A method for preparing original meat flavoring by ultra-high pressure synergistic pyrolysis and enzymatic hydrolysis

Abstract

The present invention discloses a method for preparing an original meat-flavored flavoring by ultra-high pressure coordinated pyrolysis and enzymatic hydrolysis, as well as the corresponding flavoring. Conventional enzymatic hydrolysis is first performed, followed by ultra-high pressure coordinated heating hydrolysis and enzymatic hydrolysis. Compared with conventional heating hydrolysis, the degree of meat protein hydrolysis is increased by more than 100%, and compared with enzymatic hydrolysis, the degree of meat protein hydrolysis is increased by more than 50%. Enzymatic hydrolysis is a differentiated hydrolysis, while ultra-high pressure and heating are non-differentiated hydrolysis. Ultra-high performance liquid chromatography (ULHPLC) analysis and other modern instrumental analysis techniques revealed that after conventional enzymatic hydrolysis, the product formed by ultra-high pressure combined with pyrolysis and enzymatic hydrolysis has an amino acid composition similar to that formed by conventional pyrolysis (thermal extraction), but with a significantly increased content. This effectively enhances the taste and aroma intensity of the meat-flavored flavoring, resulting in a product with a natural meat aroma and a harmonious, rich, umami taste, greatly improving meat utilization and reducing raw material costs.

Description

Technical Field

[0001] The invention belongs to the field of food processing and relates to a method for preparing original meat flavoring by ultra-high pressure coordinated pyrolysis and enzymatic hydrolysis, and corresponding flavoring. Background Art

[0002] The flavor of meat products is the most prominent flavor in Chinese savory foods. Traditional Chinese cooking primarily utilizes heating processes such as stewing, boiling, roasting, frying, stir-frying, and deep-frying to degrade or react with fat and protein components in the meat, generating flavor. Of these, 60% of volatile flavor components in meat products come from fat oxidation. Fat typically undergoes auto-oxidation at 60°C. Unsaturated fatty acids can oxidize on their own, and oxidation becomes more pronounced when hydrolyzed into free fatty acids. The primary product is fatty acid peroxides. These products, while inherently non-meat-flavoring, subsequently react rapidly to form aldehydes, ketones, and acids, which contribute to the characteristic meat aroma. Protein degradation can also produce numerous small molecules, such as peptides, amino acids, and ammonia, many of which are important flavor precursors in meat products. Furthermore, complex Maillard reactions among fat, sugar, and protein degradation products simultaneously form the various aroma components of meat. In traditional cooking, these three components contribute to the pleasant, integrated taste and aroma of meat, ensuring its authentic flavor, nutritional value, and food safety.

[0003] In the production of existing meat flavorings, meat protein hydrolysis and thermal extraction are commonly used. These two existing hydrolysis technologies have the following problems: 1. Meat protein hydrolysis produces bitter peptides and hydrophobic amino acids after protein hydrolysis, resulting in a low degree of hydrolysis. After the enzymatic reaction reaches a certain equilibrium, the degree of hydrolysis is difficult to increase, and most of the protein remains in the form of protein, unable to participate in the reaction as a flavor precursor. This results in low meat protein utilization and waste of protein resources. 2. Thermal extraction is generally performed under normal or high pressure, resulting in a degree of hydrolysis of less than 10%, resulting in low protein utilization. This hydrolysis method also wastes protein resources. 3. The meat flavorings produced by these two methods have a low aroma intensity and a weak taste. In actual application, to achieve the same effect, the amount of meat flavoring must be increased, which indirectly increases the cost of downstream customers' products. 4. Furthermore, the rich substances contained in the meat are not fully decomposed, resulting in low nutritional value. 5. The hydrolysis time required to achieve a high degree of hydrolysis with existing technologies is relatively long. The use of conventional enzymatic hydrolysis followed by ultra-high pressure combined with thermal decomposition and enzymatic hydrolysis technology can significantly reduce the hydrolysis time and improve production efficiency.

[0004] Patent CN103054032A describes a method for preparing chicken bone nutrient solution, which uses microwave-assisted ultra-high pressure technology to extract chicken bone extract. Ultra-high pressure at 100-120MPa can only destroy the quaternary structure of the protein and has no effect on its secondary or tertiary structure. Moreover, this destruction is reversible, and the protein is not damaged. The dissolved substance still exists in the form of protein, which has little effect on the increase of polypeptide substances and free amino acids. Moreover, microwave-assisted extraction technology can only show a certain degradation ability when combined with a certain temperature. In contrast, when using ultra-high pressure and microwave technology, the main target product dissolved in the patent is protein, not polypeptide or free amino acid. In addition, acetic acid is used for soaking in this process, which actually uses acid hydrolysis technology to hydrolyze the dissolved protein. Acid hydrolysis technology can easily cause the tryptophan in the protein to be destroyed during the protein hydrolysis process. Tryptophan is one of the essential amino acids required by the human body. Therefore, the nutrient solution prepared by this method is not conducive to the reflection of its nutritional value. Moreover, acetic acid is a weak acid and has relatively little effect on protein hydrolysis. The sample is prepared by centrifugation, and most of the unhydrolyzed protein is centrifuged and discarded, resulting in a waste of protein resources. It is also not in line with the zero-carbon emission concept currently advocated by the country and causes pollution to the environment. Furthermore, the temperature and time of high-temperature sterilization in the preparation process of the nutrient solution are not enough. The chicken bones may contain high-temperature resistant bacteria. The product is prone to spoilage during the later storage process, affecting its shelf life.

[0005] CN112890210A discloses adding 0.2-1% of protease by weight of the slurry to the slurry, adjusting the pH to 7-8 with sodium carbonate at a concentration of 1-3 mol / L, and performing ultra-high pressure enzymolysis at an ultra-high pressure of 300-500 MPa and a temperature of 45-60°C for 30-60 minutes to obtain an enzymatic solution; boiling the enzymatic solution for 5-10 minutes to inactivate the enzyme, cooling it, and centrifuging it at a speed of 3000-5500 rpm for 15-30 minutes, discarding the precipitate, and taking the supernatant; the paper "Effect of Ultra-High Pressure Enzymolysis on the Flavor of Blue Clam Enzymatic Hydrolysate" (Hu Xianjie, master's thesis of Bohai University) proposes a pressure of 250 MPa, a pressurization time of 60 minutes, an enzymatic solution temperature of 50°C, and an enzymatic solution time of 4 hours. The article "Effect of Ultrahigh Pressure on the Flavor and Protein Structure of Blue Clam Enzyme Hydrolysate" (Journal of Food Science and Technology, Bu Ying et al., February 2021) discloses that blue clam meat was added to water in a 1:1 mass ratio, the pH was adjusted to 7.0 with a 0.2 mol / L NaOH solution, and a composite protease and flavor protease (1:1 mass ratio) were added at 0.2% of the blue clam meat mass. The hydrolysate was then heated in a 50°C water bath for 60 minutes at 150, 200, 250, and 300 MPa, respectively. The hydrolysate was then inactivated at 100°C for 100 minutes after 4 hours of enzymatic hydrolysis. The hydrolysate was then cooled, filtered, centrifuged (8178 rpm for 20 minutes), and the supernatant was collected and frozen at -40°C for later use. The reference document uses the supernatant to determine amino acid nitrogen, which inflates the calculated degree of hydrolysis. Furthermore, the centrifugation process wastes unhydrolyzed protein, resulting in a waste of resources.

