Natural compound efficient antibacterial preservative and preparation process thereof
By using natural compound high-efficiency antibacterial preservatives in antibacterial preservatives, propolis silk composites are prepared by using the combination technology of folic acid, TG enzyme and NHS and carboxymethyl chitosan, the propolis silk composites are prepared, which solves the problem of insufficient antibacterial preservatives in meat, and achieves better antibacterial preservative effects and extended food shelf life.
Patent Information
- Application Number
- CN202411797649.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-09
- Publication Date
- 2025-02-28
AI Technical Summary
Existing antibacterial preservatives are difficult to effectively antibacterial and antibacterial in meat, resulting in insufficient antiseptic capacity.
The preparation process of natural compound high-efficiency antibacterial preservatives is adopted. By combining folic acid, TG enzyme and NHS with carboxymethyl chitosan, a load solution is prepared, and propolis and silk are prepared into a composite copolymer through the load solution to form a propolis silk composite.
It improves the antibacterial and antiseptic capacity of food, extends the shelf life of food, and reduces the amount of preservatives added to food, making it healthier.
Smart Images
Figure CN119522931A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of preservatives, and in particular to a natural compound high-efficiency antibacterial preservative and a preparation process thereof. Background Art
[0002] Antimicrobial preservatives are an indispensable type of additive in many industries such as food, medicine, and cosmetics. Their main function is to prevent the growth of microorganisms, extend the shelf life of products, and ensure product safety and quality.
[0003] Common antibacterial preservatives include chemical preservatives such as benzoic acid and its salts, sorbic acid and its salts, and natural preservatives such as vitamin C, tea polyphenols and garlic extract. These preservatives are mostly used in beverages and dairy products, while the main preservatives for meat products are nitrites and film-forming liquids prepared from various natural materials. The antibacterial principle of this film-forming liquid is to isolate the air, thereby preventing the reproduction of microorganisms, but the bactericidal ability is insufficient. Therefore, when the meat itself carries a small amount of microorganisms, this preservative in the form of a film-forming liquid is difficult to be antibacterial, resulting in insufficient antiseptic ability. Therefore, the present invention provides a natural composite high-efficiency antibacterial preservative and a preparation process thereof to solve the above-mentioned existing problems. Summary of the invention
[0004] In view of the shortcomings of the prior art, the object of the present invention is to provide a natural compound high-efficiency antibacterial preservative and a preparation process thereof.
[0005] A preparation process of a natural compound high-efficiency antibacterial preservative comprises the following steps: S1: Preparation of loading solution The folic acid, TG enzyme and NHS are mixed and added to anhydrous DMSO to obtain a mixed solution, and the mixed solution is then added to a PBS buffer of carboxymethyl chitosan to prepare a carboxymethyl chitosan complex, and the carboxymethyl chitosan complex is dissolved in deionized water to obtain a composite solution, and the pH value of the composite solution is adjusted, and stirred to obtain a loading solution; S2: Preparation of silk solution The silk is added into a sodium carbonate solution to prepare degummed silk, a malic acid aqueous solution is weighed, and then the degummed silk is added, and the degummed silk is centrifuged, and the supernatant is taken after centrifugation to obtain a silk solution; S3: Preparation of natural compound high-efficiency antibacterial preservatives Add NaOH solution to deionized water, then add propolis powder until the propolis powder is dissolved to obtain a propolis solution, then add an injection into the load liquid, adjust the pH value, stir to obtain a propolis suspension, add the silk solution dropwise to the propolis suspension, stir, ultrasonicate, centrifuge, collect solids to obtain a propolis silk complex, mix the propolis silk complex, citric acid and acetic acid to obtain a natural composite high-efficiency antibacterial preservative.
[0006] Furthermore, step S1 of preparing a loading liquid comprises the following steps: S1.1: Folic acid, TG enzyme and NHS are mixed in a molar ratio of 1: (2-3): (2-3) and added to 15-20 parts by volume of anhydrous DMSO, stirred in the dark at 25-28°C for 8-9 hours to obtain a mixed solution, and then the mixed solution is added to 1% carboxymethyl chitosan in PBS buffer and stirred for 16-18 hours. After stirring, the pH value of the system is adjusted to 9-9.5 with NaOH solution, dialyzed with PBS solution for 70-72 hours, and then dialyzed with deionized water for 70-72 hours, and freeze-dried to obtain a carboxymethyl chitosan complex; S1.2: dissolving the carboxymethyl chitosan complex in deionized water at a solid-liquid ratio of 2-3 mg / mL and stirring for 20-24 hours to obtain a composite solution; S1.3: Adjust the pH value of 20-25 parts by volume of the composite solution to 12-12.5 with NaOH solution, and stir at 1000-1200 r / min for 1-1.5 h to obtain a loading solution.
