CN111991368B - Preparation method of tea polyphenol coated soy protein isolate-quaternary ammonium salt chitosan microcapsule - Google Patents
Preparation method of tea polyphenol coated soy protein isolate-quaternary ammonium salt chitosan microcapsule Download PDFInfo
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Abstract
The invention discloses a preparation method of a soybean protein isolate-quaternary ammonium salt chitosan microcapsule with a tea polyphenol coating, which comprises the steps of firstly mixing calcium chloride and a sodium carbonate solution to generate calcium carbonate particles as a template for preparing the microcapsule, then sequentially adding the soybean protein isolate and the quaternary ammonium salt chitosan solution into a calcium carbonate dispersion liquid in a layer-by-layer self-assembly mode to obtain the chitosan-soybean protein isolate microcapsule with a multilayer wrapping structure, after the microcapsule is centrifugally washed, removing the calcium carbonate template by using an EDTA solution, and adding the tea polyphenol solution for coating to finally obtain the target microcapsule. The microcapsule is used for embedding and in-vitro release of puerarin. The results show that: the microcapsule with tea polyphenol coating has slow release effect on the release of puerarin under different pH conditions, and can improve the bioavailability of puerarin.
Description
Technical Field
The invention relates to the technical field of biological materials, in particular to a preparation method of a tea polyphenol coated soy protein isolate-quaternary ammonium salt chitosan microcapsule.
Background
The puerarin comes from the traditional Chinese medicinal material radix puerariae which is homologous in medicine and food, has various physiological activities, is sold in the market as a functional health-care component, but is seriously stimulated to the stomach and has lower bioavailability when being directly taken.
The soybean protein is a high-quality and low-cost plant protein, can meet the requirements of human nutrition supply and growth and development, is widely applied to the technical field of food processing, has electronegativity under physiological conditions due to the isoelectric point of about 4.5, can form an electrostatic complex with positively charged biomacromolecules so as to self-assemble into microcapsules to embed and deliver food functional molecules, and currently, a great deal of research is carried out on the delivery of the protein in the aspect of functional molecules.
Chitosan is used as the only natural cationic polysaccharide in the nature, has various physiological activities such as bacteriostasis, anti-inflammation, immunoregulation and the like, has the characteristics of safety, no toxicity and edibility, and is widely applied to the field of food additives. The quaternary ammonium salt chitosan obtained by carrying out structural modification on chitosan has good biological safety and good performances of antibacterial property, film forming property, cation adsorption, moisture absorption and retention, flocculation property, antistatic property and the like. The antibacterial performance of the chitosan antibacterial agent is superior to that of chitosan and other chitosan derivatives.
The principle that chitosan and soy protein isolate can form polysaccharide-protein complex under physiological conditions through electrostatic interaction has been utilized, and intensive research has been conducted on the entrapment delivery of bioactive components in the fields of biomedicine and food functions.
The current research on the drug delivery of polysaccharide-protein complex still faces the problem of high leakage rate of drug encapsulation during the delivery process, how to improve the encapsulation rate of the drug and the stability during the release process still needs to be improved through further research, and in the field of food technology, the safety of the encapsulated material is also the first problem to be considered. At present, the method for preparing the biological macromolecular microcapsules by layer self-assembly through a calcium carbonate template method has been widely researched, but the research on coating loading of the prepared microcapsules is only carried out. The experiment adopts natural product active molecular tea polyphenol to carry out secondary coating on the microcapsule, preliminarily characterizes the structure and the physical and chemical properties of the microcapsule, verifies the drug-loading properties of the microcapsule, such as encapsulation, loading, release and the like of the mode molecular drug, develops a biological microcapsule with the characteristic of PH response, and provides experimental basis for developing a drug delivery system taking biological macromolecules as a matrix.
Disclosure of Invention
Aiming at the problems, the invention provides a preparation method of a tea polyphenol coated soy protein isolate-quaternary ammonium salt chitosan microcapsule.
