Bacteriostatic combination preparation containing egg white lysozyme and its synthesis process

Abstract

The application relates to the field of biological medicine, and discloses an antibacterial composite preparation containing egg white lysozyme and a synthesis process thereof. The antibacterial composite preparation contains the following raw materials in parts by weight: modified carrageenan 20-50 parts, egg white lysozyme 0.5-1 part, collagenase liposome 0.01-0.05 part, glycerol 1-2 parts and deionized water 40-100 parts. The modified carrageenan is prepared by covalently connecting nano zinc oxide and carrageenan. Soybean lecithin, cholesterol, chloroform and diethyl ether are used as an oil phase, and collagenase and a phosphate buffer solution are used as an aqueous phase. The oil phase and the aqueous phase are subjected to water bath ultrasonic treatment, and then high-temperature treatment is conducted to gelatinize the carrageenan, so as to prepare the antibacterial composite preparation containing egg white lysozyme. The antibacterial composite preparation has good antibacterial and anti-inflammatory effects and can promote wound healing.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, in particular to a bacteriostatic combination preparation containing egg white lysozyme and a synthesis process thereof. BACKGROUND

[0002] In daily work and life, as the first barrier of the human body, the skin is inevitably damaged by bumps, scratches and other damages. In serious cases, not only do you need to take anti-inflammatory and other drug preparations, but also you need to use external preparations to coat the wound to promote wound healing and avoid infection. Gel dressings, as one of the external preparations, have been widely used in the field of wound treatment. The gel dressings currently used mainly allow the wound to scab and dry on its own, which takes a long time to heal and is prone to cause bacteria to grow at the wound site, making it difficult to meet people's needs for skin damage recovery. Therefore, it is of great significance to develop new antibacterial dressings.

[0003] Lysozyme is a non-toxic and side-effect-free antibacterial material, commonly used as a green natural food preservative. In clinical medicine, it has high antibacterial performance and can accelerate wound healing. In daily chemical industry, lysozyme can be used in cosmetics to repair wounds and scars, and has the functions of anti-infection, anti-virus, mild sterilization and elimination of swelling. In addition, lysozyme has strong specificity, low cytotoxicity, and is similar in nature to normal physiological substances in the body, making it easier to be absorbed by the body. It can be used as an antibacterial agent alone. At present, the use of lysozyme as an antibacterial agent combined with dressings has become a research hotspot. Patent No. CN201710552708.1 discloses a lightwood-lysozyme anti-infection dressing and its preparation method and application. Lysozyme is fully adhered to the lightwood's porous diameter in the form of dopamine glue, providing a relatively stable release microenvironment, so that the dressing has good antibacterial performance. However, lysozyme is a non-broad-spectrum antibacterial agent, mainly acting on gram-positive bacteria, and having little effect on gram-negative bacteria, which to some extent limits the application of lysozyme as an antibacterial agent. Therefore, other antibacterial agents need to be used to compound with lysozyme to make the dressing have more excellent antibacterial performance. SUMMARY

[0004] The purpose of the present application is to provide a bacteriostatic combination preparation containing egg white lysozyme and a synthesis process thereof, which solves the following technical problems:

[0005] (1) The problem of the gel dressing's poor effect in promoting wound healing is solved.

[0006] (2) The problem of the gel dressing's poor antibacterial performance, which cannot eliminate bacteria around the wound and is prone to cause infection at the wound site, is solved.

[0007] (3) The use of egg white lysozyme as a single antibacterial agent for dressing has poor effect on gram-negative bacteria.

[0008] The object of the present application can be achieved by the following technical solutions:

[0009] The antibacterial combined preparation containing egg white lysozyme comprises the following raw materials in parts by weight: modified carrageenan 20-50 parts, egg white lysozyme 0.5-1 part, collagenase liposome 0.01-0.05 part, glycerol 1-2 parts, and deionized water 40-100 parts.

[0010] The modified carrageenan is prepared by covalent bonding of nano zinc oxide and carrageenan.

[0011] The collagenase liposome is prepared by using soybean lecithin, cholesterol, chloroform and ether as the oil phase, and collagenase and phosphate buffer solution as the water phase, and then performing water bath ultrasonic treatment.

[0012] Further, the carrageenan is k-type carrageenan.