[0006] Ultra-high pressure is a new type of food processing technology. It refers to the use of pressure above 100 MPa to compress the volume of meat protein in a liquid medium. The extremely high static pressure generated by ultra-high pressure changes the protein tissue structure, tenderizing or maturing it, and causing changes in the non-covalent bonds such as hydrogen bonds, ionic bonds and hydrophobic bonds that form the three-dimensional structure of protein molecules, changing its spatial structure and causing certain irreversible changes, which greatly increase the degradability of the protein and the dissolution of flavor substances. However, different pressures have different effects on proteins. Generally, pressures between 100 MPa and 200 MPa can disrupt the quaternary structure of proteins, and the changes in the protein are reversible. However, when the pressure exceeds 200 MPa, the tertiary structure of the protein undergoes irreversible changes, destroying the van der Waals forces, hydrogen bonds, hydrophobic bonds, and electrostatic interactions that maintain the tertiary structure of the protein. When the pressure is greater than 700 MPa, the secondary structure of the protein is destroyed, while at pressures below this level, the secondary structure of the protein is not damaged at all. At the same time, under certain ultrahigh pressure and temperature conditions, the secondary, tertiary, and quaternary structures of the protein molecules change, exposing reactive sites that are easily exposed to the reaction donor, significantly increasing the efficiency of catalytic hydrolysis. However, the utilization rate of nutrients in ultrahigh pressure reactions is currently insufficient, and there is an urgent need to improve its utilization rate. Summary of the Invention

[0007] (1) Meat extract: The meat-water ratio is 5:1 to 1:10, with the total of meat and water as 100%. Endo- and exo-enzymes are added for enzymatic hydrolysis. The amount of endo- and exo-enzymes added is 0.01% to 0.5%, and the amount of exo-enzymes added is 0.01% to 0.4%. Enzymatic hydrolysis is carried out at 55 to 65°C for more than 30 minutes, and then ultra-high pressure is simultaneously combined with thermal hydrolysis and enzymatic hydrolysis. The ultra-high pressure pressure is 500 MPa to 1000 MPa, the ultra-high pressure medium temperature is 30 to 100°C, and the ultra-high pressure time is 10 minutes to 12 hours.

[0008] (2) Meat extract 40-80 parts, reducing sugar 1-10 parts, amino acid mixture 1-20 parts, fat 1-20 parts, edible salt 10-20 parts, MSG 5-10 parts, spices 1-10 parts, organic acid 0.1-1 part, emulsifier 0.01-1 part, thickener 0.1-10 parts, flavor base 0.01-5 parts.

[0009] Raw materials: including meat, fat, enzymes, organic acids, edible salt, amino acids, reducing sugars and others;

[0010] Furthermore, the meat is selected from one or more animal meats such as beef, chicken, and mutton, and the meat is crushed into minced meat; the fat is selected from one or more oils such as butter, chicken bone oil, and chicken fat oil; the organic acid is selected from one or more of citric acid, lactic acid, etc.; the amino acid is selected from one or more of cysteine, methionine, aspartic acid, phenylalanine, etc.; the reducing sugar is selected from one or more of glucose, xylose, galactose, etc.; the emulsifier is selected from one or more of sucrose fatty acid esters, monostearate glycerol, etc.; the thickener is selected from one or more of xanthan gum, acetylated distarch adipic acid, etc.; the endoenzyme is one or more of alkaline protease, papain, etc., and the exoenzyme is flavor protease, etc.

[0011] The ultrahigh pressure is preferably 500-600 MPa, the ultrahigh pressure medium temperature is 70-100° C., preferably 75° C., and the ultrahigh pressure time is 30 min-6 h, preferably 2 h.

[0012] When the meat is chicken, the most suitable ultra-high pressure is 600MPa, and when the meat is beef, the most suitable ultra-high pressure is 500MPa.

[0013] Beneficial effects of the present invention

[0014] (1) Ultrahigh pressure can change the tertiary structure of proteins. The tertiary structure of proteases is the basis for the formation of enzyme active centers. After treatment with ultrahigh pressure technology, it is beneficial to the release of more active sites of the enzyme and the improvement of its activity. The combination of ultrahigh pressure and thermal decomposition and enzymatic decomposition has a more obvious effect on improving the degree of hydrolysis.

[0015] (2) This patent adopts the technology of first performing conventional enzyme-catalyzed hydrolysis and then performing ultra-high pressure coordinated heating hydrolysis and enzyme-catalyzed hydrolysis. Compared with the simple thermal hydrolysis technology, the hydrolysis degree of meat protein is increased by more than 100%, and compared with the simple enzyme-catalyzed hydrolysis technology, the hydrolysis degree of meat protein is increased by more than 50%. At the same time, this method can well maintain the nutritional value of the product. Through ultra-high performance liquid phase analysis technology and other modern instrumental analysis technologies, it is found that after conventional enzymatic hydrolysis, the product formed by ultra-high pressure combined with thermal hydrolysis and enzymatic hydrolysis has an amino acid composition similar to that formed by conventional thermal hydrolysis, but the content is significantly increased, especially the content of flavor amino acids is significantly increased. On the basis of using the same amount of meat flavor extract, the content of its Maillard reaction precursor is significantly increased, which promotes the reaction equilibrium to develop in the direction of reaction substrate, which is conducive to the generation of more reaction substrates, and significantly increases the intensity of meat flavor. The product has the aroma of natural meat and a coordinated and thick umami taste, which greatly improves the utilization rate of meat and reduces the cost of raw materials.

[0016] (3) Moreover, the combination of ultra-high pressure hydrolysis, heating hydrolysis and enzyme-catalyzed hydrolysis can well ensure the configuration and structure of amino acids without producing harmful substances. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 It is a production process flow chart. DETAILED DESCRIPTION

[0018] The above content of the present invention will be further described in detail below in conjunction with specific embodiments, but it should not be understood that the scope of the above subject matter of the present invention is limited to the following embodiments.

[0019] Example 1:

[0020] Determination of ultra-high pressure intensity:

[0021] Recipe: 72 parts of meat extract, 2 parts of amino acid mixture (alanine: cysteine: methionine: glycine, arginine = 1:2:0.5:1:0.1), 3 parts of sugar (xylose: glucose = 1:1), 10 parts of table salt, 4.6 parts of MSG, 5 parts of edible chicken oil, 2 parts of spices (ginger: green onion: pepper = 8:6:0.5), 0.1 part of organic acid (succinic acid: lactic acid = 2:1), 0.3 part of emulsifier (sucrose fatty acid ester), 0.5 part of thickener (xanthan gum: guar gum: gum arabic = 2:1:1), and 0.5 part of fragrance base.

[0022] Preparation Method: To prepare the meat extract, first mince the chicken breast using a meat grinder. Next, place the chicken breast puree and water in a 3:1 ratio into a three-necked flask. Add 0.3% papain (based on a meat-to-water ratio of 100 parts) and 0.04% flavor protease (based on a meat-to-water ratio of 100 parts) to the flask. Enzymatic hydrolysis is carried out at 60°C for 1 hour. The mixture is then placed in an ultrahigh-pressure reactor, maintaining pressures of 200 MPa, 300 MPa, 400 MPa, 500 MPa, 600 MPa, and 700 MPa at 75°C for 2 hours. After the ultrahigh pressure reaction, a meat extract is prepared, and samples are collected to measure the degree of hydrolysis. Amino acids, sugar, salt, chicken fat, and spices are then added in appropriate proportions. Stir at 100°C for 1 hour, then the mixture is brought to 50°C. The flavor base is added, mixed thoroughly, and samples are collected for sensory tasting.

[0023] The amino acid nitrogen content of the sample was determined according to the national standard method, and the degree of hydrolysis was calculated. The specific results are shown in Table 1.

[0024] Table 1 Effect of different ultrahigh pressures on the hydrolysis degree of chicken breast

[0025] 200MPa 300MPa 400MPa 500MPa 600MPa 700MPa Degree of hydrolysis (%) 19.1 20.2 22.2 28.2 30.3 27.6

[0026] As the pressure increases, the degree of hydrolysis tends to gradually increase. When the pressure is low, ultra-high pressure mainly destroys non-covalent bonds such as oxygen bonds between protein molecules, causing the protein structure to unfold, the interaction between the hydrophilic groups of the side chains and water to be strengthened, and the hydration effect to be enhanced, thereby increasing the solubility of the protein; as the pressure continues to increase, the structure of the protein unfolds further, and the hydrophobic groups buried inside the molecules are exposed. At the same time, the originally stretched molecules may also aggregate, which ultimately leads to a decrease in the solubility of the protein.

