Grass carp myofibrillar protein / rutin complex and method for preparing emulsion gel

Abstract

This invention relates to the field of food technology and discloses a grass carp myofibrillar protein / rutin complex, wherein the mass ratio of grass carp myofibrillar protein to rutin is 1:0.01-0.3. This complex is mixed with an aqueous phase and an oil phase under high-speed shear, and after standing, an emulsion gel is obtained. This invention improves the utilization rate and application value of freshwater fish protein. Furthermore, the emulsion gel prepared using the grass carp myofibrillar protein / rutin complex has high stability. Using food-grade materials, it offers advantages such as high safety, wide application range, simple preparation method, and low cost.

Description

Technical Field

[0001] This invention relates to the field of food technology, and more specifically to a method for preparing an emulsion gel based on a grass carp myofibrillar protein / rutin complex. Background Technology

[0002] Emulsion gels are semi-solid food systems with gel networks filled with oil droplets. They possess typical mechanical properties and can effectively encapsulate hydrophilic / hydrophobic active substances, effectively protecting and sustaining their bioactive functions while meeting diverse emulsion needs. They are currently widely used in cheese, ice cream, yogurt, emulsified meat products, etc. Currently, due to clean labeling requirements, off-flavors and potential toxicity, and limitations on the application of synthetic surfactants in the food industry, natural surfactants such as phospholipids, proteins, and polysaccharides are extensively studied. However, information on protein-stabilized emulsion gels and their potential applications in actual foods, especially animal proteins, is scarce.

[0003] Currently, grass carp is mainly consumed fresh, and research on its high-value utilization, especially the development of novel high-value functional foods, is limited. Myofibrillar protein, as a major component of muscle protein, plays a crucial role in the quality of processed products due to its functional properties. However, some natural proteins struggle to exhibit good functional characteristics, directly impacting their effective utilization and failing to meet the diverse needs of consumers and the food industry.

[0004] Myofibrillar proteins, due to their amphiphilic nature, can adsorb onto the surface of fat particles, forming an interfacial protein film that immobilizes fat particles and prevents oil droplet aggregation, making them a highly promising food-grade emulsion stabilizer. Compared to particles from other biological sources, proteins have advantages such as high nutritional value, easy availability, and biocompatibility, thus offering multiple advantages in the production of stable, biodegradable, and food-grade Pickering emulsions. However, myofibrillar proteins are highly susceptible to oxidative denaturation during storage, altering their gelling, emulsifying, and solubilizing properties, becoming a significant factor severely restricting their high-value utilization. Therefore, current research mainly focuses on improving their functional properties through chemical modification (phosphorylation, glycosylation, acylation, etc.) and physical modification (ultrasound, ultra-high pressure, low-temperature plasma, irradiation, etc.). However, these methods often suffer from drawbacks such as uncontrollable processing, time-consuming and labor-intensive processes, environmental pollution, and the reaction products can easily trigger a series of safety issues. Summary of the Invention

[0005] This invention provides a grass carp myofibrillar protein / rutin complex, and a method for preparing an emulsion gel based on the complex.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] A grass carp myofibrillar protein / rutin complex, comprising grass carp myofibrillar protein and rutin, wherein the mass ratio of grass carp myofibrillar protein to rutin is 1:0.01-0.3.

[0008] Preferably, the mass ratio of grass carp myofibrillar protein to rutin is 1:0.02-0.2; in a preferred embodiment of the present invention, the mass ratio is 1:0.02-0.12.

[0009] The grass carp myofibril protein is extracted from the white meat on the back of the grass carp.

