Preparation method of flavor amino acid-glucose Maillard reaction product based on oyster meat

Abstract

The invention relates to the technical field of food science, and discloses a preparation method of a flavor amino acid-glucose Maillard reaction product based on oyster meat, which comprises the following steps: pretreating oyster meat, cleaning, mixing with distilled water according to a ratio of 1: 5, and mashing into slurry; then adding compound protease to carry out enzymolysis reaction, and carrying out enzyme deactivation and post-treatment after enzymolysis; the optimal process parameters are determined by optimizing Maillard reaction conditions through a single-factor test and an orthogonal test; mixing the oyster enzymatic hydrolysate with glucose under optimized conditions to perform Maillard reaction, cooling after the reaction is finished, performing centrifugal separation, and taking supernate as a product; according to the preparation method disclosed by the invention, the Maillard reaction product with good antioxidant activity and sensory evaluation is prepared by optimizing enzymolysis parameters and controlling Maillard reaction conditions, and the prepared Maillard reaction product is used as an additive to be applied to the initial stage of oyster meat pulping; and the quality of the oyster enzymatic hydrolysate obtained subsequently is directly improved by inhibiting lipid oxidation.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of food science and technology, in particular to a preparation method of flavor amino acid-glucose Maillard reaction products based on oyster meat. BACKGROUND

[0002] In the field of food processing, the preparation of flavor substances by Maillard reaction has become a research hotspot. Oyster meat is rich in nutrients such as protein, but its macromolecular protein structure is not conducive to the release and utilization of effective components. The Maillard reaction requires a rich supply of amino acids and other substrates. How to efficiently decompose the protein in oyster meat to provide sufficient substrates, optimize the reaction conditions to obtain products with good flavor and antioxidant properties, and ensure the microbial safety of the products and extend the shelf life are problems that need to be solved in this field. There is room for improvement in the existing technology in terms of enzyme hydrolysis of oyster meat, regulation of Maillard reaction conditions, and sterilization and preservation of products. Based on this, the present application aims to solve the above problems by using complex protease to hydrolyze oyster meat, precisely controlling Maillard reaction parameters, and introducing a pasteurization step, to prepare flavor amino acid-glucose Maillard reaction products with excellent flavor and functionality and safety and stability. SUMMARY

[0003] (I) Technical problems solved

[0004] In view of the deficiencies of the prior art, the present application provides a preparation method of flavor amino acid-glucose Maillard reaction products based on oyster meat, which has the advantages of efficient enzyme hydrolysis, optimized reaction conditions and ensured microbial safety, and solves the problems of insufficient release of effective components in oyster meat, poor flavor and antioxidant properties of Maillard reaction products, and short shelf life of products.

[0005] (II) Technical solutions

[0006] To achieve the above-mentioned purpose, the present application provides the following technical solutions: a preparation method of flavor amino acid-glucose Maillard reaction products based on oyster meat, comprising the following preparation steps:

[0007] S1, pretreatment of oyster meat: after removing the internal organs and impurities from fresh oyster meat, the oyster meat is washed clean in flowing clean water, and the washed oyster meat is mixed with distilled water at a ratio of 1:5 and added to a high-speed tissue crusher, and the oyster meat is completely crushed under the set operating conditions to form a uniform oyster meat slurry;

[0008] S2, complex protease hydrolysis: complex protease is added to the oyster meat slurry, and the enzyme hydrolysis reaction is carried out under neutral and constant temperature conditions. After hydrolysis, the temperature is raised, the enzyme is inactivated in boiling water, and finally the oyster hydrolysate is treated;

[0009] S3, design and optimize Maillard reaction conditions: the post-processing of oyster enzymatic hydrolysate by single factor test and orthogonal test to optimize the Maillard reaction conditions, determine the process parameters range: lysine and glucose mass ratio of 0.5-1.5:0.5-1.5, pH value is 7-9, reaction temperature is 100-130℃, reaction time is 60~100min, under the process parameter range of Maillard reaction conditions, finally get the best process parameters of Maillard reaction;

[0010] S4, Maillard reaction implementation: under the optimized conditions, the oyster enzymatic hydrolysate and glucose are mixed according to the set range of mass ratio, heated in neutral system at the set temperature, cooled to room temperature after reaction, centrifugal separation, and the supernatant is taken as the Maillard reaction product;

[0011] S5, sterilization: the Maillard reaction product is sterilized by pasteurization method;

[0012] S6, quality detection and packaging: after quality detection, the Maillard reaction product is aseptically packaged, and the aseptically packaged Maillard reaction product is used as a high-efficiency additive in the subsequent batch of oyster beating process S1 step.

