CN113248756B - Slow-release antibacterial rice bran protein composite membrane and preparation method and application thereof - Google Patents
Slow-release antibacterial rice bran protein composite membrane and preparation method and application thereof Download PDFInfo
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
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- C08J5/18—Manufacture of films or sheets
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- C08H—DERIVATIVES OF NATURAL MACROMOLECULAR COMPOUNDS
- C08H1/00—Macromolecular products derived from proteins
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Abstract
The invention discloses a sustained-release antibacterial rice bran protein composite membrane and a preparation method and application thereof, wherein the composite membrane is prepared by the following method: the rice bran protein is succinylated and modified by succinic anhydride to obtain rice bran acylated protein, the rice bran acylated protein and carboxymethyl chitosan are used as main film base material, glycerol is used as plasticizer, Tween 20 is used as dispersant, and simultaneously the added rice bran nanogel loaded with curcumin are combined according to a specific proportion, and the raw material components are mutually synergistic, so that the composite film has excellent antibacterial property and mechanical property, and the sustained and controlled release effect of the antibacterial agent in the composite film is realized. As the film base material is a food-borne raw material, the film has the characteristics of edibility, environmental protection, low energy consumption and the like, is an effective novel food packaging film with wide application prospect, and has profound influence on the development of the field of food packaging.
Description
Technical Field
The invention relates to the technical field of food packaging, in particular to a sustained-release antibacterial rice bran protein composite membrane and a preparation method and application thereof.
Background
At present, most of food packages are non-degradable plastic products, and have great challenges for living environment. With the push of the international plastic banning law, degradable packaging materials and natural non-toxic edible packaging materials become research hotspots in the field of food packaging. Many researchers began to select natural polymer materials as film-forming base materials to prepare edible films to replace traditional plastic packages: on one hand, the research for preparing safe, edible, environment-friendly and degradable biological films by taking natural high molecular materials as base materials is gradually started; on the other hand, the introduction of antibacterial packages has made food quality more reliable, and the transfer of antibacterial agents originally added to foods to food packaging materials has become the mainstream of research in the field of food packaging. Therefore, the antibacterial biodegradable material with the sustained and controlled release characteristics is researched and developed, and is applied to food packaging to realize real-time and long-acting antibacterial, so that a new idea can be provided for novel food antibacterial packaging.
The protein-based antibacterial film is a food preservative film which is prepared by adding an antibacterial agent, a cross-linking agent, an emulsifying agent and the like into protein and has the functions of barrier, antibiosis, degradability and packaging. However, the special matrix composition and structure of the protein film cause the protein film to have poor water resistance, and the controlled release function of the additive is poor, so that the application of the protein film in the field of food packaging is limited. Research shows that the protein can be prepared into a composite membrane with polysaccharide, lipid, starch and the like through physical blending, chemical crosslinking and electrostatic and hydrophobic interaction, and the composite membrane can be beneficial to protein membranes and mechanical properties and physicochemical properties, such as good flexibility, tensile strength, air permeability, moisture permeability, slow release property and the like, and has wide application prospects and profound research significance. The carboxymethyl chitosan is prepared by introducing carboxymethyl (-CH) on chitosan molecular skeleton 2 COOH) to a polymer having an-NH-group 2 and-COOH) product, has extremely high solubility in water, stable chemical properties, good moisture absorption and retention and film forming properties, but the carboxymethyl chitosan film has poor mechanical properties and strong water solubility, so that the carboxymethyl chitosan film has limited application as food packaging, and can form a stable structure by combining with rice bran protein to further improve the mechanical strength, stability and other properties of the composite film.