[0027] The samples were rinsed with water and subjected to sensory evaluation. The specific evaluation results are shown in Table 2:

[0028] Table 2 Sensory evaluation results of meat flavorings prepared using chicken breast extracts at different ultrahigh pressures

[0029] 200MPa 300MPa 400MPa 500MPa 600MPa 700MPa Color 14 14 14 14 14 14 Appearance status 14 14 14 14 14 14 aroma 18 19 21 26 29 23 taste 30 33 35 38.1 39.1 38.1 Total score 76 80 84 92.1 96.1 89.1

[0030] Sensory evaluation data show that as pressure increases, the aroma intensity of the solution gradually increases, the raw odor decreases, and the aroma of cooked egg white emerges, with a richer aroma. At low pressure, the taste is primarily fresh, with other sensations playing a supporting role. As pressure increases, a complex taste emerges, with a gradually intensified sense of freshness, meatiness, and smoothness. The freshness is combined with other tastes, making it less abrupt than at low pressure, reflecting a cohesive, complex taste. Based on sensory evaluation and monitoring data, 600 MPa was selected as the optimal condition for chicken processing.

[0031] Example 2:

[0032] Ultra-high pressure determination experiment:

[0033] Recipe: 72 parts of meat extract, 3 parts of amino acid mixture (glutamic acid: cysteine: leucine: glycine, arginine: aspartic acid = 1:2:0.5:1:0.1:1), 2 parts of sugar (ribose: xylose: glucose = 1:2:2), 11 parts of table salt, 5 parts of MSG, 4 parts of edible butter, 1 part of spice (ginger: aniseed: cinnamon = 7:1:0.5), 0.1 part of organic acid (citric acid: malic acid = 1:1), 0.5 part of emulsifier (sodium caseinate), 0.7 part of thickener (acetylated distarch adipate: xanthan gum = 9:1), 0.7 part of flavoring base.

[0034] Preparation Method: To prepare the meat extract, first mince the beef using a meat grinder. Next, add the beef puree and water in a 2.5:1 ratio into a three-necked flask. Add 0.3% alkaline protease (based on a meat-to-water ratio of 100 parts) and 0.04% flavor protease (based on a meat-to-water ratio of 100 parts) to the flask. Enzymatic hydrolysis is performed at 58°C for 1 hour. The mixture is then placed in an ultrahigh-pressure reactor, maintained at pressures of 200MPa, 300MPa, 400MPa, 500MPa, 600MPa, and 700MPa, and at 75°C for 2 hours. After the ultrahigh pressure reaction, a meat extract is prepared, and samples are taken to measure the degree of hydrolysis. To the meat extract used in the recipe, amino acids, sugar, salt, chicken fat, and spices are added in appropriate proportions. Stir at 100°C for 1 hour, then the mixture is stirred to 55°C. The flavor base is added, mixed thoroughly, and samples are collected for sensory tasting.

[0035] The amino acid nitrogen content of the sample was determined according to the national standard method, and the degree of hydrolysis was calculated. The specific results are shown in Table 3.

[0036] Table 3 Effect of different ultrahigh pressure on the hydrolysis degree of beef

[0037] 200MPa 300MPa 400MPa 500MPa 600MPa 700MPa Degree of hydrolysis (%) 17.8 18.1 21.0 25.2 24.5 22.0

[0038] As the pressure increases, the degree of hydrolysis tends to gradually increase. When the pressure is low, ultra-high pressure mainly destroys non-covalent bonds such as hydrogen bonds between protein molecules, causing the protein structure to unfold, the interaction between the hydrophilic groups of the side chains and water to be strengthened, and the hydration effect to be enhanced, thereby increasing the solubility of the protein; as the pressure continues to increase, the structure of the protein unfolds further, and the hydrophobic groups buried inside the molecules are exposed. At the same time, the originally stretched molecules may also aggregate, which ultimately leads to a decrease in the solubility of the protein and a decrease in the degree of hydrolysis.

[0039] The samples were rinsed with water and subjected to sensory evaluation. The specific evaluation results are shown in Table 4:

[0040] Table 4 Sensory evaluation results of meat flavorings prepared using beef extracts under different ultra-high pressures

[0041] 200MPa 300MPa 400MPa 500MPa 600MPa 700MPa Color 14 14 14 14 14 14 Appearance status 14 14 14 14 14 14 aroma 17 18 20 27 25 23 taste 30 31 34 38 36 35 Total score 75 77 83 93 89 85

[0042] Sensory evaluation data show that as pressure increases, the aroma intensity of the solution gradually increases, the raw odor decreases, and a cooked egg white aroma emerges, with a richer aroma. At low pressure, the taste is primarily fresh, with other sensations playing a supporting role. As pressure increases, a complex taste emerges, with freshness, meatiness, and a lingering texture gradually intensifying. The freshness is combined with other tastes, making it less abrupt than at low pressure, resulting in a cohesive, complex taste. Based on sensory evaluation and monitoring data, 500 MPa was selected as the optimal condition for beef processing.

[0043] Comparative Example 1:

[0044] Recipe: Same as that of Example 1.

[0045] Preparation Method: To prepare the meat extract, first mince the chicken breast using a meat grinder. Next, place the chicken breast puree and water in a 3:1 ratio into a three-necked glass bottle. Stir and heat to 60°C. Stop heating and add 0.3% papain (based on a meat-to-water ratio of 100 parts) and 0.04% flavor protease (based on a meat-to-water ratio of 100 parts). Maintain enzymatic hydrolysis at 60°C and atmospheric pressure for 3 hours. After enzymatic hydrolysis, a meat extract is prepared and sampled for hydrolysis degree measurement. Next, take the meat extract used in the recipe, add amino acids, sugar, salt, chicken fat, and spices in appropriate proportions, and stir at 100°C for 1 hour. Then, bring the mixture to 50°C, add the flavor base, mix thoroughly, and sample for sensory tasting.

[0046] Comparative Example 2:

[0047] Formula: Same as that of Example 1

[0048] Preparation Method: Prepare the meat extract: First, mince the chicken breast using a meat grinder. Then, mix the chicken breast puree with water in a 3:1 ratio. Place the mixture in an ultra-high pressure autoclave at a pressure of 600 MPa and a temperature of 75°C for 3 hours. Sample the mixture to determine the degree of hydrolysis. Next, add the meat extract to the formula, along with amino acids, sugar, salt, chicken fat, and spices in appropriate proportions. Stir at 100°C for 1 hour. Then, bring the mixture to 50°C, add the flavor base, stir thoroughly, and sample for sensory tasting.

[0049] Comparative Example 3:

[0050] Recipe: Same as that of Example 1.

[0051] Preparation Method: Prepare the meat extract: First, mince the chicken breast using a meat grinder. Then, mix the chicken breast puree with water in a 3:1 ratio. Heat the mixture in a three-necked flask at 100°C for 3 hours. Sample the mixture to determine the degree of hydrolysis. Next, add the formulated meat extract to the appropriate proportions of amino acids, sugar, salt, chicken fat, and spices. Stir at 100°C for 1 hour. Then, bring the mixture to 50°C, add the flavor base, stir thoroughly, and sample for sensory tasting.

[0052] Comparative Example 4:

[0053] Recipe: Same as that of Example 1.

