Beef essence preparation method and system based on dual enzymolysis-Maillard reaction synergistic interaction

Abstract

The invention provides a beef essence preparation method and system based on dual enzymolysis-Maillard reaction synergistic interaction. The preparation method comprises the following steps: (1) carrying out first-step enzymolysis, namely adding compound protease into minced beef, regulating the pH value of a system to 6.5-7.0, controlling the temperature to be 45-50 DEG C, and carrying out enzymolysis reaction for 2-3 hours; (2) carrying out second-step enzymolysis, adding lipase derived from aspergillus oryzae, adjusting the pH value of the reaction system to 7.5-8.0, raising the temperature to 55-60 DEG C, and carrying out enzymolysis reaction for 1-2 hours; (3) performing enzyme deactivation treatment to obtain enzymatic hydrolysate; and (4) mixing the enzymatic hydrolysate with reducing sugar, thiamine and a yeast extract, and carrying out a Maillard reaction. The beef ingredients can be fully decomposed through double enzymolysis, and sufficient flavor precursor substances are provided for reaction; the Maillard reaction is intelligently regulated and controlled, more flavor substances are generated, and the flavor of the essence is remarkably improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of food flavor additives, and particularly relates to a method and system for preparing beef essence based on the synergistic enhancement of double enzymatic hydrolysis-Maillard reaction. Background Art

[0002] As an important flavor enhancer in the food industry, meat essence plays an indispensable role in various processed foods. Its core preparation technologies mainly include enzymatic hydrolysis, thermal reaction (Maillard reaction), and blending processes. In the existing technical field, numerous patents have disclosed the preparation methods of essence by traditional enzymatic hydrolysis combined with Maillard reaction. However, a series of problems that cannot be ignored have emerged in the actual application of these traditional preparation methods.

[0003] First of all, traditional methods often use a single enzymatic hydrolysis system to decompose meat proteins, making it difficult for the decomposition process to proceed fully. Due to the specific action of enzymes, a single enzyme cannot act on all sites of meat proteins comprehensively, resulting in insufficient production of flavor precursors. Flavor precursors are important raw materials for subsequent Maillard reactions, and the shortage of their quantity directly affects the flavor richness and intensity of the final essence product, making the product flavor monotonous and unable to meet consumers' demands for rich taste.

[0004] In addition, traditional heating methods lack precise temperature and time control means during the Maillard reaction. The Maillard reaction is extremely sensitive to temperature and time. If the temperature is too high or the reaction time is too long, it is extremely easy to produce burnt and charred flavors, seriously damaging the original flavor of the essence; conversely, if the temperature is too low or the reaction time is insufficient, the production of flavor substances will decrease, resulting in a weak flavor of the essence, unable to meet the standards of high-quality essence, and difficult to form competitiveness in the market.

[0005] Furthermore, in traditional preparation processes, the method of manually regulating reaction parameters (such as temperature, pH value) has many drawbacks. The skill levels of operators vary, and various errors will inevitably occur during the operation process. These factors will all affect the accuracy of reaction parameters. Even for the same formula, there will be quality differences between different batches of products due to parameter fluctuations. This problem of unstable quality seriously restricts the market competitiveness of essence products and the development of industrial large-scale production.

[0006] On the other hand, in order to make up for flavor defects, some traditional preparation methods rely too much on chemically synthesized essence for blending. However, with the continuous improvement of consumers' health awareness and the increasing attention to food health and safety, such products that do not conform to the trend of clean labels are gradually losing their market advantages. Consumers are more inclined to choose natural and additive-free food essence products, which poses a severe market challenge to traditional preparation methods.

[0007] Therefore, developing a new method for preparing high - efficiency, stable and natural beef essence and its supporting system has become an important topic that urgently needs to be overcome in the field of food flavor additives, and is of great significance for promoting the development of the food industry. Summary of the Invention

[0008] Starting from the technical problems described above, the present invention aims to solve the defects of traditional beef essence preparation technology and provide innovative preparation methods and systems. On the one hand, through the synergistic effect of double enzymatic hydrolysis and Maillard reaction, combined with an intelligent control system, the enzymatic hydrolysis efficiency and reaction controllability are improved, more flavor substances are precisely generated, the flavor intensity of beef essence is enhanced, and the naturalness is improved. On the other hand, an intelligent system is used to precisely control the reaction parameters to ensure the stable quality of products in different batches.

[0009] Through in - depth and meticulous research, the present invention has been completed.

[0010] Specifically, according to one aspect of the present invention, a method for preparing beef essence based on the synergistic effect of double enzymatic hydrolysis - Maillard reaction is provided. The preparation method includes: (1) Conduct the first enzymatic hydrolysis, wherein compound protease is added to minced beef, the pH value of the system is adjusted to 6.5 - 7.0, the temperature is controlled at 45 - 50 °C, and the enzymatic hydrolysis reaction is carried out for 2 - 3 hours, wherein the compound protease includes papain and flavor protease; (2) Conduct the second enzymatic hydrolysis, wherein lipase derived from Aspergillus oryzae is added to the product obtained in step (1), the pH value of the reaction system is adjusted to 7.5 - 8.0, the temperature is raised to 55 - 60 °C, and the enzymatic hydrolysis reaction is carried out for 1 - 2 hours; (3) Raise the temperature of the reaction system to 85 - 90 °C and maintain it for 10 - 20 minutes for enzyme inactivation treatment to obtain an enzymatic hydrolysate; (4) Mix the enzymatic hydrolysate obtained in step (3) with reducing sugar, thiamine, and yeast extract to obtain a mixture, adjust the pH value of the mixture to 5.0 - 5.5 and carry out the Maillard reaction at a temperature of 110 - 150 °C.

[0011] According to another aspect of the present invention, a system for the method for preparing beef essence based on the synergistic effect of double enzymatic hydrolysis - Maillard reaction described above is provided. The system includes: An enzymatic hydrolysis reaction module, which is equipped with: A multi - parameter sensor, which is used to real - time monitor the temperature, pH value, and conductivity during the reaction process; An automatic enzyme addition device, which is connected to the multi-parameter sensor and can control the addition amount and addition time of enzymes according to the reaction process and parameter changes monitored by the multi-parameter sensor; A Maillard reaction kettle, which integrates an online near-infrared spectrometer and a programmable logic controller (PLC controller). The online near-infrared spectrometer is used to monitor the changes of Maillard reaction products in real time and transmit the monitoring data to the PLC controller; the PLC controller analyzes and processes the monitoring data and automatically adjusts the temperature and pH value of the Maillard reaction kettle.