Secondly, the interaction between membrane matrixes also affects the water resistance and antibacterial property of the protein membrane. Antibacterial agents, cross-linking agents, emulsifiers and the like in the protein film influence the water resistance and antibacterial property of the protein film due to the interaction of intermolecular static electricity, hydrophobicity, hydration, hydrogen bonds and the like. The reason may be that the poor compatibility between these substances and proteins results in large particle diameters in the deposition solution, which affects the interfacial adhesion and compactness of the membrane. In order to further reduce the particle diameter of the film-forming solution and improve the interaction among the components, researchers prepare the matrix components into nanoparticles, and then adopt a blending mode to carry out casting film forming, so that the water resistance of the composite film can be obviously improved.
With respect to sustained and controlled release techniques, researchers have suggested that controlled release techniques such as nanocapsules may delay the release of antibacterial agents such as potassium sorbate. The contact of the antibacterial component in the nano capsule and food needs to be subjected to secondary migration, namely, the antibacterial component migrates into the membrane matrix from the inside of the nano capsule and then migrates into the food from the membrane matrix, so that the sustained and controlled release effect is realized. Researches find that the rapeseed protein nanogel has good water absorption and swelling capacity, and can resist different pH values and ionic strengths, and freeze-drying and diluting capacities. According to the characteristics of water absorption swelling and antibacterial agent controlled release of the nano antibacterial gel, the rice bran protein is prepared into the nano gel and then is mixed with the rice bran protein, so that the water resistance and antibacterial property of the acylated rice bran protein basement membrane are hopefully and synergistically enhanced.
At present, researches on the preparation of the antibacterial film by combining modified rice bran protein and nanogel are few at home and abroad, and the application of the antibacterial film in food packaging materials is yet to be developed. The research of using the byproduct rice bran protein in rice processing for the sustained and controlled release antibacterial film is high-value utilization of low-value rice bran products, can reduce environmental burden and relieve white pollution, can reasonably utilize natural resources, and has certain research significance and ecological value.
Disclosure of Invention
The invention aims to provide a sustained-release antibacterial rice bran protein composite membrane, and a preparation method and application thereof.
To achieve the purpose, the invention provides the following scheme:
a sustained-release antibacterial rice bran protein composite membrane is prepared from the following raw materials: the Rice bran Acylated protein, the Carboxymethyl chitosan and the auxiliary agent, wherein the mass ratio of the Rice bran Acylated protein (ARBP) to the Carboxymethyl chitosan (CMCS) is 2: 1-8: 1.
Further, the auxiliary agent is plasticizer-glycerol; dispersing agent-Tween 20, nanometer additive-testa oryzae protein nanogel (ARBPNG); antibacterial agent-Curcumin (CUR) and solvent-water.
Furthermore, the mass ratio of the two raw materials of the composite membrane, namely ARBP and CMCS, is 1: 1-4: 1, namely the mass ratio of the rice bran acylated protein to the carboxymethyl chitosan is 1: 1-4: 1.
The invention also provides a preparation method of the slow-release antibacterial rice bran protein composite membrane, which mainly comprises the following steps:
(1) adding the rice bran acylated protein into distilled water, and fully stirring until the rice bran acylated protein is completely dissolved to prepare a rice bran acylated protein solution;
(2) adding carboxymethyl chitosan into distilled water, and fully stirring until the carboxymethyl chitosan is completely dissolved to prepare a carboxymethyl chitosan solution;
(3) mixing the rice bran acylated protein solution prepared in the step (1) with the carboxymethyl chitosan solution prepared in the step (2) under magnetic stirring, adjusting the pH to 6-8, heating in a water bath for 20-40 min, cooling, gradually dripping glycerin and tween 20 into the solution, and uniformly stirring at 20-25 ℃ to obtain a mixed solution;
(4) adding the rice bran protein nanogel into the mixed solution prepared in the step (3) under magnetic stirring, and fully and uniformly stirring to prepare a composite film forming solution;
(5) and (4) pouring the composite film-forming solution prepared in the step (4) on a mould after vacuum degassing, and demoulding to obtain the slow-release antibacterial rice bran protein composite film.