[0054] Preparation Method: To prepare the meat extract: First, mince the chicken breast using a meat grinder. Next, place the chicken breast puree and water in a 3:1 ratio into a three-necked glass bottle. Stir and heat to 60°C. Stop heating and add 0.3% papain (based on a meat-to-water ratio of 100 parts). Maintain enzymatic hydrolysis at 60°C and atmospheric pressure for 3 hours. After enzymatic hydrolysis, a meat extract is prepared and sampled for hydrolysis degree. Next, take the meat extract for the recipe, add amino acids, sugar, salt, chicken fat, and spices in appropriate proportions, and stir at 100°C for 1 hour. Then, bring the mixture to 50°C, add the flavor base, mix thoroughly, and sample for sensory tasting.

[0055] Comparative Example 5:

[0056] Recipe: Same as that of Example 1.

[0057] Preparation Method: To prepare the meat extract: First, mince the chicken breast using a meat grinder. Next, place the chicken breast puree and water in a 3:1 ratio into a three-necked glass bottle. Stir and heat to 60°C. Stop heating and add 0.04% flavor protease (calculated based on a meat-to-water ratio of 100 parts). Maintain enzymatic hydrolysis at 60°C and atmospheric pressure for 3 hours. After enzymatic hydrolysis, a meat extract is prepared and sampled for hydrolysis degree. Next, take the meat extract used in the recipe, add amino acids, sugar, salt, chicken fat, and spices in appropriate proportions, and stir at 100°C for 1 hour. Then, bring the mixture to 50°C, add the flavor base, mix thoroughly, and sample for sensory tasting.

[0058] Comparative Example 6:

[0059] Formula: Same as that of Example 1

[0060] Preparation Method: Prepare the meat extract: First, mince the chicken breast using a meat grinder. Then, mix the chicken breast puree with water in a 3:1 ratio. Place the mixture in an ultra-high pressure autoclave without heating for enzymatic hydrolysis. Maintain a pressure of 600 MPa and a temperature of 60°C. Add 0.3% papain (based on a meat-to-water ratio of 100 parts) and 0.04% flavor protease (based on a meat-to-water ratio of 100 parts). Press for 3 hours, then sample and measure the degree of hydrolysis. Next, take the meat extract for the recipe, add amino acids, sugar, salt, chicken fat, and spices in appropriate proportions, and stir at 100°C for 1 hour. Then, stir the mixture to 50°C, add the flavor base, stir evenly, and sample for sensory tasting.

[0061] Comparative Example 7:

[0062] Formula: Same as that of Example 2.

[0063] Preparation Method: Prepare the meat extract: First, mince the beef using a meat grinder. Next, place the beef puree and water in a 2.5:1 ratio into a three-necked flask. Add 0.3% alkaline protease (based on a meat-to-water ratio of 100 parts) and 0.04% flavor protease (based on a meat-to-water ratio of 100 parts) to the flask. Enzymatic hydrolysis is performed at 58°C for 3 hours to prepare the meat extract. Samples are then taken to measure the degree of hydrolysis. Next, the meat extract is added to the formula, along with amino acids, sugar, salt, chicken fat, and spices in appropriate proportions. Stir at 100°C for 1 hour, then the mixture is stirred to 50°C. The flavor base is added and mixed thoroughly. Samples are then taken for sensory tasting.

[0064] Comparative Example 8:

[0065] Formula: Same as that of Example 2.

[0066] Preparation Method: To prepare the meat extract: First, mince the beef using a meat grinder. Next, mix the beef puree with water in a ratio of 2.5:1. Place the mixture in an ultrahigh-pressure reactor, maintain a pressure of 500 MPa and a temperature of 75°C, and press for 3 hours. After the ultrahigh pressure reaction, a meat extract is prepared, and a sample is taken to measure the degree of hydrolysis. Next, take the meat extract used in the recipe, add amino acids, sugar, salt, chicken fat, and spices in appropriate proportions, and stir at 100°C for 1 hour. Then, bring the mixture to 50°C, add the flavor base, stir thoroughly, and sample for sensory tasting.

[0067] Comparative Example 9:

[0068] Formula: Same as that of Example 2.

[0069] Preparation Method: Prepare the meat extract: First, mince the beef using a meat grinder. Next, place the beef puree and water in a 2.5:1 ratio into a three-necked flask and heat at 100°C for 3 hours. This will create the meat extract, and sample the material to determine the degree of hydrolysis. Next, take the meat extract for the recipe and add amino acids, sugar, salt, chicken fat, and spices in appropriate proportions. Heat at 100°C with stirring for 1 hour. Then, bring the mixture to 50°C, add the flavor base, stir thoroughly, and sample for sensory tasting.

[0070] Comparative Example 10:

[0071] Formula: Same as that of Example 2.

[0072] Preparation Method: Prepare the meat extract: First, mince the beef using a meat grinder. Next, place the beef puree and water in a 2.5:1 ratio into a three-necked flask. Add 0.3 parts alkaline protease (based on a meat-to-water ratio of 100 parts) to the flask. Enzymatic hydrolysis is performed at 58°C for 3 hours to prepare the meat extract. Samples are then taken to measure the degree of hydrolysis. Next, the meat extract is added to the formula, along with amino acids, sugar, salt, chicken fat, and spices in appropriate proportions. Stir at 100°C for 1 hour. The mixture is then brought to 50°C, and the flavor base is added. Mix thoroughly and sample for sensory tasting.

[0073] Comparative Example 11:

[0074] Formula: Same as that of Example 2.

[0075] Preparation Method: Prepare the meat extract: First, mince the beef using a meat grinder. Next, place the beef puree and water in a 2.5:1 ratio into a three-necked flask. Add 0.4% flavor protease (based on a meat-to-water ratio of 100 parts) to the flask. Enzymatic hydrolysis is performed at 58°C for 3 hours to prepare the meat extract. Samples are then taken to measure the degree of hydrolysis. Next, the meat extract is added to the formula, along with amino acids, sugar, salt, chicken fat, and spices in appropriate proportions. Stir at 100°C for 1 hour. The mixture is then brought to 50°C, and the flavor base is added. Mix thoroughly and sample for sensory tasting.

[0076] Comparative Example 12:

[0077] Formula: Same as that of Example 2.

[0078] Preparation Method: To prepare the meat extract, first mince the beef using a meat grinder. Next, mix the beef puree and water in a ratio of 2.5:1. Place the mixture in an ultrahigh-pressure reactor without heating for enzymatic hydrolysis. Maintain a pressure of 500 MPa and a temperature of 58°C. Add 0.3% alkaline protease (based on a meat-to-water ratio of 100 parts) and 0.04% flavor protease (based on a meat-to-water ratio of 100 parts) and press for 3 hours. After ultrahigh pressure, a meat extract is prepared and sampled for hydrolysis degree measurement. Next, take the meat extract for the recipe, add amino acids, sugar, salt, chicken fat, and spices in appropriate proportions, and stir at 100°C for 1 hour. Then, bring the mixture to 50°C, add the flavor base, stir thoroughly, and sample for sensory tasting.

[0079] Sensory evaluation method:

[0080] The sensory evaluation panel consisted of 9 people. They evaluated the prepared spices in terms of color, appearance, aroma and taste. The specific operation was as follows: the samples were rinsed with 1% water and 0.2% edible salt were added. The samples were then compared and evaluated. The total score was 100 points, where:

[0081] The full score for color is 15 points. Normal is 14-15 points, above normal is 12-13.9 points, acceptable is 9.7-11.9 points, fair is 7.5-9.6 points, average is 6-7.4 points, slightly poor is 3-5.9 points, poor is 1-2.9 points, and very poor is 0 points.

[0082] The maximum score for appearance condition is 15 points. Normal scores 14-15 points, above normal scores 12-13.9 points, acceptable scores 9.7-11.9 points, fair scores 7.5-9.6 points, average scores 6-7.4 points, slightly poor scores 3-5.9 points, poor scores 1-2.9 points, and very poor scores 0 points.