[0013] Further, the covalent bonding comprises the following steps:

[0014] A: Nano zinc oxide is added to a mixed solvent of methanol and N,N-dimethylformamide, ultrasonic dispersion is performed, and then the product is transferred to an oil bath pot, carboxyethyl silatrane sodium salt is added under stirring, the temperature in the oil bath pot is increased to 50-60 DEG C, reflux reaction is performed in a nitrogen atmosphere for 6-18 h, heating is stopped, the solid product is centrifuged and separated after the product is cooled, the solid product is washed to neutral with ethanol, hydrochloric acid and deionized water, and vacuum drying is performed to obtain carboxyl functionalized zinc oxide;

[0015] B: Carboxyl functionalized zinc oxide is added to N,N-dimethylformamide, ultrasonic treatment is performed for 20-40 min, carrageenan is added, mechanical stirring is performed, a composite catalyst is continuously added, the mixture is uniformly mixed, and then the mixture is placed in a water bath pot at 15-35 DEG C, stirring reaction is performed for 24-48 h, the solid product is collected by centrifugation after the reaction is completed, washing is performed, and vacuum drying is performed to obtain modified carrageenan.

[0016] Further, in step A, the volume ratio of the methanol and N,N-dimethylformamide is 3-4:1.

[0017] Further, in step B, the composite catalyst is 4-dimethylamino pyridine and N,N-dicyclohexyl carbodiimide in a mass ratio of 1:8-10.

[0018] By the technical scheme, the surface of the nano zinc oxide is rich in hydroxyl groups, which can undergo condensation reaction with Si-OH in the structure of sodium carboxyethyl silanetriol, so as to modify the carboxyl group on the surface of the nano zinc oxide, and carboxyl functionalized zinc oxide is prepared. Under the composite catalysis of 4-dimethylaminopyridine and N,N-dicyclohexyl carbodiimide, the carboxyl group can further undergo esterification condensation reaction with the hydroxyl group in the structure of carrageenan, so as to chemically bond the nano zinc oxide and the carrageenan in a covalent bond connection mode, improve the interface performance between each other, and be beneficial to the uniform dispersion of the nano zinc oxide, thereby avoiding the influence of the agglomeration problem on the photocatalytic antibacterial performance of the nano zinc oxide.

[0019] Further, the synthesis process of the collagenase liposome comprises the following steps:

[0020] S1: collagenase is added to phosphate buffer solution, stirred uniformly, configured into a buffer solution, and an aqueous phase is formed;

[0021] S2: soybean lecithin and cholesterol are dissolved in a mixed solvent of chloroform and diethyl ether with a volume ratio of 1:1-2, stirred uniformly, and an oil phase is formed. The aqueous phase prepared in step S1 is added to the oil phase, and the water bath kettle is placed at 35-38℃, and ultrasonic treatment is carried out until a stable water / oil emulsion is formed. The organic solvent is removed by rotary evaporation, and the phosphate buffer solution is added to the rotary evaporation flask, and the rotation is continued for 4-8h to obtain a collagenase liposome suspension. The suspension is filtered using a microporous filter membrane to remove large particle impurities, and the collagenase liposome is obtained.

[0022] Further, in step S1, the mass concentration of the buffer solution is 1-2g / L.

[0023] Further, in step S2, the mass ratio of the soybean lecithin and cholesterol is 1:0.2-0.6.

[0024] By the above technical scheme, the soybean lecithin, cholesterol, chloroform and diethyl ether are used as the oil phase, and the collagenase and the phosphate buffer solution are used as the water phase to prepare the collagenase liposome. The liposome has a bilayer structure composed of phospholipid molecules, and as a carrier of collagenase, not only has a sustained release effect, but also can promote the collagenase to penetrate the sebum and enter the deep layer of the skin by using the lipophilic function of the liposome.

[0025] A synthesis process of an antibacterial combined preparation containing egg white lysozyme comprises the following steps:

[0026] (1) Modified carrageenan is added to deionized water, ultrasonically dispersed uniformly, and egg white lysozyme is added. The mixture is stirred uniformly at a rotation speed of 80-100rpm to obtain a premix ①;

[0027] (2) The collagenase liposome and glycerol are placed in a blender and stirred uniformly to obtain a premix ②;

[0028] (3) The premix 2 is added into the premix 1, stirred uniformly, transferred into an oil bath pot at 50-60 DEG C, fully stirred until a gelatinous bacteriostatic combination preparation containing egg white lysozyme is formed.

