Method for extracting high-concentration seafood flavor raw material from kelp and clam

Abstract

The invention discloses a method for extracting a high-concentration seafood-flavor raw material from kelp and clam, which comprises the following steps of: crushing the kelp and clam, adding a proper amount of distilled water and protein compound enzyme into the kelp and clam, and performing enzymolysis under a certain enzymolysis condition; the preparation method comprises the following steps: taking papain, pancreatin, neutral protease and flavourzyme as enzymolysis combined liquid, performing enzymolysis and enzyme deactivation on the raw material homogenate, inoculating saccharomyces cerevisiae suspension into the enzymolysis liquid according to the inoculation amount of 5%, performing fermentation and fishy smell removal to obtain homogenate fermentation liquid, performing centrifugal filtration on the homogenate fermentation liquid, and performing ultrafiltration on the homogenate fermentation liquid. The extraction rate of the seafood flavor raw materials in the kelp and the clam is further improved through the excellent enzymolysis capacity of the enzymolysis combined liquid, so that the more thorough raw material utilization rate is achieved, the enzymolysis process of the compound enzyme is simple and easy to achieve, and the raw materials do not need to be subjected to other pretreatment in the using process.

Description

Technical Field

[0001] The invention relates to the technical field of seafood flavor raw material extraction, in particular to a method for extracting high-concentration seafood flavor raw materials from kelp and clams. Background Art

[0002] Kelp is one of the most abundant large algae. It is not only rich in polysaccharides, proteins, vitamins, minerals and other nutrients with development potential, but also contains alginate, alginate, mannitol and other substances with specific functions in the food industry. These unique nutrients have enabled it to be developed into various foods with high added value, becoming one of the most important research hotspots in recent years. It is worth mentioning that the large amount of seafood flavor substances in kelp makes it the most common ingredient in seasonings and is widely added to a variety of products. Clams are delicious, high in protein and low in fat. They are not only rich in polysaccharides, peptides, taurine, glycosaminoglycans and other bioactive substances, but also have a comprehensive and reasonable amino acid composition, which can meet the basic needs of the human body for 20 essential amino acids. Antioxidant peptides can also be prepared from marine biological raw materials such as clams, which also have the functions of scavenging free radicals, inhibiting lipid and protein oxidation and protecting cells. In addition, the various flavor components in clams, such as flavor nucleotides, organic acids and betaine, make clams rich in unique seafood flavor.

[0003] Although kelp and clam meat have rich nutritional value and very promising development potential, there are currently two problems: first, the strong fishy smell of both has brought great challenges to the food industry, especially to the extraction process of seafood flavor substances, limiting the application of extracts in subsequent processed foods; second, the existing extraction methods cannot guarantee complete high-concentration extraction, resulting in unnecessary cost losses. Summary of the invention

[0004] The present invention provides a method for extracting high-concentration seafood flavor raw materials from kelp and clams, which can effectively solve the problem that the strong fishy smell substances in kelp and clam meat mentioned in the above background technology bring great challenges to the food industry, especially to the extraction process of seafood flavor substances, which limits the application of the extracts in subsequent processed foods; secondly, the existing extraction methods cannot guarantee complete high-concentration extraction, resulting in unnecessary cost losses.

[0005] To achieve the above object, the present invention provides the following technical solutions: a method for extracting high-concentration seafood flavor raw materials from kelp and clams, wherein the kelp and clams are processed to extract high-concentration seafood flavor raw materials;

[0006] The specific steps include:

[0007] S1. Use kelp and clams commonly sold in the market, wash the selected fresh kelp and clam meat, and blend them with a homogenizer;

[0008] S2, after the kelp and the clam are crushed, an appropriate amount of distilled water and protein complex enzyme are added thereto, and enzymolysis is performed under certain enzymolysis conditions;

[0009] S3, after the enzymatic hydrolysis is completed, the enzymatic hydrolyzate is placed at 100°C for 20 minutes to inactivate the enzyme, and the enzymatic hydrolyzate is obtained after the enzyme inactivation treatment;

[0010] S4, inoculating the saccharomyces cerevisiae suspension into the enzymatic hydrolysate at an inoculation rate of 5% to ferment and remove the fishy smell, to obtain a homogenized fermentation liquid, and then ultrafiltering it after centrifugal filtration;

[0011] S5. After centrifugal ultrafiltration, the upper layer of hydrolyzed protein liquid is taken to detect and analyze the quality of the homogenized fermentation liquid;

[0012] S6. Then, the homogenized fermentation liquid is subjected to microwave-vacuum freeze-drying treatment, and after drying, it is packaged to obtain a finished product.

[0013] According to the above technical solution, in S2, during the enzymatic hydrolysis process, specifically includes:

[0014] S201, filtering the raw material homogenate after fermentation, and performing enzymolysis on the filter residue under different pH conditions under the same material-liquid ratio, enzyme activity and single enzyme composition ratio;

[0015] S202, filtering the raw material homogenate after fermentation, and performing enzymolysis on the filter residue under different temperature conditions under the same material-liquid ratio, enzyme activity and single enzyme composition ratio;

[0016] S203, filtering the raw material homogenate after fermentation, and performing enzymolysis on the filter residue under the same material-liquid ratio, enzyme activity and single enzyme composition ratio for different time conditions;

[0017] S204, filtering the raw material homogenate after fermentation, and performing enzymolysis on the filter residue under different enzyme addition conditions under the same material-liquid ratio, enzyme activity and single enzyme composition ratio;

[0018] S205, filtering the raw material homogenate after fermentation, and performing enzymatic hydrolysis on the filter residue under different enzymatic hydrolysis conditions under the same material-liquid ratio, enzyme activity and single enzyme composition ratio.