[0083] The maximum score for aroma is 30 points. No foreign smell, strong, pure (or pleasant) aroma: 29.3-30 points; no foreign smell, relatively strong, pure (or pleasant) aroma: 27-29.2 points; no foreign smell, acceptable: 24-26.9 points; acceptable: 21-23.9 points; passing: 18-20.9 points; failing: less than 18 points.

[0084] The maximum score for taste is 40 points. Pure and refreshing scores 39.1-40 points, relatively pure and palatable scores 36-39 points, acceptable scores 32-35.9 points, fair scores 28-31.9 points, passing scores 24-27.9 points, and failing scores below 24 points.

[0085] Through sensory evaluation, the scores of each sample are as follows: See Table 5

[0086] Table 5: Sensory evaluation table of Examples 1-2 and Comparative Examples 1-12

[0087] Example 1 Example 2 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 Comparative Example 7 Comparative Example 8 Comparative Example 9 Comparative Example 10 Comparative Example 11 Comparative Example 12 Color 14 14 13 12 10 13 12 13 13 12 9 12 11 12 Appearance status 14 14 14 13 12 14 14 14 14 12 11 13 13 13 aroma 29 27 25 10 9 20 15 22 23 9 8 18 14 20 taste 39.1 38 25 15 13 20 18 21 24 14 12 19 18 21 Total score 96.1 93 73 50 44 67 59 69 70 47 40 62 56 66

[0088] Determination of degree of hydrolysis

[0089] Free amino nitrogen content was determined according to the food standard GB5009.239-2016 for the determination of amino acid nitrogen in foods. Total nitrogen content was determined using the Kjeldahl method described in GB5009.5-2010 for the determination of protein in foods. The degree of hydrolysis reflects protein utilization. A higher degree of hydrolysis indicates a more extensive protein hydrolysis process, making it more readily absorbed and utilized. Conversely, a lower degree of hydrolysis results in a greater proportion of the protein remaining as protein, resulting in a lower protein utilization rate.

[0090] Degree of hydrolysis % = (content of free amino nitrogen in the extract - content of free amino nitrogen in the blank) / content of total nitrogen

[0091] Table 6 Hydrolysis degree determination table of Examples 1-2 and Comparative Examples 1-12

[0092] Example 1 Example 2 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 Comparative Example 7 Comparative Example 8 Comparative Example 9 Comparative Example 10 Comparative Example 11 Comparative Example 12 Degree of hydrolysis (%) 30.3 25.2 17.8 10.5 5.2 13 6 15.1 15.5 8.8 5 11.9 5.8 13.3

[0093] By measuring the degree of hydrolysis of the extract, it was found that the patent of the present invention uses conventional enzymatic hydrolysis and then ultra-high pressure combined with thermal decomposition and enzymatic hydrolysis to hydrolyze the meat. The degree of hydrolysis is the highest and the utilization rate of protein is higher. The hydrolysis degree is higher than that of conventional double-enzyme compound enzymatic hydrolysis, thermal decomposition alone, single enzyme enzymatic hydrolysis and ultra-high pressure combined with thermal decomposition and enzymatic hydrolysis alone. It improves the utilization rate of protein, increases the precursor substances participating in the Maillard reaction, and is more conducive to the development of the reaction towards the increase of flavor substances.

[0094] In order to test the difference in amino acid composition between Example 1 of the present invention and comparative examples 1, 2, 3, 4, 5, and 6 of the extract prepared by the conventional method, and the difference in amino acid composition between Example 2 and comparative examples 7, 8, 9, 10, 11, and 12 of the extract prepared by the conventional method, the following experiments were conducted:

[0095] The free amino acid content in the extract was determined by ultra-high performance liquid chromatography (UPLC), and it was found that the amino acid content increased significantly, as shown in the following table:

[0096] Experimental conditions: Instrument system: ACQUITY UPLC I-Class + TUV ultra-high performance liquid chromatography system

[0097] Chromatographic column: AccQ-Tag Ultra C18 (1.7 um, 2.1×100㎜), mobile phase: mobile phase A: AccQ Eluent A: water = 1:9; mobile phase B: AccQ Eluent B; gradient elution; injection volume: 1uL; column temperature: 50℃; sample temperature: 15℃.

[0098] Table 7 Comparison of amino acid composition between Example 1 and Comparative Examples 1-6

[0099] Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 Amino acid name Amino acid content (umol / mL) Amino acid content (umol / mL) Amino acid content (umol / mL) Amino acid content (umol / mL) Amino acid content (umol / mL) Amino acid content (umol / mL) Amino acid content (umol / mL) Histidine 5.80 3.00 1.76 0.00 0.79 0.34 2.10 Serine 12.10 7.00 3.64 1.83 2.84 2.11 4.91 Arginine 19.30 10.00 3.93 2.22 7.45 1.51 7.96 Glycine 24.00 15.70 12.16 0.02 11.53 7.99 10.45 Aspartic acid 8.00 7.40 2.52 5.49 3.22 1.51 5.75 glutamate 16.90 10.00 4.80 3.14 7.03 2.20 7.75 Threonine 10.20 7.00 3.05 1.94 3.61 1.46 5.46 Alanine 17.00 12.00 6.25 4.60 6.57 4.13 9.19 Proline 4.10 2.00 2.03 1.43 1.39 1.13 1.55 Cystine 4.40 3.00 2.83 0.25 5.08 1.61 2.22 Lysine 17.80 11.00 3.26 1.43 7.62 1.94 8.72 Tyrosine 7.30 4.00 1.57 1.43 5.58 0.76 2.96 Methionine 10.70 4.00 1.80 1.22 6.59 0.92 3.06 Valine 7.8 5.00 2.68 2.45 4.17 1.70 3.96 Isoleucine 8.30 4.00 2.06 1.84 1.27 1.21 3.30 Leucine 29.00 17.40 4.01 2.86 9.77 3.04 12.05 Phenylalanine 11.20 6.00 1.84 1.02 6.21 1.02 4.54 Total 213.9 128.5 60.19 33.17 90.72 34.58 95.93 Essential amino acid content 100.8 48.0 18.70 12.76 39.24 11.29 41.09 Non-essential amino acid content 101.4 73.5 37.09 18.73 40.82 20.92 49.66 Semi-essential amino acid content 11.7 7.00 4.40 1.68 10.66 2.37 5.18

[0100] Table 8 Comparison of amino acid composition between Example 2 and Comparative Examples 7-12 of extracts prepared by traditional methods

[0101] Example 2 Comparative Example 7 Comparative Example 8 Comparative Example 9 Comparative Example 10 Comparative Example 11 Comparative Example 12 Amino acid name Amino acid content (umol / mL) Amino acid content (umol / mL) Amino acid content (umol / mL) Amino acid content (umol / mL) Amino acid content (umol / mL) Amino acid content (umol / mL) Amino acid content (umol / mL) Histidine 3.23 1.87 0 1.19 1.52 0.98 1.47 Serine 7.23 4.22 3.14 2.62 3.96 2.12 3.85 Arginine 24.70 16.33 14.77 12.98 12.68 9.43 12.76 Glycine 5.74 2.83 2.69 2.45 2.71 1.88 2.26 Aspartic acid 3.25 1.73 1.72 1.74 1.57 1.19 1.28 glutamate 3.54 0.48 0.76 0.76 0.48 0.42 1.11 Threonine 8.33 2.68 1.14 1.02 2.24 1.04 3.93 Alanine 11.66 4.42 1.11 0.65 2.94 0.99 4.84 Proline 10.51 7.20 4.89 4.94 6.26 4.20 5.40 Cystine 10.01 9.59 2.03 1.29 6.11 2.53 4.95 Lysine 3.69 1.45 0.53 0.41 1.80 0.41 1.95 Tyrosine 14.56 5.47 0.96 0.45 4.15 0.79 7.54 Methionine 14.65 6.47 0.23 0.80 7.48 1.42 7.32 Valine 9.07 7.31 0.65 0 6.12 032 4.33 Isoleucine 9.18 2.62 1.29 0.48 1.54 1.00 4.79 Leucine 25.17 11.00 3.54 0.72 5.74 2.63 11.67 Phenylalanine 25.46 10.74 1.88 0.47 7.52 2.18 13.23 Total 189.98 96.41 41.32 32.99 74.83 33.52 92.68 Essential amino acid content 110.11 42.27 9.26 3.90 32.44 9.00 47.22 Non-essential amino acid content 55.30 39.08 29.07 27.35 32.13 21.2 32.97 Semi-essential amino acid content 24.57 15.06 2.99 1.74 10.26 3.32 12.49

[0102] Through comparison, it was found that conventional heating enzymatic hydrolysis followed by ultra-high pressure combined with thermal decomposition and enzymatic hydrolysis greatly improved the hydrolysis degree of meat protein and increased the amino acid content. The precursors of the Maillard reaction are mainly amino acids. Through the above data, it was found that the content of amino acids increased significantly, which increased the precursors participating in the Maillard reaction, thereby making the reaction move towards the substrate and generating more aroma substances, thereby enhancing the aroma intensity of the meat flavoring.