[0010] Preferably, the preparation method is as follows:

[0011] (1) Take fresh grass carp back meat and chop it into small pieces;

[0012] (2) Add 2-6 times the volume of phosphate extract (20 mmol / L K2HPO4 / KH2PO4, 0.1 mol / L NaCl, 2 mmol / L MgCl2, 1 mmol / L EDTA, pH 7.0) to the minced fish meat, homogenize and centrifuge at low temperature;

[0013] (3) Take the precipitate and repeat step (2) 1-3 times;

[0014] (4) Take the precipitate and add 2-6 times the volume of high-salt buffer solution (preferably 0.5-1 mol / L NaCl), homogenize, and centrifuge at low temperature to obtain the supernatant;

[0015] (5) Add 8-12 times the volume of distilled water, deionized water or purified water to the supernatant of step (4), centrifuge at 10000-20000 rpm for 15-30 min at 0-4℃, and take the precipitate as grass carp myofibril protein.

[0016] In steps (2)-(3), preferably, the amount of phosphate extract used is 4 times the volume.

[0017] In step (4), preferably, the amount of high-salt buffer solution used is 4 times the volume.

[0018] In steps (2)-(4), the homogenization conditions are: homogenize at 3000-5000 rpm for 45-90s, preferably homogenize at 4000 rpm for 60s; the low-temperature centrifugation conditions are: centrifuge at 4000-6000 rpm for 10-25min at 0-4℃, preferably centrifuge at 5000 rpm for 15min at 4℃.

[0019] In step (5), preferably, the volume ratio of the supernatant to distilled water, deionized water or purified water is 1:10.

[0020] The myofibrillar protein extracted in step (5) can be further freeze-dried to obtain grass carp myofibrillar protein particles, which can be stored for later use. Preferably, the myofibrillar protein is freeze-dried at -65°C for 60 h.

[0021] Grass carp myofibrillar protein and rutin can be mixed to obtain a grass carp myofibrillar protein / rutin complex. The mixture can be in the form of a powder or an aqueous solution.

[0022] The above-mentioned complexes can be used to prepare emulsifiers, emulsion gels, or emulsion gel stabilizers.

[0023] An emulsifier or emulsion gel stabilizer containing the above-mentioned grass carp myofibrillar protein / rutin complex. Its dosage form may be a powder, an aqueous solution, or an emulsion.

[0024] A method for preparing an emulsion gel includes the following steps: mixing the above-mentioned complex with an oil phase and an aqueous phase under high-speed shear mixing. After mixing, allowing it to stand for 5-20 minutes, typically 10 minutes, allows an emulsion gel to form.

[0025] The conditions for high-speed shearing are mixing at 10,000-50,000 rpm for 0.5-30 min, preferably at 10,000-20,000 rpm for 1-10 min. In one embodiment of the present invention, mixing is performed at 10,000 rpm for 1 min.

[0026] The oil phase is edible oil, including edible oil, animal fat, and synthetic triglycerides, such as medium-chain triglycerides (MCT) or long-chain triglycerides (LCT); in one embodiment of the invention, the oil phase is MCT. The aqueous phase has a pH of 5-9, preferably 6-8; in a preferred embodiment of the invention, pH=7. In one embodiment of the invention, the aqueous phase contains NaCl.

[0027] Alternatively, the oil phase and aqueous phase may be food, food ingredients, semi-finished food products, pharmaceuticals, pharmaceutical ingredients, or semi-finished pharmaceutical products containing water and oil.

[0028] The ratio of myofibrillar protein in the complex to the amount used is 0.01-0.05 kg / L. In a preferred embodiment of the present invention, the ratio is 0.02 kg / L.

[0029] The volume ratio of the oil phase to the water phase is 0.1-1:1, preferably 0.3-0.8:1; in a preferred embodiment of the present invention, the volume ratio of the oil phase to the water phase is 0.5:1.

[0030] An emulsion gel is prepared by the method described above.

[0031] Preferably, medium-chain triglycerides (MCTs) are used as the oil phase. This emulsion gel is edible and can be used as a carrier to load functional active substances, such as drugs and nutrients, for the preparation of nutritional products or pharmaceutical carriers.