[0007] Furthermore, step S2 of preparing the silk solution comprises the following steps: S2.1: adding 7.5-9 parts by weight of silk to 10-11 parts by weight of sodium carbonate solution, heating to 100-110° C., stirring at a speed of 30-50 r / min, stirring for 1-1.5 hours, then filtering to remove the filtrate, standing for 24-28 hours to air-dry, to obtain degummed silk; S2.2: Weigh 32-35 parts by mass of malic acid aqueous solution, add 5-8 parts by mass of degummed silk, heat to 60-65°C and stir for 4-4.5 hours, then dialyze for 40-44 hours, and then concentrate to one quarter of the original volume. Centrifuge the concentrated solution at 0-4°C, 4000-4500r / min for 40-45min, take the supernatant after centrifugation to obtain a silk solution.
[0008] Furthermore, step S3 prepares a natural compound high-efficiency antibacterial preservative, comprising the following steps: S3.1: Add 3-3.5M NaOH solution to 10-15 parts by mass of deionized water to adjust the pH value to 12-13, then add 2-3 parts by mass of propolis powder until the propolis powder is dissolved to obtain a propolis solution, then add the injection into the loading liquid, the mass ratio of the propolis solution to the loading liquid is (1-3): (5-10), and adjust the pH value to 6.5-7 with 1-1.5M HCl solution, stir at 1000-1200 r / min for 1-1.5h to obtain a propolis suspension; S3.2: Take 1-2 parts by volume of silk solution and add dropwise to 9-10 parts by volume of propolis suspension, stir at 800-900 r / min for 5-10 min, use cell disruptor to sonicate for 30-40 s, then place in fume hood for 12-14 h, centrifuge at 12000-13000 r / min for 15-20 min, collect solids, wash with deionized water 2-3 times, and obtain propolis silk complex; S3.3: Mix 1-3 parts by mass of propolis silk complex, 0.5-1 parts by mass of citric acid and 1-2 parts by mass of acetic acid for 10-15 minutes to obtain a natural composite high-efficiency antibacterial preservative.
[0009] Furthermore, the concentration of the sodium carbonate solution in step S2.1 is 0.02M.
[0010] Furthermore, the ultrasonic mode of the cell disruptor in step S3.2 is ultrasonic 2s on and 2s off.
[0011] A natural compound high-efficiency antibacterial preservative is prepared by the preparation process of the natural compound high-efficiency antibacterial preservative.
[0012] Compared with the prior art, the present invention has at least the following beneficial effects: 1. The present invention prepares a loading liquid by compounding folic acid, TG enzyme and NHS and mixing them with carboxymethyl chitosan. TG enzyme is a natural enzyme that can catalyze the cross-linking reaction between protein molecules to form a more stable structure. This cross-linking effect can enhance the texture and stability of food. NHS is an activating reagent that can activate the carboxyl group to make it easier to react with the amine group to form a stable amide bond. The TG enzyme is activated by folic acid, so that the TG enzyme and NHS play a combined role, thereby improving the efficiency and stability of protein cross-linking, so that the loading liquid can load propolis and silk at the same time to form a propolis silk complex.
[0013] 2. The present invention prepares a composite copolymer of propolis and silk through a loading liquid. Silk fibers can be used as a skeleton so that propolis can be maintained in the loading liquid in a three-dimensional structure. Propolis has good antibacterial and antiseptic functions. Propolis in a three-dimensional structure can contact food as much as possible, thereby improving the antibacterial and antiseptic capabilities and extending the shelf life of food.
[0014] 3. After the silk is degummed once with sodium carbonate, the degummed silk is further degummed by immersing it in malic acid. Moreover, the malic acid can be combined with sericin for hydrolysis to generate amino antioxidants. The antioxidants can neutralize free radicals and slow down the oxidation rate. The antioxidant components obtained by changing the environment with malic acid have better antiseptic effect after being added, which can reduce the amount of preservatives added in food and is healthier. BRIEF DESCRIPTION OF THE DRAWINGS
[0015] The accompanying drawings, which are incorporated herein and constitute a part of the specification, illustrate embodiments of the present disclosure and, together with the description, further serve to explain the principles of the present disclosure and to enable those skilled in the relevant art to make and use the present disclosure.
