A method and system for preparing beef flavor based on synergistic enhancement of double enzymatic hydrolysis and Maillard reaction

Abstract

The present invention provides a method and system for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction. The preparation method comprises: (1) performing a first enzymatic hydrolysis, wherein a composite protease is added to ground 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; (2) performing a second enzymatic hydrolysis, wherein a lipase derived from Aspergillus oryzae is added, the pH value of the reaction system is adjusted to 7.5-8.0, the temperature is increased to 55-60°C, and the enzymatic hydrolysis reaction is carried out for 1-2 hours; (3) performing an enzyme inactivation treatment to obtain an enzymatic hydrolyzate; and (4) mixing the enzymatic hydrolyzate with reducing sugar, thiamine, and yeast extract and carrying out a Maillard reaction. In the present invention, the dual enzymatic hydrolysis can fully decompose the beef components, providing sufficient flavor precursors for the reaction; the Maillard reaction is intelligently regulated to generate more flavor substances, significantly improving the flavor of the flavor.

Description

Technical Field

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

[0002] Meat flavoring, as an important flavor enhancer in the food industry, plays an indispensable role in various processed foods. Its core production technologies primarily encompass enzymatic hydrolysis, thermal reaction (Maillard reaction), and blending processes. Numerous patents disclose traditional methods for preparing flavors using a combination of enzymatic hydrolysis and the Maillard reaction. However, these traditional methods present a number of significant challenges in practical application.

[0003] First, traditional methods often use a single enzymatic system to break down meat proteins, making the breakdown process difficult to complete. Due to the specificity of enzymes, a single enzyme cannot fully target every site in the meat protein, resulting in insufficient production of flavor precursors. Flavor precursors are crucial raw materials for the subsequent Maillard reaction, and their shortage directly impacts the flavor richness and intensity of the final flavor product, resulting in a monotonous flavor that fails to meet consumer demand for a richer taste.

[0004] Furthermore, traditional heating methods lack precise temperature and time control during the Maillard reaction. The Maillard reaction is extremely sensitive to both temperature and time. Excessively high temperatures or prolonged reaction times can easily produce a burnt odor, severely damaging the flavor profile of the fragrance. Conversely, excessively low temperatures or insufficient reaction time can reduce the production of flavoring compounds, resulting in a weak flavor that fails to meet high-quality standards and struggles to compete in the market.

[0005] Furthermore, the traditional preparation process, which relies on manual control of reaction parameters (such as temperature and pH), presents numerous drawbacks. Operator skill levels vary, and errors inevitably occur during operation, all of which impact the accuracy of reaction parameters. Even with the same formula, quality can vary between batches due to parameter fluctuations. This inconsistent quality severely hinders the market competitiveness of flavor products and the development of large-scale industrial production.

[0006] On the other hand, some traditional preparation methods rely excessively on chemically synthesized flavors to compensate for flavor deficiencies. However, as consumers' health awareness continues to rise and their focus on food safety and health increases, these products, which do not align with the clean label trend, are gradually losing their market advantage. Consumers are increasingly opting for natural, additive-free food flavors, posing a significant market challenge to traditional preparation methods.

[0007] Therefore, the development of an efficient, stable and natural new beef flavor preparation method and supporting system has become an important issue that needs to be tackled in the current field of food flavor additives, and is of great significance for promoting the development of the food industry. Summary of the Invention

[0008] Based on the technical issues outlined above, the present invention aims to address the shortcomings of traditional beef flavor preparation technology by providing an innovative preparation method and system. On one hand, by synergizing dual enzymatic hydrolysis with the Maillard reaction, combined with an intelligent control system, this method improves enzymatic hydrolysis efficiency and reaction controllability, precisely generating more flavor compounds, enhancing the flavor intensity of the beef flavor, and improving its naturalness. On the other hand, the intelligent system precisely controls reaction parameters, ensuring consistent product quality across batches.

[0009] The present invention has been completed after intensive and careful research.

[0010] Specifically, according to one aspect of the present invention, a method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction is provided, the preparation method comprising:

[0011] (1) performing a first enzymatic hydrolysis step, wherein a composite protease is added to the ground 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 composite protease comprises papain and flavor protease;

[0012] (2) performing a second enzymatic hydrolysis step, 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 increased to 55-60° C., and the enzymatic hydrolysis reaction is carried out for 1-2 hours;

[0013] (3) raising the temperature of the reaction system to 85-90°C and maintaining it for 10-20 minutes to inactivate the enzyme to obtain an enzymatic hydrolyzate;

[0014] (4) The enzymatic hydrolyzate obtained in step (3) 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 a Maillard reaction is performed at a temperature of 110-150° C.

[0015] According to another aspect of the present invention, a system for the above-mentioned method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction is provided, the system comprising:

[0016] An enzymatic reaction module, wherein the enzymatic reaction module is equipped with:

[0017] A multi-parameter sensor for real-time monitoring of temperature, pH value, and conductivity during the reaction process;

[0018] An automatic enzyme adding device, connected to the multi-parameter sensor, capable of controlling the amount and time of enzyme addition according to the reaction progress and parameter changes monitored by the multi-parameter sensor;

[0019] The Maillard reactor is integrated with an online near-infrared spectrometer and a programmable logic controller (PLC). The online near-infrared spectrometer is used to monitor changes in 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 reactor.

[0020] Compared with the prior art in this field, the advantages of the present invention are:

[0021] 1. Existing enzymatic hydrolysis methods have low efficiency and insufficient production of flavor precursors, resulting in a bland flavor in the final product. The present invention utilizes a dual enzymatic hydrolysis process, with compound protease and lipase performing separate enzymatic hydrolysis steps to generate more flavor peptides and free fatty acids, providing a rich source of raw materials for the Maillard reaction. Compared to traditional methods, the present invention produces a greater variety of Maillard reaction products, resulting in a richer, more complex, and more layered flavor.