[0032] This invention utilizes the characteristics of protein emulsion gels, selecting rutin to prepare a grass carp myofibrillar protein / rutin complex using a physical method. This complex is then added to the raw materials, and a one-step high-speed shearing method is used to achieve the green and efficient construction of a grass carp myofibrillar protein / rutin composite emulsion gel. The resulting emulsion gel exhibits high safety and good stability. This composite gel is characterized by rich nutrition, high safety, and strong stability, with low production costs, and can be widely used in the food industry. It provides a new reference for food emulsion gel products and meets diverse consumer needs.

[0033] Compared with the prior art, the advantages of this invention are:

[0034] (1) Grass carp is an excellent farmed fish with abundant resources and high economic value. This invention uses myofibrillar protein extracted from fish meat as raw material to prepare emulsion, instead of being limited to conventional whey protein, soy protein, etc., which can greatly improve the utilization rate of fish meat in the processing process. This not only helps to increase the production value of fish, but also expands the application range of emulsion gel. The prepared emulsion gel has high stability.

[0035] (2) The myofibrillar protein emulsion gel prepared by the present invention is rich in nutrients and has high stability.

[0036] (3) The emulsion gel prepared by the method of the present invention is a cold solid gel. Compared with the traditional protein emulsion gel preparation method, it does not require heating of the emulsion, the process is simpler, and it is more suitable for some materials that are unstable in a hot environment, thus expanding its application range.

[0037] (4) The present invention uses food-grade materials, which are highly safe and have a wide range of applications. It has the advantages of simple preparation method, short production cycle, safe processing method and low cost. Attached Figure Description

[0038] Figure 1 The emulsions prepared in Examples 1-5.

[0039] Figure 2 The condition of the emulsions prepared in Examples 1-5 after being inverted for 3 hours. Detailed Implementation

[0040] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0041] In the following embodiments, the method for extracting myofibrillar proteins is as follows:

[0042] (1) After killing the fresh grass carp, remove the red skin and tendons, take the white meat from the back and cut it at 3000 r / min for 4 min, stopping for 30 s every 1 min, and set the fish meat aside.

[0043] (2) Add four times the volume of phosphate extract (20 mmol / L K2HPO4 / KH2PO4, 0.1 mol / L NaCl, 2 mmol / L MgCl2, 1 mmol / L EDTA, pH 7.0) to the minced fish, homogenize at 4000 r / min for 60 s, and centrifuge at 5000 r / min and 4 ℃ for 15 min; discard the supernatant;

[0044] (3) Repeat step (2) twice with the remaining precipitate;

[0045] (4) Add the obtained precipitate to 4 times the volume of high-salt buffer (0.6 mol / L NaCl, pH 7.0), homogenize at 4000 r / min for 60 s, centrifuge (5000 r / min, 15 min, 4 ℃), and take the supernatant;

[0046] (5) Add 10 times the volume of distilled water to the supernatant of step (4), centrifuge at 10000 r / min for 20 min to obtain the precipitate, freeze dry at -65℃ for 60 h to obtain pure grass carp myofibril protein particles.

[0047] Example 1

[0048] A method for preparing an emulsion gel based on grass carp myofibrillar protein / rutin complex includes the following steps: first, an aqueous phase containing grass carp myofibrillar protein / rutin complex is prepared, and then it is mixed with an oil phase under high-speed shearing.

[0049] (i) Take 2.0 g of grass carp myofibrillar protein particles, disperse them in 50 mM PBS buffer (containing 0.6 M NaCl, pH 7.0) and dilute to 100 mL. Stir at 300 rpm for 50 min to ensure complete dissolution.

[0050] (ii) Add 2.0% (w / w, myofibrillar protein) of rutin (0.04 g) to the grass carp myofibrillar protein solution prepared in step (i), mix thoroughly, and react at 4 ℃ for 3 h to obtain a rutin and grass carp myofibrillar protein composite solution.

[0051] (iii) Mix MCT with the above myofibrillar protein solution at an oil-to-water volume ratio of 0.5 (v / v) and shear at 10000 r / min for 1 min. After standing for 10 min, an emulsion gel can be obtained.