[0019] According to the above technical scheme, in S201, the material-liquid ratio is 1:2, the papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 5-9, the enzymatic hydrolysis temperature is 50°C-60°C, the enzymatic hydrolysis time is 3-5h, and the enzyme addition amount is 0.2%-0.4%;

[0020] In the S202, the material-liquid ratio is 1:2, the composite enzyme includes papain: pancreatin: neutral protease: flavor protease = 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 6-8, the enzymatic hydrolysis temperature is 45°C-65°C, the enzymatic hydrolysis time is 3-5h, and the enzyme addition amount is 0.2%-0.4%;

[0021] In the S203, the material-liquid ratio is 1:2, the ratio of papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 6-8, the enzymatic hydrolysis temperature is 50°C-60°C, the enzymatic hydrolysis time is 2-6h, and the enzyme addition amount is 0.2%-0.4%;

[0022] In the S204, the material-liquid ratio is 1:2, the ratio of papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 6-8, the enzymatic hydrolysis temperature is 50°C-60°C, the enzymatic hydrolysis time is 3-5h, and the enzyme addition amount is 0.1%-0.5%;

[0023] In S205, response surface optimization test is used to optimize relevant process parameters of enzymatic hydrolysis, and the relevant process parameters include enzymatic hydrolysis pH, enzymatic hydrolysis temperature, enzymatic hydrolysis time and material-liquid ratio, wherein the material-liquid ratio is 1:2, and the ratio of papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, and the enzyme activity is 20000U / g.

[0024] According to the above technical scheme, in S205, specifically, a BBD experimental design composite model is selected to perform a four-factor three-level response surface analysis test, in which the enzymatic pH (A), enzymatic temperature (B), enzymatic time (C) and enzyme addition amount (D) are used as independent variables, and the hydrolysis degree (Y) is used as the response value. The response surface test is performed using Design Expert 8.0.6 software, and then the response surface results are subjected to multivariate regression fitting analysis;

[0025] Design Expert 8.0.6 software was used to perform multiple regression fitting on the experimental model. The multiple regression equation of the hydrolysis degree (Y) and the enzymatic pH (A), enzymatic temperature (B), enzymatic time (C) and enzyme addition amount (D) was as follows:

[0026] Y=29.60-0.50A-0.33B+0.75C+0.92D-0.50AB+1.00AC-1.50AD+0.000BC+0.000BD+1.25CD-4.84A 2 -5.59B 2 -2.47C 2 -2.22D 2 .

[0027] According to the above technical solution, in S4, the fermentation time is 3 days, the fermentation temperature is 37° C., and the fermentation speed is 220 R / min.

[0028] According to the above technical scheme, in S5, in the specific detection and analysis process, 1 mL of the hydrolyzed protein solution is taken and fixed to 50 mL, 1 mL of the diluent is taken into a test tube, and 1 mL of distilled water and 1 mL of ninhydrin colorimetric agent are added, mixed and placed in a boiling water bath for 15 min, and a blank experiment is performed at the same time;

[0029] The standard curve regression equation is: y = 0.0229x-0.0077, R 2 =0.9994, the content of peptide bonds in the hydrolyzed protein solution (mmol / g) was calculated using the standard curve;

[0030] The following formula is used to calculate the size of DH:

[0031]

[0032] In the formula: DH is the degree of hydrolysis (%), h is the number of peptide bonds cleaved per gram of protein after hydrolysis (mmol / g), h1 is the number of peptide bonds per gram of raw protein (mmol / g), and kelp and clam peptides are taken as 8.0.

[0033] According to the above technical solution, in S6, the pre-freezing temperature is -20°C and the time is 4 hours, the freeze-drying temperature is -55°C and the time is 24 hours, and the vacuum degree is maintained at 5Pa.

[0034] Compared with the prior art, the present invention has the following beneficial effects: the present invention has a scientific and reasonable structure and is safe and convenient to use:

[0035] 1. The protein complex enzyme of the present invention only contains papain, pancreatin, neutral protease and flavor protease, all of which are food raw materials, without chemical additives, safe ingredients, green and environmentally friendly, simple preparation method of the protein complex enzyme, easy operation, and only need to be added to the kelp and clam meat homogenate, without other processing operations, while the protein complex enzyme is based on the excellent properties of pancreatin, and the kelp and clam meat homogenate is treated with papain, pancreatin, neutral protease and flavor protease four enzyme combinations, based on the excellent properties of pancreatin, the synergistic effect between the components enables the protein complex enzyme to well improve the enzymolysis degree of the homogenate, so as to solve the problem of low extraction rate of seafood flavor substances, and there is no substance with strong odor among the various materials of the protein complex enzyme, which will not affect the final extract.