In the step (1), the preparation of the rice bran acylated protein specifically comprises the following steps:
adding succinic anhydride (namely succinic anhydride) accounting for 10-15% of the weight of the rice bran protein into the rice bran protein aqueous solution for reaction to obtain modified rice bran protein, dialyzing at low temperature of 0-10 ℃ for 48 hours, and freeze-drying to prepare the rice bran acylated protein.
The mass concentration of the rice bran acylated protein in the rice bran acylated protein solution is 1-5%, and the preferred mass concentration is 3%.
In the step (2), the mass concentration of the carboxymethyl chitosan in the carboxymethyl chitosan solution is 1-5%, and preferably 3%.
In the step (3), the mass ratio of the rice bran acylated protein to the carboxymethyl chitosan is 1: 1-4: 1, the water bath temperature is 60-80 ℃, the addition amount of glycerol is 20-25% of the total mass of the rice bran acylated protein and the carboxymethyl chitosan, and the addition amount of Tween 20 is 5-6% of the total mass of the rice bran acylated protein) and the carboxymethyl chitosan.
In the step (4), the preparation of the rice bran protein nanogel specifically comprises the following steps:
dissolving curcumin in dehydrated ethanol, dropwise adding the solution into the rice bran acylated protein aqueous solution under magnetic stirring, heating in a water bath (80-95 ℃), cooling for 20-40 min to room temperature, centrifuging to remove free curcumin in the suspension, filtering, collecting filtrate, and freeze-drying to obtain the rice bran protein nanogel.
The addition amount of the rice bran protein nanogel is 3-10% of the total mass of the rice bran acylated protein and the carboxymethyl chitosan in the step (3).
In the step (5), vacuum degassing is performed for 5-10 min, the reverse modulus is 25g, and the method for preparing the composite membrane is a tape casting method.
The demoulding operation comprises the following steps: and (3) drying the die in a ventilation cabinet for 12-36 hours, then taking off the die, and placing the die in a dryer with relative humidity RH being 45-65% and at 15-35 ℃ for balancing for 36-60 hours. Most preferably, the mold is removed after drying in a fume hood for 24 hours and equilibrated in a desiccator with a relative humidity RH of 55% at 25 ℃ for 48 hours.
The prepared composite membrane has the characteristic of sustained and controlled release of antibacterial agent curcumin.
The prepared composite membrane is subjected to liquid culture experiment, and the antibacterial activity of the composite membrane is evaluated by a turbidimetric method.
The prepared composite film is evaluated according to the release amount of the antibacterial agent in three food simulation systems at different temperatures.
Compared with the prior art, the edible antibacterial composite film provided by the invention has the characteristics of simple operation and convenient preparation, has good antibacterial property and mechanical property, can be completely degraded, has certain sustained and controlled release performance, realizes antibacterial long-acting property and timeliness, effectively prolongs the shelf life of food, has wide raw material sources and excellent biocompatibility, safety and degradability, provides a new idea for the development of the food packaging industry, and has a far-reaching development prospect.
According to the invention, rice bran acylated protein is obtained by succinic anhydride succinylation modification of rice bran protein, the rice bran acylated protein and carboxymethyl chitosan are used as main film substrate raw materials, glycerol is used as a plasticizer, Tween 20 is used as a dispersing agent, and simultaneously, the added rice bran nanogel loaded with curcumin is combined according to a specific proportion, and the raw material components are mutually synergistic, so that the composite film has excellent antibacterial property and mechanical property, and the sustained and controlled release effect of the antibacterial agent in the composite film is realized. As the film base material is a food-borne raw material, the film has the characteristics of high safety, environmental protection, low energy consumption and the like, is an effective novel food packaging film with wide application prospect, and has profound influence on the development of the field of food packaging.