[0022] 2. Some traditional preparation methods rely too heavily on chemically synthesized flavors, which is inconsistent with the clean label trend. This invention uses no synthetic flavors and is produced entirely from natural ingredients, meeting consumer demand for healthy, safe, and natural food flavors.

[0023] 3. The traditional preparation process involves manual control of reaction parameters, which is easily affected by operator skill and operational errors, leading to large quality variations between batches. This invention utilizes an intelligent control system to precisely control key parameters such as temperature and pH during the reaction process. The relative standard deviation (RSD) of flavor components between batches is less than 5%, effectively ensuring product quality stability and providing reliable support for industrial large-scale production.

[0024] 4. Existing production systems may suffer from insufficient scalability and compatibility. The intelligent production system of this invention adopts a modular design concept. Each module works independently yet collaboratively, meeting the production needs of food companies of different sizes and is suitable for industrial-scale production of beef flavor. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1A schematic flow chart of a method for preparing beef flavor based on synergistic enhancement of double enzymatic hydrolysis and Maillard reaction according to one embodiment of the present invention is shown;

[0026] Figure 2 A schematic diagram of the structure of a system for preparing beef flavor based on synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction according to one embodiment of the present invention is shown;

[0027] 1. Enzymatic reaction module; 2. Multi-parameter sensor; 3. Automatic enzyme adding device; 4. Maillard reactor; 5. Online near-infrared spectrometer; 6. PLC controller; 7. Post-processing module. DETAILED DESCRIPTION

[0028] It should be understood that those skilled in the art can conceive of other various embodiments and can modify them according to the teachings of this specification without departing from the scope or spirit of the present disclosure. Therefore, the following specific embodiments are not intended to be limiting.

[0029] Unless otherwise indicated, all numbers used in this specification to express feature sizes, quantities, and physicochemical properties should be understood as being modified in all instances by the term "about." Therefore, unless otherwise indicated, the numerical parameters listed in the above description are approximate values, and those skilled in the art will be able to appropriately change these approximate values ​​to obtain the desired properties using the teachings disclosed herein. The use of numerical ranges expressed as endpoints includes all numbers within that range and any range within that range.

[0030] The present invention focuses on overcoming the difficulties in traditional beef flavor preparation technology, and is committed to achieving the goal of significantly improving flavor quality and production efficiency. In terms of technical issues, traditional preparation methods have the problem of low enzymatic hydrolysis efficiency. Single enzymatic hydrolysis is difficult to fully decompose meat protein, resulting in a lack of flavor precursors, which limits the flavor richness of the final product. The control accuracy of the Maillard reaction is poor, and traditional heating methods cannot accurately control temperature and time, which can easily cause burnt smells or insufficient generation of flavor substances. Moreover, the manual control of reaction parameters results in poor product quality stability and significant differences between batches, which is difficult to meet the requirements of industrial large-scale production. At the same time, some traditional methods rely too much on synthetic flavors, which does not meet consumers' demand for natural foods.

[0031] This invention utilizes dual enzymatic hydrolysis and the Maillard reaction for synergistic benefits. The intelligent control system precisely controls temperature, timing, and parameters, improving enzymatic hydrolysis efficiency and enabling the Maillard reaction to efficiently generate more flavor compounds. This significantly enhances the meaty aroma score and increases the naturalness of the product. Furthermore, it ensures consistent product quality across batches. The modular design of the production system is suitable for industrial-scale production, enhancing the product's market competitiveness.

[0032] Specifically, according to one aspect of the present invention, a method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction is provided, the preparation method comprising:

[0033] (1) performing a first enzymatic hydrolysis step, wherein a composite protease is added to the ground 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 composite protease comprises papain and flavor protease;

[0034] (2) performing a second enzymatic hydrolysis step, 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 increased to 55-60° C., and the enzymatic hydrolysis reaction is carried out for 1-2 hours;

[0035] (3) raising the temperature of the reaction system to 85-90°C and maintaining it for 10-20 minutes to inactivate the enzyme to obtain an enzymatic hydrolyzate;

[0036] (4) The enzymatic hydrolyzate obtained in step (3) 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 a Maillard reaction is performed at a temperature of 110-150° C.

[0037] Figure 1 The schematic flow chart of the method for preparing beef flavor based on the synergistic enhancement of double enzymatic hydrolysis and Maillard reaction according to one embodiment of the present invention is shown. Figure 1 As shown in the figure, the method for preparing beef flavor based on the synergistic enhancement of double enzymatic hydrolysis and Maillard reaction according to the present invention comprises, in sequence: a first step of enzymatic hydrolysis, a second step of enzymatic hydrolysis, enzyme inactivation treatment and Maillard reaction.

[0038] Specifically, in the first enzymatic hydrolysis step (1), a composite protease is added to the ground 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. The composite protease includes papain and flavor protease. In this step, papain and flavor protease work synergistically to efficiently decompose beef protein into small molecular peptides and amino acids, providing rich substrates for the subsequent Maillard reaction.

[0039] According to certain technical solutions of the present invention, specific types of papain include acidic papain, neutral papain, and alkaline papain. In the beef enzymatic hydrolysis process of the present invention, neutral papain and flavor protease are combined in a specific ratio to form a composite protease that functions in the slightly acidic pH range of 6.5-7.0. Neutral papain works optimally at a near-neutral pH, typically between 6.0-7.5, and can stably and efficiently break down beef protein into small peptides and amino acids.

[0040] According to certain technical solutions of the present invention, the flavor protease can be selected from one or more of an endo-acting flavor protease and an exo-acting flavor protease. The endo-acting flavor protease acts on the peptide bonds within beef proteins, randomly severing peptide chains and breaking long-chain proteins into shorter peptide segments. This rapidly degrades large proteins in the beef during the initial enzymatic hydrolysis process, increasing the number and variety of peptide segments in the system. The exo-acting flavor protease acts on the ends of peptide chains, hydrolyzing peptide bonds one by one to release amino acids. After the endo-acting flavor protease breaks down the large proteins into short peptides, the exo-acting flavor protease further acts on the peptide ends to release free amino acids. These free amino acids participate in the Maillard reaction to form various flavor compounds.