[0029] Through the technical scheme, the modified carrageenan can be ionized in the aqueous solution, and has a large amount of sulfate in the structure, thus has a negative charge; the isoelectric point of the egg white lysozyme is 10.8, and thus has a positive charge in the aqueous solution; therefore, the modified carrageenan and the egg white lysozyme can be attracted and combined through electrostatic adsorption, to form a drug-loaded system taking the modified carrageenan as a carrier and the lysozyme as an antibacterial agent; the collagenase liposome and the humectant glycerol are mixed, and then high-temperature stirring is performed; the carrageenan forms a gelatinous bacteriostatic combination preparation containing egg white lysozyme.

[0030] The beneficial effects of the present application are as follows:

[0031] (1) The present application uses a chemical bond connection mode to bond the nano zinc oxide and the carrageenan, to prepare the modified carrageenan; the modified carrageenan is taken as a carrier to load the egg white lysozyme through electrostatic adsorption, to form a drug-loaded system; under the electrostatic action, the drug-loaded system can realize the controlled release of the egg white lysozyme, to achieve a slow release effect, which is helpful to make the egg white lysozyme have a long-acting antibacterial effect; the collagenase liposome and the glycerol are mixed, and then a gelatinous bacteriostatic combination preparation containing egg white lysozyme is formed; the gelatinous bacteriostatic combination preparation is taken as a gelatinous dressing; the nano zinc oxide has a photocatalytic antibacterial effect, and has a strong killing effect on gram-negative bacteria, which can solve the problem that the egg white lysozyme has a poor effect on gram-negative bacteria, so as to endow the gelatinous dressing with excellent antibacterial properties.

[0032] (2) The present application uses the collagenase as a wound healing promoter, which is added into the gelatinous dressing; the collagenase can play a role in promoting wound healing through itself and collagen degradation products, and can also play a role in inflammation removal while selectively removing necrotic skin tissue; the collagenase is embedded by using the liposome with amphiphilic properties, so as to not only achieve a slow release effect of the collagenase, reduce the replacement frequency of the gelatinous dressing, and reduce the pain of the patient during dressing change, but also improve the activity and stability of the collagenase after embedding; in addition, the liposome has a lipophilic function, which can carry the collagenase to penetrate the sebum layer and enter the deep layer of the skin tissue, to achieve a targeted release effect, and is more conducive to skin tissue regeneration and wound healing.

[0033] Of course, any product implementing the present application does not necessarily need to achieve all the advantages described above. BRIEF DESCRIPTION OF DRAWINGS

[0034] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following will briefly introduce the drawings needed to be used in the description of the embodiments. Obviously, the drawings in the following description only show some of the embodiments of the present application, and all other drawings that can be obtained by those skilled in the art without creative efforts are within the scope of protection of the present application.

[0035] Figure 1 The UV analysis test chart of the modified carrageenan prepared for the embodiment 1 of the present application. DETAILED DESCRIPTION

[0036] The technical solutions in the embodiments of the present application will be described clearly and completely in the following with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative efforts are within the scope of protection of the present application.

[0037] Embodiment 1:

[0038] I. Preparation of modified carrageenan

[0039] A: 1g of nano-zinc oxide was added into a mixed solvent of methanol and N,N-dimethylformamide with a volume ratio of 3:1, and after ultrasonic dispersion, it was transferred to an oil bath pot, 2mL of sodium carboxyethylsilanetriol was added under stirring, the temperature in the oil bath pot was increased to 60℃, and the reaction was carried out under reflux for 12h in a nitrogen atmosphere. After stopping heating, the solid product was separated by centrifugation, and the solid product was washed to neutral with ethanol, hydrochloric acid and ionized water, and vacuum dried to obtain carboxyl-functionalized zinc oxide;

[0040] B: 0.2g of carboxyl-functionalized zinc oxide was added into N,N-dimethylformamide and ultrasonically dispersed for 30min, 10g of k-type carrageenan was added and mechanically stirred to be uniform, 0.1g of 4-dimethylaminopyridine and 0.8g of N,N-dicyclohexyl carbodiimide composite catalyst were further added, and after mixing, it was placed in a water bath pot at 20℃ and stirred for 36h. After the reaction was completed, the solid product was collected by centrifugation, washed, and vacuum dried to obtain modified carrageenan. The modified carrageenan was characterized by using a UV1800 type ultraviolet visible spectrophotometer, and the test results are shown in Figure 1 From Figure 1 it can be seen that the modified carrageenan has strong ultraviolet absorption at 380nm wavelength. Since the carrageenan itself does not have ultraviolet absorption performance, it can be inferred that the ultraviolet absorption is caused by the nano-zinc oxide in the modified carrageenan, which confirms that the carrageenan is successfully modified.