[0016] Figure 1 The present invention is a flow chart of the preparation process of the natural compound high-efficiency antibacterial preservative used in the embodiments of the present invention. DETAILED DESCRIPTION
[0017] The following is a detailed description of a natural compound high-efficiency antibacterial preservative and its preparation process provided by the present invention in conjunction with the accompanying drawings and specific embodiments. At the same time, it is explained here that in order to make the embodiments more detailed, the following embodiments are the best and preferred embodiments, and those skilled in the art may also adopt other alternatives to implement some known technologies; and the accompanying drawings are only for a more specific description of the embodiments, and are not intended to specifically limit the present invention.
[0018] Embodiment 1: A preparation process of a natural compound high-efficiency antibacterial preservative, such as Figure 1 As shown, the following steps are included: S1: Preparation of loading solution S1.1: Folic acid, TG enzyme and NHS were mixed in a molar ratio of 1:2:2 and added to 15 parts by volume of anhydrous DMSO. The mixture was stirred at 25°C in the dark for 8 hours to obtain a mixed solution. The mixed solution was then added to 1% carboxymethyl chitosan in PBS buffer and stirred for 16 hours. After stirring, the pH value of the system was adjusted to 9 with NaOH solution, dialyzed with PBS solution for 70 hours, and then dialyzed with deionized water for 70 hours, and freeze-dried to obtain a carboxymethyl chitosan complex. S1.2: dissolving the carboxymethyl chitosan complex in deionized water at a solid-liquid ratio of 2 mg / mL and stirring for 20 h to obtain a composite solution; S1.3: Adjust the pH value of 20 parts by volume of the composite solution to 12 with NaOH solution, and stir at 1000 r / min for 1 h to obtain a loading solution.
[0019] S2: Preparation of silk solution S2.1: 7.5 parts by weight of silk were added to 10 parts by weight of a sodium carbonate solution having a concentration of 0.02 M, and the mixture was heated to 100° C. and stirred at a speed of 30 r / min for 1 h, and then the filtrate was removed by filtration, and the mixture was allowed to stand for 24 h to be air-dried to obtain degummed silk; S2.2: Weigh 32 parts by mass of malic acid aqueous solution, add 5 parts by mass of degummed silk, heat to 60°C and stir for 4 hours, then dialyze for 40 hours, and then concentrate to one-fourth of the original volume. Centrifuge the concentrated solution at 0°C and 4000 r / min for 40 minutes, take the supernatant after centrifugation, and obtain a silk solution; S3: Preparation of natural compound high-efficiency antibacterial preservatives S3.1: Add 3M NaOH solution to 10 parts by mass of deionized water to adjust the pH value to 12, then add 2 parts by mass of propolis powder until the propolis powder is dissolved to obtain a propolis solution, then add the injection into the loading liquid, the mass ratio of the propolis solution to the loading liquid is 1:5, and adjust the pH value to 6.5 with 1M HCl solution, stir at 1000 r / min for 1 hour to obtain a propolis suspension; S3.2: Take 1 volume of silk solution and add it dropwise to 9 volume of propolis suspension, stir at 800 r / min for 5 min, use cell disruptor to sonicate for 30 s, the sonication mode of cell disruptor is 2 s on and 2 s off, then place in fume hood to stand for 12 h, centrifuge at 12000 r / min for 15 min, collect solids, wash twice with deionized water, and obtain propolis silk complex; S3.3: Mix 1 part by mass of propolis silk composite, 0.5 part by mass of citric acid and 1 part by mass of acetic acid for 10 minutes to obtain a natural composite high-efficiency antibacterial preservative.
[0020] Embodiment 2: A preparation process of a natural compound high-efficiency antibacterial preservative, such as Figure 1 As shown, the following steps are included: S1.1: Folic acid, TG enzyme and NHS were mixed in a molar ratio of 1:2:2 and added to 15 parts by volume of anhydrous DMSO. The mixture was stirred at 28°C in the dark for 9 hours to obtain a mixed solution. The mixed solution was then added to 1% carboxymethyl chitosan in PBS buffer and stirred for 18 hours. After stirring, the pH value of the system was adjusted to 9.5 with NaOH solution, dialyzed with PBS solution for 72 hours, and then dialyzed with deionized water for 72 hours, and freeze-dried to obtain a carboxymethyl chitosan complex. S1.2: dissolving the carboxymethyl chitosan complex in deionized water at a solid-liquid ratio of 2 mg / mL and stirring for 24 h to obtain a composite solution; S1.3: Adjust the pH value of 20 parts by volume of the composite solution to 12.5 with NaOH solution, and stir at 1200 r / min for 1.5 h to obtain a loading solution.