[0036] 2. The present invention uses papain, pancreatin, neutral protease and flavor protease as the enzymatic hydrolysis combination liquid, and the raw material homogenate is enzymatically hydrolyzed and enzyme-killed, and then fermented with brewer's yeast to remove the fishy smell, and the enzymatic hydrolysis liquid is obtained after centrifugal filtration. The excellent enzymatic hydrolysis ability of the enzymatic hydrolysis combination liquid further improves the extraction rate of seafood flavor raw materials in kelp and clam, so as to achieve a more thorough raw material utilization rate. The enzymatic hydrolysis process of the composite enzyme is simple and easy to implement, and no other pretreatment of the raw materials is required during use;

[0037] In addition, microwave-vacuum freeze-drying technology is used to reduce the problem of low extraction rate of seafood flavor materials in kelp and clams, and minimize the loss of seafood flavor substances during extraction. Brewer's yeast can almost completely remove specific fishy substances in kelp and clams, reducing other aroma components except seafood flavor substances. The enzymatic hydrolysis method is simple, efficient and widely used. Appropriate complex enzymes can make the enzymatic hydrolysis of seafood substances more complete and the extraction of seafood flavor substances more thorough. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] The accompanying drawings are used to provide further understanding of the present invention and constitute a part of the specification. They are used to explain the present invention together with the embodiments of the present invention and do not constitute a limitation of the present invention.

[0039] In the attached picture:

[0040] Figure 1 is a flow chart of the steps of the extraction method of the present invention;

[0041] Figure 2 Schematic diagram of the effect of different enzymatic hydrolysis pH on the degree of protein hydrolysis of raw material homogenate in the present invention;

[0042] Figure 3 Schematic diagram of the effect of different enzymatic hydrolysis temperatures on the degree of protein hydrolysis of raw material homogenate in the present invention;

[0043] Figure 4 Schematic diagram of the effect of different enzymatic hydrolysis times on the degree of protein hydrolysis of the raw material homogenate in the present invention

[0044] Figure 5 Schematic diagram of the effect of different enzyme addition amounts on the degree of protein hydrolysis of raw material homogenate in the present invention;

[0045] Figure 6 It is the interaction response surface and contour map of various factors with hydrolysis degree as response in the present invention. DETAILED DESCRIPTION

[0046] The preferred embodiments of the present invention are described below in conjunction with the accompanying drawings. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0047] Example 1: Figure 1 As shown, the present invention provides a technical solution, a method for extracting high-concentration seafood flavor raw materials from kelp and clams, and processing kelp and clams to extract high-concentration seafood flavor raw materials;

[0048] The specific steps include:

[0049] S1. Use kelp and clams commonly sold in the market, wash the selected fresh kelp and clam meat, and blend them with a homogenizer;

[0050] S2, after the kelp and the clam are crushed, an appropriate amount of distilled water and protein complex enzyme are added thereto, and enzymolysis is performed under certain enzymolysis conditions;

[0051] S3, after the enzymatic hydrolysis is completed, the enzymatic hydrolyzate is placed at 100°C for 20 minutes to inactivate the enzyme, and the enzymatic hydrolyzate is obtained after the enzyme inactivation treatment;

[0052] S4, inoculating the saccharomyces cerevisiae suspension into the enzymatic hydrolysate at an inoculation rate of 5% to ferment and remove the fishy smell, to obtain a homogenized fermentation liquid, and then ultrafiltering it after centrifugal filtration;

[0053] S5. After centrifugal ultrafiltration, the upper layer of hydrolyzed protein liquid is taken to detect and analyze the quality of the homogenized fermentation liquid;

[0054] S6. Then, the homogenized fermentation liquid is subjected to microwave-vacuum freeze-drying treatment, and after drying, it is packaged to obtain a finished product.

[0055] Based on the above technical solution, in S2, during the enzymatic hydrolysis process, the fermented raw material homogenate is filtered, and under the same material-liquid ratio, enzyme activity and single enzyme composition ratio, the filter residue is enzymatically hydrolyzed at enzymatic hydrolysis pH of 5, 6, 7, 8 and 9 respectively;

[0056] The material-liquid ratio is 1:2, the ratio of papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, the enzyme activity is 20000U / g, the enzymolysis temperature is 55°C, the enzymolysis time is 4h, and the enzyme addition amount is 0.3%;

[0057] Based on the above technical solution, in S4, the fermentation time is 3 days, the fermentation temperature is 37°C, and the fermentation speed is 220R / min.

[0058] Based on the above technical scheme, in S5, in the specific detection and analysis process, 1 mL of the hydrolyzed protein solution was taken and fixed to 50 mL, 1 mL of the dilution solution was taken into a test tube, and 1 mL of distilled water and 1 mL of ninhydrin colorimetric agent were added, mixed and placed in a boiling water bath for 15 min, and a blank experiment was performed at the same time;

[0059] The standard curve regression equation is: y = 0.0229x-0.0077, R 2 =0.9994, the content of peptide bonds in the hydrolyzed protein solution (mmol / g) was calculated using the standard curve;

[0060] The following formula is used to calculate the size of DH:

[0061]

[0062] In the formula: DH is the degree of hydrolysis (%), h is the number of peptide bonds cleaved per gram of protein after hydrolysis (mmol / g), h1 is the number of peptide bonds per gram of raw protein (mmol / g), and kelp and clam peptides are taken as 8.0.

[0063] Based on the above technical solution, in S6, the pre-freezing temperature is -20°C, the time is 4 hours, the freeze-drying temperature is -55°C, the time is 24 hours, and the vacuum degree is maintained at 5Pa.