The technical scheme of the invention is as follows: a preparation method of a soy protein isolate-chitosan microcapsule with a tea polyphenol coating comprises the following specific operation steps:
respectively preparing calcium chloride and sodium carbonate into solutions, then mixing and stirring, standing for 15min, carrying out vacuum filtration to remove supernatant, washing with purified water, collecting particle products, and carrying out freeze drying to obtain calcium carbonate particles;
step (1.2), adding calcium carbonate particles into the isolated soy protein solution, stirring, carrying out vacuum filtration on the suspension, and centrifugally washing the obtained particles;
step (1.3), adding the particles obtained by centrifugal washing into a quaternary ammonium salt chitosan solution, stirring, carrying out vacuum filtration on the suspension, and centrifugally washing the particles;
step (1.4), repeating the steps (1.2) and (1.3), adding the obtained sample into puerarin solution after the required polymer coating layers are deposited on the surface of the calcium carbonate template, stirring overnight, and then centrifuging and washing the precipitate to obtain the puerarin-loaded soy protein isolate-quaternary ammonium salt chitosan microcapsule;
step (1.5), collecting and respectively adding the soy protein isolate-quaternary ammonium salt chitosan microcapsules loaded with puerarin into tea polyphenol solutions with different concentration gradients, carrying out centrifugal washing after stirring to remove the tea polyphenol solution remained on the surfaces of the capsules, and carrying out freeze drying on the centrifuged sample to prepare dried tea polyphenol coated microcapsules;
and (1.6) dispersing the dried tea polyphenol coated microcapsule in an EDTA solution to remove CaCO3 particles, stirring, washing, centrifuging, and washing centrifugally to finally obtain the puerarin-loaded quaternary ammonium salt chitosan-soybean protein isolate hollow microcapsule with the tea polyphenol coated layer.
Further, in the step (1.1), the concentration of the calcium chloride and the sodium carbonate is 0.2-0.4mol/L, the reaction temperature is 20-40 ℃, the stirring time is 1-10min, and the standing time is 10-60 min.
Further, in the step (1.2), the material-to-liquid ratio of the calcium carbonate to the soy protein isolate solution is 1:50-100(w/v), the concentration of the soy protein isolate is 0.5-3mg/ml, the stirring time is 1-3h, the solvent used in the centrifugal washing process is purified water, the centrifugal rotation speed is 8000-.
Further, in the step (1.3), the material-liquid ratio of the precipitate obtained in the step (1.2) to the quaternary ammonium salt chitosan solution is 1:50-100(w/v), the concentration of the quaternary ammonium salt chitosan is 0.5-3mg/ml, the stirring time is 0.5-3h, the solvent used in the centrifugal washing process is purified water, the centrifugal rotation speed is 8000-.
Further, in the step (1.4), the ratio of the obtained precipitate to the puerarin solution is 1:50-100(w/v), the concentration of the puerarin solution is 1-4mg/ml, the stirring speed is 100-.
Further, in the step (1.5), the concentration of the tea polyphenol solution is 1-10mg/ml, and the material-liquid ratio of the microcapsule to the tea polyphenol solution is 1:50-100(w/v), stirring speed of 100-300rpm, stirring time of 1-6h, each centrifugation time of 5-10min, and centrifugation speed of 8000-10000 rpm.
Further, in the step (1.6), the ratio of the material to the liquid is 1:50-100(w/v), the concentration of the EDTA solution is 0.1-0.5mol/l, the stirring time is 0.5-3h, the centrifugation speed is 8000-10000rpm, and the centrifugation time is 5-10 min.
The invention takes calcium carbonate as a microcapsule template, selects cationic biomacromolecule quaternary ammonium salt chitosan and natural anionic protein soybean protein isolate protein as two electrostatic binding components of the microcapsule, sequentially forms 4 layers of protein-polysaccharide coatings on the surface of the calcium carbonate template by a preparation method of LBL (layer-by-layer self-assembly), and finally removes the calcium carbonate template in EDTA solution. The prepared microcapsule is further incubated with tea polyphenol and puerarin to obtain the hollow microcapsule with puerarin loading and tea polyphenol coating.
The invention realizes the above purposes by the following technical scheme: calcium carbonate templates are prepared by taking calcium chloride and sodium carbonate solution as raw materials, and calcium carbonate particles are obtained by stirring, standing, precipitating, washing and drying. Then, the quaternary ammonium salt chitosan and the soybean protein isolate are sequentially coated on the surface of the calcium carbonate through oscillation incubation and centrifugal washing to form a multilayer structure. The microcapsule is incubated with EDTA, tea polyphenol and puerarin, washed, dialyzed and freeze dried to obtain the puerarin-loaded macro protein isolate-quaternary ammonium salt chitosan hollow microcapsule with tea polyphenol coating. The encapsulation efficiency of the puerarin is 25-40%, the loss rate of the puerarin is 10-20% after the puerarin passes through the tea polyphenol coating, and the loading rate of the puerarin after the template is removed is 15-20%
The invention has the beneficial effects that: the soybean protein isolate-quaternary ammonium salt polysaccharide microcapsule with the tea polyphenol coating prepared by the invention has obvious slow release effect on the release of puerarin through in-vitro simulated release tests, and the capsule preparation material is safe, non-toxic and good in biocompatibility.
