Soy sauce containing high protein and preparation method thereof

Abstract

The invention relates to the field of marine food manufacturing, in particular to high-protein-content soy sauce and a preparation method thereof, and the high-protein-content soy sauce comprises the following raw materials: laminarin, modified anthocyanin, compound protein, docosahexaenoic acid, astaxanthin, taurine, carrageenan, beta-amylase, soybeans, salt and purified water. The laminarin, the modified anthocyanin and the compound protein play a role through a synergistic effect, the compound protein in the soy sauce is wrapped by the modified anthocyanin to form a compound, a laminarin film is tightly attached to the outer layer of the compound formed by the modified anthocyanin, and the wrapping capacity and the structural stability of the compound are further enhanced; meanwhile, the oxidation process of fat and protein in the soy sauce is slowed down, the modified anthocyanin and a laminarin film interact to maintain the protein content in the soy sauce, laminarin, the modified anthocyanin and compound protein form a dynamic balance system, the nutritional value and quality of the soy sauce are improved, and the soy sauce with high protein content is provided.

Description

Technical Field

[0001] The invention relates to the field of marine food manufacturing, in particular to a high-protein soy sauce and a preparation method thereof. Background Art

[0002] As a common seasoning in life, soy sauce plays an important role in cooking and diet. The soy sauce currently on the market contains a variety of nutrients such as amino acids and organic acids, but its protein content is relatively low. Protein is an essential nutrient for the human body and is essential for maintaining life activities, building body tissues and organs, and performing various physiological functions. The low protein content in soy sauce means that its nutritional value is relatively low and cannot meet the body's full demand for protein. For example, athletes and fitness enthusiasts need more protein to support muscle repair and growth during high-intensity training; pregnant and lactating women need additional protein during pregnancy and lactation to support fetal development and milk secretion; the body functions of the elderly gradually decline with age, and they need more protein to maintain a healthy state of the body. In addition, if ordinary people ignore protein intake for a long time, it will lead to a series of health problems. For example, protein is an important component for building muscles and bones. Lack of protein may lead to muscle atrophy, osteoporosis and other problems. At the same time, protein is also involved in the body's immune response and cell repair processes. Lack of protein may reduce the body's immunity and increase the risk of disease. Therefore, increasing the protein content in soy sauce is not only to meet the special needs of specific groups of people for protein, but also to enhance the overall nutritional value of soy sauce, so that it can provide more nutritional support for the human body while meeting the seasoning needs. Finally, soy sauce with a high protein content can not only enrich the functionality of soy sauce, but also meet consumers' growing demand for healthy condiments and promote innovation and development in the condiment industry.

[0003] (1) Technical issues to be solved

[0004] The purpose of the present invention is to provide a high-protein soy sauce and a preparation method thereof. The formula of soy sauce fermentation is optimized, and the advantage of seafood being high in protein is combined, so that the protein in seafood is combined with the soy sauce fermentation process, so as to achieve a significant increase in the protein content in the soy sauce, which not only enriches the nutritional components of the soy sauce, but also gives the soy sauce a unique seafood flavor. Finally, a soy sauce product having multiple flavors and being rich in high-quality protein is provided.

[0005] (2) Technical solution

[0006] To achieve the above object, on the one hand, the present invention provides a high-protein soy sauce, comprising the following raw materials in parts by weight: 10-20 parts of kelp polysaccharide, 5-15 parts of carrageenan, 1-3 parts of β-amylase, 20-40 parts of complex protein, 20-40 parts of docosahexaenoic acid, 5-15 parts of astaxanthin, 10-20 parts of taurine, 100-200 parts of soybeans, 20-30 parts of salt, and 200-400 parts of purified water;

[0007] The high-protein soy sauce also includes:

[0008] Modified anthocyanins;

[0009] The modified anthocyanin and the composite protein are in a weight ratio of 1:(3-6);

[0010] The modified anthocyanidin is prepared by encapsulating the anthocyanidin in a lipid bilayer structure, and the modified anthocyanidin has a particle size of 10 to 20 μm and a specific surface area of ​​20 to 30 m 2 / g.

[0011] Furthermore, the preparation method of the modified anthocyanidin comprises:

[0012] S11. Add 50 to 60 g of blueberry to 75% to 80% ethanol solution under heating and stirring, heat to 60 to 70 ° C, stir for 4 to 5 hours, stir evenly until completely dissolved to obtain a blueberry extract;

[0013] S12. 5 to 6 g of soybean lecithin and 10 to 15 g of cholesterol are mixed under stirring at room temperature, and then a 75% to 80% ethanol solution is added to promote the reaction to obtain a liposome mixture;

[0014] S13. After the liposome mixture is cooled to room temperature, the blueberry extract is slowly added to the liposome mixture, stirred evenly, and the pH is adjusted to 6 to 7 with a phosphate buffer to obtain a first mixture;

[0015] S14. Add the first mixed solution and 75% to 80% ethanol solution to a rotary evaporator and then evaporate until membrane lipids are formed on the inner wall of the bottle on the rotary evaporator, continue to add 75% to 80% ethanol solution, and perform ultrasonic treatment to form an emulsion, then place the bottle on the rotary evaporator in a 30 to 40° C. water bath to remove the emulsion gel on the bottle, thereby obtaining a first liposome;

[0016] S15. The pH of the first liposome is adjusted to 6-7 with phosphate buffer, and then anthocyanin liposomes of 200-220 nm are obtained using a high-pressure homogenizer, which are freeze-dried at -4 to -10°C, and modified anthocyanins are obtained after drying for 8 to 12 hours.

[0017] Furthermore, the mass ratio of soybean, β-amylase and carrageenan is 1:(0.5-2):(1-1.5).