Drawings
FIG. 1 is a diagram showing the influence of ARBP/CMCS composite membranes with different ratios on the growth curve of Escherichia coli;
FIG. 2 is a graph showing the release profile of CUR in 3% acetic acid solution in composite membranes at different ARBPNG addition levels;
FIG. 3 is a graph showing the release profile of CUR in 50% ethanol solution in composite membranes at different ARBPNG addition levels;
FIG. 4 is a graph showing the release profile of CUR in 95% ethanol solution in composite membranes at different ARBPNG addition levels;
FIG. 55% NG-ARBP/CMCS composite membrane release pattern of CUR under different temperature and pH.
FIG. 6 scanning electron micrograph of composite membrane prepared in example 1, A is ARBP/CMCS composite membrane scanning electron micrograph; b is a scanning electron microscope image of the 5 percent NG-ARBP/CMCS composite membrane.
Detailed Description
The present invention will be further described with reference to the following specific examples and the accompanying drawings, but the embodiments of the present invention are not limited thereto.
In the following examples, unless otherwise specified, all starting materials used are commercially available and all methods used are conventional procedures well known to those skilled in the art.
Example 1
This example is provided to illustrate a method for preparing a sustained/controlled release antibacterial rice bran protein composite membrane.
A method for preparing a sustained-release antibacterial rice bran protein composite membrane, wherein raw materials of a membrane substrate are carboxymethyl chitosan (purchased from sigma company) and rice bran acylated protein, an antibacterial agent is curcumin (purchased from Meclin company), auxiliaries are glycerol and Tween 20 and are purchased from Meclin company, and rice bran protein nanogel is prepared in a laboratory, and specifically comprises the following steps:
(a) acylation modification of rice bran protein: firstly, adding a rice bran protein solution (2%) into a phosphate buffer solution, stirring at a constant temperature, simultaneously dropwise adding succinic anhydride (15%), continuously stirring at the constant temperature (40 ℃) after the dropwise adding is finished, maintaining the pH value at 8.0 by using 1.0M NaOH, reacting for 30min, dialyzing after the reaction is finished (the cut-off molecular weight of a dialysis bag is 3.5kD, 4 ℃ and 24h), freezing for 12h in a refrigerator at the temperature of minus 80 ℃, then transferring to a freeze drier at the temperature of minus 40 ℃ for freeze drying for about 48h to obtain the acylation modified rice bran protein in a gray sponge shape, and crushing to obtain the acylation modified rice bran protein powder.
(b) Construction of the nano antibacterial gel: the nano antibacterial gel is prepared by adopting a self-assembly method. Curcumin was dissolved in deionized ethanol, and the solution was added dropwise to 10mL of an acylated rice bran protein solution (pH 6.8, 1mg/mL) under magnetic stirring to a concentration of 30% (w/w). The mixture was heated in a water bath environment (90 ℃, 30min) and then rapidly cooled to room temperature of 25 ℃. The nanogel suspension was centrifuged to remove free curcumin, and the filtrate was collected using a 0.1 μm filter (Millipore, USA) and freeze-dried to give a rice bran protein nanogel.
A preparation method of a sustained-release antibacterial rice bran protein composite membrane comprises the following steps:
(1) 3g of rice bran acylated protein powder prepared above (i.e., the acylated modified rice bran protein powder in step (a)) was dissolved in 100mL of distilled water under magnetic stirring at 500r/min to prepare a 3% concentration ARBP solution.
(2) CMCS solution with a concentration of 3% was prepared by dissolving 3g of carboxymethyl chitosan powder in 100mL of distilled water under magnetic stirring at 500 r/min.
(3) Under the magnetic stirring of 500r/min, the ARBP solution and the CMCS solution which are prepared are mixed according to the mass ratio of 1:1, 2:1, 3:1 and 4:1, the total mass of the ARBP and the CMCS is 3g, the pH value is adjusted to 7, the mixture is rapidly cooled to the room temperature of 25 ℃ after being heated in water bath at the temperature of 80 ℃ for 30min, and then 600mg of glycerol and 150mg of Tween 20 are slowly added into the just-prepared composite solution according to the speed of 8 mL/h.