[0012] Compared with the prior art in the field, the advantages of the present invention are as follows: 1. In the prior art, the enzymatic hydrolysis efficiency is low and the formation of flavor precursor substances is insufficient, resulting in a weak flavor of the final product. The present invention adopts a double enzymatic hydrolysis process, in which compound protease and lipase are used for stepwise enzymatic hydrolysis to generate more flavor peptides and free fatty acids, providing rich raw materials for the Maillard reaction. The types of Maillard reaction products of the present invention are increased compared with the traditional method, and the flavor is more intense, complex and rich in layers; 2. Some traditional preparation methods rely too much on the blending of chemical synthetic flavors, which does not conform to the trend of clean labels. The present invention does not add any synthetic flavors throughout the process and completely uses natural raw materials for production, meeting the needs of consumers for healthy, safe and natural food flavors; 3. In the traditional preparation process, the reaction parameters are manually regulated, which is easily affected by the skill level and operation errors of the operators, resulting in large quality differences between batches. The present invention applies an intelligent regulation system to achieve precise control of key parameters such as temperature and pH value during the reaction process. The relative standard deviation (RSD) of flavor components between different batches of products is <5%, effectively ensuring the stability of product quality and providing a reliable guarantee for large-scale industrial production; 4. The production systems of the prior art may have problems of insufficient scalability and compatibility. The intelligent production system of the present invention adopts a modular design concept, and each module is independent and works in cooperation with each other, which can meet the production needs of food enterprises of different scales and is suitable for large-scale industrial production of beef flavor. Description of the Drawings

[0013] Figure 1 Shows a schematic flow chart of a method for preparing beef flavor based on the synergistic effect of double enzymatic hydrolysis-Maillard reaction according to an embodiment of the present invention; Figure 2 Shows a schematic structural diagram of a system for a method for preparing beef flavor based on the synergistic effect of double enzymatic hydrolysis-Maillard reaction according to an embodiment of the present invention; 1 Enzymatic hydrolysis reaction module; 2 Multi-parameter sensor; 3 Automatic enzyme addition device; 4 Maillard reaction kettle; 5 On-line near-infrared spectrometer; 6 PLC controller; 7 Post-treatment module. Detailed implementation mode

[0014] It should be understood that, without departing from the scope or spirit of the present disclosure, those skilled in the art can conceive of various other implementation schemes according to the teachings of this specification and can modify them. Therefore, the following detailed implementation modes do not have a restrictive meaning.

[0015] Unless otherwise specified, all numbers representing characteristic dimensions, quantities, and physical and chemical properties used in this specification should be understood to be modified by the term "about" in all cases. Therefore, unless there is a contrary statement, the numerical parameters listed in the above specification are approximate values, and those skilled in the art can appropriately change these approximate values to obtain the required characteristics by using the teachings disclosed herein. The use of numerical ranges expressed by endpoints includes all numbers within that range and any range within that range.

[0016] The present invention focuses on overcoming the problems of traditional beef essence preparation technology and is committed to achieving the goal of significantly improving the quality and production efficiency of the essence. In terms of technical problems, the traditional preparation method has a low enzymatic hydrolysis efficiency. Single enzymatic hydrolysis is difficult to fully decompose meat proteins, resulting in a lack of flavor precursor substances and limiting the flavor richness of the final product. The control accuracy of the Maillard reaction is poor. The traditional heating method cannot accurately control the temperature and time, and it is easy to cause burnt smell or insufficient generation of flavor substances. Moreover, manual adjustment of reaction parameters leads to poor product quality stability and significant differences between batches, making it difficult to meet the requirements of industrial large-scale production. At the same time, some traditional methods rely too much on synthetic flavors, which do not meet the needs of consumers for natural foods.

[0017] The present invention synergistically enhances the effect through double enzymatic hydrolysis and Maillard reaction. The intelligent control system accurately controls the temperature, time, and parameters, improves the enzymatic hydrolysis efficiency, enables the Maillard reaction to efficiently generate more flavor substances, greatly enhances the score of meaty aroma intensity, and the product has a high naturalness. In addition, it ensures the stable quality of products in different batches. The modular design of the production system is suitable for industrial large-scale production, enhancing the market competitiveness of the product.

[0018] Specifically, according to one aspect of the present invention, a method for preparing beef essence based on the synergistic enhancement of double enzymatic hydrolysis-Maillard reaction is provided. The preparation method includes: (1) Perform the first enzymatic hydrolysis, wherein add compound protease to minced beef, adjust the pH value of the system to 6.5-7.0, control the temperature at 45-50 °C, and perform the enzymatic hydrolysis reaction for 2-3 hours, wherein the compound protease includes papain and flavor protease; (2) Perform the second enzymatic hydrolysis. Add lipase derived from Aspergillus oryzae to the product obtained in step (1), adjust the pH value of the reaction system to 7.5 - 8.0, raise the temperature to 55 - 60 °C, and carry out the enzymatic hydrolysis reaction for 1 - 2 hours. (3) Raise the temperature of the reaction system to 85 - 90 °C and maintain it for 10 - 20 minutes for enzyme inactivation treatment to obtain the enzymolysis solution. (4) Mix the enzymolysis solution obtained in step (3) with reducing sugar, thiamine, and yeast extract to obtain a mixture, adjust the pH value of the mixture to 5.0 - 5.5, and carry out the Maillard reaction at a temperature of 110 - 150 °C.

[0019] Figure 1 Shows a schematic flow chart of a method for preparing beef flavor based on the synergistic effect of double enzymatic hydrolysis - Maillard reaction according to an embodiment of the present invention. As Figure 1 shown, the method for preparing beef flavor based on the synergistic effect of double enzymatic hydrolysis - Maillard reaction according to the present invention sequentially includes: the first enzymatic hydrolysis, the second enzymatic hydrolysis, enzyme inactivation treatment, and Maillard reaction.

[0020] Specifically, in the first enzymatic hydrolysis step (1), add compound protease to minced beef, adjust the pH value of the system to 6.5 - 7.0, control the temperature at 45 - 50 °C, and carry out the enzymatic hydrolysis reaction for 2 - 3 hours. Among them, the compound protease contains papain and flavor protease. In this step, papain and flavor protease act synergistically to efficiently decompose beef protein into small - molecule peptides and amino acids, providing rich substrates for the subsequent Maillard reaction.

[0021] According to certain technical solutions of the present invention, the specific types of papain include acidic papain, neutral papain, and basic papain. In the beef enzymatic hydrolysis process of the present invention, neutral papain and flavor protease form a compound protease in a specific ratio and act in a slightly acidic range of pH 6.5 - 7.0. The most suitable pH for neutral papain to exert its function is close to neutral, usually between 6.0 - 7.5, and it can stably and efficiently decompose beef protein into small - molecule peptides and amino acids.

[0022] According to certain technical solutions of the present invention, the flavor protease can be selected from one or more of endo - type flavor protease and exo - type flavor protease. The endo - type flavor protease can act on the peptide bonds inside beef proteins, randomly cut the peptide chains, decompose long - chain proteins into shorter peptide segments, and can rapidly degrade the macromolecular proteins in beef initially in the early stage of beef enzymolysis, increasing the number and types of peptide segments in the system. The exo - type flavor protease acts on the ends of peptide chains, hydrolyzes peptide bonds one by one to release amino acids. When the endo - type flavor protease decomposes macromolecular proteins into short peptides, the exo - type flavor protease further acts from the ends of the peptide chains to release free amino acids, and these free amino acids participate in the formation of various flavor substances in the Maillard reaction.

