CN113812506B - Modified silver carp muscle protein, processing method and application thereof - Google Patents
Modified silver carp muscle protein, processing method and application thereof Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
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- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
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- A—HUMAN NECESSITIES
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- A23J3/04—Animal proteins
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- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
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- A—HUMAN NECESSITIES
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- A61K35/60—Fish, e.g. seahorses; Fish eggs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- A61P17/16—Emollients or protectives, e.g. against radiation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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 the silver carp tissue blocks and preparing into mixed homogenate; raising the pH value of the mixed homogenate to 10.5-11.5, and stirring to fully dissolve; performing first solid-liquid separation and collecting supernatant; adjusting the pH value of the supernatant collected by the first solid-liquid separation to 5.0-5.5; performing solid-liquid separation for the second time, and collecting precipitate to obtain silver carp muscle protein; dissolving silver carp muscle protein extracted by an alkali extraction method in a salt solution, and carrying out ultrasonic treatment for 3-30 min under the condition of 100-1000W of power to obtain the modified silver carp muscle protein. After the silver carp muscle protein extracted by the alkali extraction method is modified by ultrasonic waves, the synergistic effect is achieved, the synchronous improvement of the solubility, the emulsifying activity, the emulsifying stability, the water holding capacity and the gel forming effect of the silver carp muscle protein is realized, and the comprehensive processing performance of the protein is remarkably improved.
Description
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 important components of the human diet that help build, repair, and maintain the structure and composition of the body, while also providing the food with the desired color, flavor, texture, these properties being largely dependent on the function of the protein. In animal meat proteins, fish proteins have more prominent advantages in terms of essential amino acid composition, digestibility and the like compared with livestock and poultry meat proteins; unsaturated fatty acids rich in fish meet the current healthy diet trend; meanwhile, myofibrillar protein in fish protein has good emulsifying property, gel property, water-holding capacity and other processing characteristics, and the functional characteristics are also key for determining the quality of related protein products.
Protein solubility, which is a prerequisite and basis for the expression of all functional properties of proteins, is of vital importance in meat processing, as it is closely related to the expression of many other functions of proteins (emulsifying, foaming, gelling, etc.), and increasing protein solubility during processing is a non-negligible important part of the food process.
The emulsifying property is specifically divided into emulsifying activity and emulsifying stability, and refers to the ability of muscle protein to destroy fat into small particles and stabilize the oil-water system, and particularly, the good emulsifying property in emulsified meat emulsion products can form a muscle protein film between the interface of the water phase and the oil phase of the system and can prevent the mutual combination of the fat particles, so that the muscle protein film is a key factor for controlling the quality and storing stability of the processed product of the emulsified meat emulsion.
Protein gel properties can be expressed in terms of gel strength and gel water holding capacity. Meat protein gel is the ability of muscle protein to structurally unfold and aggregate under heat induction, and then form a three-dimensional network structure with viscoelasticity, and the gel property is a basic attribute of meat emulsion product quality. Gel strength means the pressure required by the gel to be punctured by external force, and the good gel strength after optimization can endow the protein thermal gel product with tough but not firm, loose and elastic but not soft texture; the water retention is the capability of maintaining original moisture and adding moisture of meat and meat protein products thereof under the action of external force, and the excellent gel water retention can ensure that the meat protein products maintain stable moisture content and water activity in the processes of processing, transporting, selling and storing, thereby maintaining the stable quality of the products.
At present, in domestic deep processing application of fish and aquatic products, the consumer market of minced fillet products is huge, wherein minced fillet occupies the main stream, and the reason is that freshwater fish has loose meat quality compared with the freshwater fish, and the processing performance (solubility, emulsifying property and gel property) of the fish protein is poorer, so that the development of the domestic freshwater fish water product processing product is limited; meanwhile, research discovers that myosin in fish muscle protein can perform self-assembly under normal processing conditions, and partial functional groups and amino acid side chains in the protein are embedded in a non-covalent bond (hydrophobic interaction, electrostatic interaction, hydrogen bond, van der Waals force and the like) mode, so that the processing functional characteristics of fish muscle protein solubility, emulsifying property, gel property, water retention and the like 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 in China is rich in resources, and the silver carp is one of four freshwater fish, commonly called silver carp, which is easy to raise, fast in growth, large in individual, high in nutrition, high in biological value, and the protein content can reach 18%. The annual output of the silver carp is two in the forefront of China, the resource reserve of the silver carp is huge, and the silver carp has good development prospect, but compared with other 'three large freshwater fishes', the silver carp has strong earthy smell and rich bone spurs, the silver carp is loose in meat quality and lacks elasticity and other inherent characteristics, so that the production added value of the silver carp is lower, the potential of the consumer market is limited, and meanwhile, the extraction rate and the processing characteristic of the silver carp muscle protein are indirectly influenced by endogenous enzymes in the silver carp fish meat, so that the silver carp resource has huge waste. The silver carp muscle protein mainly contains myofibrillar protein, myoplasma protein and matrix protein, wherein the myofibrillar protein accounts for 60% -70% of total protein, is a main component of silver carp protein, and plays a main structural and functional role in food processing of meat and meat products.
In conclusion, the method has profound significance in further improving the protein structure and the protein processing performance to meet the requirements of more various food processing modes and wide practical application on the basis of efficiently extracting low-value silver carp or silver carp processing byproducts to obtain fish protein, is beneficial to the richness of silver carp muscle protein products, improves the utilization rate of silver carp resources, and promotes the development of freshwater fish industry in China.
At present, researches and application reports on rapid extraction of silver carp muscle protein and remarkable improvement of silver carp muscle protein processing characteristics are freshly reported. The invention tries to find a method for efficiently extracting and modifying the high added value silver carp muscle protein based on low-value silver carp, which can meet the demands of food production enterprises and consumers.
