CN113080468A - Slow-digestion casein nano micelle powder and preparation method thereof - Google Patents

Slow-digestion casein nano micelle powder and preparation method thereof Download PDF

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CN113080468A
CN113080468A CN202110396218.3A CN202110396218A CN113080468A CN 113080468 A CN113080468 A CN 113080468A CN 202110396218 A CN202110396218 A CN 202110396218A CN 113080468 A CN113080468 A CN 113080468A
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费颖
李舒宇
程慧君
樊启磊
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Suzhou Langbang Nutrition Technology Co ltd
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Abstract

The invention discloses slowly digestible casein nano micelle powder and a preparation method thereof, and belongs to the field of nutrition and health. The casein nano micelle powder takes casein, calcium salt and citric acid as initial suspension, a specific three-dimensional network structure is formed under ultrasonic crushing, the self-assembly of nano micelles is realized by matching with space charge migration and collision induced by radio frequency, and the novel casein nano micelle powder with uniform and tiny particle size is prepared after freeze dehydration and drying. The novel casein nano micelle powder prepared by the method has obvious hydrolysis resistance to pepsin and trypsin, so that the novel casein nano micelle powder has the characteristic of continuously providing protein and energy, can maintain the muscle state and slowly release encapsulated hydrophobic functional food or medicine, and provides a new solution for body-building people, obesity people and disease treatment.

Description

Slow-digestion casein nano micelle powder and preparation method thereof
Technical Field
The invention relates to slowly digestible casein nano micelle powder and a preparation method thereof, belonging to the field of nutrition and health.
Background
With the development of science and technology and the improvement of the knowledge level of people, the close relationship between diet and health is valued by people, and the opportunities and challenges facing the food industry and the medical field are followed, including the development of new protein supplements and protein carriers in the medical field.
Research shows that casein can continuously provide protein for athletes, help athletes inhibit protein decomposition, and release amino acids into blood, so as to improve metabolism rate and promote muscle protein synthesis. In addition, because the hydrophilic and hydrophobic residues in the casein polypeptide chain are distributed in clusters, the casein molecule has a blocky amphiphilic structure. By utilizing the natural conformation, the casein serves as a nano capsule, the solubility and the stability of the hydrophobic nutrient substance are improved by the targeted delivery of the hydrophobic nutrient substance, the hydrophobic nutrient substance can be slowly released in vivo, the bioavailability in vivo of the hydrophobic nutrient substance is enhanced, and the like.
However, casein structures are susceptible to hydrolysis by proteases due to the presence of high levels of proline, but low levels of cysteine residues. Although casein molecules have a blocky amphiphilic structure and phosphorylation centers, which can spontaneously form spherical casein micelles by self-association and interaction with colloidal calcium phosphate, studies have shown that in vitro digestion simulation experiments, the addition of a certain amount of pepsin in simulated gastric juice at pH 2.5, a large portion of casein is digested and decomposed within 120min of the study. Meanwhile, because a large amount of hydrogen ions exist in gastric juice, nano-micelles formed by casein are gradually dissociated along with the increase of acidity, kappa-casein 'hair layers' on the surfaces of the micelles gradually collapse, and the spatial distribution of the casein in the internal structures of the micelles changes, so that the casein micelles cannot maintain the spatial structures of the casein micelles, and the casein micelles are more easily hydrolyzed by protease.
Some studies on the improvement of the stability and the slowing of the digestion rate of casein have been reported, and multiple emulsion aggregation method, polysaccharide grafting method and the like are commonly used for enhancing the resistance of casein to protease. For example, chenghao (functional factor co-embedding research based on different carriers of casein/zein, university of south of the Yangtze river, 2020) and the like prepare the nano compound which takes casein as a core and realizes the embedding of alpha-tocopherol by adding an emulsifier, and the nano compound has a good slow release effect, however, the casein digestion rate is still high, and the prepared particle size is large, so that the production application cannot be expanded. Min Min (preparation of glycosylated casein, performance and application research of the glycosylated casein, Zhejiang university of industry and commerce, 2017) and the like adopt chitosan and carrageenan to graft casein under the condition of ultrasonic dry heat to obtain the glycosylated casein, however, the digestion rate of the nanocapsule in simulated gastric juice is still high, nearly 60% of capsanthin pigment is released within 90Min, and the resistance of the casein to protease is weak.