[0021] S2: Preparation of silk solution S2.1: 7.5 parts by weight of silk was added to 10 parts by weight of sodium carbonate solution with a concentration of 0.02 M, and heated to 110° C., while stirring at a speed of 50 r / min for 1.5 h, and then the filtrate was filtered out, and the mixture was allowed to stand for 28 h and air-dried to obtain degummed silk; S2.2: Weigh 32 parts by mass of malic acid aqueous solution, add 5 parts by mass of degummed silk, heat to 65°C and stir for 4.5 hours, then dialyze for 44 hours, and then concentrate to one-fourth of the original volume. Centrifuge the concentrated solution at 4°C and 4000 r / min for 45 minutes, and take the supernatant after centrifugation to obtain a silk solution; S3: Preparation of natural compound high-efficiency antibacterial preservatives S3.1: Add 3M NaOH solution to 10 parts by mass of deionized water to adjust the pH value to 13, then add 2 parts by mass of propolis powder until the propolis powder is dissolved to obtain a propolis solution, then add the injection into the loading liquid, the mass ratio of the propolis solution to the loading liquid is 1:5, and adjust the pH value to 7 with 1M HCl solution, stir at 1200 r / min for 1.5 hours to obtain a propolis suspension; S3.2: Take 1 volume of silk solution and add it dropwise to 9 volume of propolis suspension, stir at 900 r / min for 10 min, use cell disruptor to sonicate for 30 s, the sonication mode of cell disruptor is 2 s on and 2 s off, then place in fume hood to stand for 12 h, centrifuge at 13000 r / min for 20 min, collect solids, wash with deionized water twice, and obtain propolis silk complex; S3.3: Mix 1 part by mass of propolis silk composite, 0.5 part by mass of citric acid and 1 part by mass of acetic acid for 15 minutes to obtain a natural composite high-efficiency antibacterial preservative.
[0022] Embodiment 3: A preparation process of a natural compound high-efficiency antibacterial preservative, such as Figure 1 As shown, the following steps are included: S1.1: Folic acid, TG enzyme and NHS were mixed in a molar ratio of 1:3:3 and added to 20 parts by volume of anhydrous DMSO. The mixture was stirred at 25°C in the dark for 8 hours to obtain a mixed solution. The mixed solution was then added to 1% carboxymethyl chitosan in PBS buffer and stirred for 16 hours. After stirring, the pH value of the system was adjusted to 9 with NaOH solution, dialyzed with PBS solution for 70 hours, and then dialyzed with deionized water for 70 hours, and freeze-dried to obtain a carboxymethyl chitosan complex. S1.2: dissolving the carboxymethyl chitosan complex in deionized water at a solid-liquid ratio of 3 mg / mL and stirring for 20 h to obtain a composite solution; S1.3: Adjust the pH value of 25 parts by volume of the composite solution to 12 with NaOH solution, and stir at 1000 r / min for 1 h to obtain a loading solution.
[0023] S2: Preparation of silk solution S2.1: 9 parts by weight of silk were added to 11 parts by weight of a sodium carbonate solution having a concentration of 0.02 M, and heated to 100° C., while stirring at a speed of 30 r / min for 1 h, and then the filtrate was removed by filtration, and the mixture was allowed to stand for 24 h to be air-dried to obtain degummed silk; S2.2: Weigh 35 parts by weight of malic acid aqueous solution, add 8 parts by weight of degummed silk, heat to 60°C and stir for 4 hours, then dialyze for 40 hours, and then concentrate to one-fourth of the original volume. Centrifuge the concentrated solution at 0°C and 4000 r / min for 40 minutes, take the supernatant after centrifugation, and obtain a silk solution; S3: Preparation of natural compound high-efficiency antibacterial preservatives S3.1: Add 3M NaOH solution to 10 parts by mass of deionized water to adjust the pH value to 12, then add 3 parts by mass of propolis powder until the propolis powder is dissolved to obtain a propolis solution, then add the injection into the loading liquid, the mass ratio of the propolis solution to the loading liquid is 3:10, and adjust the pH value to 6.5 with 1M HCl solution, stir at 1000 r / min for 1 hour to obtain a propolis suspension; S3.2: Take 2 parts by volume of silk solution and add dropwise to 10 parts by volume of propolis suspension, stir at 800 r / min for 5 min, use cell disruptor to sonicate for 30 s, the sonication mode of cell disruptor is 2 s on and 2 s off, then place in fume hood to stand for 12 h, centrifuge at 12000 r / min for 15 min, collect solids, wash twice with deionized water, and obtain propolis silk complex; S3.3: Mix 3 parts by mass of propolis silk composite, 1 part by mass of citric acid and 2 parts by mass of acetic acid for 10 minutes to obtain a natural composite high-efficiency antibacterial preservative.