[0041] II. Preparation of collagenase liposomes

[0042] S1: 0.15 g collagenase was added into 100 mL phosphate buffer, stirred uniformly, configured into a buffer solution with a mass concentration of 1.5 g / L, to form an aqueous phase;

[0043] S2: 1 g of soybean lecithin and 0.5 g of cholesterol were dissolved in a mixed solvent of chloroform and ether with a volume ratio of 1:1, stirred uniformly to form an oil phase, the oil phase was added with the aqueous phase prepared in step S1, placed in a 37℃ water bath, and ultrasonically treated until a stable water / oil emulsion was formed, the organic solvent was removed by rotary evaporation, phosphate buffer solution was added into the rotary evaporation flask, and the rotation was continued for 6 h to obtain a collagenase liposome suspension, which was filtered using a microporous filter to remove large particle impurities, to obtain collagenase liposomes, 1 g of collagenase liposomes was added into 5 mL of petroleum ether and mixed uniformly, centrifuged, 2 mL of supernatant was collected, and the mass concentration of collagenase in the supernatant was measured, and the formula was used to calculate the encapsulation efficiency of collagenase, wherein T is the encapsulation efficiency of collagenase, M is the added amount of collagenase, mg; V is the volume of the collected liposome suspension, mL; C is the mass concentration of collagenase in the supernatant, mg / mL; through testing, C is 17.8 mg / mL, and calculation can obtain T of 76.3%, that is, the encapsulation efficiency of collagenase in the collagenase liposome is 76.3%.

[0044] III. Preparation of an antibacterial combined preparation containing egg white lysozyme

[0045] (1) 20 parts of modified carrageenan were added into 40 parts of deionized water, ultrasonically dispersed uniformly, 0.5 parts of egg white lysozyme was added, and stirred uniformly at a rotation speed of 80 rpm to obtain a premix ①;

[0046] (2) 0.01 parts of collagenase liposomes and 1 part of glycerol were placed in a blender and stirred uniformly to obtain a premix ②;

[0047] (3) The premix ② was added into the premix ①, stirred uniformly, and transferred into a 50℃ oil bath, and stirred sufficiently until a gel was formed, to obtain an antibacterial combined preparation containing egg white lysozyme.

[0048] Example 2:

[0049] Preparation of an antibacterial combined preparation containing egg white lysozyme

[0050] (1) 40 parts of modified carrageenan were added into 80 parts of deionized water, ultrasonically dispersed uniformly, 0.8 parts of egg white lysozyme was added, and stirred uniformly at a rotation speed of 90 rpm to obtain a premix ①;

[0051] (2) 0.04 parts of collagenase liposomes and 1.5 parts of glycerol were placed in a blender and stirred uniformly to obtain a premix ②;

[0052] (3) Add premix ② to premix ①, stir evenly, transfer to an oil bath at 55°C, and stir thoroughly until a gel is formed to obtain an antibacterial combination preparation containing egg white lysozyme.

[0053] The preparation method of modified carrageenan and collagenase liposomes is the same as that in Example 1.

[0054] Example 3:

[0055] Preparation of antibacterial combination preparation containing egg white lysozyme

[0056] (1) Add 50 parts of modified carrageenan to 100 parts of deionized water, disperse uniformly by ultrasonication, add 1 part of egg white lysozyme, and stir uniformly at a speed of 100 rpm to obtain premix ①;

[0057] (2) Place 0.05 parts of collagenase liposomes and 2 parts of propylene glycol in a blender and stir evenly to obtain a premix ②;

[0058] (3) Add premix ② to premix ①, stir evenly, transfer to an oil bath at 60°C, and stir thoroughly until a gel is formed to obtain an antibacterial combination preparation containing egg white lysozyme.

[0059] The preparation method of modified carrageenan and collagenase liposomes is the same as that in Example 1.