Disclosure of Invention
Technical problems:
silver carp is known as one of four large freshwater fishes in China, has huge fish resource reserves, but the silver carp itself has rich bone spurs and strong earthy smell, the meat quality is also lack of elasticity, the expansion of the consumer market of the silver carp is greatly limited, endogenous protease such as calpain, cathepsin and myofibrillar serine protease in the fish meat is greatly limited, protein is degraded and fish meat is degraded in the conventional fish meat rinsing process, meanwhile, the multi-spur structure of the silver carp difficult to process and the higher production cost of the conventional protein rinsing method make the production of a plurality of aquatic enterprises prohibitive, so the efficient extraction of muscle protein in the low-value silver carp and the application of the muscle protein in the low-value silver carp in the development and production of high-value fish protein resources are particularly important.
Aiming at the defects of the prior art, the technical concept of the invention is to provide a modified protein which is widely applied to an emulsified minced fillet food system and is prepared by using silver carp or silver carp processing byproducts as main raw materials, using an alkaline offset extraction technology to efficiently obtain silver carp muscle protein and then assisting low-frequency high-intensity ultrasonic treatment to further improve the processing functional property of silver carp muscle protein. The invention can provide technical support for widening the development of the deep processing industry of freshwater fish, especially silver carp.
The technical scheme is as follows:
the invention is based on the extraction of fish muscle protein of silver carp or silver carp processing byproducts, and further carries out the modification of processing performance through low-frequency high-intensity ultrasound, so as to develop the multifunctional protein with good solubility, emulsifying property, gel property and water retention property, and provide technical support for the application of silver carp muscle protein in emulsified minced fillet food system products.
The present invention has been made in view of the above-mentioned and/or existing problems and research directions in the production and processing of fish muscle proteins.
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 silver carp muscle protein, and provide a concept for efficiently extracting the silver carp muscle protein and improving the structure thereof so as to improve the processing performance.
The invention aims at providing a processing method of modified silver carp muscle protein, which comprises the following steps:
extracting silver carp muscle protein by an alkali extraction method: mashing fresh silver carp tissue blocks and preparing mixed homogenate; raising the pH value of the mixed homogenate to 10.5-11.5, and stirring to fully dissolve; performing first solid-liquid separation and collecting supernatant; adjusting the pH value of the supernatant collected by the first solid-liquid separation to 5.0-5.5; performing solid-liquid separation for the second time, and collecting precipitate to obtain silver carp muscle protein;
ultrasonic wave modified silver carp muscle protein: dissolving the silver carp muscle protein extracted by an 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 or can be freeze-dried for preservation.
As one embodiment of the present invention, the ultrasonic power is preferably 100 to 700W.
As one embodiment of the present invention, the ultrasonic time is preferably 3 to 20 minutes.
In one embodiment of the invention, in the step of extracting the silver carp muscle protein by the alkali extraction method, the pH value of the mixed homogenate is increased to 11.5, the mixed homogenate is fully dissolved by stirring, and the pH value of the supernatant collected by the first solid-liquid separation is adjusted to 5.3.
As one embodiment of the invention, the reagent used for improving the pH value is preferably 2.0mol/L NaOH solution, so as to deviate from the isoelectric point of the silver carp muscle protein, lead the surface of the silver carp muscle protein to have net negative charge, lead the silver carp muscle protein to generate electrostatic repulsive force with high charge density, and improve the hydration capability and the solubility.
In one embodiment of the present invention, the reagent used for lowering the pH is one of an organic acid such as malic acid, tartaric acid, citric acid, or an inorganic acid such as hydrochloric acid or sulfuric acid, and the concentration is 1.0 to 2.0mol/L. The purpose is to reduce the pH of the solution to isoelectric point, at which time the muscle protein surface lacks net charge, and the electrostatic repulsive force is weaker than other non-covalent forces (hydrophobic interactions, hydrogen bonding forces, van der Waals forces) to reduce the protein solubility, making the protein easier to recover by centrifugation. Preferably 2.0mol/L HCl solution.
In one embodiment of the invention, in the step of extracting the chub muscle protein by the alkali extraction method, fresh chub tissue blocks are smashed and made into mixed homogenate specifically as follows: mixing fresh silver carp tissue blocks with ice water, mashing, grinding, homogenizing to obtain mixed homogenate.
As one embodiment of the invention, the silver carp tissue block is at least one of a silver carp block after simple scale removal and bone removal or a by-product after silver carp production and processing.
As one embodiment of the invention, the mixing proportion of the silver carp tissue blocks and the ice water in the preparation and homogenization step is 1:9, and the homogenization conditions are 15000-20000 rpm, and the homogenization is carried out for 1-2 min. Aims to fully crush the tissues of the silver carp, break the bone spurs and connective tissues of the silver carp, fully contact and dissolve muscle proteins with ice water and provide a solution environment for the subsequent alkaline migration.
In one embodiment of the present invention, both the first solid-liquid separation and the second solid-liquid separation are performed by centrifugal filtration.
As one embodiment of the invention, the first solid-liquid separation adopts centrifugal rotation speed of 8000-9000 rpm, the control temperature of 0-7 ℃ and the centrifugation for 20min. The aim is to remove insoluble material by high speed centrifugation: the chub bone spur, scales, neutral fat and the like, and improves the protein extraction rate and purity.
As one implementation mode of the invention, the second solid-liquid separation adopts the centrifugal rotation speed of 5000-6000 rpm, the temperature is controlled to be 0-7 ℃, and the centrifugal speed is controlled to be 15min. The purpose is to recover soluble silver carp muscle protein without cell membrane and fat.
As one implementation mode of the invention, the ultrasonic treatment protein solution in the ultrasonic modified silver carp muscle protein is 50-100 g, the ultrasonic treatment power is 400W, and the ultrasonic treatment time is 10min.