However, for the related solutions such as the multiple emulsion aggregation method and the polysaccharide grafting method, for example, the polysaccharide grafting method grafts proteins with the components that are not easily digested by pepsin such as polysaccharide, so as to form a polysaccharide-protein complex, in the presence of gastric acid, the casein micelles with the existing structure still cause dissolution of calcium ions, so as to cause collapse of the nano-micelles, so that the nano-micelles are more easily hydrolyzed by protease, and the technical problems that casein is easily hydrolyzed by protease in gastric juice and the digestion rate is higher are not effectively improved.
In summary, in order to improve the stability of casein and slow down the digestion rate of casein, research and development of slowly digested casein nano micelle powder is an urgent need in the fields of food nutrition, medical care and the like.
Disclosure of Invention
The purpose of the invention is as follows: aiming at the problems, in order to overcome the defects in the prior art, the slowly digestible casein nano micelle powder and the preparation method thereof are developed, the prepared casein nano micelle powder can resist protease hydrolysis, and has a slow release effect and a slow decomposition characteristic in the casein digestion process; the casein nano micelle has uniform particle size, no toxic or side effect and good industrial application prospect.
In view of the above, the problems of casein as a protein carrier are: is easy to be hydrolyzed by protease in gastric juice and has high digestion rate. In contrast, according to the invention, the forming route of the nano-micelle is reasonably designed again, acidic calcium salt of any one or a combination of more of calcium phosphate, calcium carbonate, calcium lactate and calcium gluconate is added into casein-phosphate buffer solution, citric acid is used for adjusting the pH value of the solution, and casein and calcium ions form a compact three-dimensional network structure through phosphorylation centers, calcium bridging, electrostatic interaction and the like. Meanwhile, under the action of ultrasound, the hydrophobic cavity inside the nano micelle becomes more compact, and the pores of the micelle become smaller. In addition, the invention further adopts a radio frequency device to process the casein nano-micelle, utilizes the high-frequency alternating electromagnetic wave of the radio frequency to induce the oscillation and migration of cations such as calcium ions, hydrogen ions and the like in the casein micelle, on one hand, the exposed proline generates certain degree of thermal denaturation to trigger the self-assembly of the nano-micelle, thereby generating resistance to the hydrolysis of protease, and on the other hand, the hydrated particle size of the nano-micelle is reduced to be in a state of uniform and tiny particle size. Finally, the aqueous phase is subjected to freeze drying, normal pressure drying or reduced pressure drying, and the obtained casein nano-micelle can generate a shielding effect on the hydrolysis of protease, namely the digestion rate in gastric juice is slowed down. The casein nano-micelle prepared by the principle and having slow decomposition and slow release effects has uniform particle size, no toxic or side effect and good industrial application prospect.
The technical scheme is as follows: the invention adopts the following technical scheme.
The invention aims to provide a preparation method of slowly digestible casein nano micelle powder, in particular to a preparation method of casein nano micelle powder capable of resisting protease hydrolysis, and the specific scheme is as follows:
(1) adding casein, calcium salt and citric acid into a phosphate buffer solution, and performing ultrasonic treatment on the stirred mixed solution;
(2) performing radio frequency treatment on the mixed solution obtained in the step (1), stirring and standing;
(3) and (3) carrying out solid-liquid separation on the solution obtained in the step (2), and drying the obtained water phase to obtain the casein nano micelle powder.
In one embodiment of the invention, the casein in step (1) is derived from any one or more of α s 1-casein, α s 2-casein, β -casein and κ -casein. The alpha s 1-casein and alpha s 2-casein are casein in common cow milk and are easy to obtain.
In one embodiment of the present invention, the calcium salt in step (1) includes any one or more of calcium phosphate, calcium carbonate, calcium lactate, and calcium gluconate.
In one embodiment of the invention, the mass ratio of the casein to the calcium salt in the step (1) is 10 (1-5), and the total concentration of the casein and the calcium salt in the mixed solution is 2% -4.5% (w/v).
In one embodiment of the invention, the mass ratio of casein to citric acid in the step (1) is 20 (1-10).
In one embodiment of the present invention, the phosphate buffer in step (1) has a pH of 7.4 and a concentration of 1 to 100mM,
in one embodiment of the present invention, the concentration of the phosphate buffer in step (1) is 20 to 80 mM.
In one embodiment of the present invention, the ultrasonic treatment conditions in the step (1) are: the ultrasonic power is 50-200w, and the ultrasonic time is 5-20 min.