[0041] According to some technical solutions of the present invention, the weight ratio of papain and flavor protease in the composite 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 and flavor protease is 1:4 to 1:5, in the first step enzymatic hydrolysis process, the two have the best synergistic effect, can decompose beef protein to the greatest extent, and generate abundant small molecule peptides and amino acids. The commercially available products of papain include the papain purchased from Nanning Pangbo Bioengineering Co., Ltd. and the papain of Sigma-Aldrich Co., Ltd. The commercially available products of flavor protease include the flavor protease products purchased from Flavourzyme 500MG of Novozymes A / S, Denmark and Shanghai Source Leaf Biotechnology Co., Ltd.

[0042] According to the technical solution of the present invention, the weight percentage of the composite protease relative to the ground beef is 0.5%-2%. Within this range, the synergistic effect of the composite protease can be fully exerted to efficiently decompose beef protein while taking into account cost and product quality.

[0043] 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, the temperature is raised to 55-60°C, and the enzymatic hydrolysis reaction is carried out for 1-2 hours. In the second enzymatic hydrolysis of ground beef, the lipase derived from Aspergillus oryzae can specifically decompose the fat in the beef and convert it into free fatty acids, adding a unique fatty flavor to the beef flavor. This type of lipase is a specific lipase that is highly specific for specific fatty acid ester bonds and can accurately act on certain ester bonds in beef fat and selectively decompose triglycerides of 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 the beef flavor and enriching the flavor level of the flavor. Commercially available lipases derived from Aspergillus oryzae that can be used in the present invention include: Lipase L0777 from Sigma-Aldrich, Lipozyme RM IM from Beijing Gao Ruinuo Technology Co., Ltd., Lipozyme TL IM from Novozymes, Denmark, and Aspergillus oryzae lipase from Shanghai Yuanye Biotechnology Co., Ltd.

[0044] According to the technical solution of the present invention, the weight percentage of Aspergillus oryzae lipase relative to the product obtained from the first enzymatic hydrolysis step is between 0.1% and 0.3%. Within this range, the lipase's specific ability to break down fats is fully utilized, generating sufficient free fatty acids to provide rich flavor precursors for the Maillard reaction, while also preventing excessive fat hydrolysis caused by excessive addition, which could produce off-flavors or affect subsequent processes and product quality.

[0045] In the enzyme inactivation step (3), the temperature of the reaction system is raised to 85-90°C and maintained for 10-20 minutes to perform enzyme inactivation to obtain an enzymatic hydrolyzate. Enzyme inactivation can terminate the activity of the enzyme, preventing the enzyme from continuing to act in subsequent reactions and affecting product quality. For example, an enzyme inactivation treatment at 85°C for 15 minutes can effectively inactivate the enzyme without causing excessive damage to the flavor precursors in the enzymatic hydrolyzate.

[0046] In the Maillard reaction step (4), the enzymatic hydrolyzate is mixed with a reducing sugar, thiamine, and yeast extract to obtain a mixture, the pH 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, the reducing sugar is a mixture of xylose and glucose in a weight ratio of 5:1-2:1, preferably 4:1-2:1, and more preferably 3:1-2:1.

[0047] According to certain technical solutions of the present invention, thiamine, as a catalyst for the Maillard reaction, can reduce the activation energy of the reaction, allowing the Maillard reaction to proceed more rapidly under relatively mild conditions. After the enzymatic hydrolyzate is combined with reducing sugars, thiamine, and yeast extract to form a Maillard reaction system, thiamine accelerates the reaction rate between the reducing sugars and amino acids and other substances produced by the enzymatic hydrolysis of beef. According to certain technical solutions of the present invention, during the dynamic temperature-controlled Maillard reaction, thiamine continuously catalyzes the reaction at each stage of the gradual temperature increase from 110°C to 150°C, reducing reaction time and improving production efficiency. This ensures the production of sufficient flavor compounds within a reasonable timeframe, laying the foundation for the rich flavor of the final beef essence. Furthermore, thiamine's participation in the Maillard reaction can promote the production of a variety of flavor compounds. It interacts with other components in the reaction system, altering the reaction pathway and product distribution. During the reaction, thiamine undergoes complex chemical reactions with reducing sugars and amino acids to produce compounds with unique flavors, such as thiazole and pyrazine.

[0048] 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 abundant amino acids, sugars, and other substances for the reaction, promote the production of flavor substances, and enhance the flavor quality of beef flavor. Commercially available yeast extracts that can be used in the present invention include Angel's TL36 powdered yeast extract and Lesaffre yeast extract products.

[0049] According to certain technical solutions of the present invention, the Maillard reaction utilizes a gradient temperature ramp, proceeding sequentially through three temperature stages: 110°C, 130°C, and 150°C. Different temperature stages correspond to different reaction rates and pathways. This precise temperature setting avoids the difficulty of precise temperature control with traditional heating methods, allowing the Maillard reaction to proceed in an orderly manner, reducing over- or underreactions and ensuring ideal reaction conditions. In the initial stage at 110°C, the reaction initiates relatively gently, facilitating the initial reaction of temperature-sensitive flavor precursors and preventing their destruction due to excessive temperatures. As the temperature gradually increases to 130°C and 150°C, the reaction rate accelerates, promoting more complex chemical reactions and satisfying the temperature requirements of the different Maillard reaction stages, streamlining the overall reaction process. Different temperature stages facilitate the production of different flavor substances. At 110°C, the reaction primarily promotes the formation of basic flavor compounds. When the temperature rises to 130°C, the reaction becomes more intense, producing more compounds with unique flavors. At 150°C, it promotes the formation of specialized flavor compounds that contribute significantly to the flavor of beef essence, such as pyrazines and furans. This gradient heating pattern enriches the variety of Maillard reaction products, increasing them by 30% compared to traditional single-temperature reactions, significantly enhancing the flavor complexity of beef essences. This heating pattern can also increase the content of key flavor compounds. For example, the production of key flavor compounds such as 2-methyl-3-furanthiol and furanone during the gradient heating Maillard reaction increases by 40% compared to traditional methods.