(4) And (b) respectively adding the prepared rice bran protein nanogel (ARBPNG) containing the antibacterial agent curcumin (namely the rice bran protein nanogel obtained in the step (b)) into the composite solution according to the mass fractions of 3%, 5% and 10% (the total mass of the ARBP and the CMCS is 3g), and stirring at the room temperature of 25 ℃ and at the speed of 500r/min for 8 hours to prepare different NG-ARBP/CMCS composite film-forming solutions.
(5) And (5) carrying out vacuum degassing on the different composite film forming solutions obtained in the step (4) for 5min, weighing 25g, pouring into a mold, naturally air-drying for 24h under a fume hood at the temperature of 25 ℃, and then uncovering the film to obtain the sustained-release antibacterial rice bran protein composite films containing ARBPNG in different proportions.
Example 2
This example illustrates the performance characterization of a sustained/controlled release antibacterial rice bran protein composite membrane.
After the controlled-release antibacterial rice bran protein composite membrane prepared in the example 1 is stored for 48 hours at the temperature of 25 ℃ and the humidity of 55%, various performance measurements are carried out (pure rice bran acylated protein membrane is used as a reference).
1. Measurement of film thickness
And measuring the thickness of the film by using a micrometer caliper, respectively measuring the thicknesses, calculating the average value, namely the film thickness, randomly taking 5 points on the film to be measured according to a certain rule, and obtaining the result of 0.001 mm.
2. Determination of Tensile Strength (TS) and elongation at Break (EB)
The mechanical properties of the sustained and controlled release antibacterial rice bran protein composite membrane were analyzed by a tensile tester (Labthink C610M, Shandong). The film to be tested is made into a rectangular strip with the thickness of 15mm multiplied by 150mm, the distance between probes is ensured to be 60mm in each test, the test speed is 100mm/min, each sample is repeated three times, and the average value is taken.
3. Water solubility assay (WS)
Cutting the composite membrane into 1cm × 4cm rectangles, drying at 105 deg.C to constant weight recording mass m 1 Then placing the mixture into a conical flask, adding 40mL of distilled water, placing the mixture at room temperature for 24 hours, drying the mixture at 105 ℃ for 24 hours, and recording the weight as m 2 . The water solubility is calculated as follows:
solubility (%) ═ m 1 -m 2 )/m 2 ；
4. Determination of optical Properties
The optical properties of the composite films were measured by colorimeter and the readings are expressed in CIE1976 chromaticity space L (dark → light: 0 → 100), a (green- → red +), b (blue- → yellow +). The total color difference is denoted as Δ E. Wherein Ls, as and bs are the values of a standard whiteboard, 94.22, -1.32 and-0.59, respectively.
The specific test results of the TS, EB and WS performances are shown in Table 1, the TS value of the pure ARBP membrane is minimum, the TS and EB of the composite membrane can be effectively improved by adding different ARBP and CMCS, the TS and EB of the composite membrane show the trend of increasing first and then decreasing along with the increase of the ARBP proportion, when w (ARBP) and w (CMCS) are 3:1, the TS and EB of the composite membrane reach the maximum value in the same group, the TS (5.03MPa) of the composite membrane is increased by 104.6 percent compared with the TS (43.33 percent) of the pure ARBP membrane, and the EB value of the composite membrane is increased by 31.8 percent compared with the EB (43.33 percent) of the pure ARBP membrane, which probably results in that the ARBP is uniformly dispersed and the film forming liquid is uniform and stable, so that the ductility and the tensile property of the composite membrane are enhanced; on the other hand, the polar group (-OH, -NH) in ARBP 2 ) Can be combined with polar groups (-OH, -NH) in CMCS molecules 2 ) The intermolecular interaction such as hydrogen bond is formed, so that the composite film has strong acting force, the tensile strength is increased, and the intermolecular connection is tight and the toughness is good, so that the composite film has strong strengthAnd EB. However, as the ratio of ARBP continues to increase, the TS and EB of the composite membrane decrease, probably because at high levels ARBP tends to agglomerate in the matrix affecting the network structure of the protein molecules and the compatibility with CMCS becomes poor, resulting in a decrease in the TS and EB properties of the membrane. Meanwhile, the water solubility of the composite membrane is also affected by the proportion of ARBP, and the water solubility of the composite membrane is gradually reduced along with the increase of the proportion of ARBP and is better than that of WS of a pure ARBP membrane, which may be caused by the fact that the microstructure of the composite membrane is affected by intermolecular hydrogen bond, electrostatic attraction and the like formed by the combination of CMCS and ARBP, such as the formation of a highly stable network structure, and the solubility of the composite membrane is reduced.