Drawings
FIG. 1 is a flow chart of the architecture of the present invention;
FIG. 2 is a potential diagram of the particle size of soy protein isolate-chitosan microcapsules with a tea polyphenol coating in accordance with the present invention;
FIG. 3 is a CLSM image of soy protein isolate-chitosan microcapsules with a tea polyphenol coating in accordance with the present invention;
FIG. 4 is an SEM image of isolated soy protein-chitosan microcapsules with a tea polyphenol coating in accordance with the present invention;
FIG. 5 is a graph showing in vitro release of soy protein isolate-chitosan microcapsules having a tea polyphenol coating in accordance with the present invention.
Detailed Description
In order to more clearly illustrate the technical solution of the present invention, the following detailed description is made with reference to the accompanying drawings:
as depicted in fig. 1; a preparation method of a soy protein isolate-chitosan microcapsule with a tea polyphenol coating comprises the following specific operation steps:
respectively preparing calcium chloride and sodium carbonate into solutions, then mixing and stirring, standing for 15min, carrying out vacuum filtration to remove supernatant, washing with purified water, collecting particle products, and carrying out freeze drying to obtain calcium carbonate particles;
step (1.2), adding calcium carbonate particles into the isolated soy protein solution, stirring, carrying out vacuum filtration on the suspension, and centrifugally washing the obtained particles;
step (1.3), adding the particles obtained by centrifugal washing into a quaternary ammonium salt chitosan solution, stirring, carrying out vacuum filtration on the suspension, and centrifugally washing the particles;
step (1.4), repeating the steps (1.2) and (1.3), adding the obtained sample into puerarin solution after the required polymer coating layers are deposited on the surface of the calcium carbonate template, stirring overnight, and then centrifuging and washing the precipitate to obtain the puerarin-loaded soy protein isolate-quaternary ammonium salt chitosan microcapsule;
step (1.5), collecting and respectively adding the soy protein isolate-quaternary ammonium salt chitosan microcapsules loaded with puerarin into tea polyphenol solutions with different concentration gradients, carrying out centrifugal washing after stirring to remove the tea polyphenol solution remained on the surfaces of the capsules, and carrying out freeze drying on the centrifuged sample to prepare dried tea polyphenol coated microcapsules;
and (1.6) dispersing the dried tea polyphenol coated microcapsule in an EDTA solution to remove CaCO3 particles, stirring, washing, centrifuging, and washing centrifugally to finally obtain the puerarin-loaded quaternary ammonium salt chitosan-soybean protein isolate hollow microcapsule with the tea polyphenol coated layer.
Further, in the step (1.1), the concentration of the calcium chloride and the sodium carbonate is 0.2-0.4mol/L, the reaction temperature is 20-40 ℃, the stirring time is 1-10min, and the standing time is 10-60 min.
Further, in the step (1.2), the material-to-liquid ratio of the calcium carbonate to the soy protein isolate solution is 1:50-100(w/v), the concentration of the soy protein isolate is 0.5-3mg/ml, the stirring time is 1-3h, the solvent used in the centrifugal washing process is purified water, the centrifugal rotation speed is 8000-.
Further, in the step (1.3), the material-liquid ratio of the precipitate obtained in the step (1.2) to the quaternary ammonium salt chitosan solution is 1:50-100(w/v), the concentration of the quaternary ammonium salt chitosan is 0.5-3mg/ml, the stirring time is 0.5-3h, the solvent used in the centrifugal washing process is purified water, the centrifugal rotation speed is 8000-.
Further, in the step (1.4), the ratio of the obtained precipitate to the puerarin solution is 1:50-100(w/v), the concentration of the puerarin solution is 1-4mg/ml, the stirring speed is 100-.
Further, in the step (1.5), the concentration of the tea polyphenol solution is 1-10mg/ml, and the material-liquid ratio of the microcapsule to the tea polyphenol solution is 1:50-100(w/v), stirring speed of 100-300rpm, stirring time of 1-6h, each centrifugation time of 5-10min, and centrifugation speed of 8000-10000 rpm.
Further, in the step (1.6), the ratio of the material to the liquid is 1:50-100(w/v), the concentration of the EDTA solution is 0.1-0.5mol/l, the stirring time is 0.5-3h, the centrifugation speed is 8000-10000rpm, and the centrifugation time is 5-10 min.
