CN113812506A - Modified silver carp muscle protein, processing method and application thereof - Google Patents

Modified silver carp muscle protein, processing method and application thereof Download PDF

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CN113812506A
CN113812506A CN202111073664.7A CN202111073664A CN113812506A CN 113812506 A CN113812506 A CN 113812506A CN 202111073664 A CN202111073664 A CN 202111073664A CN 113812506 A CN113812506 A CN 113812506A
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silver carp
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蒋将
金斐
熊幼翎
崔雅茹
张文斌
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Jiangnan University
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Abstract

The invention belongs to the field of animal protein processing, and particularly relates to modified silver carp muscle protein, a processing method and application thereof. The invention discloses a processing method of modified silver carp muscle protein, which comprises the following steps: mashing tissue blocks of chubs and preparing mixed homogenate; increasing the pH value of the mixed homogenate to 10.5-11.5, and stirring to fully dissolve; carrying out solid-liquid separation for the first time and collecting supernatant; adjusting the pH value of the supernatant collected by the first solid-liquid separation to 5.0-5.5; carrying out solid-liquid separation for the second time and collecting the precipitate to obtain the muscle protein of the silver carp; dissolving the silver carp muscle protein extracted by the alkali extraction method in a salt solution, and carrying out ultrasonic treatment for 3-30 min under the condition that the power is 100-1000W to obtain the modified silver carp muscle protein. The chub muscle protein extracted by the alkali extraction method has a synergistic effect after being modified by ultrasonic waves, so that the solubility, the emulsifying activity, the emulsifying stability, the water holding capacity and the gel forming effect of the chub muscle protein are synchronously improved, and the comprehensive processing performance of the protein is obviously improved.

Description

Modified silver carp muscle protein, processing method and application thereof
Technical Field
The invention belongs to the technical field of animal protein processing, and particularly relates to modified silver carp muscle protein, a processing method and application thereof.
Background
Animal proteins are an important part of the human diet and help to build, repair and maintain the structure and composition of the body, while also providing the food with the desired color, flavor, texture, which properties depend largely on the function of the protein. Among the animal meat proteins, the fish protein has more outstanding advantages in the aspects of essential amino acid composition, digestibility and the like compared with the livestock meat protein; the fish meat is rich in unsaturated fatty acid which also accords with the current healthy diet trend; meanwhile, myofibrillar protein in the fish protein has the processing characteristics of good emulsifying property, gel property, water retention property and the like, and the functional characteristics are also the key for determining the quality of related protein products.
Protein solubility, which is crucial in meat processing because it is closely related to the expression effects of many other functions of protein (emulsifying, foaming, gelling, etc.), is a prerequisite and basis for the expression of all functional properties of protein, and it is an important ring in food processing to improve protein solubility.
The emulsifiability is specifically divided into emulsifying activity and emulsifying stability, which means the ability of muscle protein to destroy fat into small particles and stabilize the oil-water system, and particularly, the good emulsifying property in the emulsified meat emulsion product forms a muscle protein film between the interface of the water phase and the oil phase of the system and prevents the fat particles from being combined with each other, which is a key factor for controlling the quality and stabilizing the storage of the emulsified meat emulsion processed product.
Protein gel properties can be expressed in terms of gel strength and gel water holding capacity. The meat protein gel is the capability of the muscle protein to expand and aggregate under the induction of heat and then form a three-dimensional network structure with viscoelasticity, and the gel property is the basic attribute of the quality of meat emulsion products. The gel strength represents the pressure required for the gel to be punctured by an external force, and the good gel strength after optimization can endow the protein thermal gel product with tough texture without hardness, elasticity and soft collapse; the water retention property is the capacity of keeping the original water content and adding water content of meat and meat protein products thereof under the action of external force, and the excellent gel water retention property ensures that the meat protein products keep stable water content and water activity in the processes of processing, transportation, selling and storage, thereby maintaining the stable quality of the products.
At present, in the deep processing application of domestic fish and aquatic products, the consumption market of minced fillet products is huge, wherein the minced fillet products of sea fish occupies the mainstream, and the reasons for the consumption are that the meat quality of freshwater fish is looser than that of the sea fish, the fish protein processing performance (solubility, emulsibility and gel property) is poorer, and the development of domestic freshwater fish and aquatic product processing products is limited; meanwhile, researches show that myosin in the fish muscle protein can perform self-assembly under normal processing conditions, part of functional groups and amino acid side chains in the protein are embedded in a non-covalent bond (hydrophobic interaction, electrostatic interaction, hydrogen bonds, Van der Waals force and the like) form, the processing functional characteristics of solubility, emulsifying property, gel property, water holding property and the like of the fish muscle protein are limited, and the industrial processing, utilization and new product research and development of the fish muscle protein are influenced to a certain extent.
The freshwater fish resource is rich in China, the silver carp is particularly praised as one of four kinds of freshwater fish, commonly called silver carp, and the silver carp is easy to raise, fast to grow, large in individual, high in nutrition, high in protein content up to 18% and high in biological value. The annual output of the silver carps is two in China all year round, the resource storage is huge, and the silver carps have good development prospects, but compared with other 'three freshwater fishes', the silver carps have low production added values due to inherent characteristics that the silver carps have strong earthy smell and abundant bone spurs, the meat quality is relatively loose and lacks elasticity, and the like, so that the consumption market potential of the silver carps is limited, meanwhile, endogenous enzymes in the silver carps can indirectly influence the extraction rate and the processing characteristics of muscle proteins of the silver carps, and therefore, the silver carps have huge waste. The silver carp muscle protein mainly contains myofibrillar protein, sarcoplasmic protein and matrix protein, wherein the myofibrillar protein accounts for about 60-70% of the total protein, is the main component of the silver carp protein, and plays a main structural and functional role in the food processing of meat and meat products.
In conclusion, on the basis of efficiently extracting the low-value silver carp or silver carp processing by-products to obtain the fish protein, the method has profound significance for further improving the protein structure and improving the protein processing performance to meet more various food processing modes and wide practical application, is favorable for enriching the muscle protein product of the silver carp, improves the utilization rate of the silver carp resource and promotes the development of the freshwater fish industry in China.
At present, there are few reports about research and application of rapidly extracting the muscle protein of the silver carp and obviously improving the processing characteristic of the muscle protein of the silver carp. The invention seeks to find a method for efficiently extracting and modifying chub muscle protein with high added value based on low-value chubs, which can meet the requirements of food production enterprises and consumers.