[0052] Example 2

[0053] (i) Take 1.0 g of grass carp myofibrillar protein particles, disperse them in 50 mM PBS buffer (containing 0.6 M NaCl, pH 7.0) and dilute to 50 mL. Stir at 300 rpm for 60 min to dissolve them completely.

[0054] (ii) Add 4.0% (w / w, / myofibrillar protein) of rutin (0.04 g) to the prepared grass carp myofibrillar protein solution, and place at 4℃ for 3 h to obtain rutin-grass carp myofibrillar protein composite solution.

[0055] (iii) Mix MCT with the above myofibrillar protein solution at an oil-to-water volume ratio of 0.5 (v / v) and shear at 10,000 r / min for 1 min. After standing for 10 min, an emulsion gel can be obtained.

[0056] Example 3

[0057] (i) Take 1.0 g of grass carp myofibrillar protein particles, disperse them in 50 mM PBS buffer (containing 0.6 M NaCl, pH 7.0) and dilute to 50 mL. Stir at 300 rpm for 60 min to dissolve them completely.

[0058] (ii) Add 6.0% (w / w, / myofibrillar protein) of rutin (0.06 g) to the prepared grass carp myofibrillar protein solution, and place at 4°C for 3 h to obtain grass carp myofibrillar protein / rutin composite solution.

[0059] (iii) Mix MCT with the above myofibrillar protein solution at an oil-to-water volume ratio of 0.5 (v / v) and shear at 10,000 r / min for 1 min. After standing for 10 min, an emulsion gel can be obtained.

[0060] Example 4

[0061] (i) Take 1.0 g of grass carp myofibrillar protein particles, disperse them in 50 mM PBS buffer (containing 0.6 M NaCl, pH 7.0) and dilute to 50 mL. Stir at 300 rpm for 60 min to dissolve them completely.

[0062] (ii) Add 8.0% (w / w, / myofibrillar protein) of rutin (0.08 g) to the prepared grass carp myofibrillar protein solution, and place at 4℃ for 3 h to obtain grass carp myofibrillar protein / rutin composite solution.

[0063] (iii) Mix MCT with the above myofibrillar protein solution at an oil-to-water volume ratio of 0.5 (v / v) and shear at 10,000 r / min for 1 min. After standing for 10 min, an emulsion gel can be obtained.

[0064] Example 5

[0065] (i) Take 4.0 g of grass carp myofibrillar protein particles, disperse them in 50 mM PBS buffer (containing 0.6 M NaCl, pH 7.0) and dilute to 200 mL. Stir at 300 rpm for 55 min to fully dissolve them.

[0066] (ii) Take 100 mL of grass carp myofibrillar protein solution, add 12.0% (w / w, / myofibrillar protein) of rutin (0.24 g), and place at 4℃ for 3 h to obtain a rutin and grass carp myofibrillar protein complex solution.

[0067] (iii) Mix MCT with the above myofibrillar protein complex solution at an oil-to-water volume ratio of 0.5 (v / v) under high-speed shearing at 10000 r / min for 1 min, and let stand for 10 min to obtain emulsion gel.

[0068] Comparative Example

[0069] Take another 100 mL of grass carp myofibrillar protein solution (2 g / 100 mL), mix it with MCT at an oil-to-water volume ratio of 0.5 (v / v), and mix under high-speed shear at 10000 r / min for 1 min.

[0070] Images of the emulsion gels obtained in Examples 1-5 and the comparative product after standing for 0 days are shown below. Figure 1As shown in the figure, the samples from left to right represent emulsions obtained when MCT is used as the oil phase, the oil-to-water ratio is 0.5 (v / v), and the ratio of rutin to grass carp myofibrillar protein complex is 0, 2.0, 4.0, 6.0, 8.0, and 12.0% (w / w, rutin / myofibrillar protein), and the ratio of grass carp myofibrillar protein to the aqueous phase is 2 g / 100 mL. The figure shows that a stable emulsion was formed.