[0023] According to certain technical solutions of the present invention, the weight ratio of papain to flavor protease in the compound protease is 1:1 to 1:10, preferably 1:3 to 1:6, more preferably 1:4 to 1:5. For example, when the weight ratio of papain to flavor protease is 1:4 to 1:5, in the first - step enzymolysis process, their synergistic effect is the best, which can decompose beef proteins to the greatest extent and generate rich small - molecule peptides and amino acids. Commercially available products of papain include papain purchased from Nanning Pangbo Bio - engineering Co., Ltd. and papain from Sigma - Aldrich. Commercially available products of flavor protease include Flavourzyme 500MG from Novozymes A / S of Denmark and flavor protease products from Shanghai Yuanye Bio - technology Co., Ltd.

[0024] According to the technical solutions of the present invention, the weight percentage ratio of the compound protease relative to minced beef is 0.5% - 2%. Within this range, it can not only give full play to the synergistic effect of the compound protease to efficiently decompose beef proteins, but also take into account the cost and product quality.

[0025] In the second enzymatic hydrolysis step (2), lipase derived from Aspergillus oryzae is added to the product obtained from the first enzymatic hydrolysis. The pH value of the reaction system is adjusted to 7.5 - 8.0, and the temperature is raised to 55 - 60 °C for an enzymatic hydrolysis reaction for 1 - 2 hours. In the second enzymatic hydrolysis of minced beef, the lipase derived from Aspergillus oryzae can specifically decompose the fat in beef, converting it into free fatty acids, adding a unique fatty flavor to beef essence. Such lipases are specific lipases, highly specific to specific fatty acid ester bonds, capable of precisely acting on certain ester bonds in beef fat and selectively decomposing triglycerides with specific structures. In the beef enzymatic hydrolysis process of the present invention, the specific lipase can preferentially act on triglycerides containing unsaturated fatty acids, hydrolyzing them to generate free unsaturated fatty acids. These unsaturated fatty acids can further participate in complex chemical reactions in subsequent reactions, contributing a unique flavor to beef essence and enriching the flavor hierarchy of the essence. Commercially available lipases derived from Aspergillus oryzae that can be used in the present invention include: Lipase L0777 from Sigma-Aldrich, Lipase Lipozyme RM IM from Beijing Gaoruino Technology Co., Ltd., Lipozyme TL IM purchased from Novozymes A / S in Denmark, and Aspergillus oryzae lipase from Shanghai Yuanye Bio-Technology Co., Ltd.

[0026] According to the technical solution of the present invention, the weight percentage ratio of the lipase derived from Aspergillus oryzae relative to the product obtained from the first enzymatic hydrolysis is 0.1% - 0.3%. Within this range, it can not only make full use of the lipase to specifically decompose fat to generate sufficient free fatty acids to provide rich flavor precursors for the Maillard reaction, but also avoid excessive hydrolysis of fat caused by excessive addition, resulting in off-flavors or affecting subsequent processes and product quality.

[0027] In the enzyme inactivation treatment step (3), the temperature of the reaction system is raised to 85 - 90 °C and maintained for 10 - 20 minutes for enzyme inactivation treatment to obtain an enzymolysis solution. Through enzyme inactivation treatment, the activity of the enzyme can be terminated to prevent the enzyme from continuing to act in subsequent reactions and affecting product quality. For example, under the enzyme inactivation treatment conditions of maintaining at 85 °C for 15 minutes, the enzyme can be effectively inactivated without causing excessive damage to the flavor precursor substances in the enzymolysis solution.

[0028] In the Maillard reaction step (4), the enzymolysis solution is mixed with reducing sugar, thiamine, and yeast extract to obtain a mixture. The pH value of the mixture is adjusted to 5.0 - 5.5 and the Maillard reaction is carried out at a temperature of 110 - 150 °C. Reducing sugar is a key reactant in the Maillard reaction. According to certain technical solutions of the present invention, xylose and glucose are used as the reducing sugar in the present invention, and they are mixed in a weight ratio of 5:1 - 2:1, preferably 4:1 - 2:1, more preferably 3:1 - 2:1.

[0029] According to certain technical solutions of the present invention, thiamine, as a catalyst for the Maillard reaction, can lower the activation energy of the reaction, enabling the Maillard reaction to proceed more rapidly under relatively mild conditions. After constructing a Maillard reaction system with the enzymatic hydrolysate, reducing sugar, thiamine, and yeast extract, thiamine promotes the reaction rate between the reducing sugar and substances such as amino acids produced by beef enzymolysis. According to certain technical solutions of the present invention, during the Maillard reaction with dynamic temperature control, at each stage of gradually heating from 110°C to 150°C, thiamine continuously exerts its catalytic effect, reducing the reaction time required, improving production efficiency, ensuring the generation of sufficient flavor substances within a reasonable time, and laying a foundation for the formation of the rich flavor of the final beef essence. In addition, thiamine participates in the Maillard reaction and can promote the production of various flavor substances. It interacts with other components in the reaction system, changing the reaction pathway and product distribution. During the reaction process, thiamine can undergo complex chemical reactions with reducing sugars and amino acids to generate compounds with special flavors such as thiazole and pyrazine.

[0030] According to certain technical solutions of the present invention, yeast extract is rich in various nutrients and plays a key role in the Maillard reaction of beef. It can provide rich substances such as amino acids and sugars for the reaction, promote the generation of flavor substances, and improve the flavor quality of beef essence. Commercially available products of yeast extract that can be used in the present invention include: Angel's TL36 powdered yeast extract and Lesaffre yeast extract products.

[0031] According to certain technical solutions of the present invention, the Maillard reaction adopts a gradient temperature increase mode and is carried out in three temperature stages of 110°C, 130°C, and 150°C in sequence. Different temperature stages correspond to different reaction rates and reaction paths. This precise temperature setting avoids the problem that the temperature is difficult to accurately control in the traditional heating method, enables the Maillard reaction to proceed orderly, reduces the occurrence of over-reaction or under-reaction, and ensures that the reaction is always under relatively ideal conditions. In the initial stage at 110°C, the reaction starts relatively mildly, which is conducive to the preliminary reaction of some flavor precursor substances that are more sensitive to temperature, and avoids the destruction of these substances due to too high temperature. As the temperature gradually rises to 130°C and 150°C, the reaction rate accelerates, which can promote more complex chemical reactions to occur, meet the temperature requirements of different stages of the Maillard reaction, and make the entire reaction process more reasonable. Different temperature stages are beneficial to the formation of different types of flavor substances. At 110°C, it mainly promotes the formation of some basic flavor substances; when the temperature rises to 130°C, the reaction is more intense, and more compounds with unique flavors will be produced; when the temperature reaches 150°C, it can promote the formation of some special flavor substances that contribute greatly to the flavor of beef essence, such as pyrazine compounds, furan compounds, etc. Through this gradient temperature increase mode, the types of Maillard reaction products are enriched. Compared with the traditional single-temperature reaction, the types of Maillard reaction products increase by 30%, greatly enhancing the flavor complexity of beef essence. This temperature increase mode can increase the content of key flavor substances. For example, the production amounts of key flavor substances such as 2-methyl-3-furanthiol and furanone are increased by 40% compared with the traditional method during the Maillard reaction with gradient temperature increase.