[0103] Amino acids are categorized as essential, semi-essential, and non-essential. Essential amino acids are essential for the human body but cannot be synthesized in the body, or their synthesis rate is far insufficient to meet human needs, requiring supplementation through dietary protein. Essential amino acids play a crucial role in maintaining normal physiological functions and metabolism. Therefore, increasing the essential amino acids significantly improves the nutritional value of the reaction extract. The data above demonstrates a significant increase in the content and percentage of essential amino acids (lysine, phenylalanine, threonine, methionine, isoleucine, leucine, and valine), thereby enhancing the nutritional value of the meat flavoring. Non-essential amino acids are those that the human body can synthesize or convert from other amino acids, and do not necessarily require ingestion from food. These include alanine, arginine, aspartic acid, and proline. A comparison reveals that the total content and total proportion of non-essential amino acids in Examples 1 and 2 show a lower trend compared to the comparative example. Semi-essential amino acids are also known as conditionally essential amino acids. Mainly refers to cysteine ​​and tyrosine, which are converted from methionine and phenylalanine in the body respectively. If these two amino acids can be directly provided in the diet, the body's need for methionine and phenylalanine can be reduced.

[0104] Application test

[0105] Example 1 and Comparative Examples 1, 2, 3, 4, 5, and 6 were selected for application and applied to the production of ham sausage. The application formula is as follows:

[0106] 40 parts of chicken breast; 0.5 parts of carrageenan; 15 parts of starch; 3 parts of salt; 23 parts of chicken bone paste; 0.2 parts of polyphosphate; 10 parts of soy protein; 0.02 parts of sodium nitrite; 0.3 parts of MSG; 1 part of spices; 0.03 parts of sodium isocyanate; 6.95 parts of sugar, 0.3 parts of flavoring.

[0107] The mixed samples were enema treated and then heated at 100°C for 30 minutes. After cooling, sensory evaluation was performed. The specific flavoring materials used in the experiment are as follows:

[0108] 1. Example 1 Flavoring Material

[0109] ② Use 90% of the extract of Example 1 to prepare the flavoring, and replace the missing 10% of the extract with water

[0110] ③ Comparative Example 1 Flavoring

[0111] ④ Comparative Example 2 Flavoring

[0112] ⑤ Comparative Example 3 Flavoring

[0113] ⑥ Comparative Example 4 Flavorings

[0114] ⑦Comparative Example 5 Flavoring

[0115] ⑧Comparative Example 6 Flavoring

[0116] The sensory evaluation results are shown in Table 9:

[0117] Table 9 Comparison of the application effects of the flavoring materials prepared in Example 1 and Comparative Examples 1-6

[0118] Example 1 90% of the extract of Example 1 was used to prepare the flavoring, and the missing 10% of the extract was replaced with water. Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 Color 14 14 14 14 14 14 14 14 Appearance status 14 14 14 14 14 14 14 14 aroma 29 27 22 18 10 20 13 21 taste 39.1 36 28 24 15 25 18 26 Total score 96.1 91 78 70 53 73 59 75

[0119] Example 2 and comparative examples 7, 8, 9, 10, 11, and 12 were selected for application and applied to the production of ham sausage. The application formula is as follows:

[0120] 40 parts of beef; 0.5 parts of carrageenan; 15 parts of starch; 3 parts of salt; 23 parts of chicken bone paste; 0.2 parts of polyphosphate; 10 parts of soy protein; 0.02 parts of sodium nitrite; 0.3 parts of MSG; 1 part of spices; 0.03 parts of sodium isocyanate; 6.95 parts of sugar, 0.3 parts of flavoring.

[0121] The mixed samples were enema treated and then heated at 100°C for 30 minutes. After cooling, sensory evaluation was performed. The specific flavoring materials used in the experiment are as follows:

[0122] 1. Embodiment 2 flavoring material

[0123] ② Use 90% of the extract of Example 2 to prepare the flavoring, and replace the missing 10% of the extract with water

[0124] ③ Comparative Example 7 Flavoring

[0125] ④ Comparative Example 8 Flavoring

[0126] 5. Comparative Example 9 Flavoring

[0127] ⑥ Comparative Example 10 Flavoring

[0128] 7. Comparative Example 11 Flavoring

[0129] ⑧Comparative Example 12 Flavoring Materials

[0130] The sensory evaluation results are shown in Table 10:

[0131] Table 10 Comparison of the application effects of the flavoring materials prepared in Example 2 and Comparative Examples 7-12

[0132] Example 2 90% of the extract of Example 2 was used to prepare the flavoring, and the missing 10% of the extract was replaced with water. Comparative Example 7 Comparative Example 8 Comparative Example 9 Comparative Example 10 Comparative Example 11 Comparative Example 12 Color 14 13 13 13 13 13 13 13 Appearance status 14 14 14 14 14 14 14 14 aroma 27 26 20 16 8 16 9 17 taste 38.1 35 26 23 13 25 14 25 Total score 93.1 88 73 65 48 68 50 69

[0133] From the data of the previous application tests, we can find that: the flavoring prepared by the meat extract prepared in Examples 1 and 2, in the later application process, even if the amount of meat extract is reduced in the later stage, the effect of the product in the later application is better than that prepared according to Examples 1 and 2, thereby reducing the cost of the product and improving the utilization rate of protein; at the same time, it can be seen from the amino acid data that the content of essential amino acids in the flavoring is significantly increased, and the nutritional value is improved.

[0134] In summary, we can see that:

[0135] 1. Example 1 of the present invention was compared with Comparative Example 1. It was found that the conventional heating enzymatic hydrolysis was first performed, and then ultra-high pressure combined with thermal decomposition and enzymatic hydrolysis was performed. The hydrolysis degree of the product was greatly improved by 70%. In addition, in the later application process, even if the amount of meat extract was reduced in the later stage, the effect of the product in the later application was better than that prepared according to Comparative Example 1, thereby reducing the cost of the product and improving the utilization rate of the protein. At the same time, it can be seen from the amino acid data that the essential amino acid content in the flavoring was significantly increased, the nutritional value was improved, and the amino acid composition trend was roughly the same.

[0136] 2. Compared with Comparative Example 2, Example 1 of the present invention has a greatly improved hydrolysis degree, which is increased by 188%. The flavoring prepared by Example 1 is superior to the flavoring prepared in Comparative Example 2 in terms of aroma, taste, color, appearance, etc., and the amino acid content is increased by 255%. Moreover, its essential amino acid content is significantly better than that in Comparative Example 2, so its nutritional value is significantly better than that in Comparative Example 2.