[0050] According to the technical solution of the present invention, preferably, the mass ratio of reducing sugar to enzymatic hydrolysate is between 5:100 and 20:100; the mass ratio of thiamine to enzymatic hydrolysate is between 0.05:100 and 0.2:100; and the mass ratio of yeast extract to enzymatic hydrolysate is between 0.5:100 and 5:100.

[0051] According to certain technical solutions of the present invention, the preparation method further includes a post-processing step (5) after step (4). Specifically, after the Maillard reaction, the mixture subjected to 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 embedding flavor molecules 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 embedding system. Sodium chloride is used to adjust the flavor, and its addition amount in food is usually low. 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 oxidation of the flavor and extend the shelf life. 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 flavors. There are many commercially available rosemary extract products on the market, such as the rosemary extract products of Shandong Yaotu Bioengineering Co., Ltd., the rosemary extract products of Shandong Lusen Biotechnology Co., Ltd., and the rosemary extract products of Hubei Rutian Bioengineering Co., Ltd. In step (5), 100 parts by weight of the mixture obtained in step (4) and subjected to the Maillard reaction are 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 an antioxidant, and dried.

[0052] According to another aspect of the present invention, a system for the above-mentioned method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction is provided, the system comprising:

[0053] An enzymatic reaction module, wherein the enzymatic reaction module is equipped with:

[0054] A multi-parameter sensor for real-time monitoring of temperature, pH value, and conductivity during the reaction process;

[0055] An automatic enzyme adding device, connected to the multi-parameter sensor, capable of controlling the amount and time of enzyme addition according to the reaction progress and parameter changes monitored by the multi-parameter sensor;

[0056] The Maillard reactor is integrated with an online near-infrared spectrometer and a PLC controller. The online near-infrared spectrometer is used to monitor the changes in the 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 reactor.

[0057] Specifically, the system for preparing beef flavor based on the dual enzymatic hydrolysis and Maillard reaction synergistic enhancement method described above includes an enzymatic hydrolysis reaction module. The enzymatic hydrolysis reaction module is equipped with a multi-parameter sensor for real-time monitoring of temperature, pH, and conductivity during the reaction process. An automatic enzyme addition device is connected to the multi-parameter sensor and can control the amount and timing of enzyme addition based on the reaction progress and parameter changes monitored by the multi-parameter sensor. The multi-parameter sensor has data storage and analysis capabilities, capable of recording temperature, pH, and conductivity curves during the reaction process and generating corresponding analysis reports to provide data support for subsequent process optimization. The enzymatic hydrolysis reaction module is also equipped with a stirring device that can adjust the stirring speed based on conductivity changes monitored by the multi-parameter sensor to ensure the uniformity of the enzymatic hydrolysis reaction. The automatic enzyme addition device is equipped with multiple independent enzyme addition channels, one for adding a composite protease and the other for adding a lipase derived from Aspergillus oryzae. Each enzyme addition channel can precisely control the amount and timing of the corresponding enzyme addition.

[0058] Furthermore, the system for preparing beef flavor based on the dual enzymatic hydrolysis and Maillard reaction synergistic enhancement method described above includes a Maillard reactor. The Maillard reactor is integrated with an online near-infrared spectrometer and a programmable logic controller (PLC). The online near-infrared spectrometer monitors changes in Maillard reaction products in real time and transmits the monitored data to the PLC controller, which analyzes and processes the monitored data and automatically adjusts the temperature and pH of the Maillard reactor. Optionally, the Maillard reactor is equipped with a pressure sensor connected to the PLC controller, which automatically adjusts the pressure of the Maillard reactor based on pressure changes monitored by the pressure sensor. The online near-infrared spectrometer quantitatively analyzes flavor compounds in the Maillard reaction products, and the PLC controller adjusts the temperature, pH, and reaction time of the Maillard reactor based on the quantitative analysis results.

[0059] In addition, the system for preparing beef flavor based on the dual enzymatic hydrolysis and Maillard reaction synergistic enhancement method described above also includes a post-processing module. This module is used to mix and dry the Maillard reaction mixture with β-cyclodextrin, gum arabic, sodium chloride, and an antioxidant. The post-processing module is connected to the Maillard reactor and can automatically receive the Maillard reaction product and perform post-processing operations. The system adopts a modular design concept, with each module working independently and collaboratively to meet the production needs of food companies of different sizes and is suitable for industrial large-scale production of beef flavor.

[0060] Figure 2 The schematic diagram of the structure of the system 100 for preparing beef flavor based on the synergistic enhancement of double enzymatic hydrolysis and Maillard reaction according to one embodiment of the present invention is shown. Figure 2 As shown in FIG, the system 100 includes:

[0061] Enzymatic reaction module 1, the enzymatic reaction module 1 is equipped with:

[0062] A multi-parameter sensor 2, which is used to monitor the temperature, pH value and conductivity during the reaction in real time;

[0063] An automatic enzyme adding device 3, which is connected to the multi-parameter sensor 2 and can control the amount and time of enzyme addition according to the reaction progress and parameter changes monitored by the multi-parameter sensor 2;

[0064] A Maillard reactor 4 is integrated with an online near-infrared spectrometer 5 and a PLC controller 6. The online near-infrared spectrometer 5 is used to monitor changes in 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 reactor 4.

[0065] The post-processing module 7 is used to mix and dry the mixture after the Maillard reaction with β-cyclodextrin, gum arabic, sodium chloride and antioxidant. The post-processing module 7 is connected to the Maillard reactor 4 and can automatically receive the Maillard reaction product and perform post-processing operations.