[0060] Comparative Example 1:

[0061] Preparation of antibacterial combination preparation containing egg white lysozyme

[0062] (1) Add 40 parts of k-type carrageenan to 80 parts of deionized water, disperse uniformly by ultrasonication, add 0.8 parts of egg white lysozyme, and stir uniformly at a speed of 90 rpm to obtain premix ①;

[0063] (2) Place 0.04 parts of collagenase liposomes and 1.5 parts of glycerol in a blender and stir evenly to obtain a premix ②;

[0064] (3) Add premix ② to premix ①, stir evenly, transfer to an oil bath at 55°C, and stir thoroughly until a gel is formed to obtain an antibacterial combination preparation containing egg white lysozyme.

[0065] The preparation method of collagenase liposomes is the same as that in Example 1.

[0066] Comparative Example 2:

[0067] Preparation of antibacterial combination preparation containing egg white lysozyme

[0068] (1) adding 40 parts of modified carrageenan into 80 parts of deionized water, uniformly dispersing by ultrasonic, adding 0.8 parts of egg white lysozyme, uniformly stirring at a rotating speed of 90 rpm to obtain a premix ①;

[0069] (2) adding 1.5 parts of glycerol into the premix ①, uniformly stirring, transferring into an oil bath pot at 55℃, fully stirring until forming a gel to obtain the bacteriostatic combination preparation containing egg white lysozyme.

[0070] The preparation method of the modified carrageenan is the same as that in Example 1.

[0071] Performance detection of the bacteriostatic combination preparations prepared in Examples 1-3 and Comparative Examples 1-2 of the present application:

[0072] a. Bacteriostatic performance detection: inoculating activated E. coli and S. aureus strains on ordinary broth agar plates, culturing at 37℃ for 24 h, diluting the bacterial bodies with nutrient broth, adding the bacteriostatic combination preparation, incubating for 1 h, taking 50 ul of each and coating on two plates, one group being cultured at 37℃ for 12 h in the dark, and the other group being cultured at 37℃ for 12 h under xenon lamp irradiation, observing the number of bacterial colonies, taking the blank control without adding the bacteriostatic combination preparation, calculating the bacteriostatic rate, and the test results are shown in the following table:

[0073] E. coli antibacterial rate (%) S. aureus antibacterial rate (%) Irradiated E. coli antibacterial rate (%) Irradiated S. aureus antibacterial rate (%) Example 1 45.3 91.9 98.5 99.1 Example 2 44.6 93.0 99.3 99.9 Example 3 42.9 92.4. 98.8 99.4 Comparative Example 1 38.1 86.8 39.0 86.2 Comparative Example 2 40.4 90.5 96.1 98.5

[0074] As shown in the above table, under the lightless condition, the bacteriostatic combination preparations prepared in Examples 1-3 and Comparative Examples 1-2 of the present application have a lower antibacterial rate on the gram-negative bacteria E. coli, but have a higher antibacterial rate on the gram-positive bacteria S. aureus. Under the light condition, the bacteriostatic combination preparations prepared in Examples 1-3 and Comparative Example 2 have a higher antibacterial rate on both E. coli and S. aureus, which is presumably because under the lightless condition, only the egg white lysozyme in the bacteriostatic combination preparation can play an antibacterial role, so the antibacterial rate on E. coli is lower. Under the light condition, the bacteriostatic combination preparation can have a synergistic effect of the photocatalytic antibacterial effect of the nano zinc oxide and the antibacterial effect of the egg white lysozyme, so that the antibacterial performance of the bacteriostatic combination preparation is stronger.

[0075] b. Wound healing promotion detection

[0076] Taking 60 mice of 20-25 g, randomly dividing them into six groups of ten, shaving a 2 cm x 2 cm area on the back of each mouse, injecting anesthetic, drawing a 1 cm diameter circular wound in the area, covering the wound surface with the bacteriostatic combination preparation, covering the wound surface of the control group with traditional cotton gauze dressing, observing the wound surface under the light condition, and calculating the wound healing rate by the formula The wound healing rate is calculated, wherein P is the wound healing rate, V0 is the original wound area, and V1 is the unhealed wound area, and the test results are as follows:

[0077]

[0078] From the above table, it can be seen that the bacteriostatic combined preparation prepared in Example 1-Example 3 of the present application has a significant effect on promoting wound healing in the early, middle and late stages. The bacteriostatic combined preparation prepared in Comparative Example 1 does not contain nano-zinc oxide, so its antibacterial performance is weak, which affects its effect on promoting wound healing. The bacteriostatic combined preparation prepared in Comparative Example 2 does not contain collagenase liposomes, and can only rely on antibacterial action to avoid bacterial infection of the wound, so it cannot promote rapid wound healing.