[0137] 3. Compared with Comparative Example 3, Example 1 of the present invention has a higher degree of hydrolysis of up to 483%, indicating that the hydrolysis method of the present invention is more conducive to improving the utilization rate of protein. Sensory evaluation shows that the flavoring prepared by the method is significantly better than the flavoring of the product prepared by heating alone, and the amino acid content is significantly increased to 545%, and the essential amino acid content is significantly better than that in Comparative Example 3. Therefore, its nutritional value is significantly better than that of Comparative Example 3.

[0138] 4. Compared with Comparative Example 4, Example 1 of the present invention has a hydrolysis degree increased by 133%, indicating that the hydrolysis method of the present invention is more conducive to improving the utilization rate of protein, and through sensory evaluation, it was found that the flavoring prepared by using it was significantly better than the flavoring of the product prepared by single endonuclease hydrolysis, and the amino acid content was significantly increased to 136%, and its essential amino acid content was significantly better than that in Comparative Example 4, so its nutritional value was significantly better than that of Comparative Example 4.

[0139] 5. Compared with Comparative Example 5, Example 1 of the present invention has a hydrolysis degree increased by 405%, indicating that the hydrolysis method of the present invention is more conducive to improving the utilization rate of protein. Sensory evaluation found that the flavoring prepared by using it is significantly better than the flavoring of the product prepared by exonuclease hydrolysis alone, and the amino acid content is significantly increased to 519%, and its essential amino acid content is significantly better than that in Comparative Example 5. Therefore, its nutritional value is significantly better than that of Comparative Example 5.

[0140] 6. Compared with Comparative Example 6, the hydrolysis degree of Example 1 of the present invention is increased by 101 % , indicating that the hydrolysis method of the present invention is more conducive to improving the utilization rate of protein, and through sensory evaluation, it was found that the flavoring prepared by using it is significantly better than the flavoring of the product prepared by exonuclease hydrolysis alone, and the amino acid content was significantly increased to 123%, and its essential amino acid content was significantly better than that in Comparative Example 5, so its nutritional value is significantly better than that of Comparative Example 6.

[0141] 7. Comparison of Example 2 of the present invention with Comparative Example 7 reveals that a combination of conventional enzymatic hydrolysis followed by ultrahigh pressure combined with thermal and enzymatic hydrolysis significantly improves the product's degree of hydrolysis, increasing by 62.5%. Furthermore, even when the amount of meat extract used is reduced in later applications, the product performs better than the product prepared according to Comparative Example 6, thereby reducing product costs and improving protein utilization. Furthermore, amino acid data demonstrates a significant increase in the essential amino acid content of the flavoring, improving its nutritional value, while maintaining a generally similar amino acid composition.

[0142] 8. Compared with Comparative Example 8, Example 2 of the present invention has a significantly increased degree of hydrolysis, reaching 186%. The flavoring prepared using Example 2 is superior to the flavoring prepared in Comparative Example 7 in terms of aroma, taste, color, and appearance. Furthermore, the amino acid content is significantly increased, reaching 360%, and the essential amino acid content is significantly higher than that in Comparative Example 7. Therefore, its nutritional value is significantly superior to that of Comparative Example 8.

[0143] 9. Compared with Comparative Example 9, Example 2 of the present invention showed a significantly higher degree of hydrolysis, reaching 404%, indicating that the hydrolysis method of the present invention is more conducive to improving protein utilization. Sensory evaluation revealed that the flavoring prepared using this method was significantly superior to that prepared using heating alone. Furthermore, the amino acid content was significantly increased by 476%, and the essential amino acid content was significantly higher than that in Comparative Example 8. Therefore, its nutritional value is significantly superior to that of Comparative Example 9.

[0144] 10. Compared with Comparative Example 10, Example 2 of the present invention has a hydrolysis degree increased by 131%, indicating that the hydrolysis method of the present invention is more conducive to improving the utilization rate of protein. Sensory evaluation found that the flavoring prepared by using it is significantly better than the flavoring of the product prepared by single endonuclease hydrolysis.

[0145] 11. Compared with Comparative Example 11, Example 2 of the present invention has a hydrolysis degree increased by 334%, indicating that the hydrolysis method of the present invention is more conducive to improving the utilization rate of protein, and through sensory evaluation, it was found that the flavoring prepared by using it is significantly better than the flavoring of the product prepared by exonuclease hydrolysis alone.

[0146] 12. Compared with Comparative Example 11, Example 2 of the present invention has a hydrolysis degree increased by 104%, indicating that the hydrolysis method of the present invention is more conducive to improving the utilization rate of protein. Sensory evaluation found that the flavoring prepared by using it is significantly better than the flavoring of the product prepared by exonuclease hydrolysis alone.

[0147] In addition, a product was prepared according to the experimental scheme of patent CN103054032A (as comparative example 13 of the present application), and its hydrolysis degree was measured and compared with that of Example 1.

[0148] Comparative Example 13:

[0149] The raw materials were the same as those in Example 1, and the preparation method was as follows: chicken breast was crushed to 6-8 mm, soaked in vinegar for 40-50 minutes, subjected to microwave-assisted ultrahigh pressure extraction, filtered through a 110-mesh sieve, centrifuged, and fat removed to obtain a chicken breast extract. The mass ratio of chicken breast: water: vinegar was 1:1:0.006. The microwave-assisted ultrahigh pressure extraction was performed at a pressure of 110 MPa, a microwave power of 950 W, and a processing time of 35 minutes. The ultrahigh pressure extracted material was allowed to stand for 32 minutes. The centrifugation speed was 7500 rpm and the centrifugation time was 5 minutes.

[0150] The chicken breast extract is vacuum concentrated, and olive oil, tea polyphenols, and sucrose fatty acid ester are added and homogenized; wherein, the chicken breast extract is 90 parts, the olive oil is 2 parts, the tea polyphenols are 0.03 parts, and the sucrose fatty acid ester is 0.6 parts; the vacuum low-temperature concentration is performed at a vacuum degree of -0.07 MPa and a temperature of 60°C, and the concentration is performed until the refractive index Brix reaches 25%; the homogenization is performed using a high-pressure homogenizer, the homogenization time is 7 minutes, and the homogenization pressure is 45 MPa; filling and sterilization: the sterilization temperature is 90°C and the sterilization time is 15 seconds.

[0151] Comparative Example 14:

[0152] The raw materials were the same as those in Example 1, and the preparation method was as follows: chicken breast was minced to 6-8 mm, soaked in vinegar for 40-50 minutes, subjected to microwave-assisted ultrahigh pressure extraction, filtered through a 110-mesh sieve, centrifuged, and fat removed to obtain a chicken breast extract. The mass ratio of chicken breast: water: vinegar was 1:1:0.006; the ultrahigh pressure extraction pressure was 110 MPa, the processing time was 35 minutes, and the ultrahigh pressure extracted material was allowed to stand for 32 minutes; the centrifugation speed was 7500 rpm, and the centrifugation time was 5 minutes;

[0153] The chicken breast extract is vacuum concentrated, and olive oil, tea polyphenols, and sucrose fatty acid ester are added and homogenized; wherein, the chicken breast extract is 90 parts, the olive oil is 2 parts, the tea polyphenols are 0.03 parts, and the sucrose fatty acid ester is 0.6 parts; the vacuum low-temperature concentration is performed at a vacuum degree of -0.07 MPa and a temperature of 60°C, and the concentration is performed until the refractive index Brix reaches 25%; the homogenization is performed using a high-pressure homogenizer, the homogenization time is 7 minutes, and the homogenization pressure is 45 MPa; filling and sterilization: the sterilization temperature is 90°C and the sterilization time is 15 seconds.