[0066] The following schemes are intended to illustrate the present disclosure by way of example and not limitation.

[0067] Scheme 1 is a method for preparing beef flavor based on the synergistic enhancement of double enzymatic hydrolysis and Maillard reaction, the preparation method comprising:

[0068] (1) performing a first enzymatic hydrolysis step, wherein a composite protease is added to the ground 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 composite protease comprises papain and flavor protease;

[0069] (2) performing a second enzymatic hydrolysis step, 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 increased to 55-60° C., and the enzymatic hydrolysis reaction is carried out for 1-2 hours;

[0070] (3) raising the temperature of the reaction system to 85-90°C and maintaining it for 10-20 minutes to inactivate the enzyme to obtain an enzymatic hydrolyzate;

[0071] (4) The enzymatic hydrolyzate obtained in step (3) 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 a Maillard reaction is performed at a temperature of 110-150° C.

[0072] Scheme 2 is a method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction according to Scheme 1, wherein the composite protease comprises the papain and the flavor protease in a weight ratio of 1:1 to 1:10, preferably 1:3 to 1:6, and more preferably 1:4 to 1:5.

[0073] Scheme 3 is a method for preparing beef flavor based on the synergistic enhancement of double enzymatic hydrolysis and Maillard reaction according to Scheme 1, wherein the papain is neutral papain.

[0074] Scheme 4 is a method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction according to Scheme 1, wherein the flavor protease is Flavourzyme 500MG purchased from Novozymes of Denmark.

[0075] Scheme 5 is a method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction according to Scheme 1, wherein the lipase derived from Aspergillus oryzae is lipase L0777 from Sigma-Aldrich or lipase Lipozyme RM IM from Beijing Gao Ruinuo Technology Co., Ltd.

[0076] Scheme 6 is a method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction according to 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, and more preferably 3:1-2:1.

[0077] Scheme 7 is a method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction according to Scheme 1, wherein the Maillard reaction in step (4) adopts a gradient temperature rising mode, wherein the Maillard reaction is carried out in three temperature stages of 110°C, 130°C and 150°C in sequence.

[0078] Scheme 8 is a method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction according to Scheme 1, wherein the preparation method further comprises, after step (4):

[0079] (5) The Maillard reaction mixture obtained in step (4) is uniformly mixed with β-cyclodextrin, gum arabic, sodium chloride and an antioxidant, and dried.

[0080] Scheme 9 is a method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction according to Scheme 8, wherein the antioxidant is one or more of vitamin C palmitate or rosemary extract.

[0081] Scheme 10 is a method for preparing beef flavor based on the synergistic enhancement of double enzymatic hydrolysis and Maillard reaction according to Scheme 8, wherein in step (5), 100 parts by weight of the mixture obtained in step (4) and subjected to the Maillard reaction are 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 an antioxidant and dried.

[0082] Scheme 11 is a system for the method for preparing beef flavor based on the synergistic enhancement of dual enzymatic hydrolysis and Maillard reaction according to any one of Schemes 1 to 10, the system comprising:

[0083] An enzymatic reaction module, wherein the enzymatic reaction module is equipped with:

[0084] A multi-parameter sensor for real-time monitoring of temperature, pH value, and conductivity during the reaction process;

[0085] An automatic enzyme adding device, connected to the multi-parameter sensor, capable of controlling the amount and time of enzyme addition according to the reaction progress and parameter changes monitored by the multi-parameter sensor;

[0086] The Maillard reactor is integrated with an online near-infrared spectrometer and a PLC controller. The online near-infrared spectrometer is used to monitor the changes in the 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 reactor.

[0087] Scheme 12 is a system according to Scheme 11, wherein the enzymatic hydrolysis reaction module is further equipped with a stirring device, which can adjust the stirring speed according to the conductivity changes monitored by the multi-parameter sensor to ensure the uniformity of the enzymatic hydrolysis reaction.

[0088] Scheme 13 is a system according to Scheme 11, wherein the automatic enzyme addition device is provided with a plurality of independent enzyme addition channels, respectively used for adding the composite protease and the lipase derived from Aspergillus oryzae, and each enzyme addition channel can control the addition amount and addition time of the corresponding enzyme.

[0089] Option 14 is a system according to Option 11, wherein the Maillard reactor 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 reactor according to the pressure changes monitored by the pressure sensor.

[0090] Option 15 is a system according to Option 11, wherein the online near-infrared spectrometer is capable of quantitatively analyzing flavor substances in the Maillard reaction product, and the PLC controller adjusts the temperature, pH value and reaction time of the Maillard reactor according to the quantitative analysis results.

[0091] Option 16 is a system according to Option 11, wherein the system further includes a post-processing module, which is used to mix and dry the mixture that has undergone the Maillard reaction with β-cyclodextrin, gum arabic, sodium chloride and an antioxidant. The post-processing module is connected to the Maillard reactor and can automatically receive the Maillard reaction product and perform post-processing operations.

[0092] The present invention will be described in more detail below with reference to the following examples. It should be noted that these descriptions and examples are intended to facilitate understanding of the present invention, but are not intended to limit the present invention.

[0093] Example

[0094] In the present invention, unless otherwise specified, all reagents used were commercially available products and were used directly without further purification.

[0095] Test Method

[0096] According to the methods described in detail below, the beef flavors prepared in the following examples and comparative examples were tested for flavor, important flavor substance content and stability.