Example (b):
firstly, preparing a quaternary ammonium salt chitosan-soy protein isolate microcapsule with a tea polyphenol coating:
(1) mixing a 0.3M sodium carbonate solution and a 0.3M calcium chloride solution in equal volume, stirring at the rotating speed of 600rpm for 1min, stopping stirring, standing for 30min, then carrying out vacuum filtration to remove supernatant, collecting precipitate to obtain calcium carbonate spherical particles, carrying out suction filtration washing for 3 times by using distilled water, and carrying out freeze drying after washing is finished;
(2) adding the freeze-dried calcium carbonate particles into a soybean protein isolate solution of 1mg/ml according to a material-to-liquid ratio of 1:100(w/v), stirring at the room temperature at the rotating speed of 200rpm for 1h, stopping stirring, carrying out vacuum filtration on a suspension, adding purified water into the obtained precipitate, and carrying out centrifugal washing for 3 times; a small amount of sample was dispersed in purified water to measure the Zeta potential.
(3) Adding the obtained precipitate into 1mg/ml quaternary ammonium salt chitosan solution according to the feed-liquid ratio of 1:100(w/v), stirring at the rotating speed of 200rpm at room temperature for 30min, stopping stirring, carrying out vacuum filtration on the suspension, adding purified water into the obtained precipitate, and carrying out centrifugal washing for 3 times; taking a small amount of samples, and measuring the Zeta potential in purified water;
(4) repeating the steps (2) and (3) until the number of the polymer coating layers required by the deposition of the surface of the calcium carbonate template is reached, adding the obtained sample into the puerarin solution according to the material-liquid ratio of 1:50(w/v), stirring at the rotating speed of 300rpm for overnight, centrifuging the precipitate at the rotating speed of 10000rpm for 10min, and repeatedly washing with purified water for three times to obtain the puerarin-loaded soybean protein isolate-quaternary ammonium salt chitosan microcapsule;
(5) collecting the microcapsules, respectively adding the microcapsules into tea polyphenol solutions with different concentration gradients (2.5/5/10mg/ml) according to a material-liquid ratio of 1:100(w/v), stirring at a rotating speed of 300rpm for 2h, centrifuging at 8000rpm for 5min after stirring is finished, washing for 3 times to remove the tea polyphenol solution remained on the surfaces of the capsules, freeze-drying a sample obtained by centrifuging, and preparing the dried tea polyphenol coated microcapsules after 48 h;
(6) and (3) mixing the tea polyphenol coated soy protein isolate-quaternary ammonium salt chitosan according to a material-liquid ratio of 1:100(w/v) was dispersed in 0.25M EDTA solution to remove CaCO3 particles, and after stirring for 1h, washing and centrifuging were performed, and after repeating the removal process twice, washing with purified water by centrifugation three times (1000rpm, 5 min); finally obtaining the quaternary ammonium salt chitosan-soybean protein isolate hollow microcapsule carrying the puerarin tea polyphenol coating.
Secondly, characterization and structure of the quaternary ammonium salt chitosan-soy protein isolate microcapsule:
(1) and the particle size and the potential of the quaternary ammonium salt chitosan-soy protein isolate microcapsule are as follows:
research shows that when the microcapsule taking calcium carbonate as a template is prepared by a layer-by-layer self-assembly method, the size and distribution of the particle size and the change of Zeta potential electrical property can both show the deposition condition of the macromolecular polyelectrolyte on the surface of the calcium carbonate; taking a 10mg microcapsule sample, adding 1ml of purified water, uniformly dispersing, and measuring the particle size and distribution of the microcapsule by using a particle size analyzer through a DLS dynamic light scattering method; adding a sample into a potential cell, and measuring the potential distribution on the surface of the capsule by a potentiometer; as shown in fig. 2, as the number of layers of polyelectrolyte deposited on the surface of the microcapsule increases, the particle size of the whole particle tends to increase, and the distribution coefficient of the particle size of PDI increases, indicating that the particle size distribution of the microcapsule gradually begins to be uneven and the dispersibility decreases; zeta potential measurement results show that the surface of the calcium carbonate template presents electropositivity, which is consistent with the reports in the literature, and the potential changes in positive and negative electrical property in sequence along with the alternate coating of the soybean protein isolate and the quaternary ammonium salt chitosan with opposite charges, which shows that the polyelectrolyte is electrostatically bonded on the surface of the microcapsule.