[0154] The data was measured and the degree of hydrolysis was measured according to the hydrolysis degree detection method of this patent. It was found that the degree of hydrolysis was very low, as shown in Table 11 below:

[0155] Table 11 Comparison of hydrolysis degree between Example 1 and Comparative Examples 13 and 14

[0156] Example 1 Comparative Example 13 Comparative Example 14 Degree of hydrolysis (%) 30.3 4 3.5

[0157] . By measuring the hydrolysis degree data, it was found that the utilization rate of its protein was very low, and most of the protein still existed in the form of protein and was not hydrolyzed into polypeptides and amino acids, which was far from the experimental scheme of the present invention. In addition, it was centrifuged to centrifuge out and discard the protein that was not dissolved in water, resulting in a waste of resources and environmental pollution, and increased the cost of the nutrient solution. At the same time, the types and contents of free amino acids in its solution were determined using an ultra-high performance liquid chromatography method (the specific method is the same as above), and it was found that the amino acid content in its solution was significantly reduced compared with the extract of the present invention, and the content of its essential amino acids was also significantly less than that of the extract of the present invention. Specific data are shown in Table 12 below:

[0158] Table 12 Comparison of amino acid content between Example 1 and Comparative Examples 13-14

[0159] Example 1 Comparative Example 13 Comparative Example 14 Amino acid name Amino acid content (umol / mL) Amino acid content (umol / mL) Amino acid content (umol / mL) Histidine 5.80 0.31 0.27 Serine 12.10 1.37 1.11 Arginine 19.30 0.21 0.16 Glycine 24.00 9.20 7.6 Aspartic acid 8.00 1.17 1.01 glutamate 16.90 2.48 2.01 Threonine 10.20 0.98 0.80 Alanine 17.00 2.82 2.30 Proline 4.10 1.10 0.82 Cystine 4.40 0.88 0.75 Lysine 17.80 1.36 1.12 Tyrosine 7.30 0.40 0.35 Methionine 10.70 0.48 0.40 Valine 7.8 0.99 0.80 Isoleucine 8.30 0.71 0.61 Leucine 29.00 1.04 0.75 Phenylalanine 11.20 0.55 0.40 Total 213.9 26.04 21.36 Essential amino acid content 100.8 6.11 4.98 Non-essential amino acid content 101.4 18.65 15.28 Semi-essential amino acid content 11.7 1.28 1.1

[0160] The data on hydrolysis degree and amino acid content show that the extract prepared by the present invention is much better than the nutrient solution product prepared in Comparative Example 13. The protein utilization rate of the present invention is high, the degree of hydrolysis is high, and the content of essential amino acids and semi-essential amino acids is significantly higher than that of the nutrient solution prepared in Comparative Example 13. Applying it to the product is more conducive to digestion and absorption by the human body.

[0161] CN112890210A mentioned in the background technology, which mentions direct use of ultra-high pressure enzymatic hydrolysis for a protein powder and its preparation method, does not carry out selective enzymatic hydrolysis on the raw material product in advance. Moreover, the pressure involved in this patent is relatively small, focusing on the impact on the activity of the enzyme, and affecting the protein through the change of enzyme activity. Our patent first adopts conventional enzymatic hydrolysis (differential hydrolysis), and on the basis of differentiated hydrolysis, adopts non-differentiated hydrolysis (ultra-high pressure, heating) combined with differentiated hydrolysis (because the enzyme is not inactivated after the first stage of normal pressure enzymatic hydrolysis, entering the second ultra-high pressure heating hydrolysis stage, the enzyme still has a certain activity and will produce enzymatic hydrolysis). Through the first and second entire processes, more abundant polypeptides and uniform amino acids will be produced, making the taste closer to the feeling of home-made stewed soup.

[0162] Regarding the “Effect of Ultra-High Pressure Enzymolysis on the Flavor of Blue Clam Hydrolysate and Research on Ultra-High Pressure Assisted Enzymolysis Process of Blue Clam” mentioned in the background technology, it is mentioned that ultra-high pressure, temperature and enzymolysis are carried out simultaneously for enzymolysis. Its problem is the same as that of patent CN112890210A. The raw material product is not differentiated enzymolyzed in advance. Moreover, the pressure involved in this article is relatively small, and it focuses on the effect on enzyme activity, which affects the protein through the change of enzyme activity. It also has an enzyme inactivation process, while the present invention does not carry out enzyme inactivation treatment, and it selects the supernatant for subsequent experiments. At the same time, it is centrifuged after the ultra-high pressure enzymolysis treatment, and a large part of the protein that has not been completely enzymolyzed is centrifuged and separated. In this process, the unhydrolyzed protein has an adsorption effect and will adsorb some soluble amino acids and polypeptides, resulting in a waste of resources. The present invention adopts the overall experiment. Regarding the background technology mentioned in the article "The Effect of Ultra-High Pressure on the Flavor and Protein Structure of Blue Clam Enzyme Hydrolysate", it is mentioned that the protein is pressed under high pressure and then enzymatically hydrolyzed. In essence, non-differentiated hydrolysis is performed first and then differentiated hydrolysis. The supernatant is used for subsequent experiments, which is completely inconsistent with the experimental idea of ​​the present invention. The present invention adopts a holistic experiment to make full use of protein resources and avoid waste of protein.

[0163] The above are only specific embodiments of the present invention, but the technical features of the present invention are not limited thereto. Any simple changes, equivalent substitutions, or modifications based on the present invention to solve substantially the same technical problems and achieve substantially the same technical effects are included within the scope of protection of the present invention.

Claims

1. A method for preparing concentrated original meat flavoring by utilizing conventional heating enzymatic hydrolysis followed by ultrahigh pressure combined with thermal hydrolysis and enzymatic hydrolysis, characterized in that: The steps include: (1) preparing a meat extract: the meat-water ratio is 5:1 to 1:10, the total of meat and water is 100%, and endo-enzyme and exo-enzyme are added for enzymatic hydrolysis, wherein the amount of endo-enzyme added is 0.01% to 0.5%, and the amount of exo-enzyme added is 0.01% to 0.4%, and the enzymatic hydrolysis is carried out at 55 to 65°C for more than 30 minutes, and then ultra-high pressure is simultaneously combined with thermal hydrolysis and enzymatic hydrolysis, the ultra-high pressure pressure is 500MPa to 600MPa, the ultra-high pressure medium temperature is 70 to 100°C, and the ultra-high pressure time is 30 minutes to 6 hours; wherein the endo-enzyme is one or more of alkaline protease and papain, and the exo-enzyme is flavor protease; (2) Preparation of flavoring: 40-80 parts of meat extract, 1-10 parts of reducing sugar, 1-20 parts of amino acid mixture, 1-20 parts of fat, 10-20 parts of edible salt, 5-10 parts of monosodium glutamate, 1-10 parts of spices, 0.1-1 part of organic acid, 0.01-1 part of emulsifier, 0.1-10 parts of thickener, keep warm and stir, then cool the material, add 0.01-5 parts of flavor base, stir evenly to obtain concentrated original meat flavoring.

2. The method according to claim 1, wherein: The meat is selected from one or more of beef, chicken and mutton.

3. The method according to claim 1, wherein: The fat is selected from one or more of butter, chicken bone oil and chicken fat.

4. The method according to claim 1, wherein: The organic acid is selected from one or more of citric acid and lactic acid.

5. The method according to claim 1, wherein: The amino acid mixture is selected from multiple species of cysteine, methionine, aspartic acid and phenylalanine.

6. The method according to claim 1, wherein: The reducing sugar is selected from one or more of glucose, xylose and galactose.

7. The method according to claim 1, wherein: The emulsifier is selected from one or more of sucrose fatty acid esters and glyceryl monostearate.

8. The method according to claim 1, wherein: The thickener is selected from one or more of xanthan gum and acetylated distarch adipate.

9. The method according to claim 1, wherein: When the meat is chicken, the ultrahigh pressure is 600 MPa.

10. The method according to claim 1, wherein: When the meat is beef, the ultrahigh pressure is 500 MPa.

11. A concentrated original meat flavoring, characterized in that The method according to any one of claims 1 to 10 is used to prepare the product.

Images