In one embodiment of the present invention, the radio frequency processing device applied in step (2) is a radio frequency device equipped with a hot air auxiliary system, and the resonance frequency of the radio frequency device is one of 13.56MHz, 27.12MHz and 40.68 MHz.
In one embodiment of the invention, the radio frequency output power in the step (2) is 1.0-8.0 kW.
In one embodiment of the present invention, the inter-plate distance of the radio frequency device in the step (2) is 40-120 mm.
In one embodiment of the present invention, the container of the radio frequency device in the step (2) is a thermoplastic basket with good electrical insulation.
In one embodiment of the invention, the temperature of the hot air auxiliary in the radio frequency device in the step (2) is 50-55 ℃.
In one embodiment of the present invention, the rf processing time in the step (2) is 100-.
In one embodiment of the present invention, the drying in the step (3) includes any one of freeze drying, atmospheric drying or reduced pressure drying.
It is another object of the present invention to provide a slowly digestible casein micelle powder prepared using the above method.
The invention also aims to apply the slowly digestible casein micelle powder to the fields of preparing common foods, health-care foods, special medical formula foods, drug carriers and the like.
The invention has the beneficial effects that:
(1) according to the method, acidic calcium salts such as calcium phosphate, calcium carbonate, calcium lactate and calcium gluconate are adopted, the pH value of a casein aqueous solution is adjusted by adding citric acid, and the three-dimensional network structure of the casein nano-micelle is redefined under the assistance of ultrasound, so that a hydrophobic cavity in the casein nano-micelle becomes more compact, and the pores of the micelle are reduced;
(2) the casein nano-micelle is treated by high-frequency alternating-current electromagnetic waves, cation oscillation migration such as calcium ions, hydrogen ions and the like in the micelle is induced, proline exposed on the surface of the micelle generates certain thermal denaturation, the nano-micelle is initiated to be self-assembled again, and the novel slow-digestion casein nano-micelle with small hydrated particle size and uniform structure is formed and has certain digestion resistance in the protease hydrolysis process;
(3) the raw materials of the slow-digestion casein nano micelle powder prepared by the invention are all food grade, and the slow-digestion casein nano micelle powder can be applied to the food fields of common food, health food, special medical formula food and the like and the medicine fields of drug carriers and the like. It can provide protein continuously, also can provide anti-hydrolysis characteristic, maintain muscle state, slowly release encapsulated hydrophobic functional food or drug, and provide new solution for body-building people, obese people and disease treatment.
Drawings
Fig. 1 is a schematic diagram of the preparation of slowly digestible casein nano micelle powder, wherein the solid-liquid separation and drying processes are not shown in the figure, but the understanding of ultrasonic and radio frequency treatment is not affected.
Detailed Description
The invention provides casein nano micelle powder capable of resisting protease hydrolysis and a preparation method thereof, and the specific scheme of the preparation method is as follows:
(1) dissolving casein and calcium salt in phosphate buffer solution with a certain volume, adding a certain amount of citric acid, magnetically stirring for 5-10 min, and treating the mixed solution by using ultrasound;
(2) putting the mixed solution obtained in the step (1) into a radio frequency device with a treatment cavity for radio frequency treatment, continuously stirring for 15-20 min, and standing;
(3) and (3) carrying out solid-liquid separation on the mixed solution obtained in the step (2), removing redundant undissolved calcium salt, and drying the water phase to obtain the casein nano micelle powder product.
The following description of the preferred embodiments of the present invention is provided for the purpose of better illustrating the invention and is not intended to limit the invention thereto.
Example 1
A slowly digestible casein nano micelle powder and its preparation method are provided. The method comprises the following specific steps:
(1) 10g of alpha s 1-casein, 10g of alpha s 2-casein and 2g of calcium phosphate are taken from cow milk, dispersed in 500mL of 20mM phosphate buffer solution with pH7.4, 5g of citric acid is added, magnetic stirring is carried out for 5min, and then ultrasonic treatment is carried out for 5min at 100 w.
(2) Slowly adding the solution obtained in the step (1) into a thermoplastic plastic basket with good electrical insulation, wherein the radio frequency working frequency is 27.12MHz, the power is set to be 5kW, the distance between polar plates is 45mm, the auxiliary temperature in a radio frequency treatment device is set to be 55 ℃, and the treatment time is 300 s. Followed by magnetic stirring for 15 min.
(3) And (3) separating the water phase in the step (2), and freeze-drying the water phase to obtain the slow-digestion casein nano micelle powder.