Disclosure of Invention
The technical problem is as follows:
silver carp is praised as one of four kinds of fresh water fish in China, and has huge fish resource reserves, but the silver carp has abundant bone spurs and strong soil fishy smell, the meat quality is lack of elasticity, the expansion of the consumer market is greatly limited, endogenous proteases in the fish meat such as calpain, cathepsin and myofibril combined serine protease degrade protein and deteriorate the fish meat in the conventional fish meat rinsing process, and meanwhile, the difficultly processed silver carp multi-spur structure and the high production cost of the conventional protein rinsing method make many aquatic enterprises look against, so that the efficient extraction of muscle protein in low-value silver carp and the development and production of high-value fish protein resources are particularly important.
Aiming at the defects of the prior art, the technical idea of the invention is to provide a modified protein which is prepared by using silver carp or silver carp processing by-products as main raw materials, efficiently obtaining silver carp muscle protein by using an alkaline offset extraction technology, and then assisting low-frequency high-intensity ultrasonic treatment to further improve the processing functional property of the silver carp muscle protein so as to obtain the modified protein which can be widely applied to an emulsified minced fillet food system. The invention can provide technical support for widening the development of the freshwater fish, particularly the silver carp deep processing industry.
The technical scheme is as follows:
the invention develops a multifunctional protein with good solubility, emulsifying property, gel property and water binding property based on the extraction of the fish muscle protein of the silver carp or the silver carp processing by-product and the modification of the processing property by low-frequency high-intensity ultrasound, and provides technical support for the application of the silver carp muscle protein to the emulsified minced fillet food system product.
The present invention has been made in view of the above and/or problems and research directions existing in the existing fish muscle protein production processes.
Therefore, one of the purposes of the invention is to overcome the defects of difficult extraction, poor protein processing performance and the like of the existing chub muscle protein and provide a thought for efficiently extracting the chub muscle protein and improving the structure of the chub muscle protein to improve the processing performance of the chub muscle protein.
One of the purposes of the invention is to provide a processing method of modified silver carp muscle protein, which comprises the following steps:
extracting the muscle protein of the silver carp by an alkali extraction method: mashing fresh chub tissue blocks and preparing mixed homogenate; increasing the pH value of the mixed homogenate to 10.5-11.5, and stirring to fully dissolve; carrying out solid-liquid separation for the first time and collecting supernatant; adjusting the pH value of the supernatant collected by the first solid-liquid separation to 5.0-5.5; carrying out solid-liquid separation for the second time and collecting the precipitate to obtain the muscle protein of the silver carp;
ultrasonically modifying the muscle protein of the silver carp: dissolving the silver carp muscle protein extracted by the alkali extraction method in a salt solution, and carrying out ultrasonic treatment for 3-30 min under the condition of 100-1000W to obtain the modified silver carp muscle protein. The obtained modified protein can be directly used for production and processing and can also be stored by freeze-drying.
As one embodiment of the invention, the ultrasonic power is preferably 100-700W.
In one embodiment of the present invention, the ultrasound time is preferably 3 to 20 min.
In one embodiment of the invention, in the step of extracting the muscle protein of the silver carp by the alkali extraction method, the pH value of the mixed homogenate is increased to 11.5, the mixed homogenate is stirred to be fully dissolved, and the pH value of the supernatant collected by the first solid-liquid separation is adjusted to 5.3.
As one embodiment of the present invention, the pH is raised by using a reagent preferably 2.0mol/L NaOH solution, so as to deviate from the isoelectric point of the silver carp muscle proteins, to make the surface of the silver carp muscle proteins have a net negative charge, and the high charge density causes electrostatic repulsion between the silver carp muscle proteins, and to improve the hydration ability and solubility.
In one embodiment of the present invention, the reagent used for lowering the pH is one of organic acids such as malic acid, tartaric acid and citric acid, or inorganic acids such as hydrochloric acid and sulfuric acid, and the concentration is 1.0 to 2.0 mol/L. The aim is to reduce the pH of the solution to the isoelectric point, at which point the muscle protein surface lacks net charge and the electrostatic repulsion force is weaker than other non-covalent forces (hydrophobic interactions, hydrogen bonding forces, van der waals forces) to reduce protein solubility, making the protein more easily recovered by centrifugation. A2.0 mol/L HCl solution is preferred.
In the step of extracting the muscle protein of the silver carp by the alkali extraction method, the fresh silver carp tissue block is mashed and made into a mixed homogenate, and the method specifically comprises the following steps: mixing fresh tissue blocks of silver carp with ice water, mashing, grinding, and homogenizing to obtain mixed homogenate.
In one embodiment of the present invention, the chub tissue mass is at least one of a chub mass after simple scaling, bone removal, or a byproduct after production and processing of the chub.
In one embodiment of the invention, the mixing ratio of the silver carp tissue mass and the ice water in the step of preparing the mixed homogenate is 1:9, the homogenization condition is 15000-20000 rpm, and the homogenization is 1-2 min. The method aims to fully crush chub tissues, break down bone spurs and connective tissues of the chub, fully contact and dissolve muscle proteins and ice water, and provide a solution environment for subsequent alkaline excursion.
In one embodiment of the present invention, the first solid-liquid separation and the second solid-liquid separation are both performed by a centrifugal filtration method.
As one embodiment of the invention, the first solid-liquid separation is carried out at a centrifugal speed of 8000-9000 rpm, at a controlled temperature of 0-7 ℃ for 20 min. The aim was to remove insoluble material by high speed centrifugation: chub bony spur, scale and neutral fat, etc., and can raise protein extracting rate and purity.
As one embodiment of the invention, the second solid-liquid separation is carried out at a centrifugal speed of 5000-6000 rpm, at a temperature of 0-7 ℃ for 15 min. The purpose is to recover soluble chub muscle protein substantially free of cell membranes and fat.
As one embodiment of the invention, 50-100 g of ultrasonic treatment protein solution in the ultrasonic modified silver carp muscle protein is obtained, the ultrasonic treatment power is 400W, and the ultrasonic treatment time is 10 min.