[0097] Flavor test

[0098] A team of 30 people with experience in food sensory evaluation was selected. Members must have a keen sense of taste and smell, and no olfactory or taste disorders. Before the test, the samples were placed at room temperature (25°C ± 2°C) for 30 minutes to balance and minimize the impact of temperature on the flavor. The sensory evaluation was carried out in a special sensory testing room, which should be quiet, odorless, and with soft and uniform light. Each evaluator was equipped with an independent testing room to avoid mutual interference. During the test, the indoor temperature was maintained at 25°C ± 2°C, and the relative humidity was controlled at 50% ± 5%. The evaluators evaluated the samples one by one according to the specified flavor characteristics (richness of meat aroma, richness of flavor, and coordination of taste). Then, the scoring data of all evaluators for each sample were collected, and the mean and standard deviation of each flavor index were calculated. Through statistical analysis, the differences in the beef flavors of the various embodiments and comparative examples were compared to judge the performance of the product of the present invention in terms of flavor.

[0099] Evaluation levels of the results obtained:

[0100] Excellent (9-10 points): This product excels in terms of meaty aroma, richness of flavor, and harmonious taste. It has a strong meaty aroma and a rich and harmonious flavor.

[0101] Good (7-8 points): indicates that the product performs well, with a certain meaty aroma and flavor, but there is room for improvement in some aspects;

[0102] Acceptable (5-6 points): indicates that the product is at an average level, with some obvious deficiencies in flavor, etc.

[0103] Poor (below 5 points): indicates that the product has major defects in flavor and cannot meet the market demand for high-quality beef flavor.

[0104] Content of important flavor substances

[0105] The present invention adopts gas chromatography-mass spectrometry (GC-MS) to analyze the content of key flavor substances (2-methyl-3-furanthiol and furanone) to evaluate the flavor quality of beef flavor.

[0106] Specifically, accurately weigh appropriate amounts of samples and place them in different sample bottles. Add dichloromethane to the sample bottles to extract the flavor substances in the essence. Vibrate the sample bottles thoroughly, 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 samples with the speed set to 5000-8000 rpm for 10-15 minutes to separate the extract from the solid impurities. Finally, take the supernatant and filter it with an organic phase filter membrane to remove any possible tiny particulate impurities to obtain a pure sample solution to be tested.

[0107] Gas chromatography-mass spectrometry (GC-MS) was used to detect the flavor compounds 2-methyl-3-furanthiol and furanone in the samples. Specifically, an HP-5MS capillary column (30 m × 0.25 mm × 0.25 μm) was selected, which offers excellent separation performance for a wide range of flavor compounds. The column temperature was initially set at 40°C for 3-5 minutes to ensure effective separation of low-boiling-point flavor compounds. The temperature was then increased to 250°C at a rate of 5-10°C / minute and held for 5-10 minutes to fully separate high-boiling-point flavor compounds. The inlet temperature was set between 250-280°C to ensure rapid sample vaporization upon entry into the column. Nitrogen was used as the carrier gas at a flow rate of 1-2 mL / minute. Mass spectrometry was performed using an electron impact ionization (EI) source with an electron energy of 70 eV and a scan range of 35-500 m / z. The treated sample solution is injected into the GC-MS using an autosampler at an injection volume of 1-2 μL. The chromatogram and mass spectrum are then recorded and analyzed. By comparing the chromatograms and mass spectra with a database of mass spectra of reference substances, the identity of key flavor compounds in the sample, such as 2-methyl-3-furanthiol and furanone, is determined. The relative content of each key flavor compound in the sample is calculated using peak area normalization.

[0108] Evaluation levels of the results obtained:

[0109] Excellent (9-10 points): The content of key flavor compounds is increased by 50% or more compared to traditional methods, and the proportions of key flavor compounds are harmonious, indicating that the product has a rich and unique flavor and has a significant advantage in flavor quality;

[0110] Good (7-8 points): The content of key flavor substances increased by 30%-49%, the flavor substance ratio was basically reasonable, the product flavor was significantly improved, and the product had high quality.

[0111] Acceptable (5-6 points): The content of key flavor substances increased by 10%-29%, and the flavor was improved to a certain extent, but there were still some deficiencies, and it was at a medium level;

[0112] Poor (less than 5 points): The content of key flavor substances increased by less than 10% or decreased, and the product flavor was not significantly improved or was poor, which made it difficult to meet the market demand for high-quality beef flavor.

[0113] stability

[0114] Samples were placed in a constant temperature and humidity chamber at 40°C and 75% relative humidity to simulate the harsh environmental conditions the products might encounter during actual storage. During the accelerated testing period, samples were collected and tested at the first, second, third, fourth, fifth, and sixth months. Each test was evaluated using sensory evaluation methods to assess flavor characteristics such as meaty aroma intensity, flavor richness, and mouthfeel harmony.

[0115] Evaluation levels of the results obtained:

[0116] Excellent (9-10 points): During the 6-month accelerated testing period, the product's flavor indicators, such as meaty aroma intensity, flavor richness, and taste harmony, showed little change, and the content of key flavor compounds remained stable.

[0117] Good (7-8 points): The flavor index of the product changes slightly during the accelerated test, but does not affect the overall flavor quality;

[0118] Acceptable (5-6 points): The product has a certain degree of flavor deterioration, with a decrease in the richness of the meat aroma, the richness of the flavor, or the harmony of the taste.

[0119] Poor (less than 5 points): The product exhibits significant flavor deterioration during the accelerated test period, with the richness of the meat aroma significantly weakened, the flavor becoming monotonous, and the taste coordination becoming poor.

[0120] Example 1 (E1)

[0121] Select fresh beef and remove impurities such as fascia and fat. Grind the beef into a uniform mince. Accurately weigh the ground beef and water, using a 1:3 beef to water ratio, and place them in a homogenizer. The high-speed agitation in the homogenizer thoroughly mixes the beef and water, forming a uniform slurry.