[0064] Example 2: Figure 1 As shown, the present invention provides a technical solution, a method for extracting high-concentration seafood flavor raw materials from kelp and clams, and processing kelp and clams to extract high-concentration seafood flavor raw materials;

[0065] The specific steps include:

[0066] S1. Use kelp and clams commonly sold in the market, wash the selected fresh kelp and clam meat, and blend them with a homogenizer;

[0067] S2, after the kelp and the clam are crushed, an appropriate amount of distilled water and protein complex enzyme are added thereto, and enzymolysis is performed under certain enzymolysis conditions;

[0068] S3, after the enzymatic hydrolysis is completed, the enzymatic hydrolyzate is placed at 100°C for 20 minutes to inactivate the enzyme, and the enzymatic hydrolyzate is obtained after the enzyme inactivation treatment;

[0069] S4, inoculating the saccharomyces cerevisiae suspension into the enzymatic hydrolysate at an inoculation rate of 5% to ferment and remove the fishy smell, to obtain a homogenized fermentation liquid, and then ultrafiltering it after centrifugal filtration;

[0070] S5. After centrifugal ultrafiltration, the upper layer of hydrolyzed protein liquid is taken to detect and analyze the quality of the homogenized fermentation liquid;

[0071] S6. Then, the homogenized fermentation liquid is subjected to microwave-vacuum freeze-drying treatment, and after drying, it is packaged to obtain a finished product.

[0072] Based on the above technical solution, in S2, during the enzymatic hydrolysis process, the fermented raw material homogenate is filtered, and under the same material-liquid ratio, enzyme activity and single enzyme composition ratio, the filter residue is enzymatically hydrolyzed at enzymatic hydrolysis temperatures of 45°C, 50°C, 55°C, 60°C and 65°C respectively;

[0073] The material-liquid ratio is 1:2, the ratio of papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 7, the enzymatic hydrolysis time is 4h, and the enzyme addition amount is 0.3%;

[0074] Based on the above technical solution, in S4, the fermentation time is 3 days, the fermentation temperature is 37°C, and the fermentation speed is 220R / min.

[0075] Based on the above technical scheme, in S5, in the specific detection and analysis process, 1 mL of the hydrolyzed protein solution was taken and fixed to 50 mL, 1 mL of the dilution solution was taken into a test tube, and 1 mL of distilled water and 1 mL of ninhydrin colorimetric agent were added, mixed and placed in a boiling water bath for 15 min, and a blank experiment was performed at the same time;

[0076] The standard curve regression equation is: y = 0.0229x-0.0077, R 2 =0.9994, the content of peptide bonds in the hydrolyzed protein solution (mmol / g) was calculated using the standard curve;

[0077] The following formula is used to calculate the size of DH:

[0078]

[0079] In the formula: DH is the degree of hydrolysis (%), h is the number of peptide bonds cleaved per gram of protein after hydrolysis (mmol / g), h1 is the number of peptide bonds per gram of raw protein (mmol / g), and kelp and clam peptides are taken as 8.0.

[0080] Based on the above technical solution, in S6, the pre-freezing temperature is -20°C, the time is 4 hours, the freeze-drying temperature is -55°C, the time is 24 hours, and the vacuum degree is maintained at 5Pa.

[0081] Example 3: Figure 1 As shown, the present invention provides a technical solution, a method for extracting high-concentration seafood flavor raw materials from kelp and clams, and processing kelp and clams to extract high-concentration seafood flavor raw materials;

[0082] The specific steps include:

[0083] S1. Use kelp and clams commonly sold in the market, wash the selected fresh kelp and clam meat, and blend them with a homogenizer;

[0084] S2, after the kelp and the clam are crushed, an appropriate amount of distilled water and protein complex enzyme are added thereto, and enzymolysis is performed under certain enzymolysis conditions;

[0085] S3, after the enzymatic hydrolysis is completed, the enzymatic hydrolyzate is placed at 100°C for 20 minutes to inactivate the enzyme, and the enzymatic hydrolyzate is obtained after the enzyme inactivation treatment;

[0086] S4, inoculating the saccharomyces cerevisiae suspension into the enzymatic hydrolysate at an inoculation rate of 5% to perform fermentation and deodorization to obtain a homogenized fermentation liquid, and then ultrafiltration is performed after centrifugal filtration;

[0087] S5. After centrifugal ultrafiltration, the upper layer of hydrolyzed protein liquid is taken to detect and analyze the quality of the homogenized fermentation liquid;

[0088] S6. Then, the homogenized fermentation liquid is subjected to microwave-vacuum freeze-drying treatment, and after drying, it is packaged to obtain a finished product.

[0089] Based on the above technical solution, in S2, during the enzymatic hydrolysis process, the fermented raw material homogenate is filtered, and under the same material-liquid ratio, enzyme activity and single enzyme composition ratio, the filter residue is enzymatically hydrolyzed under the conditions of enzymatic hydrolysis time of 2h, 3h, 4h, 5h and 6h respectively;

[0090] The material-liquid ratio is 1:2, the ratio of papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 7, the enzymatic hydrolysis temperature is 55°C, and the enzyme addition amount is 0.3%;

[0091] Based on the above technical solution, in S4, the fermentation time is 3 days, the fermentation temperature is 37°C, and the fermentation speed is 220R / min.

[0092] Based on the above technical scheme, in S5, in the specific detection and analysis process, 1 mL of the hydrolyzed protein solution was taken and fixed to 50 mL, 1 mL of the dilution solution was taken into a test tube, and 1 mL of distilled water and 1 mL of ninhydrin colorimetric agent were added, mixed and placed in a boiling water bath for 15 min, and a blank experiment was performed at the same time;

[0093] The standard curve regression equation is: y = 0.0229x-0.0077, R 2 =0.9994, the content of peptide bonds in the hydrolyzed protein solution (mmol / g) was calculated using the standard curve;

[0094] The following formula is used to calculate the size of DH:

[0095]

[0096] In the formula: DH is the degree of hydrolysis (%), h is the number of peptide bonds cleaved per gram of protein after hydrolysis (mmol / g), h1 is the number of peptide bonds per gram of raw protein (mmol / g), and kelp and clam peptides are taken as 8.0.