[0071] The image shows the emulsion gel after being inverted for 3 hours. Figure 2 As shown, the sample arrangement order is as follows: Figure 1 The results showed that a myofibrillar protein emulsion could be prepared by using a composite solution with 2 g / 100 mL protein and rutin added at a ratio of 2.0-12.0% (w / w, rutin / myofibrillar protein) as the aqueous phase, MCT as the oil phase, and an oil-water ratio of 0.5. Furthermore, the emulsion prepared using the rutin-added composite solution as the aqueous phase was stable after being inverted for 3 hours, exhibiting an emulsion gel-like consistency. The emulsion prepared without added rutin did not form an emulsion gel.

[0072] In the examples, when the grass carp myofibrillar protein content in the aqueous phase is 1.5 g / 100 mL or 3 g / 100 mL, the rutin addition is 6% (w / w, rutin / myofibrillar protein), and the oil-water volume ratio is 0.4 or 0.6, the basic characteristics of the resulting emulsion gel are similar to those described above.

[0073] The specific embodiments described above are merely illustrative examples of the spirit of the present invention and some experiments. Those skilled in the art can make various modifications or additions to the described specific embodiments or use similar methods to replace them, but without departing from the spirit of the present invention or exceeding the scope defined by the appended claims.

Claims

1. A grass carp myofibrillar protein / rutin complex, characterized in that, This complex is used to prepare emulsion gels containing grass carp myofibrillar protein and rutin, wherein the mass ratio of grass carp myofibrillar protein to rutin is 1:0.01-0.3; The preparation method of the grass carp myofibrillar protein is as follows: (1) Take fresh grass carp back meat and chop it into small pieces; (2) Add 2-6 times the volume of phosphate extract to the minced fish meat, homogenize at 3000-5000 rpm for 45-90 s, and centrifuge at 4000-6000 rpm for 10-25 min at 0-4℃; the phosphate extract is: 20 mmol / L K2HPO4 / KH2PO4, 0.1 mol / L NaCl, 2 mmol / L MgCl2, 1 mmol / L EDTA, pH 7.0; (3) Take the precipitate and repeat step (2) 1-3 times; (4) Take the precipitate and add 2-6 times the volume of 0.5-1 mol / L NaCl, homogenize, and centrifuge at low temperature to collect the supernatant; (5) Add 8-12 times the volume of purified water to the supernatant of step (4), centrifuge at 10000-20000rpm for 15-30min at 0-4℃, and take the precipitate as grass carp myofibril protein.

2. The grass carp myofibrillar protein / rutin complex according to claim 1, characterized in that, The mass ratio of grass carp myofibrillar protein to rutin is 1:0.02-0.

2.

3. The use of the grass carp myofibrillar protein / rutin complex according to claim 1 in the preparation of emulsion gels or emulsion gel stabilizers.

4. An emulsion gel stabilizer, characterized in that, Contains the grass carp myofibrillar protein / rutin complex as described in any one of claims 1-2.

5. A method for preparing an emulsion gel, characterized in that, The grass carp myofibrillar protein / rutin complex according to any one of claims 1-2 is mixed with an oil phase and an aqueous phase, and then subjected to high-speed shear mixing. The ratio of myofibrillar protein to aqueous phase in the grass carp myofibrillar protein / rutin complex is 0.01-0.05 kg / L; the volume ratio of oil phase to aqueous phase is 0.1-1:

1.

6. The method according to claim 5, characterized in that, The oil phase is edible oil.

7. The method according to claim 5, characterized in that, The oil phase and aqueous phase mentioned are food or pharmaceutical products containing water and oil.

8. An emulsion gel, characterized in that, Prepared by the method described in any one of claims 5-7.

9. The use of the emulsion gel of claim 8 in the preparation of a carrier for nutrients or pharmaceuticals.

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