CN116102743B - Electrostatic self-assembled protein fiber hydrogel and preparation method thereof - Google Patents
Electrostatic self-assembled protein fiber hydrogel and preparation method thereof Download PDFInfo
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- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 39
- 239000000835 fiber Substances 0.000 title claims abstract description 25
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 239000000243 solution Substances 0.000 claims abstract description 53
- 108010046377 Whey Proteins Proteins 0.000 claims abstract description 47
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 claims abstract description 46
- 102000007544 Whey Proteins Human genes 0.000 claims abstract description 44
- 235000021119 whey protein Nutrition 0.000 claims abstract description 44
- 235000018102 proteins Nutrition 0.000 claims abstract description 38
- 235000010987 pectin Nutrition 0.000 claims abstract description 31
- 229920001277 pectin Polymers 0.000 claims abstract description 31
- 239000001814 pectin Substances 0.000 claims abstract description 31
- 229940109262 curcumin Drugs 0.000 claims abstract description 26
- 239000002121 nanofiber Substances 0.000 claims abstract description 25
- 235000012754 curcumin Nutrition 0.000 claims abstract description 23
- 239000004148 curcumin Substances 0.000 claims abstract description 23
- VFLDPWHFBUODDF-UHFFFAOYSA-N diferuloylmethane Natural products C1=C(O)C(OC)=CC(C=CC(=O)CC(=O)C=CC=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-UHFFFAOYSA-N 0.000 claims abstract description 23
- 239000005862 Whey Substances 0.000 claims abstract description 11
- 239000011259 mixed solution Substances 0.000 claims abstract description 8
- 238000000034 method Methods 0.000 claims abstract description 6
- 238000003756 stirring Methods 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
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- 229910021641 deionized water Inorganic materials 0.000 claims description 10
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- 230000001105 regulatory effect Effects 0.000 claims description 5
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- 238000002156 mixing Methods 0.000 claims description 3
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- 239000002245 particle Substances 0.000 abstract description 15
- 238000002834 transmittance Methods 0.000 abstract description 6
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- 229940121375 antifungal agent Drugs 0.000 description 1
- 239000013060 biological fluid Substances 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
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- 230000029087 digestion Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 239000000416 hydrocolloid Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000000707 layer-by-layer assembly Methods 0.000 description 1
- 229930013686 lignan Natural products 0.000 description 1
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- 235000010493 xanthan gum Nutrition 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- 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
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
- C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
- C08J3/075—Macromolecular gels
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- C—CHEMISTRY; METALLURGY
- 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
- C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
- C08J2305/06—Pectin; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- 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
- C08J2489/00—Characterised by the use of proteins; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K5/00—Use of organic ingredients
- C08K5/04—Oxygen-containing compounds
- C08K5/13—Phenols; Phenolates
- C08K5/132—Phenols containing keto groups, e.g. benzophenones
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
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Abstract
The invention discloses an electrostatic self-assembled protein fiber hydrogel and a preparation method thereof. The invention adopts subcritical hydrothermal synthesis method to make whey protein isolate fibrillate to form whey protein isolate nanofiber, then the whey protein isolate nanofiber is mixed with curcumin to form curcumin-whey protein isolate nanofiber mixed solution, pectin solution is added, and static self-assembled protein fiber hydrogel is obtained by stirring. The preparation method is simple, the prepared hydrogel has smaller average particle size, moderate transmittance and high stability, and can realize efficient embedding and delivery of active substance curcumin.
Description
Technical Field
The invention belongs to the technical field of protein hydrogel preparation, and relates to an electrostatic self-assembled protein fiber hydrogel and a preparation method thereof.
Background
Curcumin (Curcumin, cur) is a hydrophobic polyphenol isolated from the roots of traditional herbs and has various physiological activities such as antioxidation, anticancer, antibacterial, antifungal and anti-inflammatory. However, curcumin has low bioavailability due to its low chemical stability, poor water solubility, and poor storage stability. To increase the water solubility, stability and bioavailability of curcumin, many researchers have encapsulated curcumin in a delivery system for gastrointestinal delivery.
Hydrogels are polymeric materials that retain a large amount of water or biological fluid within their structure, a three-dimensional crosslinked network structure that is not readily dissolvable. The natural hydrogel has poor mechanical properties, and the protein synthesized hydrogel has good mechanical properties, but has the defects of fragile texture, easy degradation in gastrointestinal digestion process and the like.