[0097] Based on the above technical solution, in S6, the pre-freezing temperature is -20°C, the time is 4 hours, the freeze-drying temperature is -55°C, the time is 24 hours, and the vacuum degree is maintained at 5Pa.

[0098] Example 4: Figure 1 As shown, the present invention provides a technical solution, a method for extracting high-concentration seafood flavor raw materials from kelp and clams, and processing kelp and clams to extract high-concentration seafood flavor raw materials;

[0099] The specific steps include:

[0100] S1. Use kelp and clams commonly sold in the market, wash the selected fresh kelp and clam meat, and blend them with a homogenizer;

[0101] S2, after the kelp and the clam are crushed, an appropriate amount of distilled water and protein complex enzyme are added thereto, and enzymolysis is performed under certain enzymolysis conditions;

[0102] S3, after the enzymatic hydrolysis is completed, the enzymatic hydrolyzate is placed at 100°C for 20 minutes to inactivate the enzyme, and the enzymatic hydrolyzate is obtained after the enzyme inactivation treatment;

[0103] S4, inoculating the saccharomyces cerevisiae suspension into the enzymatic hydrolysate at an inoculation rate of 5% to perform fermentation and deodorization to obtain a homogenized fermentation liquid, and then ultrafiltration is performed after centrifugal filtration;

[0104] S5. After centrifugal ultrafiltration, the upper layer of hydrolyzed protein liquid is taken to detect and analyze the quality of the homogenized fermentation liquid;

[0105] S6. Then, the homogenized fermentation liquid is subjected to microwave-vacuum freeze-drying treatment, and after drying, it is packaged to obtain a finished product.

[0106] Based on the above technical solution, in S2, during the enzymatic hydrolysis process, the fermented raw material homogenate is filtered, and under the same material-liquid ratio, enzyme activity and single enzyme composition ratio, the filter residue is enzymatically hydrolyzed at enzyme addition amounts of 0.1%, 0.2%, 0.3%, 0.4% and 0.5%, respectively;

[0107] The solid-liquid ratio is 1:2, the ratio of papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 7, the enzymatic hydrolysis temperature is 55°C, and the enzymatic hydrolysis time is 4h;

[0108] Based on the above technical solution, in S4, the fermentation time is 3 days, the fermentation temperature is 37°C, and the fermentation speed is 220R / min.

[0109] Based on the above technical scheme, in S5, in the specific detection and analysis process, 1 mL of the hydrolyzed protein solution was taken and fixed to 50 mL, 1 mL of the dilution solution was taken into a test tube, and 1 mL of distilled water and 1 mL of ninhydrin colorimetric agent were added, mixed and placed in a boiling water bath for 15 min, and a blank experiment was performed at the same time;

[0110] The standard curve regression equation is: y = 0.0229x-0.0077, R 2 =0.9994, the content of peptide bonds in the hydrolyzed protein solution (mmol / g) was calculated using the standard curve;

[0111] The following formula is used to calculate the size of DH:

[0112]

[0113] In the formula: DH is the degree of hydrolysis (%), h is the number of peptide bonds cleaved per gram of protein after hydrolysis (mmol / g), h1 is the number of peptide bonds per gram of raw protein (mmol / g), and kelp and clam peptides are taken as 8.0.

[0114] Based on the above technical solution, in S6, the pre-freezing temperature is -20°C, the time is 4 hours, the freeze-drying temperature is -55°C, the time is 24 hours, and the vacuum degree is maintained at 5Pa.

[0115] Example 5: Figure 1 As shown, the present invention provides a technical solution, a method for extracting high-concentration seafood flavor raw materials from kelp and clams, and processing kelp and clams to extract high-concentration seafood flavor raw materials;

[0116] The specific steps include:

[0117] S1. Use kelp and clams commonly sold in the market, wash the selected fresh kelp and clam meat, and blend them with a homogenizer;

[0118] S2, after the kelp and the clam are crushed, an appropriate amount of distilled water and protein complex enzyme are added thereto, and enzymolysis is performed under certain enzymolysis conditions;

[0119] S3, after the enzymatic hydrolysis is completed, the enzymatic hydrolyzate is placed at 100°C for 20 minutes to inactivate the enzyme, and the enzymatic hydrolyzate is obtained after the enzyme inactivation treatment;

[0120] S4, inoculating the saccharomyces cerevisiae suspension into the enzymatic hydrolysate at an inoculation rate of 5% to perform fermentation and deodorization to obtain a homogenized fermentation liquid, and then ultrafiltration is performed after centrifugal filtration;

[0121] S5. After centrifugal ultrafiltration, the upper layer of hydrolyzed protein liquid is taken to detect and analyze the quality of the homogenized fermentation liquid;

[0122] S6. Then, the homogenized fermentation liquid is subjected to microwave-vacuum freeze-drying treatment, and after drying, it is packaged to obtain a finished product.