[0018] Furthermore, the mass ratio of laminarin to carrageenan is 1:(2-4).

[0019] Furthermore, the method for preparing the composite protein comprises:

[0020] S21. Wash 20-30 g of freshwater shrimp, 40-50 g of clams, 40-50 g of sea crucian carp, and 10-30 g of oysters, peel and debone them, cut them into small pieces, and then put them into a wall-breaking machine to grind them to obtain a seafood mixture;

[0021] S22. 5 to 6 g of acetic acid buffer solution is added dropwise to the seafood mixture, followed by adding 1 to 2 g of protease and stirring, heating at 40 to 50° C. for 1 to 2 h to obtain a first mixture;

[0022] S23. The first mixture was transferred to a microwave reactor, the microwave power was set to 300W to 500W, the reaction time was 10 to 20min, and a pause of 1 to 2min was performed after each microwave, and the microwave was repeated 5 to 6 times to obtain a second mixture;

[0023] S24. The second mixture was transferred to an ultrasonic reactor, the ultrasonic power was set to 100W to 200W, the reaction time was 30 to 40min, and a pause of 1 to 2min was performed after each microwave, and the ultrasound was repeated 5 to 6 times to obtain a third mixture;

[0024] S25. The third mixture is centrifuged at a speed of 5000 to 6000 rpm for 15 to 20 min, and the resulting liquid is collected to obtain a first mixed solution;

[0025] S26. The first mixed liquid is concentrated by steam distillation for 3 to 5 hours to extract the protein in the seafood, and after cooling, it is stored in a refrigerator at 0 to -4°C to obtain a composite protein.

[0026] Furthermore, the composite protein is obtained by combining microwave-assisted extraction with ultrasound-assisted extraction of protein from seafood.

[0027] Furthermore, the types of the proteins are mainly myofibrillar proteins, troponin, myofibrillar proteins and sarcoplasmic proteins.

[0028] Furthermore, the mass ratio of the composite protein, docosahexaenoic acid and astaxanthin is (1-20):(2-4):(100-300).

[0029] On the other hand, based on the same inventive concept, the present invention also provides a method for preparing a high-protein soy sauce, which is applied to the high-protein soy sauce, comprising the following steps:

[0030] S31. Add 100-200g soybeans into a container and soak them in clean water for 6-10h, wash and drain, put them into a normal pressure steamer and steam them thoroughly without sandwich, take them out and cool them to 20-30℃, the mass ratio of soybeans, carrageenan and β-amylase strains is 100:2:3, mix them repeatedly and transfer them to the Quchi, culture them in the Qu room for 60-72h to obtain the first fermentation product;

[0031] S32. The mass ratio of salt to pure water is (10-20): 100 to obtain a saline solution, which is evenly sprayed on the first fermentation product in the Quchi, and then naturally fermented, and then docosahexaenoic acid, astaxanthin, taurine, complex protein, modified anthocyanin and kelp polysaccharide are sequentially added to obtain a second fermentation product;

[0032] S33. Wait for the second fermentation product to undergo natural fermentation, monitor the status of the second fermentation product every 3 to 4 days, ventilate and stir the product when the temperature is too high or too low during fermentation, turn the product over once a month, 3 to 5 times, and after 6 to 7 months, take the bottom liquid with a tube, heat and sterilize, cool, and filter to obtain high-protein soy sauce.

[0033] The mechanism of action of the above raw material components is as follows:

[0034] Modified anthocyanidins are formed by encapsulating anthocyanidins in liposomes. Anthocyanidins are a type of water-soluble natural pigments belonging to the flavonoids. The double-layer structure of liposomes can effectively encapsulate anthocyanidins and protect them from the influence of the external environment, such as light, oxygen and changes in acid-base conditions, thereby improving their stability. The hydrophobic tail of the modified anthocyanidin can tightly wrap hydrophobic substances such as oils and fats to prevent them from separating from soy sauce. The hydrophilic head can interact with water molecules to keep the complex stable in soy sauce. A double-layer membrane structure, i.e., a complex, can be spontaneously formed through physical forces such as hydrogen bonds and hydrophobic interactions. When the modified anthocyanidin comes into contact with nutrients (such as vitamins, minerals, amino acids, etc.), these nutrients can be encapsulated inside the modified anthocyanidin or adsorbed on its surface.

[0035] Laminaria polysaccharide is a natural polysaccharide extracted from kelp, which has good biocompatibility and stability. Laminaria polysaccharide can form gels, encapsulating nutrients such as proteins in the gel network. It can increase the stability and functionality of liposomes. In soy sauce, it can reduce the tension of the oil-water interface, promote the mixing of oil and water phases, form a stable emulsion, which is beneficial to improve the uniformity and stability of soy sauce and prevent oil-water stratification. Laminaria polysaccharide also has antioxidant properties, which can protect other ingredients in soy sauce from oxidative damage.

[0036] The composite protein is made from seafood raw materials such as shrimp, clams, sea carp and oysters through enzymatic hydrolysis and extraction. These shrimp, clams, sea carp and oysters are rich in protein, and the protein is completely retained during the extraction process, which not only retains the delicious taste of the raw materials, but also makes it easy to increase the protein content. At the same time, the composite protein also contains nutrients such as amino acids, vitamins, and minerals, which indirectly promote the growth and reproduction of microorganisms in soy sauce. These microorganisms will use the nutrients in soy sauce (such as carbon sources, nitrogen sources, etc.) to carry out metabolic activities and synthesize more protein during the fermentation process of soy sauce. The thickening and stabilizing effects of kelp polysaccharides can help liposomes disperse and prevent them from aggregating.