As one embodiment of the invention, silver carp muscle protein extracted by an alkali extraction method is dissolved in NaCl solution (0.55-0.65 mol/L) to prepare 50-100 g of silver carp muscle protein solution with the mass fraction of 2.0-4.0%, and the weight ratio of pulse interval is 3 s: 3s to 7 s: and carrying out ultrasonic treatment for 10min under the condition of ice bath environment for 7s to obtain the modified silver carp muscle protein.
As one embodiment of the invention, silver carp muscle protein extracted by an alkali extraction method is dissolved in NaCl solution (0.55-0.65 mol/L) to prepare 50g of silver carp muscle protein solution with the mass fraction of 2.0-4.0%, and the silver carp muscle protein solution is subjected to ultrasonic treatment for 10min under the conditions of 400W of power, 20kHz of frequency, 10% of amplitude, 5s:5s of pulse pause ratio and ice bath environment to obtain the modified silver carp muscle protein.
As one embodiment of the invention, the ultrasonic treatment is preferably performed by using a stainless steel probe type ultrasonic processor with the diameter of 7mm, wherein the probe is positioned 2.0cm below the central liquid level of the solution, the ultrasonic treatment is performed in an ice bath, and the temperature of the muscle protein solution of the silver carp is not more than 10 ℃ during the processing process so as to prevent the denaturation of the muscle protein.
The second purpose of the invention is to provide the modified silver carp muscle protein prepared by the processing method.
The third purpose of the invention is to provide the application of the modified silver carp muscle protein as food.
For example, the chub pure protein surimi product, the emulsified surimi product or the chub protein freeze-dried product and the like are obtained by processing the modified chub muscle protein prepared by the invention as a raw material.
As one embodiment of the invention, the method further comprises the following steps after the step of ultrasonically modifying the chub muscle protein: preparing silver carp muscle protein freeze-drying: and (3) freeze-drying the modified chub muscle protein to obtain chub muscle protein freeze-dried powder.
The third purpose of the invention is to provide the application of the modified silver carp muscle protein as feed.
The fourth purpose of the invention is to provide the application of the modified silver carp muscle protein in the production of beauty and skin care products.
The fifth purpose of the invention is to provide the application of the modified silver carp muscle protein in the preparation of health products and pharmaceutical additives.
Compared with the prior art, the invention has the beneficial effects that:
(1) the processing method has the characteristics of low cost, short time consumption, easy operation, low pollution, safety and the like. The research of the invention finds that when the pH value of homogenate is increased to be within the range of 10.5-11.5 by alkali liquor and the pH value of centrifugal clear liquid is adjusted to be within the range of 5.0-5.5 by acid liquor, the extraction rate of muscle protein of the silver carp prepared by the alkali extraction method with specific parameters is more than 70%, the solubility is more than 35%, and the emulsifying activity is 5.38m2More than g, and the emulsion stability (standing at 4 ℃ for 180min) is more than 57.14 percent. Therefore, the silver carp muscle protein obtained by the processing method of the invention obtains better solubility, emulsifying activity, emulsifying stability, gel property and water holding capacity, and can improve the meat quality and the protein processing performance of the rinsing preparation method fishPoor problem, improves the utilization rate of low-value chub resources, can better meet diversified processing technological conditions and is suitable for the development of richer emulsified minced fillet system products. Compared with livestock and poultry meat protein, the protein has more outstanding advantages in the aspects of essential amino acid composition, digestibility and the like, so that the protein can replace the traditional livestock and poultry meat protein and vegetable protein to be applied to feeds, foods, health products and pharmaceutical additives, and can also be applied to producing beauty and skin care products due to good emulsifying property.
(2) The research of the invention finds that the extraction rate, the solubility, the emulsifying activity and the emulsifying stability of the muscle protein of the silver carp can be obviously improved by optimizing the two pH values. Preferably, when the alkali liquor increases the pH value of the homogenate to 11.5 and the acid liquor adjusts the pH value of the centrifugate to 5.3, the chub muscle protein performance is optimal: the extraction rate is 89.0%, the solubility is 37.05%, and the emulsifying activity is 6.03m2The emulsion stability (standing at 4 ℃ for 180min) was 63.73%.
(3) The research of the invention finds that compared with the traditional rinsing method for extracting the muscle protein of the silver carp, the alkali extraction method with specific parameters of the invention is applied to the extraction of the muscle protein of the silver carp, the extraction rate of the muscle protein of the silver carp is obviously improved (from 42 percent to more than 70 percent), and the emulsifying activity is obviously improved (from 4.37 m)2Lifting the volume of the solution to 5.38m2More than g), the emulsion stability (standing for 180min at 4 ℃) is obviously improved (from 14 percent to more than 57 percent), and the muscle protein solubility of the silver carp is increased. The alkaline extraction method under the specific conditions adopted by the invention has obvious advantages in the aspects of efficiently extracting the silver carp muscle protein and improving the solubility, the emulsifying activity and the emulsifying stability (standing for 180min at 4 ℃) of the silver carp muscle protein product.
(4) The research of the invention discovers that the chub muscle protein extracted by the alkali extraction method has a synergistic effect after being modified by ultrasonic waves, and realizes the synchronous improvement of the solubility (the highest can reach 93.27%) and the emulsifying activity (the highest can reach 10.36 m) of the chub muscle protein2The emulsion stability (up to 95.83 percent) of the gel when standing at 4 ℃ for 30min, the emulsion stability (up to 88.65 percent) of the gel when standing at 4 ℃ for 180min, the gel strength (up to 1.037N), the gel water holding capacity (up to 68.05 percent) and the gel formingThe effect (the gel is visual and has better stereoscopic impression and better plastic property) is synchronously improved, and the comprehensive processing performance of the protein is obviously improved.
(5) The research of the invention finds that after the chub muscle protein extracted by the traditional rinsing method is modified by ultrasonic wave, the solubility is reduced to some extent compared with that before the ultrasonic wave modification treatment, the gel strength and the water holding capacity are obviously reduced, the gel is poorer in visual molding, and the molding performance is poor. Therefore, the silver carp muscle protein extracted by the traditional rinsing method cannot realize the similar synergistic effect after being modified by ultrasonic waves.
(6) The research of the invention finds that by changing the time of ultrasonic treatment, specifically, compared with the ultrasonic treatment of firstly alkali extracting and then ultrasonic modifying, the solubility, the emulsion stability (4 ℃, 180min), the gel strength and the water holding capacity of the muscle protein of the silver carp are obviously reduced, the obtained gel has extremely poor visual performance, the unexpected influence on the performance can be generated by the sequence of ultrasonic alkali extracting, the adjustment of the ultrasonic sequence can not be regarded as routine adjustment, and the specific sequence of the invention can play an unexpected technical effect.