[0024] Comparative Example 1: Comparative Example 1 is a commercially available chemical preservative, potassium sorbate.
[0025] Comparative Example 2: Comparative Example 2 is a natural preservative citric acid preservative available on the market.
[0026] Comparative Example 3: Compared with Example 1, the difference of Comparative Example 3 is that folic acid is not added in step S1.1, specifically: "S1.1: Mix TG enzyme and NHS in a molar ratio of 1:1 and add to 15 parts by volume of anhydrous DMSO, stir in the dark at 25°C for 8 hours to obtain a mixed solution, and then add the mixed solution to 1% carboxymethyl chitosan in PBS buffer, and continue stirring for 16 hours. After stirring, adjust the pH value of the system to 9 with NaOH solution, dialyze with PBS solution for 70 hours, and then dialyze with deionized water for 70 hours, and freeze-dry to obtain a carboxymethyl chitosan complex", and the other steps remain unchanged. The prepared natural composite high-efficiency antibacterial preservative is recorded as Comparative Example 3.
[0027] Comparative Example 4: Compared with Example 1, the difference of Comparative Example 4 is that TG enzyme is not added in step S1.1, specifically: "S1.1: Mix folic acid and NHS in a molar ratio of 1:2 and add to 15 parts by volume of anhydrous DMSO, stir in the dark at 25°C for 8 hours to obtain a mixed solution, and then add the mixed solution to 1% carboxymethyl chitosan in PBS buffer, and continue stirring for 16 hours. After stirring, adjust the pH value of the system to 9 with NaOH solution, dialyze with PBS solution for 70 hours, and then dialyze with deionized water for 70 hours, and freeze-dry to obtain a carboxymethyl chitosan complex", and the other steps remain unchanged. The prepared natural composite high-efficiency antibacterial preservative is recorded as Comparative Example 4.
[0028] Comparative Example 5: Compared with Example 1, the difference of Comparative Example 5 is that NHS is not added in step S1.1, specifically: "S1.1: Mix folic acid and TG enzyme in a molar ratio of 1:2 and add to 15 parts by volume of anhydrous DMSO, stir in the dark at 25°C for 8 hours to obtain a mixed solution, and then add the mixed solution to 1% carboxymethyl chitosan in PBS buffer, and continue stirring for 16 hours. After stirring, adjust the pH value of the system to 9 with NaOH solution, dialyze with PBS solution for 70 hours, and then dialyze with deionized water for 70 hours, and freeze-dry to obtain a carboxymethyl chitosan complex", and the other steps remain unchanged. The prepared natural composite high-efficiency antibacterial preservative is recorded as Comparative Example 5.
[0029] Comparative Example 6: Compared with Example 1, the difference of Comparative Example 6 is that no silk solution is added in step S3.2, specifically: "S3.2: taking 10 parts by volume of propolis suspension and stirring at 800 r / min for 5 min, ultrasonicating with a cell disruptor for 30 s, the ultrasonic mode of the cell disruptor is ultrasonicating for 2 s and stopping for 2 s, then placing in a fume hood for 12 h, and then centrifuging at 12000 r / min for 15 min, collecting solids, and washing with deionized water twice to obtain a propolis complex; S3.3: Mix 3 parts by mass of propolis complex, 1 part by mass of citric acid and 2 parts by mass of acetic acid for 10 minutes to obtain a natural compound high-efficiency antibacterial preservative. The remaining steps remain unchanged. The prepared natural compound high-efficiency antibacterial preservative is recorded as Comparative Example 6.