[0122] Transfer the prepared meat paste to Figure 2 In the enzymatic reaction module of the system for preparing beef flavor shown in the figure, a composite protease is added thereto. The weight percentage of the composite protease relative to the ground beef is 1%. The weight ratio of papain (Sigma-Aldrich) and flavor protease (Flavourzyme500MG purchased from Novozymes, Denmark) in the composite protease is 1:1. Using a pH regulator (citric acid or sodium carbonate), the pH value of the reaction system is accurately adjusted to a slightly acidic range of 6.8, and the temperature is stably controlled at 50°C by a temperature control device. Start the stirring device and stir at an appropriate speed to ensure that the enzyme and the meat slurry are fully contacted and evenly mixed. Under such conditions, the enzymatic reaction lasts for 2.5 hours.

[0123] After the first enzymatic hydrolysis step, lipase from Aspergillus oryzae (Sigma-Aldrich Lipase L0777) was added to the reaction system. The weight percentage of the lipase from Aspergillus oryzae relative to the product obtained from the first enzymatic hydrolysis step was 0.2%. The pH of the reaction system was adjusted to a weakly alkaline range of 7.5 using sodium hydroxide as a pH adjuster. Simultaneously, the temperature was raised to 55°C using a heating device. The stirring device was kept running to ensure full contact between the lipase and the reaction system. The reaction was continued for 1 hour.

[0124] After the enzymatic hydrolysis reaction is completed, the temperature of the reaction system is quickly raised to 90°C and maintained for 15 minutes to inactivate the enzyme. After the enzyme inactivation treatment, the reaction system is allowed to cool naturally to room temperature to obtain an enzymatic hydrolyzate.

[0125] Transfer the enzymatic hydrolysate to a Maillard reactor and add reducing sugar, a mixture of xylose and glucose in a 5:1 weight ratio. Thiamine and yeast extract are also added. The weight ratio of reducing sugar to enzymatic hydrolysate is 5:100; the weight ratio of thiamine to enzymatic hydrolysate is 0.05:100; and the weight ratio of yeast extract to enzymatic hydrolysate is 5:100. Stir thoroughly to ensure uniform mixing of all ingredients, thus establishing the Maillard reaction system.

[0126] The Maillard reaction's intelligent control program is then activated, gradually increasing the reaction temperature from 110°C to 130°C to 150°C. At 110°C, the reaction begins relatively gently, allowing for the initial reaction of temperature-sensitive flavor precursors and preventing their destruction due to excessive temperatures. This temperature is maintained for a period of time to allow the initial formation of basic flavor compounds. As the temperature gradually rises to 130°C, the reaction rate accelerates, and more compounds with unique flavors begin to form. Finally, the temperature is raised to 150°C, promoting the production of specific flavor compounds that contribute significantly to the beef flavor, such as pyrazines and furans.

[0127] At each temperature stage, an online near-infrared spectrometer installed within the Maillard reactor monitors the changes in reaction products in real time. This spectrometer quickly and accurately obtains structural and compositional information about the substances in the reaction system and transmits this data to a programmable logic controller (PLC). The PLC analyzes and processes this data based on pre-set programs and algorithms, automatically adjusting the temperature, pH, and reaction time of the Maillard reactor. If the production rate of a particular flavor compound begins to decline, the PLC promptly adjusts the temperature or extends the reaction time to ensure the Maillard reaction proceeds under optimal conditions, promoting the production of more flavor compounds. Furthermore, based on the inherent correlation between changes in the reaction solution's conductivity and pH, the PLC automatically adds phosphate buffer to maintain the pH of the reaction system within the range of 5.0-5.5, providing a stable acid-base environment for the Maillard reaction, ensuring smooth reaction progress and improving the efficiency and quality of flavor compound production.

[0128] After the Maillard reaction is completed, the reaction product is cooled to 60°C. Pre-prepared β-cyclodextrin, gum arabic, sodium chloride and rosemary extract (Shandong Yaotu Bioengineering Co., Ltd.) are added thereto, uniformly mixed and dried. Among them, based on 100 parts by weight of the mixture subjected to 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 parts by weight of rosemary extract are added. Finally, beef flavor 1 is obtained.

[0129] Then, the beef flavor 1 was tested according to the methods described in detail above regarding flavor, important flavor substance content and stability. The test results of beef flavor 1 are shown in Table 1 below.

[0130] Example 2 (E2)

[0131] Beef flavor 2 was prepared in a manner similar to Example 1, except that the weight ratio of papain (Sigma-Aldrich) to flavor enzyme (Flavourzyme 500MG purchased from Novozymes, Denmark) used in the first enzymatic hydrolysis step was changed from 1:1 to 1:4.

[0132] Then, the 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 beef flavor 2 are shown in Table 1 below.

[0133] Example 3 (E3)

[0134] Beef flavor 3 was prepared in a manner similar to Example 1, except that the weight ratio of xylose to glucose in the reducing sugar used in the Maillard reaction was changed from 5:1 to 3:1.

[0135] Then, the beef flavor 3 was tested according to the methods described in detail above regarding flavor, important flavor substance content and stability. The test results of beef flavor 3 are shown in Table 1 below.

[0136] Example 4 (E4)

[0137] Beef flavor 4 was prepared in a manner similar to Example 1, except that the weight ratio of papain (Sigma-Aldrich) and flavor enzyme (Flavourzyme 500MG purchased from Novozymes, Denmark) used in the first enzymatic hydrolysis was changed from 1:1 to 1:4; and the weight ratio of xylose to glucose in the reducing sugar used in the Maillard reaction was changed from 5:1 to 3:1.

[0138] Then, the beef flavor 4 was tested according to the methods described in detail above regarding flavor, important flavor substance content and stability. The test results of beef flavor 4 are shown in Table 1 below.

[0139] Example 5 (E5)

[0140] Prepare beef flavor 5 in a similar manner to Example 1, except that: Example 5 does not contain Figure 2 The method for preparing beef flavor is carried out in the system shown.

[0141] Specifically, Example 5 uses a common enzymatic hydrolysis reactor for the first and second steps of enzymatic hydrolysis. The process conditions (including pH value, temperature and time, etc.) of the first and second steps of enzymatic hydrolysis are the same as those of the first and second steps of enzymatic hydrolysis in Example 1.