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In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without inventive exercise.
Wherein:
FIG. 1 shows the change in the extraction rate of the chub muscle protein obtained in examples 1 to 5 and comparative example 1;
FIG. 2 shows the muscle protein solubilities of chubs prepared in examples 3, 6, 7 and 10;
FIG. 3 shows the silver carp muscle protein emulsifying activities obtained in examples 3, 6, 7 and 10;
FIG. 4 shows the changes of the gel strength and water holding capacity of the chub muscle protein prepared in examples 3, 6, 7 and 10;
FIG. 5 shows changes in the size of the muscle protein of chubs prepared in examples 3 and 8 to 12;
FIG. 6 shows the solubility of the muscle protein of chubs prepared in examples 3 and 8 to 12;
FIG. 7 shows the silver carp muscle protein emulsifying activities obtained in examples 3 and 8 to 12;
FIG. 8 shows the emulsion stability of chub muscle protein obtained in examples 3 and 8 to 12;
FIG. 9 shows microstructures of the silver carp muscle protein emulsions prepared in examples 3 and 8 to 12, wherein each scale is 50 μm;
FIG. 10 shows changes in the gel strength and water holding capacity of the chub muscle protein prepared in examples 3 and 8 to 12;
FIG. 11 shows the muscle protein solubilities of chubs prepared in comparative examples 1 to 3, example 3 and example 10;
FIG. 12 shows the silver carp muscle protein emulsification activities obtained in comparative examples 1 to 3, example 3, and example 10;
FIG. 13 shows the emulsion stability of chub muscle protein obtained in comparative examples 1 to 3, example 3, and example 10;
FIG. 14 shows changes in the gel strength and water holding capacity of chub muscle protein prepared in comparative examples 1 to 3, example 3 and example 10;
FIG. 15 is a visual representation of chub muscle protein gels prepared in comparative examples 1 to 3, example 3 and example 10;
fig. 16 is a process flow diagram of silver carp muscle protein processing according to the present invention.
Detailed Description
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.
Furthermore, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.
The test and characterization methods related to the embodiments and the comparative examples of the present invention are as follows:
and (3) measuring the extraction rate of the muscle protein of the silver carp:
(1) the extraction rate of the muscle protein of the silver carp prepared by the rinsing method is as follows: mixing a proper amount of fresh silver carp tissue blocks with ice water, mashing the tissue, slowly filtering the homogenate by using two layers of coarse cotton cloth to obtain a crude extract, wherein the protein content of the crude extract is marked as A; the silver carp muscle protein precipitate obtained by the rinsing method is dissolved in a salt solution (0.6mol/L NaCl, pH 7.0, 4 ℃), and the content of the extracted silver carp muscle protein is recorded as B by a biuret protein determination method.
Figure BDA0003261383930000071
(2) The extraction rate of the muscle protein of the silver carp prepared by the alkali extraction method is as follows: mixing a proper amount of fresh silver carp tissue blocks with ice water, mashing the tissue, thoroughly dissolving in an alkaline environment, centrifuging, taking supernatant, and measuring the protein content and recording as A; after isoelectric precipitation and centrifugation, silver carp muscle protein precipitate is finally obtained, dissolved in saline solution (0.6mol/L NaCl, pH 7.0, 4 ℃), and the content of extracted silver carp muscle protein is recorded as B by biuret protein determination method.
Figure BDA0003261383930000072
Protein solubility assay: diluting the chub muscle protein solution to be detected to 2.0mg/mL, centrifuging at 4 ℃ for 15min at 5000 Xg, collecting supernatant, and determining protein content by using biuret, wherein the protein solubility is expressed as the content percentage of the supernatant protein and the original chub muscle protein diluent.
Protein particle size determination:diluting the silver carp muscle protein solution to be detected to 1.0mgAnd mL, taking a proper amount of the protein, placing the mixture into a Zeta cuvette, and automatically measuring the particle size of the protein.
Preparing an emulsion:before the experiment, the chub muscle protein solution (4 ℃) needs to be cooled in advance, and then the chub muscle protein solution (the protein concentration is 1.1% w/v) to be detected and the rapeseed oil are mixed according to the volume ratio of 9:1 to obtain a mixture so as to prepare the emulsion (the final concentration of the chub muscle protein in the emulsion is 1.0% w/v). Shearing the mixture for 2min at 17500rpm by a high-speed shearing disperser (low-foaming probe) to prepare corresponding emulsion for protein emulsification activity determination, protein emulsification stability determination and protein emulsion microstructure characterization.
Protein emulsification activity assay:the emulsion to be tested was immediately transferred to a 25mL beaker, and 20. mu.L of the emulsion was taken from the bottom of the beaker and dispersed in 5mL of a 0.1% (w/w) Sodium Dodecyl Sulfate (SDS) solution. Measurement of Dispersion Absorbance A at 500nm500The Emulsion Activity Index (EAI) is calculated according to the following formula:
Figure BDA0003261383930000073
wherein C is the protein concentration (g/mL) before emulsification of 0.01,
Figure BDA0003261383930000074
the oil volume fraction (v/v) of the emulsion was 0.9, and the dilution factor was 251.
Protein emulsion stability determination:the emulsion to be tested was immediately transferred to a 25mL beaker and placed in refrigeration at 4 ℃. When the refrigeration time reached 30min and 180min, 20. mu.L of the emulsion was taken from the bottom of the beaker and dispersed in 5mL of 0.1% (w/w) SDS solution. The absorbance was measured at 500nm and the Emulsion Stability (ESI) was calculated according to the following formula:
ESI(％)＝100*At/A0
wherein A is0And AtRespectively corresponding to the light absorption values of the emulsions at a storage time of 4 ℃.
And (3) characterizing the microstructure of the protein emulsion:using ECLIPSE 80i Nikon microscopeThe freshly prepared emulsion to be tested was observed. An emulsion sample (10. mu.L) was dropped onto the slide, the cover slip was gently covered, and the image was observed with a 100-fold objective lens and recorded with a Nikon DS-Ri1 camera equipped on the microscope.