[0030] Comparative Example 7: Compared with Example 1, the difference of Comparative Example 7 is that step S3.1 is not performed, and no propolis suspension is added in step S3.2, specifically: "S3.2: 2 volumes of silk solution are taken and stirred at 800 r / min for 5 min, and then ultrasonicated for 30 s using a cell disruptor, the ultrasonic mode of the cell disruptor is ultrasonicated for 2 s and stopped for 2 s, then placed in a fume hood for 12 h, and then centrifuged at 12000 r / min for 15 min, the solid is collected, and washed twice with deionized water to obtain a propolis complex; S3.3: Mix 3 parts by mass of propolis complex, 1 part by mass of citric acid and 2 parts by mass of acetic acid for 10 minutes to obtain a natural compound high-efficiency antibacterial preservative. The remaining steps remain unchanged. The prepared natural compound high-efficiency antibacterial preservative is recorded as Comparative Example 7.
[0031] Comparative Example 8: Compared with Example 1, the difference of Comparative Example 8 is that no malic acid aqueous solution is added in step S2.2, specifically: "S2.2: Heat 5 parts by mass of degummed silk to 60°C and stir for 4 hours, then dialyze for 40 hours, and then concentrate to one quarter of the original volume, centrifuge the concentrated solution at 0°C, 4000r / min for 40 minutes, take the supernatant after centrifugation to obtain a silk solution", and the other steps remain unchanged, and the prepared natural composite high-efficiency antibacterial preservative is recorded as Comparative Example 8.
[0032] The natural composite high-efficiency antibacterial preservatives prepared in Examples 1-3 and Comparative Examples 1-8 were experimentally tested, and the antibacterial effects were tested using the filter paper method and the dilution plate colony counting method, and the size of the microbial inhibition zone (unit: mm) was measured, as shown in Table 1.
[0033] Table 1 Escherichia coli Staphylococcus aureus Bacillus subtilis Example 1 15.4 16.7 18.4 Example 2 14.7 16.1 18.7 Example 3 14.8 16.3 18.8 Comparative Example 1 11.2 13.1 13.9 Comparative Example 2 10.9 12.7 13.5 Comparative Example 3 12.3 13.6 12.2 Comparative Example 4 12.5 13.8 12.4 Comparative Example 5 12.6 11.2 12.7 Comparative Example 6 11.2 10.3 11.9 Comparative Example 7 8.9 9.2 10.8 Comparative Example 8 11.7 10.1 11.4 The diameters of the inhibition zones of Escherichia coli, Staphylococcus aureus and Bacillus subtilis in Examples 1-3 are all larger than those in the comparative examples, indicating that the examples have better antibacterial effects. Comparative Examples 1 and 2 are commercially available preservatives, and it can be seen that the antibacterial effects of the preservatives of the present invention are better than those of commercially available products and have better antiseptic capabilities.
[0034] Comparative Examples 3-5 have different antibacterial effects on different bacterial species, but are all lower than the examples. It can be seen that the combined use of folic acid, TG enzyme and NHS enables the loading liquid to load propolis and silk at the same time, thereby achieving better antibacterial and preservative effects, while the single use does not have this effect, and the antibacterial ability decreases.
[0035] The antibacterial effects of Comparative Examples 6 and 7 further decreased, indicating that the complex composed of propolis and silk can better improve the antibacterial and antiseptic capabilities, especially when propolis is not added, the antibacterial effect is significantly reduced, indicating that propolis is the main antibacterial component.
[0036] It can be seen from Comparative Example 8 that the malic acid aqueous solution also plays a role in improving the antibacterial effect, but the effect is not as significant as that of propolis.
[0037] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical ideas disclosed by the present invention shall still be covered by the claims of the present invention.
Claims
1. A preparation process of a natural composite high-efficiency antibacterial preservative, characterized in that: The steps include: S1: Preparation of loading solution The folic acid, TG enzyme and NHS are mixed and added to anhydrous DMSO to obtain a mixed solution, and the mixed solution is then added to a PBS buffer of carboxymethyl chitosan to prepare a carboxymethyl chitosan complex, and the carboxymethyl chitosan complex is dissolved in deionized water to obtain a composite solution, and the pH value of the composite solution is adjusted, and stirred to obtain a loading solution; S2: Preparation of silk solution The silk is added into a sodium carbonate solution to prepare degummed silk, a malic acid aqueous solution is weighed, and then the degummed silk is added, and the degummed silk is centrifuged, and the supernatant is taken after centrifugation to obtain a silk solution; S3: Preparation of natural compound high-efficiency antibacterial preservatives Add NaOH solution to deionized water, then add propolis powder until the propolis powder is dissolved to obtain a propolis solution, then add an injection into the load liquid, adjust the pH value, stir to obtain a propolis suspension, add the silk solution dropwise to the propolis suspension, stir, ultrasonicate, centrifuge, collect solids to obtain a propolis silk complex, mix the propolis silk complex, citric acid and acetic acid to obtain a natural composite high-efficiency antibacterial preservative.