[0123] Based on the above technical scheme, in S2, during the enzymatic hydrolysis process, the fermented raw material slurry is filtered, and the filter residue is enzymatically hydrolyzed under different enzymatic hydrolysis conditions when the solid-liquid ratio is 1:2, the composite enzyme includes papain: pancreatin: neutral protease: flavor protease = 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 7, the enzymatic hydrolysis temperature is 55°C, and the enzymatic hydrolysis time is 4h. The response surface optimization test is used to optimize the relevant process parameters of the enzymatic hydrolysis, and the relevant process parameters include enzymatic hydrolysis pH, enzymatic hydrolysis temperature, enzymatic hydrolysis time and solid-liquid ratio.

[0124] The BBD experimental design composite model was selected to conduct a response surface analysis experiment with four factors and three levels. The experiment used enzymatic pH (A), enzymatic temperature (B), enzymatic time (C) and enzyme addition amount (D) as independent variables, and hydrolysis degree (Y) as the response value. The response surface experiment was conducted using Design Expert8.0.6 software. The response surface experimental design and results are shown in Table 1. The response surface results were then subjected to multivariate regression fitting analysis, and the analysis results are shown in Table 2.

[0125] Table 1 shows the response surface analysis design and results of the effect of enzymatic hydrolysis conditions on the hydrolysis degree of raw material homogenate

[0126]

[0127]

[0128] Table 2 Analysis of variance of regression equation with hydrolysis degree as response

[0129]

[0130]

[0131] Design Expert 8.0.6 software was used to perform multiple regression fitting on the experimental model. The multiple regression equation of the hydrolysis degree (Y) and the enzymatic pH (A), enzymatic temperature (B), enzymatic time (C) and enzyme addition amount (D) was as follows:

[0132] Y=29.60-0.50A-0.33B+0.75C+0.92D-0.50AB+1.00AC-1.50AD+0.000BC+0.000BD+1.25CD-4.84A 2 -5.59B 2 -2.47C 2 -2.22D 2 .

[0133] Based on the above technical solution, in S4, the fermentation time is 3 days, the fermentation temperature is 37°C, and the fermentation speed is 220R / min.

[0134] Based on the above technical scheme, in S5, in the specific detection and analysis process, 1 mL of the hydrolyzed protein solution was taken and fixed to 50 mL, 1 mL of the dilution solution was taken into a test tube, and 1 mL of distilled water and 1 mL of ninhydrin colorimetric agent were added, mixed and placed in a boiling water bath for 15 min, and a blank experiment was performed at the same time;

[0135] The standard curve regression equation is: y = 0.0229x-0.0077, R 2 =0.9994, the content of peptide bonds in the hydrolyzed protein solution (mmol / g) was calculated using the standard curve;

[0136] The following formula is used to calculate the size of DH:

[0137]

[0138] In the formula: DH is the degree of hydrolysis (%), h is the number of peptide bonds cleaved per gram of protein after hydrolysis (mmol / g), h1 is the number of peptide bonds per gram of raw protein (mmol / g), and kelp and clam peptides are taken as 8.0.

[0139] Based on the above technical solution, in S6, the pre-freezing temperature is -20°C, the time is 4 hours, the freeze-drying temperature is -55°C, the time is 24 hours, and the vacuum degree is maintained at 5Pa.

[0140] Comparative Example 1: A portion of fresh kelp and clam meat was washed and then crushed using a homogenizer, distilled water and 0.3% protein complex enzyme were added, the solid-liquid ratio of distilled water was 1:2, the enzymolysis time was 4 hours, the enzymolysis pH was 7, the enzymolysis temperature was 55°C, and after the enzymolysis was completed, the enzymolysis solution was placed at 100°C for 20 minutes to inactivate the enzyme to obtain an enzymolysis solution;

[0141] The brewer's yeast suspension is inoculated into the enzymatic hydrolyzate at an inoculation rate of 5% and fermented for 3 days at a fermentation temperature of 37° C. and a fermentation speed of 220 R / min. The homogenized fermentation liquid obtained is centrifugally filtered and then ultrafiltered, followed by microwave-vacuum freeze drying, with a pre-freezing temperature of -20° C. and a time of 4 hours, a freeze drying temperature of -55° C. and a time of 24 hours, and a vacuum degree maintained at 5 Pa. After drying, the finished product is packaged.

[0142] Comparative Example 2: Wash a portion of fresh kelp and clam meat, blend them with a homogenizer, add distilled water, and the ratio of distilled water to liquid is 1:2;

[0143] The brewer's yeast suspension is inoculated into a homogenate at an inoculation rate of 5% and fermented for 3 days at a fermentation temperature of 37°C and a fermentation speed of 220R / min. The homogenate fermentation liquid obtained is centrifugally filtered and then ultrafiltered, followed by microwave-vacuum freeze drying, with a pre-freezing temperature of -20°C and a time of 4 hours, a freeze drying temperature of -55°C and a time of 24 hours, and a vacuum degree maintained at 5Pa. After drying, the finished product is packaged.