[0037] Carrageenan is a hydrophilic colloid, also known as kelp gum, agar, carrageenan or carrageenan. During the brewing process of soy sauce, carrageenan can absorb water and form a viscous colloid, thereby increasing the viscosity and thickness of the soy sauce, making the soy sauce more textured and tasty. Carrageenan can interact with the protein, fat and other ingredients in the soy sauce to form a stable colloid system. It not only extends the shelf life of the soy sauce, but also maintains the good appearance and taste of the soy sauce.

[0038] Astaxanthin is a ketocarotenoid with a chemical name of 3,3′-dihydroxy-4,4′-diketo-β,β′-carotene. It is fat-soluble and insoluble in water, but soluble in organic solvents such as chloroform, acetone, benzene, etc. Astaxanthin itself is not very stable, easily oxidized, and easily decomposed by light. It is a powerful antioxidant that can effectively remove free radicals and prevent cells from oxidative damage. Adding astaxanthin to soy sauce can protect the lipids and other components in the soy sauce from oxidation, thereby extending the shelf life of the soy sauce and maintaining its freshness. In addition, the antioxidant effect of astaxanthin also helps prevent the soy sauce from deteriorating due to factors such as light and temperature during storage and transportation.

[0039] Soybean is the main raw material for brewing soy sauce. It is rich in nutrients such as protein, fat, minerals and vitamins. The fat in soybean decomposes to produce higher fatty acid esters during the brewing process. These lipids give soy sauce a strong taste. Salt provides appropriate saltiness. Salt can combine with amino acids in soy sauce to produce amino acid sodium salt. This compound has umami taste and enhances the flavor of soy sauce. At the same time, salt has a certain preservative effect in soy sauce, which can inhibit the growth of harmful microorganisms and extend the shelf life of soy sauce. In addition, the concentration of salt will also affect the fermentation process of soy sauce. By adjusting the concentration of salt, the fermentation speed and flavor changes of soy sauce can be controlled. β-amylase is one of the main microorganisms in the brewing process of soy sauce. β-amylase produces heat and gases such as carbon dioxide through metabolic activities, which promotes the fermentation process of soy sauce. At the same time, it can produce a variety of enzymes, such as proteases, peptidases, amylases, etc. These enzymes play a role in hydrolyzing complex organic substances such as protein and starch in the brewing process of soy sauce to generate simple flavoring substances. The flavor substances and aroma components produced during the fermentation process of soy sauce mainly come from the metabolic activity of β-amylase. Taurine and docosahexaenoic acid mainly come from fish in seafood. Taurine and docosahexaenoic acid have a unique umami taste and can interact with other ingredients in soy sauce to enhance the taste of soy sauce.

[0040] In high-protein soy sauce, kelp polysaccharides, modified anthocyanins and complex proteins play a role through synergistic effects. During the fermentation and preparation of soy sauce, modified anthocyanins, as a polysaccharide, have unique physical and chemical properties due to their functional groups such as amino and hydroxyl groups in their molecular structure. These functional groups can interact with nutrients such as proteins, amino acids, and polysaccharides in the complex protein, and tightly wrap the nutrients such as complex proteins in the complex formed by modified anthocyanins through physical adsorption and chemical bonding. This wrapping effect not only effectively prevents the loss of nutrients such as proteins in soy sauce, but also enables the protein to be more fully absorbed and utilized by the human body when consumed. Subsequently, the surface of the complex will be tightly attached to the kelp polysaccharide film. The kelp polysaccharide is hydrophilic, and the phosphate groups face the water phase. The kelp polysaccharide molecules interact with the hydrophobic core of the complex through hydrophobic interactions to form a tight double-layer structure. In addition, the phosphate groups of the kelp polysaccharide molecules can also interact electrostatically with the amino groups of the modified anthocyanin molecules, further enhancing the stability of the double layer. Finally, the close fit of the kelp polysaccharide film effectively enhances the wrapping ability and structural stability of the complex. At the same time, the kelp polysaccharide film can prevent the rupture and dissolution of the complex, and protect the nutrients inside the complex from being affected by the external environment. The combination of the kelp polysaccharide film and the modified anthocyanin complex can provide a stable protein carrier, so that the protein molecules in the soy sauce are not easy to aggregate or precipitate, and can also maintain the transparency and texture of the soy sauce. At the same time, the complex can effectively maintain the high protein content in the soy sauce, and the hydrophobicity of the kelp polysaccharide film can prevent protein precipitation and maintain the stability of the protein in the solution. An appropriate amount of carrageenan can help adjust the pH value of the soy sauce, help maintain the stability and flavor of the soy sauce, and can also inhibit the growth of harmful microorganisms and extend the shelf life of the soy sauce. At the same time, astaxanthin can promote the dissolution and stability of certain pigments in the soy sauce, thereby improving the color saturation of the soy sauce, and can also slow down the oxidation process of fat and protein in the soy sauce. At the same time, it can promote the dissolution and absorption of certain nutrients in the soy sauce (such as amino acids, vitamins, etc.), thereby improving the utilization rate of these nutrients by the human body. Soybeans are rich in protein, fat, minerals and vitamins. The fat in soybeans breaks down during the brewing process to produce higher fatty acid esters, which give soy sauce a rich flavor. Carrageenan absorbs water, thereby increasing the viscosity and thickness of soy sauce, making the soy sauce more textured and tasty. Taurine and docosahexaenoic acid interact with other ingredients in soy sauce to enhance the nutritional value of soy sauce. Salt provides saltiness and combines with amino acids to form amino acid sodium salts, which enhance the umami taste. It also has a preservative effect, inhibits harmful microorganisms, and prolongs the shelf life. As the main microorganism, β-amylase produces enzymes such as proteases, peptidases, and amylases through metabolism, hydrolyzes proteins and starches, produces flavoring substances, and promotes the formation of flavor substances and aroma components.