[0013] Preferably, the operating conditions of the high-speed tissue crusher in S1 are: the speed is 10000-12000r / min, and the time is 3-5min.

[0014] Preferably, the compound protease in S2 is selected from one or a combination of trypsin, papain and neutral protease, and the addition amount of compound protease is between 28-32AU / kg.

[0015] Preferably, the compound protease enzymolysis conditions in S2 are: the enzymolysis reaction is carried out under the constant temperature conditions of pH value of 7-8 and temperature of 45-55℃ for 3-5 hours, and the enzyme is inactivated in 98-100℃ boiling water for 12-15min, and the chemical reaction formula is:

[0016] Protein + H2O compound protease peptide + amino acid

[0017] In the formula, protein represents the protein substrate in oyster meat, H2O represents water as the medium of hydrolysis reaction, compound protease represents one or a combination of trypsin, papain and neutral protease used, peptide represents the intermediate product produced in the enzymolysis process, and amino acid represents the small molecule amino acid finally produced by enzymolysis.

[0018] Preferably, the S2 oyster enzymatic hydrolysate post-processing: after enzyme inactivation, hot filtration through 100-200 mesh filter cloth, then using rotary evaporation method, the solid content of the obtained oyster enzymatic hydrolysate is controlled at 5%±0.5%, temporarily stored in a 2-4℃ refrigerator, and the subsequent Maillard reaction is carried out within 24 hours.

[0019] Preferably, the S3 Maillard reaction optimal process parameters are: lysine to glucose mass ratio of 1:1, pH value of 7.95, reaction temperature of 128℃, and reaction time of 82min.

[0020] Preferably, the S4 Maillard reaction is carried out according to the S3 optimized Maillard reaction conditions, after reaction, cooling to room temperature, centrifugal separation at 1-4℃, 9000-10000r / min for 10-15min, and after standing for 30-60min, taking the supernatant as the Maillard reaction product.

[0021] Preferably, the S5 Maillard reaction product is subjected to pasteurization at 80-85℃ for 10-15min.

[0022] Preferably, the S6 Maillard reaction product is subjected to quality detection of heavy metal content, microbial indicators, pesticide residues, pathogenic bacteria and coliform group.

[0023] Preferably, during the Maillard reaction, macromolecular peptides are subjected to thermal degradation to form a protein existing form containing 94%-100% free amino acids and small molecular peptides.

[0024] Compared with the prior art, the application provides a preparation method of a flavor amino acid-glucose Maillard reaction product based on oyster meat, which has the following beneficial effects:

[0025] 1、The application not only successfully prepares a Maillard reaction product with unique flavor and high antioxidant activity by optimizing oyster meat enzymatic hydrolysis and Maillard reaction conditions, but also effectively improves the content of free amino acids in oyster meat by using complex protease in the enzymatic hydrolysis process to provide rich substrates for the Maillard reaction; at the same time, by accurately controlling the parameters of the Maillard reaction, such as optimizing the ratio of lysine to glucose, pH value, reaction temperature and time, the product has excellent flavor and antioxidant properties, and the pasteurization step is introduced to effectively ensure the microbial safety of the final Maillard reaction product, thereby prolonging its shelf life, and the application innovatively applies the prepared Maillard reaction product as an additive to the initial stage of oyster meat beating, aiming to directly improve the quality (especially flavor stability) of the subsequent oyster enzymatic hydrolysate by inhibiting lipid oxidation, which provides a new and effective technical approach to improve the overall flavor and antioxidant capacity of oyster deep processing products. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 Preparation flow chart for the present application. DETAILED DESCRIPTION

[0027] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the protection scope of the present application.

[0028] Please refer to Figure 1 The preparation method of the flavor amino acid-glucose Maillard reaction product based on oyster meat comprises the following preparation steps:

[0029] S1, pretreatment of oyster meat: after selecting fresh oyster meat and removing viscera and impurities, the oyster meat is cleaned in flowing clean water, and the cleaned oyster meat is mixed with distilled water at a ratio of 1:5 (m / m) (when the mass of the oyster meat is 100 g, 500 mL of distilled water is added). The mixture is added into a high-speed tissue crusher, and the oyster meat is completely crushed under the set operating conditions to form a uniform oyster meat slurry, so as to release the cell contents and improve the subsequent enzymolysis efficiency.

[0030] S2, complex protease enzymolysis: complex protease is added to the oyster meat slurry, and the enzymolysis reaction is carried out under neutral and constant temperature conditions. After the enzymolysis, the temperature is increased, and the enzyme is inactivated in boiling water to stop the enzymolysis reaction. Finally, the obtained oyster enzymolysis liquid is post-treated to control the solid content of the oyster enzymolysis liquid.