[0032] According to the technical solution of the present invention, preferably, the mass ratio of reducing sugar to the enzymolysis solution is between 5:100 and 20:100; the mass ratio of thiamine to the enzymolysis solution is between 0.05:100 and 0.2:100; and the mass ratio of yeast extract to the enzymolysis solution is between 0.5:100 and 5:100.

[0033] According to certain technical solutions of the present invention, the preparation method further includes a post-treatment step (5) after step (4). Specifically, after the Maillard reaction, the mixture that has undergone the Maillard reaction is uniformly mixed with β-cyclodextrin, gum arabic, sodium chloride, and an antioxidant and dried. β-cyclodextrin and gum arabic are used for flavor molecule encapsulation in a mass ratio of 3:1 to 2:1. The unique cyclic structure of β-cyclodextrin can encapsulate flavor molecules, while gum arabic enhances the stability of the encapsulation system. Sodium chloride is used to adjust the flavor, and its addition amount is usually low in food. For beef flavor, the weight ratio of sodium chloride to the Maillard reaction mixture is 1 - 5 parts by weight. The antioxidant is used to inhibit the oxidation of the flavor and extend the shelf life, and the antioxidant is one or more of vitamin C palmitate or rosemary extract. In the present invention, rosemary extract can effectively inhibit the oxidation of the flavor, and there are multiple commercially available rosemary extract products on the market, such as the rosemary extract products of Shandong Yaotu Biotechnology Co., Ltd., Shandong Lusheng Biotechnology Co., Ltd., and Hubei Rutian Biotechnology Co., Ltd. In step (5), 100 parts by weight of the mixture that has undergone the Maillard reaction obtained in step (4) is uniformly mixed with 10 - 20 parts by weight of β-cyclodextrin, 10 - 20 parts by weight of gum arabic, 1 - 5 parts by weight of sodium chloride, and 0.05 - 0.1 parts by weight of the antioxidant and dried.

[0034] According to another aspect of the present invention, there is provided a system for the preparation method of beef flavor based on the synergistic effect of double enzymatic hydrolysis-Maillard reaction described above, and the system includes: An enzymatic hydrolysis reaction module, and the enzymatic hydrolysis reaction module is equipped with: A multi-parameter sensor, which is used to monitor the temperature, pH value, and conductivity in the reaction process in real time; An automatic enzyme addition device, which is connected to the multi-parameter sensor and can control the addition amount and addition time of the enzyme according to the reaction process and parameter changes monitored by the multi-parameter sensor; A Maillard reaction kettle, which is integrated with an on-line near-infrared spectrometer and a PLC controller. The on-line near-infrared spectrometer is used to monitor the changes of Maillard reaction products in real time and transmit the monitoring data to the PLC controller; the PLC controller analyzes and processes the monitoring data and automatically adjusts the temperature and pH value of the Maillard reaction kettle.

[0035] Specifically, the system for the method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction described above according to the present invention includes an enzymolysis reaction module. The enzymolysis reaction module is equipped with multi-parameter sensors for real-time monitoring of the temperature, pH value, and conductivity during the reaction process; an automatic enzyme addition device is connected to the multi-parameter sensors and can control the addition amount and addition time of the enzyme according to the reaction progress and parameter changes monitored by the multi-parameter sensors. The multi-parameter sensors have data storage and analysis functions, can record the change curves of the temperature, pH value, and conductivity during the reaction process, and generate corresponding analysis reports to provide data support for subsequent process optimization. The enzymolysis reaction module is also equipped with a stirring device, and the stirring device can adjust the stirring speed according to the conductivity change monitored by the multi-parameter sensors to ensure the uniformity of the enzymolysis reaction. The automatic enzyme addition device has multiple independent enzyme addition channels for adding compound protease and lipase derived from Aspergillus oryzae respectively, and each enzyme addition channel can accurately control the addition amount and addition time of the corresponding enzyme.

[0036] In addition, the system for the method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction described above according to the present invention includes a Maillard reaction kettle. The Maillard reaction kettle is integrated with an online near-infrared spectrometer and a PLC controller. The online near-infrared spectrometer is used for real-time monitoring of the changes in the Maillard reaction products and transmits the monitoring data to the PLC controller; the PLC controller analyzes and processes the monitoring data and automatically adjusts the temperature and pH value of the Maillard reaction kettle. Optionally, the Maillard reaction kettle is provided with a pressure sensor, and the pressure sensor is connected to the PLC controller. The PLC controller can automatically adjust the pressure of the Maillard reaction kettle according to the pressure change monitored by the pressure sensor. The online near-infrared spectrometer can quantitatively analyze the flavor substances in the Maillard reaction products, and the PLC controller adjusts the temperature, pH value, and reaction time of the Maillard reaction kettle according to the quantitative analysis results.

[0037] In addition, the system for the method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction described above according to the present invention further includes a post-treatment module. The post-treatment module is used for mixing and drying the mixture that has undergone the Maillard reaction with β-cyclodextrin, arabic gum, sodium chloride, and an antioxidant. The post-treatment module is connected to the Maillard reaction kettle and can automatically receive the Maillard reaction products and perform post-treatment operations. The system adopts a modular design concept, and each module is independent and works in coordination, which can meet the production needs of food enterprises of different scales and is suitable for industrial large-scale production of beef essence.

[0038] Figure 2 Shows a schematic structural diagram of a system 100 for the method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction according to an embodiment of the present invention. As Figure 2 shown, the system 100 includes: Enzymatic hydrolysis reaction module 1, and the enzymatic hydrolysis reaction module 1 is equipped with: Multi-parameter sensor 2, and the multi-parameter sensor 2 is used to monitor the temperature, pH value and conductivity in the reaction process in real time; Automatic enzyme addition device 3, and the automatic enzyme addition device 3 is connected to the multi-parameter sensor 2, and can control the addition amount and addition time of the enzyme according to the reaction process and parameter changes monitored by the multi-parameter sensor 2; Maillard reaction kettle 4, and the Maillard reaction kettle 4 integrates an online near-infrared spectrometer 5 and a PLC controller 6. The online near-infrared spectrometer 5 is used to monitor the changes of Maillard reaction products in real time and transmit the monitoring data to the PLC controller 6; the PLC controller 6 analyzes and processes the monitoring data and automatically adjusts the temperature and pH value of the Maillard reaction kettle 4; Post-treatment module 7, and the post-treatment module 7 is used to mix and dry the mixture that has undergone the Maillard reaction with β-cyclodextrin, gum arabic, sodium chloride and antioxidant. The post-treatment module 7 is connected to the Maillard reaction kettle 4 and can automatically receive the Maillard reaction products and perform post-treatment operations.

[0039] The following solutions are intended to illustrate the present disclosure by way of example and not limitation.