Example 2
A slowly digestible casein nano micelle powder and its preparation method are provided. The method comprises the following specific steps:
(1) dispersing 5g of alpha s 1-casein, 7g of alpha s 2-casein, 9g of beta-casein, 4g of kappa-casein and 8g of calcium carbonate in 800mL of 80mM phosphate buffer solution with pH7.4, adding 4g of citric acid, magnetically stirring for 5min, and treating for 16min by ultrasonic treatment at 200 w.
(2) Slowly adding the solution obtained in the step (1) into a thermoplastic plastic basket with good electrical insulation, wherein the radio frequency working frequency is 40.68MHz, the power is set as 8kW, the distance between polar plates is 100mm, the auxiliary temperature in a radio frequency treatment device is set as 55 ℃, and the treatment time is 400 s. Followed by magnetic stirring for 15 min.
(3) And (3) separating the water phase in the step (2), and drying at normal pressure to obtain the slowly digestible casein nano powder.
Example 3
A slowly digestible casein nano micelle powder and its preparation method are provided. The method comprises the following specific steps:
(1) 20g of commercial cow milk composite casein (a composite of various caseins) and 8g of calcium gluconate are dispersed in 900mL of 55mM phosphate buffer solution with pH7.4, 10g of citric acid is added, magnetic stirring is carried out for 5min, and ultrasonic treatment is carried out for 17min at 120 w.
(2) Slowly adding the solution obtained in the step (1) into a thermoplastic plastic basket with good electrical insulation, wherein the radio frequency working frequency is 13.56MHz, the power is set to be 8kW, the distance between polar plates is 60mm, the auxiliary temperature in a radio frequency treatment device is set to be 55 ℃, and the treatment time is 500 s. Followed by magnetic stirring for 15 min.
(3) And (3) separating the water phase in the step (2), concentrating, and drying under reduced pressure to obtain the slowly digestible casein nano powder.
To better explain the remarkable effects of the present invention, the comparative examples were added as follows:
comparative example 1
The preparation method is the same as example 1, except that the preparation method of casein micelle powder without radio frequency treatment comprises the following specific steps:
a slowly digestible casein nano micelle powder and its preparation method are provided. The method comprises the following specific steps:
(1) dispersing 10g of cow milk alpha s 1-casein, 10g of alpha s 2-casein and 2g of calcium phosphate in 500mL of 20mM phosphate buffer solution with pH of 7.4, adding 5g of citric acid, magnetically stirring for 5min, and treating for 5min by using 100w of ultrasound.
(2) And (2) separating the water phase in the step (1), concentrating, and freeze-drying to obtain the slow-digestion casein nano micelle powder.
Comparative example 2
The preparation method is the same as example 1, except that casein micelle powder without ultrasonic and radio frequency treatment is prepared, and the specific steps are as follows:
(1) 10g of alpha s 1-casein, 10g of alpha s 2-casein and 2g of calcium phosphate are taken from cow milk, dispersed in 500mL of 20mM phosphate buffer solution with pH7.4, 5g of citric acid is added, and magnetic stirring is carried out for 5 min.
(2) And (2) concentrating the solution obtained in the step (1), and freeze-drying the concentrated solution to obtain casein nano micelle powder.
Comparative example 3
Casein micelle powder was prepared by a conventional glycosylation method. The method comprises the following specific steps:
(1) taking 10g of cow milk alpha s 1-casein and 10g of alpha s 2-casein, dissolving in 20mL of phosphate buffer solution with pH7.4, stirring for 2.5h at room temperature, inserting an ultrasonic instrument probe into the position 2cm below the protein liquid surface, carrying out ice bath treatment for 30min under the condition of power 100W, collecting a sample, adding 10g of chitosan, uniformly mixing, freeze-drying for 48h, grinding the sample into powder, sieving the powder (120 meshes), placing the powder into a 25mL small beaker, placing the beaker into a reaction container (relative humidity is 80%) containing saturated potassium bromide, controlling the reaction temperature at 70 ℃, reacting for 30min, and cooling to terminate the reaction.
(2) And (2) removing the water phase of the micelle obtained in the step (1), concentrating, and freeze-drying to obtain the glycosylated casein copolymer.