Whey protein isolate nanofibers (Whey Protein Isolate Nanofiber, WPINs) are materials prepared from whey protein isolate (Whey Protein Isolate, WPI) by subcritical water equipment and have a significant positive charge. To improve the stability of whey protein-based hydrogels, some researchers have used differently charged polysaccharides and proteins to form stable polymers to prepare hydrogels. Document 1a whey protein isolate-chitosan hydrogel is prepared by crosslinking whey protein isolate and chitosan with lignan, and the release rate of the hydrogel in intestinal juice is too slow to facilitate absorption of the entrapped bioactive substance (Food Hydrocolloids,2020, 103:105619). Document 2 reports an elastic hydrogel made of whey protein isolate and xanthan gum, which has a strong high thermal stability but a consistency too high to be applied to mass production in food factories (Journal of Food Process Engineering,2021,44 (8): e 13751).
Disclosure of Invention
The invention aims to provide an electrostatic self-assembled protein fiber hydrogel and a preparation method thereof. The invention assembles whey protein isolate nanofiber with positive charges and pectin with negative charges into polymer hydrogel by utilizing an electrostatic autonomous loading technology, solves the problems of low mechanical strength and easy degradation of single protein hydrogel, and the obtained hydrogel has a stable structure and coats active substance curcumin, thereby realizing efficient embedding of curcumin, having better stability and being capable of realizing safe and stable delivery of curcumin.
The technical scheme for realizing the purpose of the invention is as follows:
the preparation method of the electrostatic self-assembled protein fiber hydrogel comprises the following specific steps:
(1) According to the mass ratio of 2-5:1, dissolving whey protein isolate in deionized water, regulating the pH value to 2.0-4.0, and filtering to obtain whey protein isolate solution;
(2) Heating whey protein isolate solution in subcritical water device for 120min to obtain Whey Protein Isolate Nanofiber (WPINs) solution;
(3) Adding an ethanol solution of curcumin with the concentration of 1-5 mg/mL into the whey protein isolate nanofiber solution according to the volume ratio of 1:40-1:50 to obtain a curcumin-whey protein isolate nanofiber mixed solution;
(4) 1, dissolving Pectin (P) in deionized water to obtain Pectin solution, wherein the mass ratio of Pectin to Pectin is 2+/-0.5%;
(5) Mixing and stirring the curcumin-whey protein isolate nanofiber mixed solution and the pectin solution for 1-5 hours according to the volume ratio of 1:3-1:4, and obtaining the electrostatic self-assembled protein fiber hydrogel.
Preferably, in step (1), the filter membrane used in the filtration has a pore size of 0.45. Mu.m.
Preferably, in step (2), the subcritical water device is heated to a temperature of 110 ℃.
Preferably, in the step (1), the mass ratio is 5 percent to 1, the pH is adjusted to 2.0, in the step (3), the volume ratio is 1:50, the ethanol solution of curcumin is 1mg/mL, in the step (4), the mass ratio is 2 percent to 1, in the step (5), the volume ratio is 1:4, and the stirring time is 2 hours.
Compared with the prior art, the invention has the following advantages:
The whey protein isolate, pectin and curcumin adopted by the invention are all biodegradable green materials, so that the method is safe and environment-friendly, and the protein fiber hydrogel is prepared by an electrostatic self-assembly mode, and is simple and easy to implement. The protein fiber hydrogel prepared by the invention has smaller average particle size (nano level), moderate transmittance and high stability, and the embedding rate of curcumin can reach 96.17% at most, thus realizing stable and efficient embedding and delivery of active substances.
Drawings
FIG. 1 is a graph showing the transmittance of the hydrogel prepared in example 1;
FIG. 2 is a graph showing the particle diameter potential of hydrogels prepared in example 2 and comparative examples 1 to 3;
FIG. 3 is a Transmission Electron Microscope (TEM) image of the hydrogels prepared in example 2 and comparative examples 1-3;
FIG. 4 is an XRD pattern of hydrogels prepared in example 2 and comparative examples 1-3;
FIG. 5 is a graph showing the thermal stability of the hydrogels prepared in example 2 and comparative examples 1 to 3;
FIG. 6 is a graph showing the entrapment rate of curcumin in the hydrogels prepared in example 3 and comparative example 4.
Detailed Description
The invention is described in further detail below with reference to specific embodiments and figures.