TABLE 1 test results of TS, EB and WS performances of ARBP/CMCS composite membranes with different proportions
The color sense of the packaging material influences the consumption acceptance degree of the packaged object, so that the color modification of the composite film plays an important role. As can be seen from Table 2, the total color difference Delta E of ARBP film is 18.64 in comparison with standard white board, while when w (ARBP): w (CMCS) is 3:1, the total color difference of composite film is reduced to 17.05, and the difference of brightness L is smaller, and the color difference of composite films formed by other ARBP and CMCS ratios is larger than that of pure ARBP film, thus obtaining that the composite film with w (ARBP): w (CMCS) of 3:1 has good optical performance and meets the requirement of food packaging.
TABLE 2 optical property test results of ARBP/CMCS composite films with different proportions
Example 3
This example illustrates the antibacterial activity of the sustained/controlled release antibacterial rice bran protein composite membrane against escherichia coli.
The composite membrane of example 2 with better overall performance, namely w (arbp): w (CMCS) 3:1, ARBPNG addition 3%, 5% and 10%, the antimicrobial experiments of this example were tested (with the composite membrane without ARBPNG as a control).
And (3) testing the antibacterial activity of the composite membrane on escherichia coli by adopting a turbidimetry method. The composite film sample is measured according to 2 multiplied by 2cm 2 The sheet was cut into thin pieces, and the pieces were put into a container containing 20mL of a peptone solution (0.1%, w/v) and having a concentration of Escherichia coli of 10 3 CFU/mL centrifuge tube. Incubating the centrifuge tube in a TS-2102C oscillator; shaking was carried out at 37 ℃ and 150rpm for 24 hours. Collecting 0.5mL of culture solution every 2h, adding the culture solution into a 96-well plate, adopting an enzyme-linked immunosorbent assay (ELISA) instrument for light absorption value under 600nm, making a blank in a membrane-free culture medium, and repeating the experiment for three times to obtain an average value.
The antibacterial result of the composite membrane is shown in figure 1, and compared with a pure ARBP membrane, the composite membrane added with ARBPNG has higher antibacterial activity. And with the increase of the addition amount of the ARBPNG, the light absorption value of the composite membrane in 24h is slowly increased, which shows that Curcumin (CUR) is firstly released into the membrane substrate through the ARBPNG and then released into a culture solution through the composite membrane, so that the slow controlled-release antibacterial effect is realized, and the antibacterial persistence of the composite membrane added with 10% of the ARBPNG is stronger than that of other composite membranes, which shows that the release time of the antibacterial agent can be effectively prolonged, the antibacterial period of the composite membrane is prolonged, and the antibacterial performance of the antibacterial agent is improved after the ARBPNG is added.
Example 4
This example is used to illustrate the sustained and controlled release effect of the sustained and controlled release antibacterial rice bran protein composite membrane in different food systems.