As one embodiment of the invention, the silver carp muscle protein extracted by the 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 silver carp muscle protein solution is prepared at the power of 400W, the frequency of 15-25 kHz, the amplitude of 10-20% and the pulse intermittent ratio of 3s:3 s-7 s: and carrying out ultrasonic treatment for 10min under the condition of 7s ice bath environment to obtain the modified silver carp muscle protein.
As one embodiment of the invention, the 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 is obtained by ultrasonic treatment for 10min under the conditions of power 400W, frequency 20kHz, amplitude 10%, pulse intermittent ratio 5s:5s and ice bath environment.
As one implementation mode of the invention, the ultrasonic treatment is preferably performed by 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 in the whole process, and the temperature of the silver carp muscle protein solution is not higher than 10 ℃ in the processing process so as to prevent meat protein denaturation.
The second aim of the invention is to provide the modified silver carp muscle protein prepared by the processing method.
The third object of the invention is to provide the application of the modified silver carp muscle protein as food.
For example, the modified silver carp muscle protein prepared by the invention is used as a raw material to be processed into a pure protein silver carp surimi product, an emulsified meat surimi product or a silver carp protein freeze-dried product and the like.
As one embodiment of the invention, the method further comprises the following steps of modifying the silver carp muscle protein by ultrasonic waves: preparation of silver carp muscle protein freeze-drying: and freeze-drying the modified silver carp muscle protein to obtain the freeze-dried silver carp muscle protein.
The third object of the invention is to provide the application of the modified silver carp muscle protein as a feed.
The fourth object 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 aim of the invention is to provide the application of the modified silver carp muscle protein in preparing health care 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 shows that when alkali liquor increases the pH value of homogenate to be within the range of 10.5-11.5 and acid liquor adjusts the pH value of centrifugal clear liquid to be within the range of 5.0-5.5, the extraction rate of the silver carp muscle protein extracted and prepared by the alkali extraction method with specific parameters is more than 70 percent, the solubility is more than 35 percent and the emulsifying activity is 5.38m 2 The emulsion stability (standing at 4 ℃ for 180 min) is more than 57.14 percent. Therefore, the silver carp muscle protein obtained by the processing method of the invention has better solubility, emulsification activity, emulsification stability, gel property and water holding capacity, can solve the problems of loose meat quality and poor protein processing performance of the fish by a rinsing preparation method, improves the utilization rate of low-value silver carp resources, can better meet diversified processing process conditions and is suitable for developing richer emulsified surimi 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 plant protein to be applied to the application of feeds, foods, health-care products and pharmaceutical additives, and can be applied to the production of beauty and skin-care products due to the good emulsifying property.
(2) According to the research of the invention, the extraction rate, the solubility, the emulsification activity and the emulsification stability of the silver carp muscle protein 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 performance of the silver carp muscle protein is optimal: the extraction rate is 89.0%, the solubility is 37.05%, and the emulsifying activity is 6.03m 2 Per g, emulsion stability (180 min at 4 ℃) was 63.73%.
(3) The research of the invention shows that compared with the traditional rinsing method applied to the extraction of the silver carp muscle protein, the alkali extraction method with specific parameters of the invention has the advantages that the extraction rate of the silver carp muscle protein is obviously improved (from 42 percent to over 70 percent), and the emulsifying activity is obviously improved (from 4.37 m) 2 The/g is lifted to 5.38m 2 And/g above), the emulsion stability (standing for 180min at 4 ℃) is obviously improved (from 14% to 57%) and the solubility of the silver carp muscle protein is increased. The alkali extraction method under specific conditions adopted by the invention has remarkable advantages in the aspects of efficiently extracting the silver carp muscle protein, improving the solubility, the emulsifying activity and the emulsifying stability (standing for 180min at 4 ℃), of the silver carp muscle protein product.
(4) According to the invention, the silver carp muscle protein extracted by an alkali extraction method is modified by ultrasonic wave, so that the synergistic effect is achieved, and the synchronous improvement of the solubility (up to 93.27%) and the emulsifying activity (up to 10.36 m) of the silver carp muscle protein is realized 2 And (g), the emulsification stability (up to 95.83 percent) after standing for 30min at 4 ℃, the emulsification stability (up to 88.65 percent) after standing for 180min at 4 ℃, the gel strength (up to 1.037N), the gel water holding capacity (up to 68.05 percent) and the gel forming effect (the gel is visual, has more stereoscopic impression and better molding property) are synchronously improved, and the comprehensive processing performance of the protein is obviously improved.
(5) According to the research of the invention, the silver carp muscle protein extracted by the traditional rinsing method is modified by ultrasonic waves, the solubility is reduced rather than before the ultrasonic wave modification treatment, the gel strength and the water holding capacity are obviously reduced, the gel visual forming is poor, and the shaping performance is poor. Therefore, the similar synergistic effect of the invention can not be realized after the silver carp muscle protein extracted by the traditional rinsing method is modified by ultrasonic waves.