[0041] (3) Beneficial effects

[0042] Compared with the prior art, the present invention has the following beneficial effects:

[0043] 1. The modified anthocyanin is used as the core encapsulating material to tightly encapsulate the nutrients such as the complex protein in the complex, while the kelp polysaccharide film serves as the outer protective barrier, which enhances the stability and antioxidant capacity of the complex. This dynamic balance system not only improves the nutritional value and quality of soy sauce, but also significantly improves the taste, flavor and color of soy sauce.

[0044] 2. Modified anthocyanins are made by wrapping liposomes in the anthocyanin shell, which has better solubility and biocompatibility. Modified anthocyanins can effectively wrap up the complex protein and other nutrients in soy sauce through physical adsorption and chemical bonding to form a stable complex structure. This complex structure can not only protect the high protein environment inside the soy sauce, but also enhance the overall nutritional value of the soy sauce;

[0045] 3. Laminaria polysaccharide is a natural polysaccharide with good biocompatibility and stability. When the laminarin film is tightly attached to the outer layer of the complex formed by modified anthocyanins, it can further enhance the encapsulation ability and structural stability of the complex. In addition, the laminarin film can also act as a barrier to slow down the oxidation process of fat and protein in soy sauce, thereby maintaining the freshness and flavor of the soy sauce;

[0046] 4. Complex protein, taurine, astaxanthin and docosahexaenoic acid all come from seafood products. Through scientific proportions, the various precious nutrients contained in marine organisms are perfectly integrated, so that the soy sauce not only maintains its original delicious taste, but also has extremely high nutritional value. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 This is a SEM image of kelp polysaccharide of Example 1 of the present invention;

[0048] Figure 2 This is the SEM image of the composite protein of Example 1 of the present invention. DETAILED DESCRIPTION

[0049] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0050] The experimental equipment and preparations of the embodiments described below are as follows: electronic balance (Sartorius, Germany), electric constant temperature water bath (Jiangsu Kedao), magnetic stirrer (Shanghai Meiyingpu), ultrasonic instrument (Shanghai Yixin), high-speed centrifuge (Guangzhou Jidi), vacuum drying oven (Shanghai Jiecheng), rotary evaporator (Shanghai Yaote), hydrothermal reactor (Shanghai Baikal), scanning electron microscope (Zeiss, Germany), specific surface area analyzer (Beijing Best Instrument Technology), freeze dryer (Shanghai Pudong Freeze Drying), pH meter (Shanghai Yidian), constant temperature incubator (Shanghai Hetian), ultrasonic vibrator (ELMA-E5K), microwave reactor (MARS6); chemicals and reagents were purchased from Sigma-Aldrich.

[0051] Example 1: This example discloses a high-protein soy sauce, comprising the following raw materials in parts by weight: 10 parts of kelp polysaccharide, 5 parts of carrageenan, 1 part of β-amylase, 20 parts of composite protein, 100 parts of soybeans, 20 parts of salt, and 200 parts of purified water. The high-protein soy sauce also includes modified anthocyanidins, wherein the weight ratio of the modified anthocyanidins to the composite protein is 1:6, and the modified anthocyanidins are prepared by encapsulating the anthocyanidins in a lipid bilayer structure, and the modified anthocyanidins have a particle size of 10 to 20 μm and a specific surface area of ​​20 to 30 m 2 / g.

[0052] In high-protein soy sauce, kelp polysaccharides, modified anthocyanins and complex proteins play a role through synergistic effects. Modified anthocyanins in soy sauce undergo physical adsorption with complex proteins and other nutrients in soy sauce through electrostatic interactions, hydrophobic interactions and hydrogen bonding interactions. The amino groups of modified anthocyanins are protonated in acidic or neutral environments to form cations, which enables them to undergo electrostatic interactions with negatively charged nutrients (such as proteins, polysaccharides, etc.). At the same time, the hydrophobic groups of modified anthocyanins also undergo hydrophobic interactions with fat-soluble nutrients. Physical adsorption is the basis for the formation of complexes by modified anthocyanins, allowing modified anthocyanins to spontaneously aggregate and form stable complexes with complex proteins and other nutrients in soy sauce. In addition, modified anthocyanins can also form more stable complexes with nutrients in soy sauce through chemical bonding, forming a variety of chemical bonds with kelp polysaccharides or other proteins, such as amino ester bonding, hydroxy ester bonding, etc. Chemical bonding can significantly enhance the stability of the complex, allowing the complex to remain intact in soy sauce for a longer time and effectively encapsulate and protect the nutrients. The addition of kelp polysaccharide can effectively improve the stability of the complex. Figure 1It is obvious that the gel network and hollow structure of kelp polysaccharide are conducive to encapsulating nutrients such as complex proteins, forming complexes better and making them more stable. Kelp polysaccharide has a hydrophilic part with hydrophilic phosphate groups inside. In soy sauce, they will automatically arrange into a double-layer structure, with the hydrophilic phosphate groups facing the water phase, encapsulating the surface of the complex, stabilizing the structure of the complex, and reducing the rupture and aggregation caused by excessive interfacial tension. And it enhances the encapsulation ability of the complex, so that the complex can accommodate more flavor substances or oils. At the same time, the phosphate group can form a hydrogen bond network with other phospholipid molecules, water molecules, proteins, etc., to enhance the internal binding force of the complex and make the complex tighter. Improve the structural strength of the complex so that it is not easy to deform or break when subjected to external force or temperature changes. Complex protein provides high content of protein in soy sauce, and protein is rich in essential amino acids for the human body. These amino acids are important components of building human protein and are essential for maintaining human health. Figure 2 The SEM picture of the complex protein shows that the complex protein is a compact spherical structure. The complex protein of this structure can move freely in the cell and participate in various biochemical reactions. In addition, the presence of protein can make the taste of soy sauce softer and rounder, reduce the monotonous salty or sour taste, and increase the sense of hierarchy. The surface of the protein combines with water molecules to reduce the free water in the soy sauce, making it thicker instead of thin and watery. A dynamic equilibrium system is formed between kelp polysaccharide, modified anthocyanin and complex protein. In the brewing of soy sauce, the modified anthocyanin tightly wraps the nutrients such as complex protein in the complex; the kelp polysaccharide film serves as an outer protective barrier to enhance the stability and antioxidant capacity of the complex. This dynamic equilibrium system not only improves the nutritional value and quality of soy sauce, but also significantly improves the taste, flavor and color of soy sauce.