[0040] Solution 1 is a method for preparing beef essence based on the synergistic effect of double enzymatic hydrolysis-Maillard reaction. The preparation method includes: (1) Perform the first enzymatic hydrolysis, wherein add compound protease to minced beef, adjust the pH value of the system to 6.5 - 7.0, control the temperature at 45 - 50 °C, and perform the enzymatic hydrolysis reaction for 2 - 3 hours, wherein the compound protease includes papain and flavor protease; (2) Perform the second enzymatic hydrolysis, wherein add lipase derived from Aspergillus oryzae to the product obtained in step (1), adjust the pH value of the reaction system to 7.5 - 8.0, raise the temperature to 55 - 60 °C, and perform the enzymatic hydrolysis reaction for 1 - 2 hours; (3) Raise the temperature of the reaction system to 85 - 90 °C and maintain it for 10 - 20 minutes for enzyme inactivation treatment to obtain an enzymatic hydrolysate; (4) Mix the enzymatic hydrolysate obtained in step (3) with reducing sugar, thiamine and yeast extract to obtain a mixture, adjust the pH value of the mixture to 5.0 - 5.5 and perform the Maillard reaction at a temperature of 110 - 150 °C.

[0041] Scheme 2 is a method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction as described in Scheme 1, wherein the compound protease comprises papain and flavourzyme in a weight ratio of 1:1 to 1:10, preferably 1:3 to 1:6, more preferably 1:4 to 1:5.

[0042] Scheme 3 is a method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction as described in Scheme 1, wherein the papain is neutral papain.

[0043] Scheme 4 is a method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction as described in Scheme 1, wherein the flavourzyme is Flavourzyme 500MG purchased from Novozymes A / S, Denmark.

[0044] Scheme 5 is a method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction as described in Scheme 1, wherein the lipase derived from Aspergillus oryzae is Lipase L0777 from Sigma-Aldrich or Lipase Lipozyme RM IM from Beijing Gaorenuo Technology Co., Ltd.

[0045] Scheme 6 is a method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction as described in Scheme 1, wherein the reducing sugar is a mixture of xylose and glucose in a weight ratio of 5:1 - 2:1, preferably 4:1 - 2:1, more preferably 3:1 - 2:1.

[0046] Scheme 7 is a method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction as described in Scheme 1, wherein the Maillard reaction in step (4) adopts a gradient heating mode, and the Maillard reaction is carried out at three temperature stages of 110 °C, 130 °C and 150 °C in sequence.

[0047] Scheme 8 is a method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction as described in Scheme 1, wherein after step (4), the preparation method further includes: (5) Uniformly mixing and drying the mixture obtained by the Maillard reaction in step (4) with β-cyclodextrin, arabic gum, sodium chloride and an antioxidant.

[0048] Scheme 9 is a method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction as described in Scheme 8, wherein the antioxidant is one or more of vitamin C palmitate or rosemary extract.

[0049] Scheme 10 is a method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction according to Scheme 8. In step (5), 100 parts by weight of the Maillard reaction mixture obtained in step (4) is uniformly mixed with 10-20 parts by weight of β-cyclodextrin, 10-20 parts by weight of arabic gum, 1-5 parts by weight of sodium chloride, and 0.05-0.1 parts by weight of antioxidant and dried.

[0050] Scheme 11 is a system for a method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction according to any one of Schemes 1-10. The system includes: An enzymolysis reaction module equipped with: A multi-parameter sensor for real-time monitoring of temperature, pH value, and conductivity during the reaction; An automatic enzyme addition device connected to the multi-parameter sensor, capable of controlling the addition amount and addition time of the enzyme according to the reaction process and parameter changes monitored by the multi-parameter sensor; A Maillard reaction kettle integrated with an on-line near-infrared spectrometer and a PLC controller. The on-line near-infrared spectrometer is used for real-time monitoring of the changes in Maillard reaction products and transmitting the monitoring data to the PLC controller; the PLC controller analyzes and processes the monitoring data and automatically adjusts the temperature and pH value of the Maillard reaction kettle.

[0051] Scheme 12 is the system according to Scheme 11, wherein the enzymolysis reaction module is further equipped with a stirring device, and the stirring device can adjust the stirring speed according to the conductivity change monitored by the multi-parameter sensor to ensure the uniformity of the enzymolysis reaction.

[0052] Scheme 13 is the system according to Scheme 11, wherein the automatic enzyme addition device is provided with a plurality of independent enzyme addition channels for adding the compound protease and the lipase derived from Aspergillus oryzae respectively, and each enzyme addition channel can control the addition amount and addition time of the corresponding enzyme.

[0053] Scheme 14 is the system according to Scheme 11, wherein the Maillard reaction kettle is provided with a pressure sensor, the pressure sensor is connected to the PLC controller, and the PLC controller can automatically adjust the pressure of the Maillard reaction kettle according to the pressure change monitored by the pressure sensor.

[0054] Scheme 15 is the system according to Scheme 11, wherein the on-line near-infrared spectrometer can perform quantitative analysis on the flavor substances in the Maillard reaction products, and the PLC controller adjusts the temperature, pH value, and reaction time of the Maillard reaction kettle according to the quantitative analysis results.

[0055] Scheme 16 is the system according to Scheme 11, wherein the system further includes a post-treatment module for mixing and drying the mixture that has undergone the Maillard reaction with β-cyclodextrin, gum arabic, sodium chloride, and an antioxidant. The post-treatment module is connected to the Maillard reaction kettle and can automatically receive the Maillard reaction product and perform post-treatment operations.

[0056] The present invention will be described in more detail below in conjunction with embodiments. It should be noted that these descriptions and embodiments are for facilitating the understanding of the present invention rather than limiting the present invention.

[0057] Embodiment In the present invention, unless otherwise indicated, the reagents used are all commercially available products and are used directly without further purification treatment.

[0058] Testing method According to the methods described in detail below, the beef flavors prepared in the following examples and comparative examples were respectively tested for flavor, content of important flavor substances, and stability.

[0059] Flavor testing Select 30 people with food sensory evaluation experience to form a panel. The members should have sensitive taste and smell and no olfactory or taste disorders. Before the test, place the samples in an environment at room temperature (25°C ± 2°C) for 30 minutes to minimize the influence of temperature on the flavor. The sensory evaluation is carried out in a dedicated sensory test room, which should be kept quiet, odorless, with soft and uniform light. Each evaluator is equipped with an independent test cubicle to avoid mutual interference. During the test, the indoor temperature is maintained at 25°C ± 2°C, and the relative humidity is controlled at 50% ± 5%. The evaluators evaluate the samples one by one according to the specified flavor characteristics (richness of meat aroma, richness of flavor, and harmony of taste). Then, collect the scoring data of each sample by all evaluators, and calculate the average value and standard deviation of each flavor index. Through statistical analysis, compare the differences in beef flavors of each example and comparative example to judge the performance of the products of the present invention in terms of flavor.