Example 1
According to the mass ratio of 5 percent to 1, the whey protein isolate is dissolved in deionized water to prepare a solution with the concentration of 5 percent, the pH value of the solution is regulated to 2.0, and the solution is filtered through a 0.45 mu m filter membrane to obtain the whey protein isolate solution. And (3) placing the whey protein isolate solution in a subcritical water device at 110 ℃ and heating for 90, 120 and 180 minutes respectively to obtain the whey protein isolate nanofiber solution. Respectively dissolving pectin into deionized water according to the mass ratio of 0-5:1 to prepare pectin solutions with mass concentrations of 0-5%. The whey protein isolate nanofiber solution and the pectin solution are mixed and stirred for 2 hours according to the volume ratio of 1:4 to obtain protein fiber hydrogel, which is respectively named WPINs-90-P-H, WPINs-120-P-H, WPINs-180-P-H series according to heating time. The transmittance of each protein fiber hydrogel was tested and as shown in fig. 1, it can be seen from the graph that as the pectin concentration increases, the transmittance of the hydrogel tends to increase and decrease, and reaches a maximum at a pectin concentration of 1%. Since embedding experiments of curcumin are required to be carried out later, the transmittance is not too high or too low, and therefore the pectin concentration of 2% is selected for the subsequent experiments.
Example 2
According to the mass ratio of 5 percent to 1, the whey protein isolate is dissolved in deionized water to prepare a solution with the concentration of 5 percent, the pH value of the solution is regulated to 2.0, and the solution is filtered through a 0.45 mu m filter membrane to obtain the whey protein isolate solution. And (3) placing the whey protein isolate solution in a subcritical water device and heating at 110 ℃ for 120min to obtain the whey protein isolate nanofiber solution. The pectin is dissolved in deionized water according to the mass ratio of 2 percent to 1 to prepare a pectin solution with the concentration of 2 percent. And mixing and stirring the whey protein isolate nanofiber solution and the pectin solution according to the volume ratio of 1:4 for 2 hours to obtain protein fiber hydrogel WPINs-120-P-H.
As a result of testing the particle size and the potential of the hydrogel, it was found from FIG. 2 that WPINs-120-P-H had a particle size of 659nm and the potential was-30 mV. As can be seen from the transmission electron microscope of FIG. 3, the present fiber state has more branches and moderate length, and the combination amount with pectin is maximum. As can be seen from the XRD pattern of FIG. 4, WPINs-120-P-H formed two distinct peaks at 2 theta, 26.7-27.1℃and 39.2-40.8℃indicating partial crystal structures of whey protein-separating nanofibers and pectin. The protein fiber hydrogel WPINs-120-P-H was heated at 30 ℃,60 ℃ and 90 ℃ for 30min, and the thermal stability was tested, and the results are shown in FIG. 5, and it can be seen that the variation range of the overall particle size at different temperatures is the smallest and the stability is the best.
Comparative example 1
This comparative example is essentially the same as example 2, except that the whey protein isolate solution is placed in a subcritical water apparatus at 110 ℃ for 90 minutes to produce a protein fiber hydrogel of WPINs-90-P-H.
As a result of testing the particle size and the potential of the hydrogel, it was found from FIG. 2 that WPINs-90-P-H had a particle size of 728nm and the potential was-17.2 mV. As can be seen from the transmission electron microscope image of FIG. 3, the WPINs-90-P-H protein fiber is curved, has few branches and has the least combination with pectin. As can be seen from the XRD pattern of FIG. 4, WPINs-90-P-H formed 1 distinct peak at 2. Theta. At 26.7-27.1 deg., indicating a partial crystal structure of whey protein-separating nanofibers. The protein fiber hydrogel WPINs-90-P-H was heated at 30 ℃, 60 ℃ and 90 ℃ for 30min, and the thermal stability was tested, and the results are shown in FIG. 5, and it can be seen from the graph that the particle size was increased and the stability was poor after heating.
Comparative example 2
This comparative example is essentially the same as example 2, except that the whey protein isolate solution is placed in a subcritical water apparatus at 110 ℃ for 180 minutes to produce a protein fiber hydrogel of WPINs-180-P-H.
As a result of testing the particle size and the potential of the hydrogel, it was found from FIG. 2 that WPINs-180-P-H had a particle size of 683nm and the potential was-24.4 mV. As can be seen from the transmission electron microscope image of FIG. 3, the fibrous state is too much branched, but the binding amount with pectin is small. As can be seen from the XRD pattern of FIG. 4, WPINs-180-P-H formed two distinct peaks at 2 theta, 26.7-27.1℃and 39.2-40.8℃indicating partial crystal structures of whey protein-separating nanofibers and pectin. The protein fiber hydrogel WPINs-180-P-H is heated at 30 ℃, 60 ℃ and 90 ℃ for 30min respectively, and the thermal stability is tested, and the result is shown in figure 5, and the particle size is increased and the stability is poor after heating.