[0053] The preparation method of the modified anthocyanidin comprises:

[0054] S11. Add 50 to 60 g of chitosan powder to 0.5% to 2% acetic acid solution under stirring, and stir evenly until completely dissolved to obtain a chitosan solution;

[0055] S12. 10 to 15 g of propylene oxide was added dropwise to the chitosan solution under heating and stirring, followed by adding 2 to 3 g of chloroacetic acid to promote the reaction, the heating temperature was 60 to 70 ° C, and stirred for 4 to 5 h to obtain a first mixed solution;

[0056] S13. After the first mixed solution is cooled to room temperature, the pH is adjusted to 6-7 with sodium hydroxide, followed by high-speed centrifugation at a rotation speed of 3000-4000 rpm for 5-10 min. The separated solid is washed with anhydrous ethanol and deionized water for 3-4 times respectively, and then vacuum dried at a drying temperature of 45-55° C. After drying for 8-12 hours, modified anthocyanins are obtained.

[0057] The mass ratio of soybean to beta-amylase is 1:0.5.

[0058] The mass ratio of laminarin to carrageenan is 1:2.

[0059] The preparation method of the composite protein comprises:

[0060] S21. Wash 20-30 g of freshwater shrimp, 40-50 g of clams, 40-50 g of sea crucian carp, and 10-30 g of oysters, peel and debone them, cut them into small pieces, and then put them into a wall-breaking machine to grind them to obtain a seafood mixture;

[0061] S22. 5 to 6 g of acetic acid buffer solution is added dropwise to the seafood mixture, followed by adding 1 to 2 g of protease and stirring, heating at 40 to 50° C. for 1 to 2 h to obtain a first mixture;

[0062] S23. The first mixture was transferred to a microwave reactor, the microwave power was set to 300W to 500W, the reaction time was 10 to 20min, and a pause of 1 to 2min was performed after each microwave, and the microwave was repeated 5 to 6 times to obtain a second mixture;

[0063] S24. The second mixture was transferred to an ultrasonic reactor, the ultrasonic power was set to 100W to 200W, the reaction time was 30 to 40min, pausing for 1 to 2min after each microwave, and the ultrasound was repeated 5 to 6 times to obtain a third mixture;

[0064] S25. The third mixture is centrifuged at a speed of 5000 to 6000 rpm for 15 to 20 min, and the resulting liquid is collected to obtain a first mixed solution;

[0065] S26. The first mixed liquid is concentrated by steam distillation for 3 to 5 hours to extract the protein in the seafood, and after cooling, it is stored in a refrigerator at 0 to -4°C to obtain a composite protein.

[0066] The composite protein is obtained by combining microwave-assisted extraction with ultrasonic-assisted extraction of protein from seafood.

[0067] The mass ratio of the composite protein, salt and pure water is 1:2:100.

[0068] A method for preparing a high-protein soy sauce, which is applied to the high-protein soy sauce, comprises the following steps:

[0069] S31. Add 100-200g soybeans to a container and soak them in water for 6-10h, wash and drain, put them into a normal pressure steamer and steam them thoroughly without sandwich, take them out and cool them to 20-30°C, the mass ratio of soybeans and β-amylase strains is 100:3, mix them repeatedly and transfer them to Quchi, culture them in Qu room, culture them for 60-72h, and obtain the first fermentation product;

[0070] S32. The mass ratio of salt and pure water is (10-20): 100, and a saline solution is obtained, and the saline solution is evenly sprayed on the first fermentation product in Quchi, and natural fermentation is allowed to proceed, and then the composite protein, modified anthocyanin and kelp polysaccharide are sequentially added to obtain a second fermentation product;

[0071] S33. Wait for the second fermentation product to undergo natural fermentation, monitor the status of the second fermentation product every 3 to 4 days, ventilate and stir the product when the temperature is too high or too low during fermentation, turn the product over once a month, 3 to 5 times, and after 6 to 7 months, take the bottom liquid with a tube, heat and sterilize, cool, and filter to obtain high-protein soy sauce.

[0072] Example 2: This example discloses a high-protein soy sauce, comprising the following raw materials in parts by weight: 10 parts of kelp polysaccharide, 10 parts of carrageenan, 2 parts of β-amylase, 30 parts of composite protein, 150 parts of soybeans, 25 parts of salt, and 300 parts of purified water. The high-protein soy sauce also includes modified anthocyanidins, wherein the weight ratio of the modified anthocyanidins to the composite protein is 1:5, and the modified anthocyanidins are prepared by encapsulating the anthocyanidins in a lipid bilayer structure, and the modified anthocyanidins have a particle size of 10 to 20 μm and a specific surface area of ​​20 to 30 m 2 / g. The preparation method of the modified anthocyanidin and the composite protein in this embodiment is consistent with that in Example 1. The preparation method of a high-protein soy sauce in this embodiment is consistent with that in Example 1.