[0031] S3, design and optimization of Maillard reaction conditions: the post-treated oyster enzymolysis liquid is used to optimize the Maillard reaction conditions through single factor test and orthogonal test, and the process parameter range is determined as follows: the mass ratio of lysine (Lys) to glucose is 0.5-1.5:0.5-1.5, the pH value is 7-9, the reaction temperature is 100-130°C, and the reaction time is 60-100 min. The Maillard reaction is carried out under the process parameter range, and finally the optimal process parameters of the Maillard reaction are obtained.

[0032] S4, implementation of Maillard reaction: under the optimized conditions, the oyster enzymolysis liquid is mixed with glucose at a set range mass ratio, heated and reacted in a neutral system at a set temperature, cooled to room temperature after the reaction is completed, centrifuged, and the supernatant is taken as the Maillard reaction product (MRPs).

[0033] S5, sterilization: the Maillard reaction product is sterilized by using the pasteurization method.

[0034] S6, quality detection and packaging: the Maillard reaction products are subjected to strict quality detection, including heavy metal content, microbial indicators (total number of colonies, pathogenic bacteria, coliform group) and pesticide residues, to ensure that all test values meet the national food safety standard limits, after the detection is qualified, the Maillard reaction products (MRPs) are subjected to sterile packaging, after the packaging is completed, sensory evaluation, antioxidant activity (such as DPPH clearance rate) determination and flavor substances (such as pyrazines, aldehydes and ketones) analysis are required to comprehensively evaluate its quality characteristics, finally, the sterile packaged Maillard reaction products will be used as a key high-efficiency additive in the subsequent batch of oyster beating process (i.e. S1 step), aiming to inhibit lipid oxidation and significantly improve the flavor quality of the final oyster enzymatic hydrolysate.

[0035] The glucose ratio and reaction temperature in the above steps S1-S6 will affect the flavor substances of the Maillard reaction products, and the deviation of the ratio or the too high / low temperature will lead to flavor deterioration, and the pH value and reaction time need to be strictly controlled, otherwise bitter taste or abnormal color will be easily produced, wherein the complex enzyme combination is superior to single enzyme, therefore, the enzymatic treatment is the key step to improve the content of free amino acids and flavor, and the lysine-glucose Maillard reaction products prepared by the method steps S1-S6 can be used as a high-efficiency additive, which is directly added to the oyster beating process (i.e. S1 step), and the application effect shows that the additive can effectively inhibit the lipid oxidation of oyster meat paste in the subsequent treatment, thereby improving the flavor quality of the final oyster enzymatic hydrolysate.

[0036] Advantages: the present application not only successfully prepares the Maillard reaction products (MRPs) with unique flavor and high antioxidant activity by optimizing the oyster meat enzymatic and Maillard reaction conditions, but also effectively improves the content of free amino acids in oyster meat by using complex protease in the enzymatic process, thereby providing rich substrates for the Maillard reaction; meanwhile, by accurately controlling the parameters of the Maillard reaction (such as the ratio of lysine to glucose, pH value, reaction temperature and time), the product has excellent flavor and antioxidant properties, and the introduction of the pasteurization step effectively ensures the microbial safety of the final Maillard reaction product, thereby prolonging its shelf life, and more importantly, the present application innovatively applies the prepared Maillard reaction products as an additive to the initial stage of oyster meat beating, aiming to directly improve the quality of the subsequent obtained oyster enzymatic hydrolysate by inhibiting lipid oxidation, and greatly improve the flavor stability, which provides a new and effective technical approach to improve the overall flavor and antioxidant capacity of oyster deep processing products.

[0037] Specifically, the operating conditions of the high-speed tissue crusher in S1 are as follows: the rotating speed is 10000-12000 r / min, and the time length is 3-5 min.

[0038] Specifically, the complex protease in S2 is selected from one or a combination of trypsin, papain, and neutral protease, and the addition amount of the complex protease is between 28-32 AU / kg, that is, for 1000 grams (1 kg) of oyster meat pulp, the number of enzyme activity units of the complex protease to be added is between 28 to 32 according to the addition amount of 28-32 AU / kg.

[0039] Specifically, the complex protease enzyme hydrolysis conditions in S2 are as follows: the enzyme hydrolysis reaction is carried out under constant temperature conditions of pH 7-8 and temperature 45-55℃ for 3-5 hours, and the enzyme is inactivated in boiling water at 98-100℃ for 12-15 min after the enzyme hydrolysis reaction, and the chemical reaction formula is as follows:

[0040] Protein + H2O complex protease peptide + amino acid

[0041] In the formula, protein represents the protein substrate in oyster meat, H2O represents water as the medium of hydrolysis reaction, complex protease represents one or a combination of trypsin, papain, and neutral protease used, peptide represents the intermediate product produced in the enzyme hydrolysis process, and amino acid represents the small molecule amino acid finally produced by enzyme hydrolysis.