(2) CLSM graph of quaternary ammonium salt chitosan-soy protein isolate microcapsules:
the surface of the calcium carbonate template has a porous structure, and when the medicine is loaded, the medicine is adsorbed by polyelectrolyte deposited on the surface of the calcium carbonate and then continuously permeates into the calcium carbonate particles; as shown in FIG. 3, after loading a water-soluble fluorescent dye rhodamine B in the microcapsule, a distribution image of the rhodamine B in the microcapsule can be observed through a laser confocal microscope at an excitation wavelength of 525 nm; the whole fluorescence image presents spherical particles, which shows that the medicine is completely permeated in the microcapsule and is distributed more uniformly, and the microcapsule can be used as an excellent carrier of water-soluble medicine molecules; by measuring the ultraviolet absorption spectrum of the rhodamine B solution, the drug loading process of the microcapsule can be dynamically monitored; the arrow direction in the figure shows the trend of the maximum ultraviolet absorption peak of the solution at 525nm within 75mins, and the intensity of the ultraviolet absorption peak is reduced because of the reduction of the concentration of the solution, which indicates that the microcapsule is successfully loaded with part of rhodamine B.
Thirdly, SEM image of quaternary ammonium salt chitosan-soy protein isolate microcapsule:
the scanning electron microscope can observe the surface microstructure of the microcapsule, and in order to further characterize and verify the structural characteristics of the prepared microcapsule, an SEM image of the microcapsule is shot; as shown in fig. 4, the microcapsules prepared by the calcium carbonate template have a regular spherical structure, and the surface smoothness and integrity of the spherical particles cannot be influenced by the layer-by-layer deposition of the quaternary ammonium salt chitosan and the isolated soy protein on the particle surface; after the calcium carbonate template is removed by EDTA solution, the surface of the microcapsule can collapse, and the spherical structure can be changed into irregular shape; then, after calcium carbonate particles of the microcapsules coated with the three tea polyphenol solutions with different concentrations are removed by an EDTA solution, the surfaces of the microcapsules become uneven, and the microcapsules are irregular particles as a whole, which indicates that the tea polyphenol coatings do not have great influence on the surface structures of the microcapsules.
Fourthly, in-vitro simulated release diagram of the quaternary ammonium salt chitosan-soy protein isolate microcapsule:
weighing equal mass of hollow microcapsules coated with tea polyphenol solutions with different concentrations (0, 2.5, 5 and 10mg/ml), adding the hollow microcapsules into 10ml of centrifuge tubes containing citric acid buffer solution with pH 2 and 8, placing the centrifuge tubes containing the microcapsules into a constant temperature shaking table at 37 ℃ to perform in-vitro release at the rotating speed of 200rpm, taking 200 mu L of solution from the centrifuge tubes at certain intervals, centrifuging (8000rpm, 5min), removing supernatant, detecting the content of puerarin in the supernatant at the ultraviolet wavelength of 250nm by using a liquid chromatography instrument and methanol and water as mobile phases to observe the release condition, and calculating to obtain the cumulative release percentage of the puerarin under different pH conditions;
the microcapsules of the tea polyphenol coating have the characteristic of pH response to the loaded medicine, and have good slow release performance on puerarin under different pH conditions compared with a blank group; under the same condition of PH, the slow release performance of the coated microcapsule to puerarin is obviously enhanced along with the increase of the concentration of the tea polyphenol solution; when the PH is 2, the microcapsules without tea polyphenol coating release 70% of puerarin accumulatively within 8h, and for tea polyphenol solutions with different concentrations, the release rate is accelerated under an acidic condition, and the release of the puerarin is slowed down along with the increase of the tea polyphenol concentration; under the alkaline condition, the release rate of the puerarin is remarkably slowed down because the polyphenol substances in the tea polyphenol spontaneously polymerize to form a compact molecular layer to cover the surface of the microcapsule, the cumulative release percentage in 8h is lower than 30 percent, and the slow-release effect is remarkable.
Finally, it should be understood that the embodiments described herein are merely illustrative of the principles of embodiments of the present invention; other variations are possible within the scope of the invention; thus, by way of example, and not limitation, alternative configurations of embodiments of the invention may be considered consistent with the teachings of the present invention; accordingly, the embodiments of the invention are not limited to the embodiments explicitly described and depicted.