[0060] Evaluation grade division of the obtained results: Excellent (9 - 10 points): It represents that the product performs excellently in terms of richness of meat aroma, richness of flavor, and harmony of taste, with a high richness of meat aroma, rich and harmonious flavor; Good (7 - 8 points): It indicates that the product performs well, has a certain meat aroma and flavor, but there is still room for improvement in some aspects; Acceptable (5 - 6 points): It indicates that the product is at a medium level and there are some obvious deficiencies in terms of flavor, etc.; Poor (below 5 points): It indicates that there are significant defects in the flavor of the product, making it difficult to meet the market demand for high-quality beef flavorings.

[0061] Content of important flavor substances The present invention uses a gas chromatography-mass spectrometry (GC-MS) instrument to analyze the contents of key flavor substances (2-methyl-3-furanthiol and furanone), thereby evaluating the flavor quality of beef flavorings.

[0062] Specifically, accurately weigh an appropriate amount of the sample and place it in different sample bottles. Add dichloromethane to the sample bottles, which serves to extract the flavor substances in the flavoring. Vigorously shake the sample bottles, and then place the sample bottles in an ultrasonic cleaner for ultrasonic treatment for a period of time to promote the dissolution and extraction of the flavor substances. After the ultrasonic treatment is completed, centrifuge the sample at a rotation speed of 5000 - 8000 revolutions per minute for 10 - 15 minutes to separate the extraction solution from the solid impurities. Finally, take the upper clear liquid and filter it through an organic phase filter membrane to remove possible minute particulate impurities, obtaining a pure sample solution to be tested.

[0063] Use a gas chromatography-mass spectrometry (GC-MS) instrument to detect the flavor substances 2-methyl-3-furanthiol and furanone in the sample. Specifically, the selected chromatographic column is an HP-5MS capillary column (30m × 0.25mm × 0.25μm), which has good separation effects for various flavor substances. Set the initial column temperature at 40°C and hold for 3 - 5 minutes to ensure the effective separation of low-boiling-point flavor substances; then increase the temperature to 250°C at a rate of 5 - 10°C per minute and hold for 5 - 10 minutes to fully separate high-boiling-point flavor substances. Set the inlet temperature at 250 - 280°C to ensure that the sample can quickly vaporize and enter the chromatographic column. Select nitrogen as the carrier gas and control the flow rate at 1 - 2 mL per minute. For the mass spectrometry part, use an electron impact ionization source (EI), with an electron energy of 70 eV, and set the scanning range at 35 - 500 m / z. Inject the processed sample solution into the GC-MS through an automatic injector, with an injection volume of 1 - 2 μL. Subsequently, record the chromatogram and mass spectrum. Analyze the obtained chromatogram and mass spectrum. By comparing with the mass spectrum database of standard substances, determine the types of key flavor substances in the sample, such as 2-methyl-3-furanthiol, furanone, etc. According to the peak area normalization method, calculate the relative content of each key flavor substance in the sample.

[0064] Evaluation grade classification of the obtained results: Excellent (9 - 10 points): The content of key flavor substances is increased by 50% or more compared with the traditional method, and the proportions of each key flavor substance are coordinated, indicating that the product has a strong and unique flavor and has significant advantages in flavor quality; Good (7 - 8 points): The content of key flavor substances increases by 30% - 49%, the proportion of flavor substances is basically reasonable, the product flavor is significantly improved, and it has high quality; Acceptable (5 - 6 points): The content of key flavor substances increases by 10% - 29%, the flavor is improved to a certain extent, but there are still some deficiencies, and it is at a medium level; Poor (below 5 points): The content of key flavor substances increases by less than 10% or decreases, the product flavor is not significantly improved or is poor, and it is difficult to meet the market demand for high - quality beef essence.

[0065] Stability Place the sample in a thermo - hygrostat with a temperature of 40°C and a relative humidity of 75% RH to simulate the relatively harsh environmental conditions that the product may encounter during actual storage. During the accelerated test, samples are taken and tested respectively at the 1st month, 2nd month, 3rd month, 4th month, 5th month, and 6th month. Each test uses sensory evaluation methods to evaluate flavor indexes such as the richness of meat aroma, flavor richness, and taste coordination of the sample.

[0066] Evaluation grade division of the obtained results: Excellent (9 - 10 points): During the 6 - month accelerated test period, flavor indexes such as the richness of meat aroma, flavor richness, and taste coordination of the product hardly change, and the content of key flavor substances remains stable; Good (7 - 8 points): During the accelerated test of the product, the flavor indexes change slightly, but it does not affect the overall flavor quality; Acceptable (5 - 6 points): The product shows a certain degree of flavor deterioration, and the richness of meat aroma, flavor richness, or taste coordination decreases; Poor (below 5 points): The product shows significant flavor deterioration during the accelerated test period, the richness of meat aroma weakens significantly, the flavor becomes single, and the taste coordination becomes poor.

[0067] Example 1 (E1) Select fresh beef and remove impurities such as fascia and fat. Grind the processed beef into uniform minced meat. Accurately weigh the minced beef and water according to the mass ratio of beef to water of 1:3, and put them into a homogenizer. Under the high - speed stirring of the homogenizer, the beef and water are fully mixed to form a uniform meat paste.

[0068] Transfer the prepared meat paste to as Figure 2In the enzymatic hydrolysis reaction module of the system for the preparation method of beef essence shown, compound protease is added thereto. The weight percentage ratio of the compound protease to the minced beef is 1%. The weight ratio of papain (from Sigma-Aldrich) to Flavourzyme (Flavourzyme 500MG purchased from Novozymes A / S, Denmark) in the compound protease is 1:1. Using a pH regulator (citric acid or sodium carbonate), the pH value of the reaction system is precisely adjusted to the weakly acidic range of 6.8, and at the same time, the temperature is stably controlled at 50 °C by a temperature control device. Start the stirring device and make the enzyme fully contact and mix evenly with the meat slurry at an appropriate stirring speed. Under such conditions, the enzymatic hydrolysis reaction lasts for 2.5 hours.

[0069] After the first-step enzymatic hydrolysis is completed, lipase derived from Aspergillus oryzae (lipase L0777 from Sigma-Aldrich) is added to the reaction system. The weight percentage ratio of the lipase derived from Aspergillus oryzae to the product obtained from the first-step enzymatic hydrolysis is 0.2%. Using sodium hydroxide as a pH regulator, the pH value of the reaction system is adjusted to the weakly alkaline range of 7.5, and at the same time, the temperature is raised to 55 °C by a heating device. Continue to start the stirring device to make the lipase fully contact with the reaction system, and the reaction lasts for 1 hour.

[0070] After the enzymatic hydrolysis reaction is completed, the temperature of the reaction system is quickly raised to 90 °C and maintained for 15 minutes for enzyme inactivation treatment. After the enzyme inactivation treatment, let the reaction system cool naturally to room temperature to obtain an enzymatic hydrolysate.

[0071] Transfer the enzymatic hydrolysate to a Maillard reaction kettle, add reducing sugar, and the reducing sugar is a mixture of xylose and glucose in a weight ratio of 5:1. At the same time, thiamine and yeast extract are added. The mass ratio of the reducing sugar to the enzymatic hydrolysate is 5:100; the mass ratio of thiamine to the enzymatic hydrolysate is 0.05:100; and the mass ratio of the yeast extract to the enzymatic hydrolysate is 5:100. Stir well to make each component mix evenly and construct a Maillard reaction system.