Protein gel preparation:a 5.0g silver carp muscle protein solution sample to be tested (diluted to 40mg/mL and degassed at 4 ℃ by centrifugation at 1000 xg for 1 min) was sucked up by a plastic pipette and placed in a small glass bottle (inner diameter × length: 16.5mm × 50mm) and after being lightly sealed with a threaded plastic cap, the glass bottle was placed in a water bath and heated from 20 ℃ to 72 ℃ at a heating rate of 0.9 ℃/min. Immediately after heating, the sample was placed in an ice-water mixture to cool for 30min, and then placed in a refrigerator at 4 ℃ overnight to prepare the corresponding protein gel for the following gel strength measurement and gel water holding capacity measurement. Before the gel properties were determined, the gel samples were taken out of the refrigerator and allowed to equilibrate at room temperature for 2 h.
And (3) measuring the gel strength:and placing the gel sample to be tested on a test platform for fixing, and measuring by using a texture analyzer at room temperature. The speed before the test is selected as the test mode is 5 mm/s; the testing speed is 1 mm/s; the speed after the test is 5 mm/s; the pressing distance is 10 mm; the initiating power is 5 g; the model of the probe is P/0.5. Gel strength is defined as the initial pressure (N) required to puncture the gel.
And (3) measuring the gel water holding capacity:separating the gel from the glass wall by using a small shovel, accurately weighing 3.0g of the gel in a centrifuge tube, centrifuging for 15min at the rotating speed of 4000 Xg, inverting the centrifuge tube for airing, wiping residual water by using dry filter paper, and calculating the weight ratio (%) of the gel after centrifugation to the gel weighed before centrifugation.
1. Extraction of chub muscle protein from chub
1.1 optimization of alkaline extraction parameters for extraction by alkaline extraction
Example 1 alkaline extraction
Mixing a proper amount of fresh silver carp tissue blocks with ice water according to a ratio of 1:9(w/w), mechanically grinding by using a tissue triturator at a rotating speed of 20000rpm, homogenizing and mincing for 2min to obtain homogenate; taking 2.0mol/L NaOH solution to increase the pH value of the homogenate to 10.5, and stirring for 20min under ice bath for full dissolution to obtain a mixture; centrifuging the mixture at 8000rpm to remove insoluble substances; adjusting the pH of the centrifugate to 5.3 by using 2.0mol/L HCl solution; then centrifuging at 6000rpm by using a centrifuge to obtain chub muscle protein precipitate.
The chub prepared in example 1 had a muscle protein extraction rate of 70.3%, a solubility of 35.4%, and an emulsifying activity of 5.38m2The emulsion stability (standing at 4 ℃ for 180min) was 57.14%.
Example 2 alkaline extraction
Mixing a proper amount of fresh silver carp tissue blocks with ice water according to a ratio of 1:9(w/w), mechanically grinding by using a tissue triturator at a rotating speed of 20000rpm, homogenizing and mincing for 2min to obtain homogenate; taking 2.0mol/L NaOH solution to increase the pH value of the homogenate to 11.0, and stirring for 20min under ice bath for full dissolution to obtain a mixture; centrifuging the mixture at 8000rpm to remove insoluble substances; adjusting the pH of the centrifugate to 5.3 by using 2.0mol/L HCl solution; then centrifuging at 6000rpm by using a centrifuge to obtain chub muscle protein precipitate.
The chub prepared in example 2 had a muscle protein extraction rate of 79.8%, a solubility of 36.2%, and an emulsifying activity of 5.84m2The emulsion stability (standing at 4 ℃ for 180min) was 60.84%.
Example 3 alkaline extraction
Mixing a proper amount of fresh silver carp tissue blocks with ice water according to a ratio of 1:9(w/w), mechanically grinding by using a tissue triturator at a rotating speed of 20000rpm, homogenizing and mincing for 2min to obtain homogenate; taking 2.0mol/L NaOH solution to increase the pH value of the homogenate to 11.5, and stirring for 20min under ice bath for full dissolution to obtain a mixture; centrifuging the mixture at 8000rpm to remove insoluble substances; adjusting the pH of the centrifugate to 5.3 by using 2.0mol/L HCl solution; then centrifuging at 6000rpm by using a centrifuge to obtain chub muscle protein precipitate.
The chub prepared in example 3 had a muscle protein extraction rate of 89.0%, a solubility of 37.5%, and an emulsifying activity of 6.12m2The emulsion stability (standing at 4 ℃ for 180min) was 63.73%.
Example 4 alkaline extraction
Mixing a proper amount of fresh silver carp tissue blocks with ice water according to a ratio of 1:9(w/w), mechanically grinding by using a tissue triturator at a rotating speed of 20000rpm, homogenizing and mincing for 2min to obtain homogenate; taking 2.0mol/L NaOH solution to increase the pH value of the homogenate to 11.5, and stirring for 20min under ice bath for full dissolution to obtain a mixture; centrifuging the mixture at 8000rpm to remove insoluble substances; adjusting the pH of the centrifugate to 5.0 by using 2.0mol/L HCl solution; then centrifuging at 6000rpm by using a centrifuge to obtain chub muscle protein precipitate.
The chub prepared in example 4 had a muscle protein extraction rate of 86.4%, a solubility of 36.9%, and an emulsifying activity of 5.86m2The emulsion stability (standing at 4 ℃ for 180min) was 61.51%.
Example 5 alkaline extraction
Mixing a proper amount of fresh silver carp tissue blocks with ice water according to a ratio of 1:9(w/w), mechanically grinding by using a tissue triturator at a rotating speed of 20000rpm, homogenizing and mincing for 2min to obtain homogenate; taking 2.0mol/L NaOH solution to increase the pH value of the homogenate to 11.5, and stirring for 20min under ice bath for full dissolution to obtain a mixture; centrifuging the mixture at 8000rpm to remove insoluble substances; adjusting the pH of the centrifugate to 5.5 by using 2.0mol/L HCl solution; then centrifuging at 6000rpm by using a centrifuge to obtain chub muscle protein precipitate.
The chub prepared in example 5 had a muscle protein extraction rate of 87.6%, a solubility of 36.7%, and an emulsifying activity of 5.83m2The emulsion stability (standing at 4 ℃ for 180min) was 59.04%.