[0042] Specifically, the oyster enzymatic hydrolysate in S2 is post-treated: after enzyme inactivation, hot filtration is carried out through 100-200 mesh filter cloth to remove unhydrolyzed protein particles, and then the rotary evaporation method is used to control the solid content of the obtained oyster enzymatic hydrolysate at 5%±0.5%, and the oyster enzymatic hydrolysate is temporarily stored in a 2-4℃ refrigerator, and the subsequent Maillard reaction is carried out within 24 hours.

[0043] Specifically, the optimal process parameters of the Maillard reaction in S3 are as follows: the mass ratio of lysine to glucose is 1:1, the pH value is 7.95, the reaction temperature is 128℃, and the reaction time is 82 min.

[0044] Specifically, the Maillard reaction in S4 is carried out according to the optimized Maillard reaction conditions in S3, and after the reaction is completed, it is cooled to room temperature, and then centrifuged at 1-4℃ and 9000-10000 r / min for 10-15 min, and after standing for 30-60 min, the supernatant is taken as the Maillard reaction product (MRPs).

[0045] Specifically, the Maillard reaction product in S5 is subjected to pasteurization at 80-85℃ for 10-15 minutes.

[0046] Specifically, the Maillard reaction product in S6 is subjected to quality detection of heavy metal content, microbial indicators, pesticide residues, pathogenic bacteria, and coliform bacteria, and the detection values are lower than the limit values specified in the national food safety standards.

[0047] Specifically, during the Maillard reaction process, the macromolecular peptides are thermally degraded to form a protein existing form containing 94% to 100% free amino acids and small molecular peptides.

[0048] The following are the process optimization group and the reference group of the unoptimized process of the Maillard reaction product prepared by the preparation method of the present application, specifically:

[0049] Table 1

[0050]

[0051]

[0052]

[0053] From the analysis of Table 1, it is obtained that:

[0054] Example 1 (optimum conditions) has the optimal comprehensive performance, the flavor substance types are many, the antioxidant activity is high, and the sensory evaluation is good, while Comparative Example 2 has no enzymatic hydrolysis and Comparative Example 7 has no sterilization due to the missing of the key step, resulting in unqualified Maillard reaction products.

[0055] Summary: The glucose ratio and the reaction temperature will affect the flavor substances of the Maillard reaction products, and the deviation of the ratio or the excessively high / low temperature will lead to flavor deterioration, and the pH value and the reaction time need to be strictly controlled, otherwise bitter taste or abnormal color will be easily produced, wherein the composite enzyme combination is superior to the single enzyme, and the enzymatic hydrolysis treatment is the key step to improve the content of free amino acids and the flavor.

[0056] In summary: the present application successfully prepares the Maillard reaction products (MRPs) with unique flavor and high antioxidant activity by optimizing the enzymatic hydrolysis and Maillard reaction conditions of oyster meat, the method uses composite protease in the enzymatic hydrolysis process to effectively improve the content of free amino acids in oyster meat, to provide rich substrates for the Maillard reaction; at the same time, by accurately controlling the parameters of the Maillard reaction (such as the ratio of lysine to glucose, pH value, reaction temperature and time), to ensure that the product has excellent flavor and antioxidant properties, and by introducing the pasteurization step, to effectively ensure the microbial safety of the final Maillard reaction product, thereby prolonging the shelf life of the Maillard reaction product.

[0057] Although the embodiments of the present application have been shown and described, it can be understood by those of ordinary skill in the art that various changes, modifications, replacements and variations can be made to these embodiments without departing from the principles and spirits of the present application, and the scope of the present application is defined by the appended claims and their equivalents.