(3) And (3) dissolving the glycosylated casein copolymer obtained in the step (2) in a phosphate buffer solution with the pH value of 7.4 to prepare a copolymer solution with the final concentration of 2mg/mL, placing the copolymer solution in a thermostatic water bath at the temperature of 25 ℃, stirring for 3 hours, and then transferring the copolymer solution to a refrigerator at the temperature of 4 ℃ for standing for 12 hours. Then carrying out ultrasonic treatment at 100W for 6min under ice bath conditions to obtain a glycosylated casein micelle solution.
(4) And (4) concentrating the glycosylated casein micelle solution obtained in the step (3), and freeze-drying to obtain glycosylated casein micelle powder.
Comparison of product characteristics of examples and comparative examples:
the degree of hydrolysis of protein is an important index for evaluating the degree of digestion of protein, and the degree of hydrolysis of protein is tested by an in vitro digestion simulation method.
First, the degree of proteolysis by pepsin hydrolysis was determined: the casein micelle powder solutions prepared in the above-mentioned specific examples and comparative examples were hydrolyzed using pepsin in an amount of 5% by mass of the total protein, and the activity was 8 × 104U/g, degree of proteolysis was determined after hydrolysis.
Subsequently, the degree of proteolysis by trypsin hydrolysis was determined: adjusting pH of the solution to 7.0, hydrolyzing with trypsin in an amount of 10% of total protein content, and activating to 3.9 × 105U/g, degree of proteolysis was determined after hydrolysis.
The test results are shown in tables 1 and 2 (blank is casein without any treatment):
TABLE 1 degree of proteolysis of different samples by pepsin
Figure BDA0003018680600000071
TABLE 2 further degree of proteolysis of different samples by trypsin
Figure BDA0003018680600000072
According to the data in the tables 1 and 2, the sample obtained by the conventional casein nano micelle preparation method is the comparative example 2, and the test result shows that the sample has almost no resistance to two proteases and is rapidly hydrolyzed under the treatment of the two proteases; while comparative example 3 is a glycosylated casein nanomicelle prepared using glycosylated casein, which is only slightly resistant to hydrolysis by pepsin and trypsin.
Compared with the comparative example and the blank group, the hydrolysis degree of the casein nano-micelle powder prepared by the embodiment 1-3 is obviously lower than that of the comparative example and the blank group by adopting the technical scheme of the invention, and the casein nano-micelle powder prepared by the embodiment 1-3 has higher resistance to pepsin and trypsin, particularly the casein nano-micelle powder prepared by the embodiment 2 has the hydrolysis degree greatly reduced by 80 percent compared with the blank group and the comparative example, which fully verifies that the treatment process of reconstructing a three-dimensional network structure by using the acidic calcium salt, performing ultrasonic densification and performing radio frequency induced self-assembly effectively improves the technical problems that the casein is easy to hydrolyze by protease in gastric juice and has higher digestion rate, and has obvious promotion effect on improving the stability of the casein and slowing down the digestion rate.
In the invention, acidic calcium salt of any one or a plurality of combinations of calcium phosphate, calcium carbonate, calcium lactate and calcium gluconate is added into casein-phosphate buffer solution, and casein and calcium ions form a compact three-dimensional network structure through phosphorylation centers, calcium bridging, electrostatic interaction and the like. Meanwhile, the hydrophobic cavity inside the nano micelle becomes more compact by ultrasound, and the pore of the micelle is reduced; in addition, radio frequency self high frequency alternating current electromagnetic waves are utilized to induce the oscillation migration of cations such as calcium ions, hydrogen ions and the like in the casein micelles, on one hand, exposed proline is subjected to certain thermal denaturation to trigger the self-assembly of the nano micelles again, so that the resistance is generated to the hydrolysis of protease, and on the other hand, the hydrated particle size of the nano micelles is reduced to be in a uniform and micro particle size state.
In conclusion, the casein nano micelle powder prepared by the invention is only partially hydrolyzed under the treatment of pepsin and trypsin, and a slowly digested casein nano micelle powder product is successfully prepared.
The above description is only of the preferred embodiments of the present invention, and it should be noted that: it will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the principles of the invention and these are intended to be within the scope of the invention.

Claims (10)

1. A preparation method of slowly digestible casein nano micelle powder is characterized by comprising the following steps:
(1) adding casein, calcium salt and citric acid into a phosphate buffer solution, and performing ultrasonic treatment on the stirred mixed solution;
(2) performing radio frequency treatment on the mixed solution obtained in the step (1), stirring and standing;
(3) and (3) carrying out solid-liquid separation on the solution obtained in the step (2), and drying the obtained water phase to obtain the casein nano micelle powder.