Release tests were performed using different food simulants to represent actual food surface conditions. Selection of 3 standard food simulations: 3% acetic acid solution (representing acidic foods such as fruit juices, carbonated beverages and jams), 50% ethanol solution (representing oil-in-water emulsions such as milk) and 95% ethanol solution (representing foods high in fat content such as cooking oils and meats). Cutting the composite film sample into 2 x 2cm 2 Tablets were soaked in 50mL of food simulant solution at room temperature. 1mL of the solution was taken out at appropriate intervals, and the amount of curcumin released was measured while 1mL of fresh food simulant solution was added to the corresponding solution. Curcumin release was measured with an ultraviolet-visible spectrophotometer (UV-2600, Shimadzu, Japan) at a wavelength of 425 nm. All measurements were repeated three times.
The release profiles of the composite membranes in four different simulated food systems are shown in fig. 2-4, the release rate of CUR in 95% ethanol solution is the lowest, the release rate of CUR in all three composite membranes is less than 45%, the release rate in 3% acetic acid solution is the fastest, and the release rate at 8h exceeds 50%, which may be due to the facts that CMCS is easily decomposed in acetic acid solution, and arbping is easily decomposed under acidic conditions, resulting in accelerated release rate of CUR, ARBP and arbping are stable in 95% ethanol solution, hardly swelling behavior occurs, CUR is not easily released in membrane matrix, and thus release rate is low. In addition, the affinity of the composite membrane with a 50% ethanol solution is high, the composite membrane can swell and permeate, the damage degree of an intercellular network structure of the membrane is high, and the early release rate of the CUR is high.
Example 5
This example illustrates the controlled release effect of the antibacterial agent at different temperatures and pH for the controlled release antibacterial rice bran protein composite membrane.
Cutting the composite membrane (5% NG-ARBP/CMCS) containing 5% ARBPNG into 2X 2cm 2 The cut membrane pieces are respectively placed in 50mL of food system simulation solution (phosphate buffer solution) with the pH values of 3.5, 5.0 and 7.4, placed in different temperature environments of 4 ℃, 25 ℃ and 40 ℃, 1mL of solution is taken out at intervals of 0.5h, the release amount of curcumin is measured, and simultaneously 1mL of fresh food system simulation solution is added into the corresponding solution. Curcumin release was measured with an ultraviolet-visible spectrophotometer (UV-2600, Shimadzu, Japan) at a wavelength of 425 nm. All measurements were repeated three times.
The temperature and pH value of the food simulation system are important factors influencing migration, diffusion and release of the CUR into the food. The release profile of CUR in a food simulant system at different temperatures of 4 deg.C, 25 deg.C, 40 deg.C, different pHs of 3.5, 5.0, 7.4 (see FIG. 5). As can be seen from FIG. 5, the initial release rate of the CUR in the composite membrane increases with the temperature of the food simulant system, and when the temperature is constant, the release rate of the CUR in the simulant liquid with pH3.5, 5.0, and 7.4 increases with the decrease of the pH. The pH value obviously influences the diffusion process, which is related to the swelling of the composite membrane and ARBPNG, and at the pH value of 3.5, the swelling property of the membrane is high, the spatial structure of the protein in the membrane becomes loose, the diffusion is accelerated, the lower the activation energy is, the membrane is easy to decompose, and the CUR has high release rate. Meanwhile, under the same pH value, the temperature has obvious influence on the release of the CUR, the composite membrane has the fastest release rate at 40 ℃, the release rate of the CUR is obviously reduced at 4 ℃, and the sensitivity of the composite membrane to the temperature is higher than the pH value.
Generally speaking, in different food systems, the CUR in the composite film presents different release rates, the release of the CUR in the composite film can be effectively delayed by adding the ARBPNG, the purpose of slow and controlled release is achieved, and the influence of the temperature and the pH on the release rate of the CUR is further explored, so that when different foods are packaged, the composite film can be selected according to the pH and the storage temperature of the different foods, the shelf life of the foods is prolonged, and the effective antibacterial effect is achieved.