(6) According to the research of the invention, the time of ultrasonic treatment is changed, and particularly, compared with the time of alkali extraction prior to ultrasonic modification, the solubility, emulsion stability (4 ℃ for 180 min), gel strength and water holding capacity of the obtained silver carp muscle protein are obviously reduced, the obtained gel is visually extremely poor, the unexpected influence of the ultrasonic alkali extraction sequence on the performance is shown, the ultrasonic sequence is regulated and can not be regarded as conventional regulation, and the specific sequence of the invention can have unexpected technical effects.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the description of the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Wherein:
FIG. 1 shows the change in the extraction rate of the muscle protein of silver carp prepared in examples 1 to 5 and comparative example 1;
FIG. 2 shows the muscle protein solubility of silver carp prepared in examples 3, 6, 7 and 10;
FIG. 3 shows the emulsifying activity of the muscle protein of silver carp prepared in examples 3, 6, 7 and 10;
FIG. 4 shows the strength and water holding capacity of the gel of the chub muscle protein prepared in examples 3, 6, 7 and 10;
FIG. 5 shows the particle size change of the chub muscle protein obtained in example 3 and examples 8 to 12;
FIG. 6 shows the solubility of the muscle protein of silver carp prepared in example 3 and examples 8 to 12;
FIG. 7 shows the emulsifying activity of the muscle protein of silver carp prepared in example 3 and examples 8 to 12;
FIG. 8 shows emulsion stability of the chub muscle protein prepared in example 3 and examples 8 to 12;
FIG. 9 shows the microstructure of the chub muscle protein emulsion prepared in example 3 and examples 8 to 12, wherein the scales are 50 μm;
FIG. 10 shows the gel strength and water holding capacity change of the chub muscle protein prepared in example 3 and examples 8 to 12;
FIG. 11 shows the solubility of the chub muscle protein obtained in comparative examples 1 to 3 and example 10;
FIG. 12 shows the emulsifying activity of the chub muscle protein obtained in comparative examples 1 to 3 and example 10;
FIG. 13 shows emulsion stability of the chub muscle protein prepared in comparative examples 1 to 3, example 3, and example 10;
FIG. 14 shows the gel strength and water holding capacity change of the chub muscle protein obtained in comparative examples 1 to 3 and example 10;
FIG. 15 is a visual illustration of the chub muscle protein gel prepared in comparative examples 1 to 3 and example 3, example 10;
FIG. 16 is a schematic flow chart of the process for processing the silver carp muscle protein.
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 other than those described herein, and persons skilled in the art will readily appreciate that the present invention is not limited to the specific embodiments disclosed below.
Further, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic can be 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 embodiment and the comparative example of the invention are as follows:
measuring the extraction rate of the silver carp muscle protein:
(1) The extraction rate of the chub muscle protein is prepared by a rinsing method: mixing a proper amount of fresh silver carp tissue blocks with ice water, mashing the tissue, slowly filtering the homogenate with two layers of cheesecloth to obtain a crude extract, wherein the protein content of the crude extract is recorded as A; and finally obtaining silver carp muscle protein precipitate by a rinsing method, dissolving the silver carp muscle protein precipitate in a salt solution (0.6 mol/L NaCl, pH 7.0,4 ℃), and measuring the content of the extracted silver carp muscle protein by using a biuret protein measurement method, wherein the content of the extracted silver carp muscle protein is recorded as B.
(2) The extraction rate of the silver carp muscle protein prepared by an alkali extraction method is as follows: mixing a proper amount of fresh silver carp tissue blocks with ice water, mashing the tissue, thoroughly dissolving the tissue blocks in an alkaline environment, centrifuging the solution, taking supernatant, measuring the protein content and marking the protein content as A; and finally obtaining silver carp muscle protein precipitate after isoelectric precipitation and centrifugation, dissolving the silver carp muscle protein precipitate in a salt solution (0.6 mol/L NaCl, pH 7.0,4 ℃), and measuring the content of the extracted silver carp muscle protein by using a biuret protein assay method, wherein the content of the extracted silver carp muscle protein is recorded as B.
Protein solubility assay: diluting the protein solution of the silver carp muscle to be detected to 2.0mg/mL, centrifuging at 4 ℃ for 15min at 5000 Xg, collecting supernatant, and measuring the protein content by using biuret, wherein the protein solubility is expressed as the content percentage of the protein of the supernatant and the original protein diluted solution of the silver carp muscle.
Protein particle size determination:diluting the protein solution of the silver carp muscle to be detected to 1.0mg/mL, taking a proper amount of the protein solution, and filling the protein solution into a Zeta cuvette to automatically determine the particle size of the protein.
Emulsion preparation:before the experiment, the silver carp muscle protein solution (4 ℃) is cooled in advance, and then the silver carp muscle protein solution to be detected (the protein concentration is 1.1% w/v) and rapeseed oil are mixed according to the volume ratio of 9:1 to obtain a mixture, so as to prepare emulsion (the final concentration of the silver carp muscle protein in the emulsion is 1.0% w/v). The foregoing mixture was sheared with a high-speed shearing disperser (low-foaming probe) at 17500rpm for 2min to prepare the corresponding emulsion for protein emulsion activity assay, protein emulsion stability assay, and protein emulsion microstructure characterization described below.
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 0.1% (w/w) Sodium Dodecyl Sulfate (SDS) solution. Measurement of the Dispersion absorbance A at 500nm 500 The Emulsion Activity Index (EAI) is according to the following formulaAnd (3) performing calculation:
wherein C is the protein concentration (g/mL) before emulsification=0.01,for the oil volume fraction of the emulsion (v/v) =0.9, the dilution factor is 251.
Protein emulsion stability assay:the emulsion to be tested was immediately transferred to a 25mL beaker and placed in a cold store at 4 ℃. When the cold storage time reached 30min and 180min, 20. Mu.L of the emulsion was dispersed in 5mL of 0.1% (w/w) SDS solution from the bottom of the beaker. The absorbance was measured at 500nm and the Emulsion Stability (ESI) was calculated according to the following formula:
ESI(％)＝100*A t /A 0
wherein A is 0 And A t The absorbance of the emulsion at 4℃storage time was respectively corresponding.
Characterization of protein emulsion microstructure:freshly prepared emulsions to be tested were observed with an ECLIPSE 80i Nikon microscope. The emulsion sample (10. Mu.L) was dropped onto a glass slide, covered gently with a cover slip, observed with a 100-fold objective lens, and the emulsion image was recorded with a Nikon DS-Ri1 camera equipped on a microscope.