[0079] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "an example", "a specific example", or "some examples" and the like means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples. In addition, different embodiments or examples described in the present specification and the features of different embodiments or examples can be combined and combined by those skilled in the art without contradiction.

[0080] The above is only an example and description of the concept of the present application, and those skilled in the art can make various modifications or supplements or use similar ways to replace the described specific embodiments, as long as they do not deviate from the concept of the present application or exceed the scope defined by the present claims, which shall be within the protection scope of the present application.

Claims

1. An antibacterial combination preparation containing egg white lysozyme, characterized in that: The method comprises the following raw materials in parts by weight: 20-50 parts of modified carrageenan, 0.5-1 parts of egg white lysozyme, 0.01-0.05 parts of collagenase liposome, 1-2 parts of glycerol, and 40-100 parts of deionized water; The modified carrageenan is prepared by connecting nano zinc oxide and carrageenan through covalent bonds; The covalent bond connection comprises the following steps: A: Add nano zinc oxide to a mixed solvent of methanol and N,N-dimethylformamide, ultrasonically disperse, and transfer to an oil bath. Add sodium carboxyethylsilanol triol while stirring. Raise the temperature of the oil bath to 50-60°C. Reflux in a nitrogen atmosphere for 6-18 hours. Stop heating, allow the product to cool, and centrifuge to separate the solid product. Wash the solid product with ethanol, hydrochloric acid, and deionized water until neutral, and vacuum dry to obtain carboxyl-functionalized zinc oxide. B: Add carboxyl-functionalized zinc oxide to N,N-dimethylformamide, sonicate for 20-40 minutes, add carrageenan, mechanically stir until uniform, continue to add composite catalyst, mix thoroughly, place in a water bath at 15-35°C, and stir for 24-48 hours. After the reaction, collect the solid product by centrifugation, wash, and vacuum dry to obtain modified carrageenan; The composite catalyst is 4-dimethylaminopyridine and N,N-dicyclohexylcarbodiimide in a mass ratio of 1:8-10; The collagenase liposomes are prepared by using soybean lecithin, cholesterol, chloroform and ether as the oil phase and collagenase and phosphate buffer solution as the water phase, and are treated with water bath ultrasound. The synthesis process of the collagenase liposome comprises the following steps: S1: Add collagenase to phosphate buffer, stir evenly, and prepare a buffer solution to form an aqueous phase; S2: Dissolve soybean lecithin and cholesterol in a mixed solvent of chloroform and ether in a volume ratio of 1:1-2, stir evenly to form an oil phase, add the aqueous phase prepared in step S1 to the oil phase, place in a water bath at 35-38°C, and ultrasonically treat until a stable water / oil emulsion is formed, remove the organic solvent by rotary evaporation, add phosphate buffer solution to the rotary evaporator, and continue rotating for 4-8 hours to obtain a collagenase liposome suspension, filter using a microporous filter membrane to remove large particle impurities, and obtain collagenase liposomes.

2. The antibacterial combination preparation containing egg white lysozyme according to claim 1, characterized in that: The carrageenan is k-type carrageenan.

3. The antibacterial combination preparation containing egg white lysozyme according to claim 1, characterized in that: In step A, the volume ratio of methanol to N,N-dimethylformamide is 3-4:

1.

4. The antibacterial combination preparation containing egg white lysozyme according to claim 1, characterized in that: In step S1, the mass concentration of the buffer solution is 1-2 g / L.

5. The antibacterial combination preparation containing egg white lysozyme according to claim 1, characterized in that: In step S2, the mass ratio of the soybean lecithin to cholesterol is 1:0.2-0.

6.

6. A method for synthesizing the antibacterial combination preparation containing egg white lysozyme according to any one of claims 1 to 5, characterized in that: The synthesis method comprises the following steps: (1) Add the modified carrageenan according to claim 1 to deionized water, disperse it evenly by ultrasonication, add egg white lysozyme, and stir evenly at a speed of 80-100 rpm to obtain a premix ①; (2) Place collagenase liposomes and glycerol in a blender and mix well to obtain a premix ②; (3) Add premix ② to premix ①, stir evenly, transfer to an oil bath at 50-60°C, and stir thoroughly until a gel is formed to obtain an antibacterial combination preparation containing egg white lysozyme.

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