[0072] Subsequently, start the intelligent regulation program for the Maillard reaction, and the reaction temperature gradually increases step by step according to the gradient of 110 °C → 130 °C → 150 °C. At 110 °C, the reaction starts relatively gently, which is beneficial for some temperature-sensitive flavor precursors to undergo preliminary reactions and avoids the destruction of these substances due to too high temperature. Maintain for a period of time to let the basic flavor substances be initially formed. As the temperature gradually rises to 130 °C, the reaction rate accelerates, and more compounds with unique flavors begin to be generated. Finally, the temperature is raised to 150 °C to promote the generation of some special flavor substances that contribute greatly to the flavor of beef essence, such as pyrazine compounds, furan compounds, etc.

[0073] In each temperature stage, the changes of the reaction products are monitored in real time by an on-line near-infrared spectrometer installed in the Maillard reaction kettle. The on-line near-infrared spectrometer can quickly and accurately obtain the structural and compositional information of the substances in the reaction system, and transmit the monitoring data to the PLC controller. The PLC controller analyzes and processes the monitoring data according to the preset programs and algorithms, and automatically adjusts the temperature, pH value and reaction time of the Maillard reaction kettle. When it is monitored that the generation rate of a certain specific flavor substance begins to decline, the PLC controller will timely adjust the temperature or extend the reaction time to ensure that the Maillard reaction is always carried out under the optimal conditions, promoting the generation of more flavor substances. At the same time, based on the inherent relationship between the change of the conductivity of the reaction solution and the pH value, the phosphate buffer solution is automatically added through the PLC control system to maintain the pH value of the reaction system within the range of 5.0 - 5.5, providing a stable acid-base environment for the Maillard reaction, ensuring the smooth progress of the reaction, and improving the generation efficiency and quality of flavor substances.

[0074] After the Maillard reaction is completed, the reaction products are cooled to 60°C. To it are added the pre-prepared β-cyclodextrin, gum arabic, sodium chloride and rosemary extract (Shandong Yaotu Biotech Co., Ltd.), and they are evenly mixed and dried. Among them, based on 100 parts by weight of the mixture that has undergone the Maillard reaction, 10 parts by weight of β-cyclodextrin, 10 parts by weight of gum arabic, 2.5 parts by weight of sodium chloride and 0.1 part by weight of rosemary extract are added. Finally, beef essence 1 is obtained.

[0075] Then, according to the methods for flavor, important flavor substance content and stability described in detail above, the beef essence 1 is tested. The test results of the beef essence 1 are shown in Table 1 below.

[0076] Example 2 (E2) Beef essence 2 is prepared in a similar manner to Example 1, except that: the weight ratio of papain (Sigma-Aldrich) to flavor protease (Flavourzyme 500MG purchased from Novozymes A / S, Denmark) used in the first-step enzymatic hydrolysis is changed from 1:1 to 1:4.

[0077] Then, according to the methods for flavor, important flavor substance content and stability described in detail above, the beef essence 2 is tested. The test results of the beef essence 2 are shown in Table 1 below.

[0078] Example 3 (E3) Beef essence 3 is prepared in a similar manner to Example 1, except that: the weight ratio of xylose to glucose in the reducing sugar used in the Maillard reaction is changed from 5:1 to 3:1.

[0079] Then, according to the methods for flavor, important flavor substance content, and stability described in detail above, the beef essence 3 is tested. The test results of the beef essence 3 are shown in Table 1 below.

[0080] Example 4 (E4) Beef essence 4 is prepared in a similar manner to Example 1, except that: the weight ratio of papain (Sigma-Aldrich) to Flavourzyme 500MG (purchased from Novozymes A / S, Denmark) used in the first-step enzymatic hydrolysis is changed from 1:1 to 1:4; and the weight ratio of xylose to glucose in the reducing sugar used in the Maillard reaction is changed from 5:1 to 3:1.

[0081] Then, according to the methods for flavor, important flavor substance content, and stability described in detail above, the beef essence 4 is tested. The test results of the beef essence 4 are shown in Table 1 below.

[0082] Example 5 (E5) Beef essence 5 is prepared in a similar manner to Example 1, except that: Example 5 is not carried out in the system for the method of preparing beef essence as Figure 2 shown.

[0083] Specifically, in Example 5, a common enzymatic hydrolysis reaction tank is used for the first-step enzymatic hydrolysis and the second-step enzymatic hydrolysis. The process conditions (including pH value, temperature, time, etc.) of the first-step enzymatic hydrolysis and the second-step enzymatic hydrolysis are the same as those of the first-step enzymatic hydrolysis and the second-step enzymatic hydrolysis in Example 1.

[0084] In Example 5, a common Maillard reaction kettle is used for the Maillard reaction at a pH value of 5.0 - 5.5 and a temperature of 130 °C. Other processes are the same as those in Example 1.

[0085] Then, according to the methods for flavor, important flavor substance content, and stability described in detail above, the beef essence 5 is tested. The test results of the beef essence 5 are shown in Table 1 below.

[0086] Comparative Example 1 (CE1) Comparative beef essence 1 is prepared in a similar manner to Example 1, except that: the first-step enzymatic hydrolysis is not carried out.

[0087] Then, according to the methods for flavor, important flavor substance content, and stability described in detail above, the comparative beef essence 1 is tested. The test results of the comparative beef essence 1 are shown in Table 1 below.

[0088] Comparative Example 2 (CE1) Comparative beef flavor 2 was prepared in a manner similar to Example 1, except that the second step of enzymatic hydrolysis was not performed.

[0089] Then, the comparative beef flavor 2 was tested according to the methods described in detail above regarding flavor, important flavor substance content and stability. The test results of the comparative beef flavor 2 are shown in Table 1 below.

[0090] Table 1 Performance test results of beef flavors prepared in Examples 1-5 and Comparative Examples 1-2 Performance Example 1 Example 2 Example 3 Example 4 Example 5 Comparative Example 1 Comparative Example 2 Flavor property 7 (Good) 9 (Excellent) 8 (Good) 9 (Excellent) 8 (Good) 6 (Acceptable) 5 (Acceptable) Content of important flavor substances 8 (Good) 8 (Good) 8 (Good) 10 (Excellent) 7 (Good) 5 (Acceptable) 6 (Acceptable) Stability 8 (Good) 9 (Excellent) 9 (Excellent) 9 (Excellent) 7 (Good) 6 (Acceptable) 6 (Acceptable) Table 1 shows the test results of the beef flavors prepared in Examples 1-5 and Comparative Examples 1-2 in terms of flavor, content of important flavor substances and stability.