Comparative analysis examples 1 to 5 can find that: when the pH value of the homogenate is increased to be within the range of 10.5-11.5 by the alkali liquor and the pH value of the centrifugal clear liquid is adjusted to be within the range of 5.0-5.5 by the acid liquor, the extraction rate of the muscle protein of the silver carp extracted and prepared by the alkali extraction method with specific parameters is 70.3% -89.0%, the solubility is 35.4-37.5%, and the emulsifying activity is 5.38-6.03 m2(ii)/g, the emulsion stability (standing at 4 ℃ for 180min) is 57.14-63.73%. By optimizing the two pH values, the extraction rate, the solubility, the emulsifying activity and the emulsifying stability of the muscle protein of the silver carp can be obviously improved. Preferably, when the lye is to be usedWhen the pH value of the homogenate is increased to 11.5 and the pH value of the centrifuged clear liquid is adjusted to 5.3 by the acid liquid, the performance of the muscle protein of the silver carp is optimal: the extraction rate is 89.0%, the solubility is 37.5%, and the emulsifying activity is 6.12m2The emulsion stability (standing at 4 ℃ for 180min) was 63.73%.
1.2 comparison of the extraction by alkaline extraction of the invention with the extraction by the traditional rinsing method
Comparative example 1 extraction by traditional rinsing method
Extracting the muscle protein of the silver carp by adopting a rinsing method: mixing a proper amount of fresh silver carp tissue blocks with ice water according to a ratio of 1:4(w/w), mechanically grinding by using a tissue triturator at a rotating speed of 20000rpm, homogenizing and chopping for 2min, slowly stirring and homogenizing for 15min, pouring into a filter containing two layers of coarse cotton cloth for dehydration, adding ice water with 4 times of volume of the obtained precipitate, repeating the steps for 2 times, adding 0.1mol/L NaCl solution (1:4, w/w) into the last precipitate, and filtering the obtained precipitate to obtain the silver carp muscle protein.
The extraction rate of the muscle protein of the silver carp prepared in the comparative example 1 is 42.0%, and the solubility of the muscle protein of the silver carp is 33.8%; the emulsifying activity was 4.37m2The emulsion stability (standing at 4 ℃ for 180min) was 14.40%.
Comparative analysis of comparative example 1 with examples 1 to 5 it can be found that: compared with the traditional rinsing method for extracting the silver carp muscle protein, the alkaline extraction method with specific parameters provided by the invention is applied to extracting the silver carp muscle protein, the silver carp muscle protein extraction rate is obviously improved (from 42.0% to more than 70.3%, see figure 1), the silver carp muscle protein solubility is increased, and the emulsifying activity is obviously improved (from 4.37 m)2Lifting the volume of the solution to 5.38m2More than g), the emulsion stability (standing for 180min at 4 ℃) is obviously improved (from 14.40 percent to more than 57.14 percent). The alkaline extraction method under the specific conditions adopted by the invention has obvious advantages in the aspects of efficiently extracting the silver carp muscle protein and improving the solubility, the emulsifying activity and the emulsifying stability (standing at 4 ℃ for 180min) of the silver carp muscle protein product.
2. Modified silver carp muscle protein
2.1 optimization of the parameters of the ultrasonic modification
The chub muscle protein raw materials of examples 6-12 were extracted by the extraction method of example 3 (the pH of the homogenate was raised to 11.5 with an alkali solution, and the pH of the centrifuged supernatant was adjusted to 5.3 with an acid solution), and the freshly extracted chub muscle protein was immediately refrigerated at low temperature for future use.
Example 6 alkali extraction + ultrasonic modification
Dissolving silver carp muscle protein in a salt solution (0.6mol/L NaCl, pH 7.0 and 4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, positioning a stainless steel probe with the diameter of 7mm below the central liquid level of the solution for 2.0cm, and carrying out ultrasonic treatment for 10min under the conditions of 100W of power, 20kHz of frequency, 10% of amplitude, 5s:5s of pulse pause ratio and ice bath environment to obtain the modified silver carp muscle protein.
Example 7 alkali extraction + ultrasonic modification
Dissolving silver carp muscle protein in a salt solution (0.6mol/L NaCl, pH 7.0 and 4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, positioning a stainless steel probe with the diameter of 7mm below the central liquid level of the solution for 2.0cm, and carrying out ultrasonic treatment for 10min under the conditions of 700W of power, 20kHz of frequency, 10% of amplitude, 5s:5s of pulse pause ratio and ice bath environment to obtain the modified silver carp muscle protein.
Example 8 alkali extraction + ultrasonic modification
Dissolving silver carp muscle protein in a salt solution (0.6mol/L NaCl, pH 7.0 and 4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, positioning a stainless steel probe with the diameter of 7mm below the central liquid level of the solution for 2.0cm, and carrying out ultrasonic treatment for 3min under the conditions of 400W of power, 20kHz of frequency, 10% of amplitude, 5s:5s of pulse pause ratio and ice bath environment to obtain the modified silver carp muscle protein.
Example 9 alkali extraction + ultrasonic modification
Dissolving silver carp muscle protein in a salt solution (0.6mol/L NaCl, pH 7.0 and 4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, locating a stainless steel probe with the diameter of 7mm below the central liquid level of the solution for 2.0cm, and carrying out ultrasonic treatment for 6min under the conditions of 400W of power, 20kHz of frequency, 10% of amplitude, 5s:5s of pulse pause ratio and ice bath environment to obtain the modified silver carp muscle protein.
Example 10 alkali extraction + ultrasonic modification
Dissolving silver carp muscle protein in a salt solution (0.6mol/L NaCl, pH 7.0 and 4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, positioning a stainless steel probe with the diameter of 7mm below the central liquid level of the solution for 2.0cm, and carrying out ultrasonic treatment for 10min under the conditions of 400W of power, 20kHz of frequency, 10% of amplitude, 5s:5s of pulse pause ratio and ice bath environment to obtain the modified silver carp muscle protein.
Example 11 alkali extraction + ultrasonic modification
Dissolving silver carp muscle protein in a salt solution (0.6mol/L NaCl, pH 7.0 and 4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, positioning a stainless steel probe with the diameter of 7mm below the central liquid level of the solution for 2.0cm, and carrying out ultrasonic treatment for 15min under the conditions of 400W of power, 20kHz of frequency, 10% of amplitude, 5s:5s of pulse pause ratio and ice bath environment to obtain the modified silver carp muscle protein.