[0142] In Example 5, a common Maillard reactor was used to carry out the Maillard reaction at a pH of 5.0-5.5 and a temperature of 130° C. The other processes were the same as those in Example 1.

[0143] Then, the beef flavor 5 was tested according to the methods described in detail above regarding flavor, important flavor substance content, and stability. The test results of beef flavor 5 are shown in Table 1 below.

[0144] Comparative Example 1 (CE1)

[0145] Comparative beef flavor 1 was prepared in a manner similar to Example 1, except that the first enzymatic hydrolysis step was not performed.

[0146] Then, the comparative beef flavor 1 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 1 are shown in Table 1 below.

[0147] Comparative Example 2 (CE1)

[0148] Comparative beef flavor 2 was prepared in a manner similar to Example 1, except that the second step of enzymatic hydrolysis was not performed.

[0149] 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.

[0150] Table 1 Performance test results of beef flavors prepared in Examples 1-5 and Comparative Examples 1-2

[0151] performance Example 1 Example 2 Example 3 Example 4 Example 5 Comparative Example 1 Comparative Example 2 Flavor 7(good) 9 (Excellent) 8(good) 9 (Excellent) 8(good) 6 (Yes) 5 (Yes) Content of important flavor substances 8(good) 8(good) 8(good) 10 (Excellent) 7(good) 5 (Yes) 6 (Yes) stability 8(good) 9 (Excellent) 9 (Excellent) 9 (Excellent) 7(good) 6 (Yes) 6 (Yes)

[0152] 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.

[0153] Regarding flavor, the flavor scores of Examples 2 and 4 reached 9 points (excellent), with excellent performance, high meaty aroma, 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), and due to the adjustment of the reducing sugar ratio (the weight ratio of xylose to glucose is 3:1), the flavor is good but there is still room for improvement. Example 1 scored 7 points (good), and all indicators are relatively balanced but the overall flavor is slightly inferior. Example 5 uses 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 and only scored 6 points (acceptable). 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 and only scored 5 points (acceptable). The flavor defect was large, highlighting the necessity of the second step of enzymatic hydrolysis.

[0154] 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.

[0155] Regarding stability, Examples 2, 3, and 4 all received a stability score of 9 (Excellent). During the 6-month accelerated test period, the flavor index showed only slight changes, thanks to the intelligent control system and reasonable formulation of the present invention. Example 1 received a score of 8 (Good), indicating good stability but slightly inferior to the previous three. Example 5 received a score of 7 (Good), but the stability was affected by the use of ordinary equipment. Comparative Examples 1 and 2 each received a score of 6 (Acceptable), indicating acceptable stability, but due to the lack of an enzymatic hydrolysis step, the long-term stability was poor.

[0156] In summary, the preparation method of the present invention, through dual enzymatic hydrolysis and intelligent control of the Maillard reaction, can effectively enhance the flavor, content of important flavor compounds, and stability of beef flavor. Optimizing the ratio of compound protease to reducing sugar significantly improves product quality. The intelligent system based on the present invention helps improve product quality and stability. Both the first and second enzymatic hydrolysis steps are essential for the formation of the flavor and production of flavor compounds in beef flavor.

[0157] Although specific embodiments have been shown and described in the present invention, it will be understood by those skilled in the art that various alternative and / or equivalent embodiments may be substituted for the specific embodiments shown and described without departing from the scope of the present invention. This application is intended to include any improvements or modifications to the specific embodiments discussed in the present invention. It will be understood by those skilled in the art that various modifications and changes may be made without departing from the scope of the present invention. Such modifications and changes are intended to fall within the scope of the present invention as defined.

Claims

1. A method for preparing beef flavor based on the synergistic enhancement of double enzymatic hydrolysis and Maillard reaction, characterized in that: The preparation method comprises: (1) performing a first enzymatic hydrolysis step, wherein a composite protease is added to the ground 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 composite protease comprises papain and flavor protease in a weight ratio of 1:4 to 1:5, and the weight percentage of the composite protease relative to the ground beef is 1%-2%; (2) performing a second enzymatic hydrolysis step, 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 increased to 55-60° C., and the enzymatic hydrolysis reaction is carried out for 1-2 hours, wherein the weight percentage of the lipase derived from Aspergillus oryzae relative to the product obtained in the first enzymatic hydrolysis step is 0.2%-0.3%; (3) raising the temperature of the reaction system to 85-90°C and maintaining it for 10-20 minutes to inactivate the enzyme to obtain an enzymatic hydrolyzate; (4) mixing the enzymatic hydrolysate obtained in step (3) with reducing sugar, thiamine and yeast extract to obtain a mixture, adjusting the pH value of the mixture to 5.0-5.5 and performing a Maillard reaction at a temperature of 110-150° C., wherein: the reducing sugar is a mixture of xylose and glucose in a weight ratio of 3:1 to 2:1; the mass ratio of the reducing sugar to the enzymatic hydrolysate is between 5:100 and 20:100; the mass ratio of the thiamine to the enzymatic hydrolysate is between 0.05:100 and 0.2:100; and the mass ratio of the yeast extract to the enzymatic hydrolysate is between 0.5:100 and 5:100, and the Maillard reaction in step (4) adopts a gradient temperature increase mode, wherein the Maillard reaction is performed in three temperature stages of 110° C., 130° C. and 150° C. in sequence; and (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 gum arabic, 1-5 parts by weight of sodium chloride, and 0.05-0.1 parts by weight of an antioxidant, and dried.

2. The method for preparing beef flavor based on the synergistic enhancement of double enzymatic hydrolysis and Maillard reaction according to claim 1, characterized in that: The papain is neutral papain.

3. The method for preparing beef flavor based on the synergistic enhancement of double enzymatic hydrolysis and Maillard reaction according to claim 1, characterized in that: The antioxidant is one or more of vitamin C palmitate and rosemary extract.

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