The present invention is not limited to the above embodiments, and various other modifications, substitutions or alterations can be made without departing from the basic technical idea of the invention by using the common technical knowledge and the conventional means in the field according to the above content of the present invention, and the present invention falls within the protection scope of the claims of the present invention.
Claims (7)
1. The preparation method of the slow-release antibacterial rice bran protein composite membrane is characterized in that the slow-release antibacterial rice bran protein composite membrane is prepared from the following raw materials: the rice bran acylated protein, the carboxymethyl chitosan and the auxiliary agent, wherein the mass ratio of the rice bran acylated protein to the carboxymethyl chitosan is 1: 2-8: 1;
the auxiliary agent is plasticizer-glycerol, dispersant-Tween 20, nano additive-rice bran protein nanogel, antibacterial agent-curcumin and solvent-water;
the preparation method comprises the following steps:
(1) adding the rice bran acylated protein into distilled water, and fully stirring until the rice bran acylated protein is completely dissolved to prepare a rice bran acylated protein solution;
the preparation method of the rice bran acylated protein comprises the following specific steps:
adding succinic anhydride accounting for 10-15% of the weight of the rice bran protein into the rice bran protein aqueous solution for reaction to obtain modified rice bran protein, dialyzing at low temperature of 0-10 ℃ for 48 hours, and freeze-drying to prepare rice bran acylated protein;
(2) adding carboxymethyl chitosan into distilled water, and fully stirring the mixture by using a magnetic stirrer until the carboxymethyl chitosan is completely dissolved to prepare a carboxymethyl chitosan solution;
(3) mixing the rice bran acylated protein solution prepared in the step (1) with the carboxymethyl chitosan solution prepared in the step (2) under magnetic stirring, adjusting the pH value to 6-8, heating in a water bath for 20-40 min, cooling, dropwise dispersing glycerol and tween-20 into the solution, and uniformly stirring at 20-25 ℃ to obtain a mixed solution;
(4) adding the rice bran protein nanogel into the mixed solution prepared in the step (3) under magnetic stirring, and fully and uniformly stirring to prepare a composite film forming solution;
(5) and (4) pouring the composite film-forming solution prepared in the step (4) on a mould after vacuum degassing, and demoulding to obtain the slow-release antibacterial rice bran protein composite film.
2. The preparation method of the slow-release antibacterial rice bran protein composite membrane according to claim 1, wherein the mass ratio of the rice bran acylated protein to the carboxymethyl chitosan is 1: 1-4: 1.
3. The preparation method of the sustained-release antibacterial rice bran protein composite membrane according to claim 1, wherein in the step (1), the mass fraction of the rice bran acylated protein in the rice bran acylated protein solution is 1-5%.
4. The preparation method of the slow-release antibacterial rice bran protein composite membrane according to claim 1, wherein in the step (2), the mass fraction of carboxymethyl chitosan in the carboxymethyl chitosan solution is 1-5%.
5. The preparation method of the sustained-release antibacterial rice bran protein composite membrane according to claim 1, wherein in the step (3), the mass ratio of the rice bran acylated protein to the carboxymethyl chitosan is 1: 1-4: 1, and the water bath temperature is 60-80 ℃.
6. The method for preparing the sustained-release antibacterial rice bran protein composite membrane according to claim 1, wherein in the step (4), the preparation of the rice bran protein nanogel specifically comprises the following steps:
dissolving curcumin in absolute ethyl alcohol, dropwise adding a curcumin solution into a rice bran acylated protein aqueous solution under magnetic stirring, heating in a water bath at the temperature of 80-95 ℃ for 20-40 min, cooling to room temperature, centrifuging to remove free curcumin in a suspension, filtering, collecting filtrate, and freeze-drying to obtain the rice bran protein nanogel.
7. The use of the controlled-release antibacterial rice bran protein composite membrane prepared by the preparation method according to any one of claims 1 to 6 as a food packaging material.
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