Protein gel preparation:5.0g of a sample of the muscle protein solution of the silver carp to be tested (diluted to 40mg/mL and degassed by centrifugation at 1000 Xg for 1min at 4 ℃) was sucked up by a plastic pipette, placed in a small glass bottle (inner diameter X length=16.5 mm X50 mm), gently sealed by a mating plastic cap with threads, placed in a water bath, and heated from 20℃to 72℃at a heating rate of 0.9℃per minute. Immediately after the heating was completed, the sample was put into an ice-water mixture and cooled for 30min, and then put into a refrigerator at 4 ℃ overnight to prepare the corresponding protein gel for the following gel strength measurement and gel water holding capacity measurement. Before measuring the gel properties, the gel sample was taken out of the refrigerator and allowed to equilibrate at room temperature for 2 hours.
Gel strength measurement:and placing the gel sample to be measured on a test platform, fixing, and measuring by using a texture analyzer at room temperature. The rate before the test is selected by the measuring mode is 5mm/s; the test rate is 1mm/s; the speed after the test is 5mm/s; the pressing distance is 10mm; the priming force is 5g; the probe model is P/0.5. Gel strength is defined as the initial pressure (N) required to puncture the gel.
Gel water holding capacity measurement:separating the gel from the glass wall by a small shovel, accurately weighing 3.0g of the gel in a centrifuge tube, centrifuging for 15min at a rotating speed of 4000 Xg, reversely airing the centrifuge tube, wiping residual water by using a dry filter paper, and calculating the weight ratio (%) of the gel after centrifugation to the gel weighed before centrifugation.
1. Extraction of silver carp muscle protein from silver carp
1.1 optimization of the extraction-alkaline extraction parameters 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 at 20000rpm by using a tissue masher, homogenizing and chopping for 2min to obtain homogenate; 2.0mol/L NaOH solution is taken to raise the pH value of homogenate to 10.5, and the homogenate is stirred for 20min under ice bath to be fully dissolved, thus obtaining a mixture; centrifuging the mixture at 8000rpm to remove insoluble substances; the pH of the centrifuged supernatant was adjusted to 5.3 using 2.0mol/L HCl solution; and then centrifuging at 6000rpm by using a centrifuge to obtain silver carp muscle protein precipitate.
The silver carp muscle protein prepared in example 1 has a protein extraction rate of 70.3%, a solubility of 35.4% and an emulsifying activity of 5.38m 2 The emulsion stability (180 min at 4 ℃) 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 at 20000rpm by using a tissue masher, homogenizing and chopping for 2min to obtain homogenate; 2.0mol/L NaOH solution is taken to raise the pH value of homogenate to 11.0, and the homogenate is stirred for 20min under ice bath to be fully dissolved, thus obtaining a mixture; centrifuging the mixture at 8000rpm to remove insoluble substances; the pH of the centrifuged supernatant was adjusted to 5.3 using 2.0mol/L HCl solution; and then centrifuging at 6000rpm by using a centrifuge to obtain silver carp muscle protein precipitate.
The silver carp muscle protein prepared in example 2 has an extraction rate of 79.8%, a solubility of 36.2% and an emulsifying activity of 5.84m 2 Per gram, emulsion stability (180 min at 4 ℃) 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 at 20000rpm by using a tissue masher, homogenizing and chopping for 2min to obtain homogenate; 2.0mol/L NaOH solution is taken to raise the pH value of homogenate to 11.5, and the homogenate is stirred for 20min under ice bath to be fully dissolved, thus obtaining a mixture; centrifuging the mixture at 8000rpm to remove insoluble substances; the pH of the centrifuged supernatant was adjusted to 5.3 using 2.0mol/L HCl solution; and then centrifuging at 6000rpm by using a centrifuge to obtain silver carp muscle protein precipitate.
The silver carp muscle protein prepared in example 3 has 89.0% of protein, 37.5% of solubility and 6.12m of emulsifying activity 2 Per g, emulsion stability (180 min at 4 ℃) 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 at 20000rpm by using a tissue masher, homogenizing and chopping for 2min to obtain homogenate; 2.0mol/L NaOH solution is taken to raise the pH value of homogenate to 11.5, and the homogenate is stirred for 20min under ice bath to be fully dissolved, thus obtaining a mixture; centrifuging the mixture at 8000rpm to remove insoluble substances; the pH of the centrifuged supernatant was adjusted to 5.0 using 2.0mol/L HCl solution; and then centrifuging at 6000rpm by using a centrifuge to obtain silver carp muscle protein precipitate.
The silver carp muscle protein prepared in example 4 has 86.4% of protein, 36.9% of solubility and 5.86m of emulsifying activity 2 Per g, emulsion stability (180 min at 4 ℃) 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 at 20000rpm by using a tissue masher, homogenizing and chopping for 2min to obtain homogenate; 2.0mol/L NaOH solution is taken to raise the pH value of homogenate to 11.5, and the homogenate is stirred for 20min under ice bath to be fully dissolved, thus obtaining a mixture; centrifuging the mixture at 8000rpm to remove insoluble substances; the pH of the centrifuged supernatant was adjusted to 5.5 using 2.0mol/L HCl solution; and then centrifuging at 6000rpm by using a centrifuge to obtain silver carp muscle protein precipitate.
The silver carp muscle protein prepared in example 5 has an extraction rate of 87.6%, a solubility of 36.7% and an emulsifying activity of 5.83m 2 Per g, emulsion stability (180 min at 4 ℃) was 59.04%.
Comparative analysis examples 1 to 5 can find that: when alkali liquor increases the pH value of homogenate to 10.5-11.5, acid liquor adjusts the pH value of the centrifugated clear liquid to 5.0-5.5, the extraction rate of the silver carp muscle protein 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 m 2 Per gram, the emulsion stability (180 min at 4 ℃) is 57.14 to 63.73 percent. The extraction rate, the solubility, the emulsification activity and the emulsification stability of the silver carp muscle protein can be obviously improved by optimizing the two pH values. Preferably, when the alkali liquor increases the pH value of the homogenate to be within 11.5, and the acid liquor adjusts the pH value of the centrifugate to be 5.3, the performance of the silver carp muscle protein is optimal: the extraction rate is 89.0%, the solubility is 37.5%, and the emulsifying activity is 6.12m 2 Per g, emulsion stability (180 min at 4 ℃) was 63.73%.