[0073] Example 3: This example discloses a high-protein soy sauce, comprising the following raw materials in parts by weight: 10 parts of kelp polysaccharide, 15 parts of carrageenan, 3 parts of β-amylase, 40 parts of composite protein, 200 parts of soybeans, 30 parts of salt, and 400 parts of purified water. The high-protein soy sauce also includes modified anthocyanidins, wherein the weight ratio of the modified anthocyanidins to the composite protein is 1:4, and the modified anthocyanidins are prepared by encapsulating the anthocyanidins in a lipid bilayer structure, and the modified anthocyanidins have a particle size of 10 to 20 μm and a specific surface area of ​​20 to 30 m 2 / g. The preparation method of the modified anthocyanidin and the composite protein in this embodiment is consistent with that in Example 1. The preparation method of a high-protein soy sauce in this embodiment is consistent with that in Example 1.

[0074] Example 4: This example discloses a high-protein soy sauce, comprising the following raw materials in parts by weight: 15 parts of kelp polysaccharide, 15 parts of carrageenan, 3 parts of β-amylase, 40 parts of composite protein, 200 parts of soybeans, 30 parts of salt, and 400 parts of purified water. The high-protein soy sauce also includes modified anthocyanidins, wherein the weight ratio of the modified anthocyanidins to the composite protein is 1:6, and the modified anthocyanidins are prepared by encapsulating the anthocyanidins in a lipid bilayer structure, and the modified anthocyanidins have a particle size of 10 to 20 μm and a specific surface area of ​​20 to 30 m 2 / g. The preparation method of the modified anthocyanidin and the composite protein in this embodiment is consistent with that in Example 1. The preparation method of a high-protein soy sauce in this embodiment is consistent with that in Example 1.

[0075] Example 5: This example discloses a high-protein soy sauce, comprising the following raw materials in parts by weight: 20 parts of kelp polysaccharide, 15 parts of carrageenan, 3 parts of β-amylase, 40 parts of composite protein, 200 parts of soybeans, 30 parts of salt, and 400 parts of purified water. The high-protein soy sauce also includes modified anthocyanidins, wherein the weight ratio of the modified anthocyanidins to the composite protein is 1:5, and the modified anthocyanidins are prepared by encapsulating the anthocyanidins in a lipid bilayer structure, and the modified anthocyanidins have a particle size of 10 to 20 μm and a specific surface area of ​​20 to 30 m 2 / g. The preparation method of the modified anthocyanidin and the composite protein in this embodiment is consistent with that in Example 1. The preparation method of a high-protein soy sauce in this embodiment is consistent with that in Example 1.

[0076] Example 6: This example discloses a high-protein soy sauce, comprising the following raw materials in parts by weight: 10 parts of kelp polysaccharide, 5 parts of carrageenan, 1 part of β-amylase, 20 parts of composite protein, 100 parts of soybeans, 20 parts of salt, and 200 parts of purified water. The high-protein soy sauce also includes modified anthocyanidins, wherein the weight ratio of the modified anthocyanidins to the composite protein is 1:5, and the modified anthocyanidins are prepared by encapsulating the anthocyanidins in a lipid bilayer structure, and the modified anthocyanidins have a particle size of 10 to 20 μm and a specific surface area of ​​20 to 30 m 2 / g. The preparation method of the modified anthocyanidin and the composite protein in this embodiment is consistent with that in Example 1. The preparation method of a high-protein soy sauce in this embodiment is consistent with that in Example 1.

[0077] Example 6: This example discloses a high-protein soy sauce, comprising the following raw materials in parts by weight: 10 parts of kelp polysaccharide, 5 parts of carrageenan, 1 part of β-amylase, 20 parts of composite protein, 100 parts of soybeans, 20 parts of salt, and 200 parts of purified water. The high-protein soy sauce also includes modified anthocyanidins, wherein the weight ratio of the modified anthocyanidins to the composite protein is 1:4, and the modified anthocyanidins are prepared by encapsulating the anthocyanidins in a lipid bilayer structure, and the modified anthocyanidins have a particle size of 10 to 20 μm and a specific surface area of ​​20 to 30 m 2 / g. The preparation method of the modified anthocyanidin and the composite protein in this embodiment is consistent with that in Example 1. The preparation method of a high-protein soy sauce in this embodiment is consistent with that in Example 1.

[0078] Example 7: This example discloses a high-protein soy sauce, comprising the following raw materials in parts by weight: 10 parts of kelp polysaccharide, 5 parts of carrageenan, 1 part of β-amylase, 20 parts of composite protein, 100 parts of soybeans, 20 parts of salt, and 200 parts of purified water. The high-protein soy sauce also includes modified anthocyanidins, wherein the weight ratio of the modified anthocyanidins to the composite protein is 1:3, and the modified anthocyanidins are prepared by encapsulating the anthocyanidins in a lipid bilayer structure, and the modified anthocyanidins have a particle size of 10 to 20 μm and a specific surface area of ​​20 to 30 m 2 / g. The preparation method of the modified anthocyanidin and the composite protein in this embodiment is consistent with that in Example 1. The preparation method of a high-protein soy sauce in this embodiment is consistent with that in Example 1.

[0079] Control group 1: This embodiment differs from embodiment 1 in that it does not contain modified anthocyanidins. This embodiment discloses a high-protein soy sauce, comprising the following raw materials in parts by weight: 10 parts of kelp polysaccharide, 5 parts of carrageenan, 1 part of β-amylase, 20 parts of composite protein, 100 parts of soybeans, 20 parts of salt, and 200 parts of purified water. The preparation method of the modified anthocyanidins and composite protein in this embodiment is consistent with that in embodiment 1. The preparation method of a high-protein soy sauce in this embodiment is consistent with that in embodiment 1.