[0091] Regarding flavor, the flavor scores of Examples 2 and 4 reached 9 points (excellent), with excellent performance, high meat flavor and rich and coordinated flavor. This is due to its optimized composite protease ratio (the weight ratio of papain to flavor protease is 1:4). Example 3 scored 8 points (good), because the reducing sugar ratio was adjusted (the weight ratio of xylose to glucose was 3:1), the flavor was good but there was still room for improvement. Example 1 scored 7 points (good), and the various indicators were relatively balanced but the overall flavor was slightly inferior. Example 5 used ordinary equipment and different Maillard reaction conditions and scored 8 points (good), indicating that the system of the present invention has room for optimization but the basic process is effective. Comparative Example 1 did not perform the first step of enzymatic hydrolysis, only 6 points (acceptable), and the flavor was obviously insufficient, proving that the first step of enzymatic hydrolysis is crucial to flavor formation. Comparative Example 2 did not perform the second step of enzymatic hydrolysis, only 5 points (acceptable), the flavor defect was large, highlighting the necessity of the second step of enzymatic hydrolysis.

[0092] Regarding the content of important flavor substances, Example 4 scored as high as 10 points (excellent), and the content of key flavor substances increased by more than 50% compared with the traditional method and the proportion was coordinated, which was due to the dual optimization of the ratio of composite protease and reducing sugar. Examples 1, 2, and 3 were all 8 points (good), and the content of key flavor substances increased by 30%-49%, and the flavor was significantly improved. Example 5 scored 7 points (good), and the increase in the content of key flavor substances was small, indicating that the intelligent system of the present invention helps to increase the content of flavor substances. Comparative Example 1 scored 5 points (acceptable), and the content of key flavor substances increased by less than 10%, and the flavor improvement was not obvious. Comparative Example 2 scored 6 points (excellent), which was slightly better than Comparative Example 1, but still at a medium level, once again reflecting the importance of two-step enzymatic hydrolysis to the generation of flavor substances.

[0093] Regarding stability, the stability scores of Examples 2, 3, and 4 are all 9 points (excellent). During the 6-month accelerated test period, the flavor indicators only changed slightly, thanks to the intelligent control system and reasonable formula of the present invention. Example 1 got 8 points (good), with good stability but slightly inferior to the former three. Example 5 got 7 points (good), using ordinary equipment, and its stability was affected. Comparative Example 1 and Comparative Example 2 got 6 points (acceptable) respectively, with acceptable stability, but due to the lack of the enzymatic hydrolysis step, the long-term stability was poor.

[0094] In summary, the preparation method of the present invention can effectively improve the flavor, content of important flavor substances, and stability of beef essence through double enzymatic hydrolysis and intelligent control of the Maillard reaction. Optimizing the ratio of compound protease and reducing sugar has a significant effect on improving the product quality. The intelligent system based on the present invention helps to improve the product quality and stability. The first-step enzymatic hydrolysis and the second-step enzymatic hydrolysis are indispensable for the flavor formation and flavor substance generation of beef essence.

[0095] Although specific embodiments have been shown and described in the present invention, those skilled in the art will understand that various alternative and / or equivalent embodiments can be used to replace the specific embodiments shown and described without departing from the scope of the present invention. This application is intended to cover any improvement or change to the specific embodiments discussed in the present invention. Those skilled in the art should understand that various modifications and changes can be made without departing from the scope of the present invention. Such modifications and changes are intended to fall within the defined scope of the present invention.

Claims

1. A preparation method of beef essence based on the synergistic effect of double enzymatic hydrolysis-Maillard reaction, characterized in that, The preparation method includes: (1) Perform the first enzymatic hydrolysis, wherein a compound protease is added to minced beef, the pH value of the system is adjusted to 6.5 - 7.0, the temperature is controlled at 45 - 50 °C, and the enzymatic hydrolysis reaction is carried out for 2 - 3 hours, wherein the compound protease contains papain and flavor protease; (2) Perform the second enzymatic hydrolysis, wherein lipase derived from Aspergillus oryzae is added to the product obtained in step (1), the pH value of the reaction system is adjusted to 7.5 - 8.0, the temperature is raised to 55 - 60 °C, and the enzymatic hydrolysis reaction is carried out for 1 - 2 hours; (3) Raise the temperature of the reaction system to 85 - 90 °C and maintain it for 10 - 20 minutes for enzyme inactivation treatment to obtain an enzymatic hydrolysate; (4) Mix the enzymatic hydrolysate obtained in step (3) with reducing sugar, thiamine, and yeast extract to obtain a mixture, adjust the pH value of the mixture to 5.0 - 5.5 and carry out the Maillard reaction at a temperature of 110 - 150 °C.

2. The method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction according to claim 1, characterized in that The compound protease contains the papain and the flavor protease in a weight ratio of 1:1 to 1:

10.

3. The preparation method of beef essence based on the synergistic effect of double enzymatic hydrolysis-Maillard reaction according to claim 1, characterized in that The papain is neutral papain.

4. The method for preparing beef essence based on the synergistic effect of double enzymatic hydrolysis-Maillard reaction according to claim 1, wherein The reducing sugar is a mixture of xylose and glucose in a weight ratio of 5:1 - 2:

1.

5. The preparation method of beef essence based on the synergistic effect of double enzymatic hydrolysis-Maillard reaction according to claim 1, wherein The Maillard reaction in step (4) adopts a gradient heating mode, wherein the Maillard reaction is carried out in three temperature stages of 110 °C, 130 °C, and 150 °C in sequence.

6. The preparation method of beef essence based on the synergistic effect of double enzymolysis-Maillard reaction according to claim 1, characterized in that The preparation method further includes after step (4): (5) Uniformly mix and dry the mixture obtained in step (4) that has undergone the Maillard reaction with β - cyclodextrin, arabic gum, sodium chloride, and an antioxidant.

7. The method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction according to claim 6, characterized in that, The antioxidant is one or more of vitamin C palmitate or rosemary extract.

8. The method for preparing beef essence based on the synergistic effect of double enzymolysis-Maillard reaction according to claim 6, wherein, In step (5), 100 parts by weight of the mixture obtained in step (4) that has undergone the Maillard reaction is uniformly mixed and dried with 10 - 20 parts by weight of β - cyclodextrin, 10 - 20 parts by weight of arabic gum, 1 - 5 parts by weight of sodium chloride, and 0.05 - 0.1 parts by weight of the antioxidant.

9. A system for the preparation method of beef essence based on the synergistic effect of double enzymolysis-Maillard reaction according to any one of claims 1-8, characterized in that, The system includes: An enzymatic hydrolysis reaction module, and the enzymatic hydrolysis reaction module is equipped with: A multi - parameter sensor, which is used to monitor the temperature, pH value, and conductivity in the reaction process in real time; An automatic enzyme addition device, which is connected to the multi - parameter sensor and can control the addition amount and addition time of the enzyme according to the reaction process and parameter changes monitored by the multi - parameter sensor; A Maillard reaction kettle, which is integrated with an on - line near - infrared spectrometer and a PLC controller. The on - line near - infrared spectrometer is used to monitor the changes of Maillard reaction products in real time and transmit the monitoring data to the PLC controller; the PLC controller analyzes and processes the monitoring data and automatically adjusts the temperature and pH value of the Maillard reaction kettle.

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