1.2 comparison of the extraction by alkali extraction method of the invention with the extraction by conventional rinsing method
Comparative example 1 extraction by conventional rinsing
Extracting silver carp muscle protein 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 and homogenizing for 2min by using a tissue triturator at 20000rpm, slowly stirring and homogenizing for 15min, pouring into a filter containing two layers of cheesecloth for dehydration, adding ice water with a volume which is 4 times that 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 to obtain the silver carp muscle protein.
Comparative example 1 preparationThe extraction rate of the prepared silver carp muscle protein is 42.0%, and the solubility of the silver carp muscle protein is 33.8%; emulsifying activity was 4.37m 2 Per g, emulsion stability (180 min at 4 ℃) was 14.40%.
Comparative analysis comparative example 1 and examples 1 to 5 can be found that: compared with the traditional rinsing method for extracting the silver carp muscle protein, the alkali extraction method with specific parameters of the invention has the advantages that 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) 2 The/g is lifted to 5.38m 2 And/g above), the emulsion stability (standing at 4 ℃ for 180 min) is obviously improved (from 14.40% to 57.14% above). The alkali extraction method under specific conditions adopted by the invention has remarkable advantages in the aspects of efficiently extracting the silver carp muscle protein, improving the solubility, the emulsifying activity and the emulsifying stability (standing for 180min at 4 ℃), of the silver carp muscle protein product.
2. Modified silver carp muscle protein
2.1 optimization of ultrasound modification parameters
The silver carp muscle protein raw materials of examples 6 to 12 are obtained by extraction in the extraction method of example 3 (alkali liquor increases the pH value of homogenate to 11.5, acid liquor adjusts the pH value of the centrifugated clear liquid to 5.3), and fresh extracted silver carp muscle protein is immediately refrigerated at low temperature for standby.
EXAMPLE 6 alkaline extraction+ultrasonic modification
The silver carp muscle protein is dissolved in a salt solution (0.6 mol/L NaCl, pH 7.0,4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, a stainless steel probe with the diameter of 7mm is positioned below the central liquid level of the solution by 2.0cm, and the silver carp muscle protein is obtained by ultrasonic treatment for 10min under the conditions of 100W power, 20kHz frequency, 10 amplitude, 5s:5s pulse intermittent ratio and ice bath environment.
EXAMPLE 7 alkaline extraction+ultrasonic modification
The silver carp muscle protein is dissolved in a salt solution (0.6 mol/L NaCl, pH 7.0,4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, a stainless steel probe with the diameter of 7mm is positioned below the central liquid level of the solution by 2.0cm, and the silver carp muscle protein is obtained by ultrasonic treatment for 10min under the conditions of power 700W, frequency 20kHz, amplitude 10%, pulse intermittent ratio 5s:5s and ice bath environment.
EXAMPLE 8 alkaline extraction+ultrasonic modification
The silver carp muscle protein is dissolved in a salt solution (0.6 mol/L NaCl, pH 7.0,4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, a stainless steel probe with the diameter of 7mm is positioned below the central liquid level of the solution by 2.0cm, and the silver carp muscle protein is obtained by ultrasonic treatment for 3min under the conditions of power 400W, frequency 20kHz, amplitude 10%, pulse intermittent ratio 5s:5s and ice bath environment.
EXAMPLE 9 alkaline extraction+ultrasonic modification
The silver carp muscle protein is dissolved in a salt solution (0.6 mol/L NaCl, pH 7.0,4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, a stainless steel probe with the diameter of 7mm is positioned below the central liquid level of the solution by 2.0cm, and the silver carp muscle protein is obtained by ultrasonic treatment for 6min under the conditions of power 400W, frequency 20kHz, amplitude 10%, pulse intermittent ratio 5s:5s and ice bath environment.
EXAMPLE 10 alkaline extraction+ultrasonic modification
The silver carp muscle protein is dissolved in a salt solution (0.6 mol/L NaCl, pH 7.0,4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, a stainless steel probe with the diameter of 7mm is positioned below the central liquid level of the solution by 2.0cm, and the silver carp muscle protein is obtained by ultrasonic treatment for 10min under the conditions of power 400W, frequency 20kHz, amplitude 10%, pulse intermittent ratio 5s:5s and ice bath environment.
EXAMPLE 11 alkaline extraction+ultrasonic modification
The silver carp muscle protein is dissolved in a salt solution (0.6 mol/L NaCl, pH 7.0,4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, a stainless steel probe with the diameter of 7mm is positioned below the central liquid level of the solution by 2.0cm, and the silver carp muscle protein is obtained by ultrasonic treatment for 15min under the conditions of power 400W, frequency 20kHz, amplitude 10%, pulse intermittent ratio 5s:5s and ice bath environment.
EXAMPLE 12 alkaline extraction+ultrasonic modification
The silver carp muscle protein is dissolved in a salt solution (0.6 mol/L NaCl, pH 7.0,4 ℃) to prepare 50g of silver carp muscle protein solution with the mass fraction of 4.0%, a stainless steel probe with the diameter of 7mm is positioned below the central liquid level of the solution by 2.0cm, and the silver carp muscle protein is obtained by ultrasonic treatment for 20min under the conditions of power 400W, frequency 20kHz, amplitude 10%, pulse intermittent ratio 5s:5s and ice bath environment.