2. The method for preparing slowly digestible casein nanomicelle powder according to claim 1, wherein the casein in step (1) comprises any one or more of α s 1-casein, α s 2-casein, β -casein and/or κ -casein.
3. The method for preparing slowly digestible casein nanomicelle powder according to claim 1, wherein the calcium salt in step (1) comprises any one or more of calcium phosphate, calcium carbonate, calcium lactate, and calcium gluconate.
4. The preparation method of the slowly digestible casein nanomicelle powder according to claim 1, wherein the mass ratio of the casein to the calcium salt in step (1) is 10 (1-5), and the total concentration of the casein and the calcium salt in the mixed solution is 2-4.5% (w/v).
5. The preparation method of the slowly digestible casein nanomicelle powder according to claim 1, wherein the mass ratio of casein to citric acid in step (1) is 20 (1-10).
6. The method for preparing slowly digestible casein nanomicelle powder according to claim 1, wherein the phosphate buffer in step (1) has a pH of 7.4 and a concentration of 1-100mM, preferably 20-80 mM.
7. The method for preparing slowly digestible casein nanomicelle powder according to claim 1, wherein the ultrasonic treatment in step (1) is performed under the following conditions: the ultrasonic power is 50-200w, and the ultrasonic time is 5-20 min.
8. The method for preparing slowly digestible casein nanomicelle powder according to claim 1, wherein the radio frequency treatment in step (2) is performed by using a radio frequency device equipped with a hot air auxiliary system, the resonance frequency of the radio frequency device is one of 13.56MHz, 27.12MHz and 40.68MHz, and the radio frequency treatment conditions are as follows: the radio frequency output power is 1.0-8.0kW, the distance between the electrode plates is 40-120mm, the hot air auxiliary temperature is 50-55 ℃, and the treatment time is 100-500 s.
9. Casein nanomicelle powder obtainable by the process according to any one of claims 1 to 8.
10. Use of the casein nano-micelle powder of claim 9 in the preparation of general food, health food, formula food for special medical use, and drug carrier.
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114947108A (en) * 2022-04-28 2022-08-30 中国农业大学 Casein micelle accurate controlled-release composition and preparation method thereof
CN115005320A (en) * 2022-05-07 2022-09-06 中国农业大学 Preparation method of casein powder with long-acting satiety and obtained product
WO2023151198A1 (en) * 2022-02-10 2023-08-17 江南大学 Casein micelle having compositional and structural features imitating human milk and preparation method therefor
WO2023230648A1 (en) * 2022-05-30 2023-12-07 All G Foods Holdings Pty Limited Compositions
US11963542B2 (en) 2022-02-10 2024-04-23 Jiangnan University Casein micelle with composition and structural features simulating human milk and preparation method thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109480291A (en) * 2018-11-19 2019-03-19 浙江工商大学 Casein-carboxymethyl chitosan glycosylation nanoparticle preparation and its application

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109480291A (en) * 2018-11-19 2019-03-19 浙江工商大学 Casein-carboxymethyl chitosan glycosylation nanoparticle preparation and its application

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
MIN YANG,ET AL.: "Effect of ultrasound pretreatment on the physicochemical properties and simulated gastrointestinal digestibility of micellar casein concentrates", 《FOODSCIENCEANDTECHNOLOGY》 *
韩丹等: "酪蛋白作为药物纳米递送载体的研究进展", 《中国新药杂志》 *
高芸芳等: "酪蛋白结构及其酶解特性研究进展", 《农产品加工(学刊)》 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023151198A1 (en) * 2022-02-10 2023-08-17 江南大学 Casein micelle having compositional and structural features imitating human milk and preparation method therefor
US11963542B2 (en) 2022-02-10 2024-04-23 Jiangnan University Casein micelle with composition and structural features simulating human milk and preparation method thereof
CN114947108A (en) * 2022-04-28 2022-08-30 中国农业大学 Casein micelle accurate controlled-release composition and preparation method thereof
CN115005320A (en) * 2022-05-07 2022-09-06 中国农业大学 Preparation method of casein powder with long-acting satiety and obtained product
CN115005320B (en) * 2022-05-07 2023-09-05 中国农业大学 Preparation method of casein powder with long-acting satiety and obtained product
WO2023230648A1 (en) * 2022-05-30 2023-12-07 All G Foods Holdings Pty Limited Compositions

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