Example 12 alkali extraction + ultrasonic modification
Dissolving silver carp muscle protein in a salt solution (0.6mol/L NaCl, pH 7.0 and 4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, positioning a stainless steel probe with the diameter of 7mm below the central liquid level of the solution for 2.0cm, and carrying out ultrasonic treatment for 20min under the conditions of 400W of power, 20kHz of frequency, 10% of amplitude, 5s:5s of pulse pause ratio and ice bath environment to obtain the modified silver carp muscle protein.
Comparative analysis of examples 6, 7 and 10 shows that when the ultrasonic treatment power is controlled within the range of 100-700W, the modification method of the specific parameters of the invention has the silver carp muscle protein solubility of 47.05-78.65% and the emulsifying activity of 8.20-9.28 m2The gel strength is 0.42-1.04N, and the water holding capacity of the gel is 53.42-60.93%. In order to obtain better emulsifying activity, gel strength and water holding capacity, achieve the effects of low energy consumption and no obvious influence on solubility, the ultrasonic power parameter is obtained by optimization: 400W. Comparative analysis examples 8 to 12 show that when the ultrasonic time is controlled within 3 to 20min, the modification method of the specific parameters of the invention has the silver carp muscle protein particle size of 258.57 to 402.60nm, the solubility of 84.40 to 93.28 percent and the emulsifying activity of 7.76 to 10.36m2G, gel strength0.87 to 1.04N, and the gel water holding capacity is 55.16 to 68.05 percent. In order to obtain smaller protein particle size, higher solubility, higher emulsifying activity and emulsifying stability, more uniform emulsion oil droplet size, higher gel strength and higher gel water holding capacity effect, the ultrasonic time parameters are optimized and obtained: for 10 min.
The optimized parameters are mainly based on the results of laboratory measurement, and in order to meet the processing requirements of large-scale industrial production, the ultrasonic power can be properly increased and/or the ultrasonic time can be prolonged along with the increase of the muscle protein treatment capacity of the silver carps. In order to meet certain special processing requirements, the ultrasonic power and time parameters can be increased or decreased according to actual requirements. In conclusion, better modification effect can be realized by ultrasonic treatment for 3-30 min under the condition of 100-1000W.
Comparative analysis of the chub muscle protein obtained by the single alkali extraction method in example 3 and the chub muscle protein obtained by alkali extraction and ultrasonic modification in examples 6-12 shows that: the muscle protein solubility of the silver carp is remarkably improved (from 37.5 percent to over 84.0 percent, see figure 6), and the emulsifying activity is remarkably improved (from 6.12 m)2Lifting the/g to 7.76m2g, see figure 7), the emulsion stability (standing at 4 ℃ for 30min) is remarkably improved (from 72.02% to 84.30% or more, see figure 8), the emulsion stability (standing at 4 ℃ for 180min) is remarkably improved (from 63.73% to 71.75% or more, see figure 8), the oil drop size, the dispersity and the stability of the emulsion are remarkably improved (see figure 9), the gel strength is remarkably improved (from 0.27N to 0.87N or more, see figure 10), the gel water holding capacity is remarkably improved (from 47.65% to 55.16% or more, see figure 10), and the functions of the alkali extraction method extraction treatment and the ultrasonic modification treatment are mutually supported, so that the dissolving capacity of the chub muscle protein is further improved, the comprehensive processing performance of the chub muscle protein is improved by the parameters of the emulsifying property and the gel property, so that the chub muscle protein product prepared by the method has obviously improved market value.
2.2 Combined comparison of different extraction methods with ultrasonic modification
Comparative example 2 rinsing method + ultrasonic modification
The chub muscle protein material of comparative example 2 was extracted by the extraction method of comparative example 1, and the freshly extracted chub muscle protein was immediately cooled and stored at low temperature for further use.
Dissolving silver carp muscle protein in a salt solution (0.6mol/L NaCl, pH 7.0 and 4 ℃), preparing 50g of silver carp muscle protein solution with the mass fraction of 4.0%, and carrying out ultrasonic treatment for 10min under the conditions of 400W of power, 20kHz of frequency, 10% of amplitude, 5s:5s of pulse pause ratio and ice bath environment by locating a stainless steel probe with the diameter of 7mm below the central liquid level of the solution, so as to obtain the rinsed modified silver carp muscle protein (refer to the ultrasonic modification method in the example 10).
Comparative analysis comparative example 1 and comparative example 2 found that: the silver carp muscle protein extracted by the rinsing method is subjected to ultrasonic modification treatment (comparative example 2), the solubility is reduced (see figure 11), the gel strength and the water holding capacity are remarkably reduced (see figure 14), and the visual gel forming is poor (see figure 15) compared with the silver carp muscle protein before ultrasonic modification treatment (comparative example 1). Comparative analysis of comparative example 2 and example 10 revealed that, on the one hand, the solubility, emulsifying activity, emulsifying stability, water holding capacity and gel forming effect of the chub muscle protein prepared by the alkali extraction method combined with the ultrasonic modification treatment were still much higher than those of the chub muscle protein prepared by the rinsing method combined with the ultrasonic modification treatment (see fig. 11 to 15). On the other hand, after the silver carp muscle protein obtained by different extraction methods is subjected to ultrasonic modification treatment, the silver carp muscle protein shows different change trends: the combination of the rinsing method and the ultrasonic wave cannot synchronously improve the solubility, the emulsifying activity, the emulsifying stability, the water holding capacity and the gel forming effect of the muscle protein of the silver carp, and the change trend of the solubility, the gel strength and the water holding capacity is different from the combination of the alkali extraction method and the ultrasonic wave. The combined use of the alkali extraction method and the ultrasonic modification is a synergistic effect, and the replacement of any treatment process may not achieve a corresponding technical effect.
As shown in fig. 14, it can be seen from the comparison of the water holding capacity data of comparative examples 1 to 2, example 3, and example 10 that the water holding capacity of the single rinsing method is slightly higher than that of the single alkali extraction method, but the combined treatment of the alkali extraction method and the ultrasonic wave is higher than that of the rinsing method and the ultrasonic wave after the same ultrasonic treatment. The gel strength data also show similar rules, which indicate that a synergistic effect exists between the alkali extraction method and the ultrasonic treatment, and the ultrasonic treatment is utilized to compensate the defects of low gel strength and low water holding capacity caused by the alkali extraction method.