Claims (7)
1. A preparation method of a soybean protein isolate-chitosan microcapsule with a tea polyphenol coating is characterized by comprising the following specific operation steps:
respectively preparing calcium chloride and sodium carbonate into solutions, then mixing and stirring, standing, carrying out vacuum filtration to remove supernate, washing with purified water, collecting particle products, and carrying out freeze drying to obtain calcium carbonate particles;
step (1.2), adding calcium carbonate particles into the isolated soy protein solution, stirring, carrying out vacuum filtration on the suspension, and centrifugally washing the obtained particles;
step (1.3), adding the precipitate obtained by centrifugal washing into a quaternary ammonium salt chitosan solution, stirring, carrying out vacuum filtration on the suspension, and carrying out centrifugal washing on the obtained precipitate;
step (1.4), repeating the steps (1.2) and (1.3), adding the obtained sample into puerarin solution after the required polymer coating layers are deposited on the surface of the calcium carbonate template, stirring overnight, and then centrifuging and washing the precipitate to obtain the puerarin-loaded soy protein isolate-quaternary ammonium salt chitosan microcapsule;
step (1.5), collecting and respectively adding the soy protein isolate-quaternary ammonium salt chitosan microcapsules loaded with puerarin into tea polyphenol solutions with different concentration gradients, carrying out centrifugal washing after stirring to remove the tea polyphenol solution remained on the surfaces of the capsules, and carrying out freeze drying on the centrifuged sample to prepare dried tea polyphenol coated microcapsules;
step (1.6), dispersing the dried tea polyphenol coated microcapsule in EDTA solution to remove CaCO3And (3) granulating, stirring, washing, centrifuging, washing, and finally obtaining the puerarin-loaded quaternary ammonium salt chitosan-soybean protein isolate hollow microcapsule with the tea polyphenol coating.
2. The method for preparing the soy isolate protein-chitosan microcapsule with tea polyphenol coating according to claim 1, characterized in that, in the step (1.1), the concentration of the calcium chloride and the sodium carbonate is 0.2-0.4mol/L, the reaction temperature is 20-40 ℃, the stirring time is 1-10min, and the standing time is 10-60 min.
3. The method for preparing the soy protein isolate-chitosan microcapsule with tea polyphenol coating as claimed in claim 1, wherein in step (1.2), the feed-liquid ratio of calcium carbonate to soy protein isolate solution is 1:50-100w/v, the soy protein isolate concentration is 0.5-3mg/ml, the stirring time is 1-3h, the solvent used in the centrifugal washing process is purified water, the centrifugal rotation speed is 8000-.
4. The method for preparing the soy isolate protein-chitosan microcapsule with tea polyphenol coating as claimed in claim 1, wherein in the step (1.3), the feed-to-liquid ratio of the precipitate obtained in the step (1.2) to the chitosan solution of quaternary ammonium salt is 1:50-100w/v, the chitosan concentration of quaternary ammonium salt is 0.5-3mg/ml, the stirring time is 0.5-3h, the solvent used in the centrifugal washing process is purified water, the centrifugal rotation speed is 8000-10000rpm, and the centrifugal time is 5-10 min.
5. The method for preparing the isolated soy protein-chitosan microcapsule with tea polyphenol coating as claimed in claim 1, wherein, in the step (1.4), the ratio of the polymer-coated microcapsule to the puerarin solution is 1:50-100w/v, puerarin solution concentration 1-4mg/ml, stirring speed 100-.
6. The method for preparing the soy isolate protein-chitosan microcapsule with tea polyphenol coating according to claim 1, wherein in the step (1.5), the concentration of the tea polyphenol solution is 1-10mg/ml, and the feed-to-solution ratio of the microcapsule obtained in the step (1.4) to the tea polyphenol solution is 1:50-100w/v, stirring speed of 100-.
7. The method for preparing the soy isolate protein-chitosan microcapsule with tea polyphenol coating as claimed in claim 1, wherein in step (1.6), the material-to-liquid ratio of the microcapsule obtained in step (1.5) to the EDTA solution is 1:50-100w/v, the concentration of the EDTA solution is 0.1-0.5mol/l, the stirring time is 0.5-3h, the centrifugation speed is 8000-.
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| CN113999403B (en) * | 2021-11-19 | 2022-08-30 | 武汉大学 | Quaternized soybean protein, quaternized soybean protein-dopamine, and preparation method and application thereof |
| CN114480210A (en) * | 2022-02-25 | 2022-05-13 | 万之源生物科技有限公司 | Preparation method of composite bacteria microcapsule for treating salinization and promoting growth of cotton and peanut |
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