2. The preparation process of a natural composite high-efficiency antibacterial preservative according to claim 1, characterized in that: Step S1: preparing a loading liquid, comprising the following steps: S1.1: Folic acid, TG enzyme and NHS are mixed in a molar ratio of 1: (2-3): (2-3) and added to 15-20 parts by volume of anhydrous DMSO, stirred in the dark at 25-28°C for 8-9 hours to obtain a mixed solution, and then the mixed solution is added to 1% carboxymethyl chitosan in PBS buffer and stirred for 16-18 hours. After stirring, the pH value of the system is adjusted to 9-9.5 with NaOH solution, dialyzed with PBS solution for 70-72 hours, and then dialyzed with deionized water for 70-72 hours, and freeze-dried to obtain a carboxymethyl chitosan complex; S1.2: dissolving the carboxymethyl chitosan complex in deionized water at a solid-liquid ratio of 2-3 mg / mL and stirring for 20-24 hours to obtain a composite solution; S1.3: Adjust the pH value of 20-25 parts by volume of the composite solution to 12-12.5 with NaOH solution, and stir at 1000-1200 r / min for 1-1.5 h to obtain a loading solution.
3. The preparation process of a natural composite high-efficiency antibacterial preservative according to claim 2, characterized in that: Step S2: preparing a silk solution, comprising the following steps: S2.1: adding 7.5-9 parts by weight of silk to 10-11 parts by weight of sodium carbonate solution, heating to 100-110° C., stirring at a speed of 30-50 r / min, stirring for 1-1.5 hours, then filtering to remove the filtrate, standing for 24-28 hours to air-dry, to obtain degummed silk; S2.2: Weigh 32-35 parts by mass of malic acid aqueous solution, add 5-8 parts by mass of degummed silk, heat to 60-65°C and stir for 4-4.5 hours, then dialyze for 40-44 hours, and then concentrate to one quarter of the original volume. Centrifuge the concentrated solution at 0-4°C, 4000-4500r / min for 40-45min, take the supernatant after centrifugation to obtain a silk solution.
4. The preparation process of a natural composite high-efficiency antibacterial preservative according to claim 3, characterized in that: Step S3 prepares a natural compound high-efficiency antibacterial preservative, comprising the following steps: S3.1: Add 3-3.5M NaOH solution to 10-15 parts by mass of deionized water to adjust the pH value to 12-13, then add 2-3 parts by mass of propolis powder until the propolis powder is dissolved to obtain a propolis solution, then add the injection into the loading liquid, the mass ratio of the propolis solution to the loading liquid is (1-3): (5-10), and adjust the pH value to 6.5-7 with 1-1.5M HCl solution, stir at 1000-1200 r / min for 1-1.5h to obtain a propolis suspension; S3.2: Take 1-2 parts by volume of silk solution and add dropwise to 9-10 parts by volume of propolis suspension, stir at 800-900 r / min for 5-10 min, use cell disruptor to sonicate for 30-40 s, then place in fume hood for 12-14 h, centrifuge at 12000-13000 r / min for 15-20 min, collect solids, wash with deionized water 2-3 times, and obtain propolis silk complex; S3.3: Mix 1-3 parts by mass of propolis silk complex, 0.5-1 parts by mass of citric acid and 1-2 parts by mass of acetic acid for 10-15 minutes to obtain a natural composite high-efficiency antibacterial preservative.
5. The preparation process of a natural composite high-efficiency antibacterial preservative according to claim 3, characterized in that: The concentration of the sodium carbonate solution in step S2.1 is 0.02M.
6. The preparation process of a natural composite high-efficiency antibacterial preservative according to claim 4, characterized in that: The ultrasonic mode of the cell disruptor in step S3.2 is ultrasonic 2s and stop 2s.
7. A natural compound high-efficiency antibacterial preservative, which is prepared by the preparation process of the natural compound high-efficiency antibacterial preservative according to any one of claims 1 to 6.