[0144] The hydrolysis degree of the kelp and clam meat homogenates after enzymatic hydrolysis in Examples 1-4 and Comparative Example 2 was measured, and the results were as follows: Figure 2-5 As shown;

[0145] exist Figure 2 By comparing Comparative Example 2 with Example 1, it can be seen that the hydrolysis degree of kelp and clam meat enzymatic hydrolysates decreases with the increase of enzymatic hydrolysis pH, wherein the hydrolysis degree of the homogenate is the highest when pH=7, while the hydrolysis degree of the homogenate in Comparative Example 2 is the lowest, indicating that the enzymatic hydrolysis of the homogenate using a protein complex enzyme can effectively improve the hydrolysis degree of the homogenate, and the degree of hydrolysis is the highest when the enzymatic hydrolysis pH is 7;

[0146] exist Figure 3 By comparing Comparative Example 2 with Example 2, it can be seen that the hydrolysis degree of the kelp and clam meat homogenate in Comparative Example 2 is the lowest, indicating that the treatment under appropriate enzymatic hydrolysis temperature conditions can effectively improve the hydrolysis degree of the kelp and clam meat homogenate, but as the enzymatic hydrolysis temperature continues to increase, the hydrolysis degree of the homogenate treated with the protein complex enzyme gradually decreases. When the enzymatic hydrolysis temperature is 55°C, the hydrolysis degree of the kelp and clam meat homogenate is the highest, indicating that the optimal enzymatic hydrolysis temperature of the protein complex enzyme treated homogenate is 55°C;

[0147] exist Figure 4By comparing Comparative Example 2 and Example 3, it can be seen that the degree of hydrolysis of the kelp and clam meat homogenates treated with the protein complex enzyme increases with the extension of the enzymatic hydrolysis time, until the enzymatic hydrolysis time reaches 4 hours, the growth rate of the hydrolysis degree begins to slow down with the extension of time, and the degree of hydrolysis of the homogenate in Comparative Example 2 is the lowest, indicating that the enzymatic hydrolysis treatment with the protein complex enzyme can effectively improve the degree of hydrolysis of the kelp and clam meat homogenates, and the degree of hydrolysis is the highest when the enzymatic hydrolysis time is 4 hours;

[0148] exist Figure 5 By comparing Comparative Example 2 with Example 4, it can be seen that the hydrolysis degree of the kelp and clam meat homogenate in Comparative Example 2 is the lowest, indicating that the appropriate degree of enzyme addition treatment can effectively improve the hydrolysis degree of the kelp and clam meat homogenate;

[0149] The hydrolysis degree of kelp and clam meat homogenate treated with protein compound enzymes increased with the increase of enzyme dosage, until the enzyme dosage reached 0.4%, the growth rate of hydrolysis degree began to slow down with the increase of enzyme dosage, indicating that protein compound enzyme enzymatic treatment can effectively improve the hydrolysis degree of kelp and clam meat homogenate, and the hydrolysis degree was the highest when the enzyme dosage was 0.4%.

[0150] As shown in Table 2, in the regression model of the hydrolysis degree of kelp and clam meat homogenate, the P value is <0.01, indicating that the regression equation of the model is extremely significant, and the lack of fit term is 0.2185>0.05, indicating that the lack of fit term is not significant and there is no lack of fit factor. The multiple correlation coefficient R 2 =0.9771, Corrected correlation coefficient Adj R 2 =0.9542, indicating that the model fits the actual situation well, the experimental error is small, and the prediction accuracy of the hydrolysis degree of kelp and clam meat homogenate is high;

[0151] In the regression model, the linear terms C, D and the interaction terms AD, CD and the quadratic term A 2 , B 2 , C 2 , D 2 Extremely significant P<0.01, the first-order term A and the interaction term AC are significant, P<0.05, the test results show that there is no simple linear relationship between the test factor and the response value;

[0152] The existing common protein complex enzymes are not suitable for the substrate and the enzymatic hydrolysis is not sufficient. When extracting seafood flavor raw materials from kelp and clams, they often face the problem of low hydrolysis, which leads to low extraction rate and low extraction concentration. The low extraction rate will cause unnecessary raw material losses for related manufacturers. Therefore, the hydrolysis degree is one of the important indicators for evaluating the extraction rate of seafood flavor raw materials in kelp and clams. Improving the hydrolysis degree has important practical significance.

[0153] The response surface diagram and contour diagram of the interaction between the two factors affecting the hydrolysis degree of kelp and clam meat homogenate are shown in the figure. Figure 6 As shown;

[0154] Depend on Figure 6 It can be seen that the contour lines of enzymatic hydrolysis pH (A) and enzymatic hydrolysis time (C) are elliptical, and the response surface is relatively steep, indicating that the interaction between AC is significant. The contour lines of enzymatic hydrolysis pH (A) and enzyme addition amount (D), enzymatic hydrolysis time (C) and enzyme addition amount (D) are also elliptical, and the response surface is steeper, indicating that the interaction between AD and CD also has a significant effect on the enzymatic hydrolysis effect of the raw material homogenate, and their interaction has a greater effect on the enzymatic hydrolysis effect of the homogenate.

[0155] The response surface diagram can evaluate the effect of the interaction of experimental factors on the response value to determine the optimal horizontal range. The steeper the slope of the response surface, the more sensitive the response value is to the change of conditions. On the contrary, the gentler the slope of the response surface, the more sensitive the response value is to the change of conditions. On the contrary, the gentler the slope of the response surface, the more sensitive the response value is to the change of treatment conditions. The optimal process parameter regression equation of the hydrolysis degree of kelp and clam meat homogenate under different enzymatic treatment conditions was fitted and optimized by Design Expert8.0.6 software. The optimal enzymatic conditions of kelp and clam meat homogenate treated with protein complex enzymes were enzymatic pH 7, enzymatic temperature 55℃, enzymatic time 4h and enzyme addition 0.4%;

[0156] According to the treatment method of comparative example 1, the hydrolysis degree of the kelp and clam meat homogenate treated under this condition reaches 29, and the effect after treatment is good.