Claims

1. A method for preparing flavor amino acids from oyster meat based on the Maillard reaction product with glucose, characterized in that, The preparation steps include the following: S1. Pre-treatment of oyster meat: Select fresh oyster meat, remove the internal organs and impurities, and wash it clean in running water. Mix the cleaned oyster meat with distilled water at a ratio of 1:5 and add it to a high-speed tissue homogenizer. Under the set operating conditions, the oyster meat is completely crushed to form a uniform oyster meat paste. S2. Complex protease hydrolysis: Complex protease is added to oyster meat paste and the enzymatic hydrolysis reaction is carried out under neutral and constant temperature conditions. After hydrolysis, the temperature is raised and the enzyme is inactivated in boiling water. Finally, the oyster hydrolysate is post-processed. S3. Design and optimize Maillard reaction conditions: The Maillard reaction conditions of the post-treated oyster enzymatic hydrolysate were optimized through single-factor experiments and orthogonal experiments. The process parameters were determined to be: lysine to glucose mass ratio of 0.5-1.5:0.5-1.5, pH value of 7-9, reaction temperature of 100-130℃, and reaction time of 60-100 min. The Maillard reaction was carried out under these process parameter ranges, and the optimal process parameters for the Maillard reaction were finally obtained. S4. Maillard reaction implementation: Under optimized conditions, oyster enzymatic hydrolysate and glucose are mixed at a set mass ratio. The mixture is heated in a neutral system at a set temperature to carry out the reaction. After the reaction is completed, the mixture is cooled to room temperature, centrifuged, and the supernatant is taken as the Maillard reaction product. S5. Sterilization: Pasteurization is used to sterilize the Maillard reaction products; S6. Quality Inspection and Packaging: After passing the quality inspection, the Maillard reaction product is aseptically packaged. This aseptically packaged Maillard reaction product is used as a high-efficiency additive in the S1 step of the oyster pulping process in subsequent batches.

2. The method for preparing flavor amino acids-glucose Maillard reaction products based on oyster meat according to claim 1, characterized in that, The operating conditions for the high-speed tissue homogenizer in S1 are: rotation speed of 10000-12000 r / min and duration of 3-5 min.

3. The method for preparing flavor amino acids-glucose Maillard reaction products based on oyster meat according to claim 1, characterized in that, The complex protease in S2 is selected from one or a combination of two of trypsin, papain, and neutral protease, and the amount of complex protease added is between 28-32 AU / kg.

4. The method for preparing flavor amino acids-glucose Maillard reaction products based on oyster meat according to claim 1, characterized in that, The enzymatic hydrolysis conditions for the complex protease in S2 are as follows: the enzymatic hydrolysis reaction is carried out for 3-5 hours under constant temperature conditions of pH 7-8 and 45-55℃, followed by enzyme inactivation in boiling water at 98-100℃ for 12-15 minutes. The chemical reaction formula is as follows: Protein + H2O complex protease peptide + amino acid In the formula, protein represents the protein substrate in oyster meat, H2O represents water as the medium for hydrolysis, complex protease represents one or a combination of two of the trypsin, papain, and neutral protease used, peptide represents the intermediate product produced during enzymatic hydrolysis, and amino acid represents the small molecule amino acid produced at the end of enzymatic hydrolysis.

5. The method for preparing flavor amino acids-glucose Maillard reaction products based on oyster meat according to claim 1, characterized in that, The post-treatment of the oyster enzymatic hydrolysate in S2 is as follows: after enzyme inactivation, the hydrolysate is filtered while hot through a 100-200 mesh filter cloth, and then the solid content of the obtained oyster enzymatic hydrolysate is controlled at 5% ± 0.5% by rotary evaporation. The hydrolysate is then temporarily stored in a refrigerator at 2-4℃ and the Maillard reaction is carried out within 24 hours.

6. The method for preparing flavor amino acids-glucose Maillard reaction products based on oyster meat according to claim 1, characterized in that, The optimal process parameters for the S3 melard reaction are: lysine to glucose mass ratio of 1:1, pH value of 7.95, reaction temperature of 128℃, and reaction time of 82 min.

7. The method for preparing flavor amino acids-glucose Maillard reaction products based on oyster meat according to claim 1, characterized in that, The Maillard reaction in S4 was carried out under the Maillard reaction conditions optimized in S3. After the reaction was completed, the mixture was cooled to room temperature and centrifuged at 1-4℃ and 9000-10000r / min for 10-15 min. After standing for 30-60 min, the supernatant was taken as the Maillard reaction product.

8. The method for preparing flavor amino acids-glucose Maillard reaction products based on oyster meat according to claim 1, characterized in that, The S5 melard reaction product is pasteurized at 80-85°C for 10-15 minutes.

9. The method for preparing flavor amino acids-glucose Maillard reaction products based on oyster meat according to claim 1, characterized in that, The S6 melilad reaction product was subjected to quality testing for heavy metal content, microbial indicators, pesticide residues, pathogenic bacteria, and coliform bacteria.

10. The method for preparing flavor amino acids-glucose Maillard reaction products based on oyster meat according to claim 1, characterized in that, During the Maillard reaction, macromolecular peptides undergo thermal degradation, forming a protein containing 94%–100% free amino acids and small peptides.

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