Comparative example 3
This comparative example was essentially the same as example 2, except that the whey protein isolate solution was not subjected to subcritical hydrothermal treatment, and the whey protein isolate solution and the pectin solution were directly mixed and stirred for 2 hours at a volume ratio of 1:4 to obtain a protein hydrogel WPI-P-H.
As a result of testing the particle size and the potential of the hydrogel, the particle size of WPI-P-H was 691nm and the potential was-21.2 mV, as shown in FIG. 2. As can be seen from the transmission electron microscope image of FIG. 3, the whey protein isolate exhibits a dot shape and a small amount of binding to pectin. As can be seen from the XRD pattern of fig. 4, there is no distinguishable peak at 2θ. The protein fiber hydrogel WPI-P-H is heated at 30 ℃, 60 ℃ and 90 ℃ for 30min respectively, and the thermal stability is tested, and the result is shown in figure 5, and the particle size is obviously increased and the stability is poor after the heating at 90 ℃.
Example 3
According to the mass ratio of 5 percent to 1, the whey protein isolate is dissolved in deionized water to prepare a solution with the concentration of 5 percent, the pH value of the solution is regulated to 2.0, and the solution is filtered through a 0.45 mu m filter membrane to obtain the whey protein isolate solution. And (3) placing the whey protein isolate solution in a subcritical water device and heating at 110 ℃ for 120min to obtain the whey protein isolate nanofiber solution. Curcumin was dissolved in ethanol to prepare a curcumin-ethanol solution at a concentration of 1 mg/mL. And adding the curcumin-ethanol solution into the whey protein isolate nanofiber solution according to the volume ratio of 1:50 to obtain a curcumin-whey protein isolate nanofiber mixed solution. The pectin is dissolved in deionized water according to the mass ratio of 2 percent to 1 to prepare a pectin solution with the concentration of 2 percent. The curcumin-whey protein isolate nanofiber mixed solution and the pectin solution are mixed and stirred for 2 hours according to the volume ratio of 1:4, so that the protein fiber hydrogel (WPINs-120-P-H-Cur) loaded with the curcumin is obtained, the embedding rate of the curcumin is 96.17%, and as shown in figure 6, the electrostatic self-assembled protein fiber hydrogel prepared by the invention can be used as a delivery carrier of bioactive substances, and the solubility and bioavailability of the bioactive substances are improved.
Comparative example 4
The comparative example was substantially the same as example 3, except that subcritical hydrothermal treatment was not performed on the whey protein isolate solution, and curcumin-whey protein isolate mixed solution and pectin solution were directly mixed and stirred for 2 hours at a volume ratio of 1:4 to obtain curcumin-loaded protein hydrogel (WPI-P-H-Cur), and the entrapment rate of curcumin was 31.02%, as shown in FIG. 6, and was low.
Claims (5)
1. The preparation method of the electrostatic self-assembled protein fiber hydrogel is characterized by comprising the following specific steps:
(1) According to the mass ratio of 2-5:1, dissolving whey protein isolate in deionized water, regulating the pH value to 2.0-4.0, and filtering to obtain whey protein isolate solution;
(2) Heating whey protein isolate solution in subcritical water device for 120min to obtain whey protein isolate nanofiber solution;
(3) Adding an ethanol solution of curcumin with the concentration of 1-5 mg/mL into the whey protein isolate nanofiber solution according to the volume ratio of 1:40-1:50 to obtain a curcumin-whey protein isolate nanofiber mixed solution;
(4) 1, dissolving pectin in deionized water according to the mass ratio of 2+/-0.5 percent to obtain pectin solution;
(5) Mixing and stirring the curcumin-whey protein isolate nanofiber mixed solution and the pectin solution for 1-5 hours according to the volume ratio of 1:3-1:4, and obtaining the electrostatic self-assembled protein fiber hydrogel.
2. The process according to claim 1, wherein in the step (1), the pore size of the filter membrane used for the filtration is 0.45. Mu.m.
3. The method according to claim 1, wherein in the step (2), the subcritical water device is heated at a temperature of 110 ℃.
4. The preparation method of claim 1, wherein in the step (1), the mass ratio is 5% to 1, the pH is adjusted to 2.0, in the step (3), the volume ratio is 1:50, the ethanol solution of curcumin is 1mg/mL, in the step (4), the mass ratio is 2% to 1, in the step (5), the volume ratio is 1:4, and the stirring time is 2h.
5. An electrostatically self-assembled protein fibrous hydrogel made by the method of any one of claims 1 to 4.
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| CN115073768A (en) * | 2022-05-07 | 2022-09-20 | 浙江工业大学 | Preparation method of functional component-loaded double-network hydrogel |
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