As can be seen from comparative analysis examples 6, 7 and 10, when the ultrasonic treatment power is controlled within the range of 100-700W, the solubility of the silver carp muscle protein is 47.05-78.65% and the emulsifying activity is 8.20-9.28 m by the modification method of the specific parameters 2 The gel strength per gram is 0.42-1.04N, and the gel water holding capacity is 53.42-60.93%. In order to obtain better emulsification activity, gel strength and water holding capacity, the effects of lower energy consumption and no obvious influence on solubility are achieved, and ultrasonic power parameters are optimized to obtain: 400W. As can be seen from comparative analysis examples 8 to 12, when the ultrasonic time is controlled within the range of 3 to 20 minutes, the particle size of the silver carp muscle protein is 258.57 to 402.60nm, the solubility is 84.40 to 93.28 percent, and the emulsifying activity is 7.76 to 10.36m by the modification method of the specific parameters 2 The gel strength per gram is 0.87-1.04N, and the gel water holding capacity is 55.16-68.05%. In order to obtain the effects of smaller particle size, higher solubility, higher emulsifying activity and emulsifying stability, more uniform emulsion oil drop size, higher gel strength and higher gel water holding capacity of protein, the ultrasonic time parameter is optimized: and 10min.
The optimized parameters are mainly based on the laboratory measurement results, 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 silver carp muscle protein treatment capacity. The ultrasonic power and time parameters may also be increased or decreased as appropriate to meet the needs of a particular process. In conclusion, the ultrasonic treatment can achieve better modification effect under the condition of 100-1000W for 3-30 min.
Comparative analysis of the silver carp muscle protein obtained by the alkali extraction method alone of example 3 and the alkali extraction + ultrasonic wave modified silver carp muscle protein of examples 6 to 12 can be found that: silver carpThe muscle protein solubility is significantly improved (from 37.5% to 84.0% above, see fig. 6), and the emulsifying activity is significantly improved (from 6.12 m) 2 The/g is lifted to 7.76m 2 Per g above, see fig. 7), emulsion stability (30 min at 4 ℃ standing) is significantly improved (from 72.02% to 84.30% above, see fig. 8), emulsion stability (180 min at 4 ℃ standing) is significantly improved (from 63.73% to 71.75% above, see fig. 8), emulsion oil drop size, dispersibility and stability are significantly improved (see fig. 9), gel strength is significantly improved (from 0.27N to 0.87N above, see fig. 10), gel water holding capacity is significantly improved (from 47.65% to 55.16% above, see fig. 10), which illustrates the mutual support between the alkaline extraction process and the ultrasonic modification process, further improves the parameters of the silver carp muscle protein in terms of dissolving capacity, emulsifying performance and gel characteristics, and improves the comprehensive processing performance of the silver carp muscle protein, thus significantly improving the market value of the silver carp muscle protein product prepared by the method.
2.2 Combined comparison of different extraction methods with ultrasound modification
Comparative example 2 rinsing method + ultrasonic modification
The silver carp muscle protein raw material of comparative example 2 is extracted by the extraction method of comparative example 1, and fresh extracted silver carp muscle protein is immediately refrigerated at low temperature for standby.
The silver carp muscle protein was dissolved in a salt solution (0.6 mol/L NaCl, pH 7.0,4 ℃) to prepare 50g of a silver carp muscle protein solution with a mass fraction of 4.0%, a stainless steel probe with a diameter of 7mm was positioned 2.0cm below the central liquid level of the solution, and the silver carp muscle protein was subjected to ultrasonic treatment under the conditions of 400W power, 20kHz frequency, 10% amplitude, 5s:5s pulse-interval ratio and ice bath environment for 10 minutes to obtain a rinsed modified silver carp muscle protein (see the ultrasonic modification method of example 10).
Comparative analysis control 1 and control 2 can be found: after the silver carp muscle protein extracted by the rinsing method is subjected to ultrasonic modification treatment (comparative example 2), the solubility of the silver carp muscle protein is reduced (see fig. 11) compared with that of the silver carp muscle protein before the ultrasonic modification treatment (comparative example 1), the gel strength and the water holding capacity of the silver carp muscle protein are obviously reduced (see fig. 14), and the gel visual forming is poor (see fig. 15). Comparative analysis comparative example 2 and example 10 can show that, on one hand, the solubility, emulsifying activity, emulsifying stability, water holding capacity and gel forming effect of the silver carp muscle protein prepared by the alkali extraction method and the ultrasonic wave modification treatment are still far higher than those of the silver carp muscle protein prepared by the rinsing method and the ultrasonic wave modification treatment (see fig. 11-15). On the other hand, the silver carp muscle protein obtained by different extraction methods shows different change trends after ultrasonic modification treatment: the combination of rinsing method and ultrasonic wave cannot realize synchronous improvement of solubility, emulsification activity, emulsification stability, water holding capacity and gel forming effect of the silver carp muscle protein, and the variation trend of the solubility, the gel strength and the water holding capacity is different from that of the combination of alkali extraction method and ultrasonic wave. The combined use of the alkali extraction method and ultrasonic modification is proved to be a synergistic effect, and a corresponding technical effect can not be realized if any treatment process is replaced.
As shown in fig. 14, the water holding capacity data of comparative examples 1 to 2, example 3 and example 10 show that the water holding capacity by the rinsing method alone is slightly higher than that by the alkali extraction method alone, but the combined treatment of the alkali extraction method and the ultrasonic wave is rather higher than that by the rinsing method and the ultrasonic wave after the same ultrasonic treatment. The gel strength data also shows a similar rule, which indicates that the synergy exists between the alkali extraction method and the ultrasonic treatment, and the ultrasonic treatment is utilized to make up the defects of lower gel strength and lower water holding capacity caused by the alkali extraction method.