[0080] Control group 2: The difference between this embodiment and embodiment 1 is that it does not contain complex protein. This embodiment discloses a high-protein soy sauce, including the following raw materials by weight: 10 parts of kelp polysaccharide, 5 parts of carrageenan, 1 part of β-amylase, 100 parts of soybeans, 20 parts of salt, and 200 parts of purified water. The high-protein soy sauce also includes modified anthocyanidins, and the weight ratio of the modified anthocyanidins to the complex protein is 1:3. The modified anthocyanidins are prepared by encapsulating anthocyanidins in a lipid bilayer structure, and the modified anthocyanidins have a particle size of 10 to 20 μm and a specific surface area of ​​20 to 30 m 2 / g. The preparation method of the modified anthocyanidin and the composite protein in this embodiment is consistent with that in Example 1. The preparation method of a high-protein soy sauce in this embodiment is consistent with that in Example 1.

[0081] Control group 3: The difference between this embodiment and embodiment 1 is that kelp polysaccharide is not contained. This embodiment discloses a high-protein soy sauce, comprising the following raw materials in parts by weight: 5 parts of carrageenan, 1 part of β-amylase, 20 parts of composite protein, 100 parts of soybeans, 20 parts of salt, and 200 parts of purified water. The high-protein soy sauce also includes modified anthocyanidins, wherein the weight ratio of the modified anthocyanidins to the composite protein is 1:3, and the modified anthocyanidins are obtained by encapsulating the anthocyanidins in a lipid bilayer structure, and the modified anthocyanidins have a particle size of 10 to 20 μm and a specific surface area of ​​20 to 30 m 2 / g. The preparation method of the modified anthocyanidin and the composite protein in this embodiment is consistent with that in Example 1. The preparation method of a high-protein soy sauce in this embodiment is consistent with that in Example 1.

[0082] Control group 4: This embodiment differs from embodiment 1 in that it does not contain kelp polysaccharide, modified anthocyanidin and complex protein. This embodiment discloses a high-protein soy sauce, comprising the following raw materials in parts by weight: 5 parts of carrageenan, 1 part of β-amylase, 100 parts of soybeans, 20 parts of salt, and 200 parts of purified water. The preparation method of a high-protein soy sauce in this embodiment is consistent with that in embodiment 1.

[0083] Effect evaluation: Sensory evaluation: Refer to GB 2717-2018 "Soy Sauce Hygiene Standard" to evaluate the sensory organs of soy sauce. Take 10mL of soy sauce mixed evenly in each experimental group and place it in a white porcelain plate with a diameter of 60-90mm. Observe the color and state under natural light, smell its odor, and use a straw to suck an appropriate amount of sample for taste. Physical and chemical evaluation: The protein in soy sauce is determined by the Kjeldahl method to determine the total nitrogen content in the sample, and then calculate the protein content. Take 2.0mL of soy sauce sample mixed evenly in each experimental group, accurately weigh it and move it into the digestion tube, add 0.3g potassium sulfate-copper sulfate and 4.0mL concentrated sulfuric acid, heat and boil in the digestion furnace, and convert organic nitrogen into inorganic nitrogen (ammonium sulfate). After digestion is completed, cool the digestion tube, add 5mL of distilled water and 10mL of sodium hydroxide, and decompose the ammonium sulfate to produce ammonia. Subsequently, the digestion tube is placed in a nitrogen analyzer for distillation, and the evaporated ammonia is collected in a reaction cup. Add 20mL of 2% boric acid solution and 5-6 drops of methylene blue-methyl red mixed indicator to the reaction cup, titrate with 0.01mol / L standard sulfuric acid, and record the volume of standard sulfuric acid consumed. Finally, calculate the nitrogen content in the sample and infer the protein content based on the volume and concentration of standard sulfuric acid consumed.

[0084] Protein content (g / 100mL) = total nitrogen content (g / 100mL) × 6.25

[0085] Table 1 shows the sensory evaluation and protein content of soy sauce in each experimental group.

[0086]

[0087]

[0088]

[0089] Table 1 shows the sensory evaluation and protein content of the soy sauce in each experimental group. It can be seen from Table 1 that there are obvious differences in the sensory evaluation and protein content of the soy sauce prepared by each experimental group. The control group shows a low protein content. By comparing the sensory evaluation and protein content of the soy sauce prepared by Examples 1 to 7 with those prepared by Control Groups 1 to 4, it can be found that, on the whole, the sensory evaluation and protein content of the soy sauce prepared by Examples 1 to 7 are better, and the protein content is 16 to 25 g / 100 mL, which is higher than the protein content of soy sauce on the market by 10 g / 100 mL, showing high protein. The sensory evaluation and protein content of the soy sauce prepared by Example 1 are both greater than those of Examples 2 to 7, which shows that the soy sauce prepared by Example 1 has a better flavor and the highest protein content, while the protein of the soy sauce prepared by Control Groups 1 to 3 is ≤9 g / 100 mL, which is at the level of ordinary soy sauce. This shows that by adding three substances, kelp polysaccharide, modified anthocyanin and composite protein, at the same time in the preparation process of soy sauce, a soy sauce with excellent high protein can be obtained.

[0090] Through the above limited experiments, the application effect of a high-protein soy sauce in Example 1 of the present invention is remarkable. By adding three substances, kelp polysaccharide, modified anthocyanin and complex protein, during the production process, the protein content in the soy sauce can be significantly increased, and a multifunctional seasoning can be provided for human health. The three form a dynamic balance system, which not only ensures the nutritional value and quality of the soy sauce, but also makes the soy sauce significantly improved in taste, flavor and color.

