chilled fresh meat or crayfish, the biofilm inhibitor provided by the invention has good specificity and strong pertinence, is particularly suitable for the requirements of antibiosis, biological membrane resistance, color protection and fresh keeping of the chilled fresh meat and the crayfish, and can effectively inhibit the growth of various food-borne pathogenic bacteria.

Further, when the biofilm inhibitor is used, the higher the concentration, the better the bacteriostatic effect is of course, but considering the side effect and cost problem caused by the concentration of the agent, the tea polyphenol, citric acid and Na in the biofilm inhibitor are preferable₂The total use concentration of EDTA is 0.1g/L, 5g/L and 0.05g/L respectively, on the basis of effectively inhibiting the biological membrane, the EDTA inhibitor has no side effect basically and lower cost, and is more beneficial to popularization and use of the biological membrane inhibitor. If the concentration of each component in the pre-prepared biofilm inhibitor is higher than the concentration range, a proper amount of sterile water can be added for dilution before use, and the sterile water can also be plain boiled water or sterile water subjected to sterilization treatment by a chemical method, a physical filtration method and the like.

In addition, the invention also provides a bacteriostatic cleaning method of food processing equipment, which uses the provided biofilm inhibitor to carry out bacteriostatic cleaning on the food processing equipment, and the biofilm inhibitor is used for inhibitingThe preparation needs to be diluted properly before use, and in one embodiment of the bacteriostatic cleaning method for the food processing equipment, the method comprises the following steps: the biofilm inhibitor is sprayed on cleaned food processing equipment, for example, the diluted biofilm inhibitor is sprayed on parts which are difficult to clean, such as food processing table tops or pipelines and the like which are cleaned conventionally, and can be used for removing biofilms; wherein the biofilm inhibitor is the biofilm inhibitor as described above. Similarly, tea polyphenols, citric acid, Na in the biofilm inhibitor are preferred in consideration of the side effects and cost associated with the concentration of the agent₂The total use concentration of EDTA is 0.1g/L, 5g/L and 0.05g/L respectively, on the basis of effectively inhibiting the biological membrane, the EDTA inhibitor has no side effect basically and lower cost, and is more beneficial to popularization and use of the biological membrane inhibitor. If the concentration of each component in the pre-prepared biofilm inhibitor is higher than the concentration range, a proper amount of sterile water can be added for dilution before use, and the sterile water can also be plain boiled water or sterile water subjected to sterilization treatment by a chemical method, a physical filtration method and the like.

The technical solutions of the present invention are further described in detail below with reference to specific examples and drawings, it should be understood that the following examples are merely illustrative of the present invention and are not intended to limit the present invention.