2.3 changing the timing of sonication
Comparative example 3 synchronous alkaline extraction and ultrasound
Mixing a proper amount of fresh silver carp tissue blocks with ice water according to a ratio of 1:9 (w/w), mechanically grinding at 20000rpm by using a tissue masher, homogenizing and chopping for 2min to obtain homogenate; then 2.0mol/L NaOH solution is taken to raise the pH value of the homogenate to 11.5, and the homogenate is stirred for 20min under ice bath to be fully dissolved, thus obtaining a mixture; subjecting the mixture to ultrasonic treatment, centrifuging the mixture at 8000rpm at high speed to remove insoluble substances; the pH of the centrifuged supernatant was adjusted to 5.3 using 2.0mol/L HCl solution; and then centrifuging at 6000rpm by using a centrifuge to obtain silver carp 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 surface of the solution, and sonicated for 10min under conditions of power 400W, frequency 20kHz, amplitude 10%, pulse-to-pulse ratio 5s:5s, and ice bath environment (sonication conditions were the same as in example 10).
Comparative analysis comparative example 3 compared with example 10, it can be found that, compared with the alkali extraction prior to ultrasonic modification in the present invention, the solubility (see fig. 11), emulsion stability (4 ℃ for 180 min) (see fig. 13), gel strength and water holding capacity of the obtained silver carp muscle protein are significantly reduced (see fig. 14), the gel visual performance of comparative example 3 is extremely poor (see fig. 15), which indicates that the ultrasonic treatment has limited improvement on the muscle protein processing performance in the alkali extraction process, the importance of ultrasonic timing is reflected, the ultrasonic alkali extraction sequence has unexpected influence on the performance, the ultrasonic sequence can not be regarded as conventional adjustment, and the specific sequence of the present invention can have unexpected technical effects.
3. Application of modified silver carp muscle protein in production of beauty and skin care products
The application of the modified silver carp muscle protein in the preparation of emulsion is taken as an example for illustration.
The modified silver carp muscle protein of examples 6-12 can be used as an emulsifier instead of vegetable protein and applied to emulsion preparation.
The alkali-extracted silver carp muscle protein after ultrasonic modification has good emulsifying activity and emulsifying stability, plays the role of interaction between fat globules and an aqueous phase interface in the emulsion forming process, reduces the interfacial tension of two phases, generates a thicker protein interface layer, and can reduce the size of fat oil drops at the same time, so that the fat globules 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 the embodiments 6-12 has a very deep research significance for replacing plant protein or artificial synthetic emulsifier with weak emulsifying property by using the modified animal protein.
4. Application of modified silver carp muscle protein as food
The application of the modified silver carp muscle protein in the preparation of protein gel is exemplified.
The granularity of the alkali-extracted silver carp muscle protein after ultrasonic modification is smaller, the formed gel structure is more uniform and compact, the arrangement is orderly and fine, and the alkali-extracted silver carp muscle protein has good elasticity and toughness. The silver carp muscle protein gel has strong integrity, is not easy to flow, has jelly-like or pudding-like texture, is white in whole and rich in luster, does not show obvious water precipitation, and shows good water holding capacity. The gel strength and water holding capacity of the alkali-extracted silver carp muscle protein modified by ultrasound according to examples 6 to 12 can be applied to meat protein gel products, in particular minced fillet products and imitation products thereof.
5. Application of modified silver carp muscle protein as feed
The modified silver carp muscle protein of examples 6-12 can be processed into silver carp muscle protein freeze-dried by freeze drying, and is applied to feed products.
6. Application of modified silver carp muscle protein in preparation of health care product and medicine additive
The modified silver carp muscle protein of examples 6-12 can be used as an emulsifier and an important nutrient component for preparing health products and pharmaceutical additives.
Claims (8)
1. A processing method of modified silver carp muscle protein is characterized by comprising the following steps:
extracting silver carp muscle protein by an alkali extraction method: mashing fresh silver carp tissue blocks and preparing mixed homogenate; raising the pH value of the mixed homogenate to 11.5, and stirring to fully dissolve; performing first solid-liquid separation and collecting supernatant; adjusting the pH value of the supernatant collected by the first solid-liquid separation to 5.3; performing solid-liquid separation for the second time and collecting precipitate to obtain silver carp muscle protein;
ultrasonic wave modified silver carp muscle protein: dissolving the silver carp muscle protein extracted by an alkali extraction method in NaCl solution with the temperature of 4 ℃, the pH value of 7.0 and the concentration of 0.6mol/L, and carrying out ultrasonic treatment for 10min under the condition of 400W of power 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 alkali extraction, fresh silver carp tissue blocks are smashed and made into mixed homogenate, specifically: mixing fresh silver carp tissue blocks with ice water, mashing, grinding, homogenizing to obtain mixed homogenate.
3. The method for processing modified silver carp muscle protein according to claim 1, wherein the ultrasonic treatment is carried out in an ice bath in the whole process, and the temperature of the silver carp muscle protein solution in the processing process is not more than 10 ℃.
4. A modified silver carp muscle protein produced by the processing method as claimed in any one of claims 1 to 3.
5. The use of the modified silver carp muscle protein as claimed in claim 4 as a food.
6. The use of the modified silver carp muscle protein as claimed in claim 4 as a feed.
7. The use of the modified silver carp muscle protein as claimed in claim 4 for producing a cosmetic skin care product.
8. The use of the modified silver carp muscle protein as claimed in claim 4 for preparing health products and pharmaceutical additives.
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| CN104351849A (en) * | 2014-10-31 | 2015-02-18 | 华南理工大学 | High-yield extraction method for fish protein by alkali dissolution method and minced fillet thereof |
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| CN109247424A (en) * | 2018-09-21 | 2019-01-22 | 天津农学院 | A kind of method and its application preparing modified protein by minced fish rinse water |
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| CN104351849A (en) * | 2014-10-31 | 2015-02-18 | 华南理工大学 | High-yield extraction method for fish protein by alkali dissolution method and minced fillet thereof |
| CN104543323A (en) * | 2015-01-13 | 2015-04-29 | 华南理工大学 | High-yield low-pollution discharge acid soluble method for extracting fish protein and minced fillet thereof |
| CN109247424A (en) * | 2018-09-21 | 2019-01-22 | 天津农学院 | A kind of method and its application preparing modified protein by minced fish rinse water |
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