[0091] Finally, it should be noted that: Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A high-protein soy sauce, characterized in that: The invention comprises the following raw materials in parts by weight: 10-20 parts of kelp polysaccharide, 5-15 parts of carrageenan, 1-3 parts of beta-amylase, 20-40 parts of complex protein, 20-40 parts of docosahexaenoic acid, 5-15 parts of astaxanthin, 10-20 parts of taurine, 100-200 parts of soybeans, 20-30 parts of salt, and 200-400 parts of purified water; The high-protein soy sauce also includes: Modified anthocyanins; The modified anthocyanin and the composite protein are in a weight ratio of 1:(3-6); The modified anthocyanin is prepared by encapsulating the anthocyanin in a lipid bilayer structure, and the modified anthocyanin has a particle size of 10-20 μm and a specific surface area of ​​20-30 m 2 / g.

2. The high-protein soy sauce according to claim 1, characterized in that: The preparation method of the modified anthocyanidin comprises: S11. Add 50-60 g of blueberry to 75%-80% ethanol solution under heating and stirring, heat to 60-70°C, stir for 4-5 hours, stir evenly until completely dissolved, to obtain a blueberry extract; S12. 5 to 6 g of soybean lecithin and 10 to 15 g of cholesterol were mixed under stirring at room temperature, and then a 75% to 80% ethanol solution was added to promote the reaction to obtain a liposome mixture; S13. After the liposome mixture was cooled to room temperature, the blueberry extract was slowly added to the liposome mixture, stirred evenly, and the pH was adjusted to 6 to 7 with phosphate buffer to obtain a first mixture; S14. Add the first mixed solution and 75% to 80% ethanol solution to a rotary evaporator and evaporate until membrane lipids are formed on the inner wall of the bottle on the rotary evaporator, continue to add 75% to 80% ethanol solution, and perform ultrasonic treatment to form an emulsion, then place the bottle on the rotary evaporator in a 30 to 40° C. water bath to remove the emulsion gel on the bottle, thereby obtaining a first liposome; S15. The pH of the first liposome is adjusted to 6-7 with a phosphate buffer, and then anthocyanin liposomes of 200-220 nm are obtained by a high-pressure homogenizer, which are freeze-dried at -4--10°C for 8-12 hours to obtain modified anthocyanins.

3. The high-protein soy sauce according to claim 1, characterized in that: The mass ratio of soybean, beta-amylase and carrageenan is 1:(0.5-2):(1-1.5).

4. The high-protein soy sauce according to claim 1, characterized in that: The mass ratio of the kelp polysaccharide to carrageenan is 1:(2-4).

5. The high-protein soy sauce according to claim 1, characterized in that: The preparation method of the composite protein comprises: S21. Wash 20-30 g of freshwater shrimp, 40-50 g of clams, 40-50 g of sea crucian carp, and 10-30 g of oysters, peel and debone them, cut them into small pieces, and then put them into a wall breaking machine to grind them to obtain a seafood mixture; S22. 5-6 g of acetic acid buffer solution is added dropwise to the seafood mixture, followed by adding 1-2 g of protease and stirring, heating at 40-50° C. for 1-2 h to obtain a first mixture; S23. The first mixture was transferred to a microwave reactor, the microwave power was set to 300W to 500W, the reaction time was 10 to 20 min, and a 1 to 2 min pause was performed after each microwave, and the microwave was repeated 5 to 6 times to obtain a second mixture; S24. The second mixture was transferred to an ultrasonic reactor, the ultrasonic power was set to 100W to 200W, the reaction time was 30 to 40 min, pausing for 1 to 2 min after each microwave, and the ultrasonic was repeated 5 to 6 times to obtain a third mixture; S25. The third mixture was centrifuged at a speed of 5000 to 6000 rpm for 15 to 20 min, and the resulting liquid was collected to obtain a first mixed solution; S26. The first mixed solution is concentrated by steam distillation for 3 to 5 hours to extract the protein in the seafood, and the mixture is cooled and stored in a refrigerator at 0 to -4°C to obtain a composite protein.

6. The high-protein soy sauce according to claim 5, characterized in that: The composite protein is obtained by microwave-assisted extraction combined with ultrasonic-assisted extraction of seafood protein, and the types of the protein are mainly myofibrillar protein, troponin, myofibrillar protein and sarcoplasmic protein.

7. The high-protein soy sauce according to claim 1, characterized in that: The mass ratio of the composite protein, docosahexaenoic acid and astaxanthin is (1-20):(2-4):(100-300).

8. A method for preparing a high-protein soy sauce, which is used to prepare a high-protein soy sauce as claimed in any one of claims 1 to 7, characterized in that: The method comprises the following steps: S31. Add 100-200g soybeans to a container and soak them in clean water for 6-10h, wash and drain, put them into a normal pressure steamer and steam them thoroughly without sandwich, take them out and cool them to 20-30°C, the mass ratio of soybeans, carrageenan and β-amylase strains is 100:2:3, mix them repeatedly and transfer them to the Quchi, culture them in the Qu room, culture them for 60-72h, and obtain the first fermentation product; S32. The mass ratio of salt to pure water is (10-20):100 to obtain a saline solution, which is evenly sprayed on the first fermentation product in the Quchi, and then naturally fermented, and then docosahexaenoic acid, astaxanthin, taurine, complex protein, modified anthocyanin and kelp polysaccharide are sequentially added to obtain a second fermentation product; S33. Wait for the second fermentation product to ferment naturally, monitor the status of the second fermentation product every 3 to 4 days, and ventilate and stir it in time when the temperature is too high or too low during fermentation. Turn it over once a month, 3 to 5 times. After 6 to 7 months, use a tube to take the bottom liquid, heat and sterilize it, cool it, and filter it to obtain high-protein soy sauce.

9. The method for preparing a high-protein soy sauce according to claim 8, characterized in that: The soy sauce has seafood flavor and has a higher protein content than the soy sauce on the market.

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