2.3 changing the timing of sonication
Comparative example 3 simultaneous alkali extraction and sonication
Mixing a proper amount of fresh silver carp tissue blocks with ice water according to a ratio of 1:9(w/w), mechanically grinding by using a tissue triturator at a rotating speed of 20000rpm, homogenizing and mincing for 2min to obtain homogenate; then taking 2.0mol/L NaOH solution to increase the pH value of the homogenate to 11.5, and stirring for 20min under ice bath for full dissolution to obtain a mixture; carrying out ultrasonic treatment on the mixture, centrifuging the mixture at a high speed under the condition of 8000rpm, and removing insoluble substances; adjusting the pH of the centrifugate to 5.3 by using 2.0mol/L HCl solution; then centrifuging at 6000rpm by using a centrifuge to obtain chub muscle protein precipitate. Wherein the parameters of the ultrasonic treatment are as follows: a stainless steel probe with a diameter of 7mm was placed 2.0cm below the central liquid level of the solution, and the solution was sonicated for 10min under conditions of 400W power, 20kHz frequency, 10% amplitude, 5 s/5 s pulse pause ratio, and ice bath environment (sonication conditions same as in example 10).
Comparative analysis on comparison between comparative example 3 and example 10 shows that, compared with the simultaneous alkali extraction and ultrasonic treatment, the solubility of the muscle protein of the silver carp (see fig. 11), the emulsion stability (4 ℃, 180min) (see fig. 13), the gel strength and the water holding capacity of the silver carp are significantly reduced (see fig. 14), and the visual performance of the gel of the comparative example 3 (see fig. 15) is extremely poor, which indicates that the improvement of the muscle protein processing performance by the ultrasonic treatment in the alkali extraction process is limited, the importance of the ultrasonic time is embodied, the performance is unexpectedly influenced by the sequence of the ultrasonic alkali extraction, the ultrasonic sequence adjustment cannot be regarded as conventional adjustment, and the specific sequence of the invention can achieve unexpected technical effects.
3. Application of modified silver carp muscle protein in production of beauty and skin care products
The application of the modified chub muscle protein in the preparation of the emulsion is taken as an example for illustration.
The modified silver carp muscle protein obtained in the embodiment 6-12 can be used as an emulsifier instead of vegetable protein, and is applied to emulsion preparation.
The alkali-extracted chub muscle protein after ultrasonic modification has good emulsifying activity and emulsifying stability, the interaction capacity of a fat globule and a water phase interface is exerted in the emulsion forming process, the two-phase interface tension is reduced, a thicker protein interface layer is generated, the size of fat oil drops can be reduced, and the fat oil drops are uniformly and stably distributed in the solution. The application of the modified silver carp muscle protein in the oil-in-water emulsion rich in animal fat in examples 6-12 has a profound research significance for replacing plant protein with weak emulsibility or artificially synthesized emulsifier by the modified animal protein.
4. Application of modified silver carp muscle protein as food
The application of the modified chub muscle protein in the preparation of the protein gel is taken as an example for illustration.
The alkaline extraction chub muscle protein subjected to ultrasonic modification has smaller granularity, and the formed gel has more uniform and compact structure, ordered and fine arrangement and good elasticity and toughness. The chub muscle protein gel is strong in integrity, not easy to flow, in the shape of jelly or pudding, white in overall color, rich in luster, free of obvious water analysis and good in water holding capacity. The alkali-extracted chub muscle protein obtained in the embodiments 6 to 12 can be applied to meat protein gel products, especially surimi products and imitation products thereof, due to the gel strength and water holding capacity of the alkali-extracted chub muscle protein.
5. Application of modified silver carp muscle protein as feed
The modified silver carp muscle protein obtained in the embodiment 6-12 can be processed into silver carp muscle protein freeze-dried through freeze drying, and is applied to feed products.
6. Application of modified silver carp muscle protein in preparation of health-care products and pharmaceutical additives
The modified silver carp muscle protein obtained in the embodiments 6-12 can be used as an emulsifier and important nutritional ingredients to be applied to preparation of health products and pharmaceutical additives.

Claims (10)

1. A processing method of modified silver carp muscle protein is characterized by comprising the following steps:
extracting the muscle protein of the silver carp by an alkali extraction method: mashing fresh chub tissue blocks and preparing mixed homogenate; increasing the pH value of the mixed homogenate to 10.5-11.5, and stirring to fully dissolve; carrying out solid-liquid separation for the first time and collecting supernatant; adjusting the pH value of the supernatant collected by the first solid-liquid separation to 5.0-5.5; carrying out solid-liquid separation for the second time and collecting precipitate to obtain the muscle protein of the silver carp;
ultrasonically modifying the muscle protein of the silver carp: dissolving the silver carp muscle protein extracted by the alkali extraction method in a salt solution, and carrying out ultrasonic treatment for 3-30 min under the condition that the power is 100-1000W to obtain the modified silver carp muscle protein.
2. The method for processing modified silver carp muscle protein according to claim 1, wherein in the step of extracting silver carp muscle protein by alkaline extraction, the pH value of the mixed homogenate is increased to 11.5, the mixed homogenate is stirred to be fully dissolved, and the pH value of the supernatant collected by the first solid-liquid separation is adjusted to 5.3.
3. The processing method of the modified silver carp muscle protein according to claim 1 or 2, wherein the power of the ultrasonic treatment in the ultrasonic modified silver carp muscle protein is 400W, and the time of the ultrasonic treatment is 10 min.
4. The method for processing modified silver carp muscle protein according to claim 1, wherein the step of extracting silver carp muscle protein by alkaline extraction comprises the following steps: mixing fresh tissue blocks of silver carp with ice water, mashing, grinding, and homogenizing to obtain mixed homogenate.
5. The method for processing modified silver carp muscle protein according to claim 1, wherein the whole process of the ultrasonic treatment is carried out in an ice bath, and the temperature of the silver carp muscle protein solution during the processing process is not more than 10 ℃.
6. The modified silver carp muscle protein prepared by the processing method of any one of claims 1 to 5.
7. Use of the modified silver carp muscle protein according to claim 6 as a food.
8. Use of the modified silver carp muscle protein according to claim 6 as a feed.
9. Use of the modified silver carp muscle protein according to claim 6 for the production of a cosmetic skin care product.
10. The use of the modified silver carp muscle protein of claim 6 in the preparation of health products and pharmaceutical additives.
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