[0157] Finally, it should be noted that the above description is only a preferred example of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for extracting high-concentration seafood flavor raw materials from kelp and clams, characterized in that: Kelp and clams are processed to extract high concentrations of seafood flavor ingredients; The specific steps include: S1. Use kelp and clams commonly sold in the market, wash the selected fresh kelp and clam meat, and blend them with a homogenizer; S2, after the kelp and the clam are crushed, an appropriate amount of distilled water and protein complex enzyme are added thereto, and enzymolysis is performed under certain enzymolysis conditions; S3, after the enzymatic hydrolysis is completed, the enzymatic hydrolyzate is placed at 100°C for 20 minutes to inactivate the enzyme, and the enzymatic hydrolyzate is obtained after the enzyme inactivation treatment; S4, inoculating the saccharomyces cerevisiae suspension into the enzymatic hydrolysate at an inoculation rate of 5% to ferment and remove the fishy smell, to obtain a homogenized fermentation liquid, and then ultrafiltering it after centrifugal filtration; S5. After centrifugal ultrafiltration, the upper layer of hydrolyzed protein liquid is taken to detect and analyze the quality of the homogenized fermentation liquid; S6. Then, the homogenized fermentation liquid is subjected to microwave-vacuum freeze-drying treatment, and after drying, it is packaged to obtain a finished product.

2. The method for extracting high-concentration seafood flavor raw materials from kelp and clams according to claim 1, characterized in that: In the S2, during the enzymatic hydrolysis process, specifically includes: S201, filtering the raw material homogenate after fermentation, and performing enzymolysis on the filter residue under different pH conditions under the same material-liquid ratio, enzyme activity and single enzyme composition ratio; S202, filtering the raw material homogenate after fermentation, and performing enzymolysis on the filter residue under different temperature conditions under the same material-liquid ratio, enzyme activity and single enzyme composition ratio; S203, filtering the raw material homogenate after fermentation, and performing enzymolysis on the filter residue under the same material-liquid ratio, enzyme activity and single enzyme composition ratio for different time conditions; S204, filtering the raw material homogenate after fermentation, and performing enzymolysis on the filter residue under different enzyme addition conditions under the same material-liquid ratio, enzyme activity and single enzyme composition ratio; S205, filtering the raw material homogenate after fermentation, and performing enzymatic hydrolysis on the filter residue under different enzymatic hydrolysis conditions under the same material-liquid ratio, enzyme activity and single enzyme composition ratio.

3. A method for extracting high-concentration seafood flavor raw materials from kelp and clams according to claim 2, characterized in that: In the S201, the material-liquid ratio is 1:2, the ratio of papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 5-9, the enzymatic hydrolysis temperature is 50°C-60°C, the enzymatic hydrolysis time is 3-5h, and the enzyme addition amount is 0.2%-0.4%; In the S202, the material-liquid ratio is 1:2, the composite enzyme includes papain: pancreatin: neutral protease: flavor protease = 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 6-8, the enzymatic hydrolysis temperature is 45°C-65°C, the enzymatic hydrolysis time is 3-5h, and the enzyme addition amount is 0.2%-0.4%; In the S203, the material-liquid ratio is 1:2, the ratio of papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 6-8, the enzymatic hydrolysis temperature is 50°C-60°C, the enzymatic hydrolysis time is 2-6h, and the enzyme addition amount is 0.2%-0.4%; In the S204, the material-liquid ratio is 1:2, the ratio of papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, the enzyme activity is 20000U / g, the enzymatic hydrolysis pH is 6-8, the enzymatic hydrolysis temperature is 50°C-60°C, the enzymatic hydrolysis time is 3-5h, and the enzyme addition amount is 0.1%-0.5%; In S205, response surface optimization test is used to optimize relevant process parameters of enzymatic hydrolysis, and the relevant process parameters include enzymatic hydrolysis pH, enzymatic hydrolysis temperature, enzymatic hydrolysis time and material-liquid ratio, wherein the material-liquid ratio is 1:2, and the ratio of papain: pancreatin: neutral protease: flavor protease in the complex enzyme is 1:1:1:1, and the enzyme activity is 20000U / g.

4. The method for extracting high-concentration seafood flavor raw materials from kelp and clams according to claim 3, characterized in that: In the S205, specifically, a BBD experimental design composite model is selected to perform a response surface analysis experiment with four factors and three levels. The experiment uses enzymatic hydrolysis pH (A), enzymatic hydrolysis temperature (B), enzymatic hydrolysis time (C) and enzyme addition amount (D) as independent variables, and hydrolysis degree (Y) as the response value. The response surface experiment is performed using Design Expert 8.0.6 software, and then the response surface results are subjected to multivariate regression fitting analysis; Design Expert 8.0.6 software was used to perform multiple regression fitting on the experimental model. The multiple regression equation of the hydrolysis degree (Y) and the enzymatic pH (A), enzymatic temperature (B), enzymatic time (C) and enzyme addition amount (D) was as follows: Y=29.60-0.50A-0.33B+0.75C+0.92D-0.50AB+1.00AC-1.50AD+0.000BC+0.000BD+1.25CD-4.84A 2 -5.59B 2 -2.47C 2 -2.22D 2 。 5. The method for extracting high-concentration seafood flavor raw materials from kelp and clams according to claim 1, characterized in that: In S4, the fermentation time is 3 days, the fermentation temperature is 37° C., and the fermentation speed is 220 R / min.

6. The method for extracting high-concentration seafood flavor raw materials from kelp and clams according to claim 1, characterized in that: In the S5, in the specific detection and analysis process, 1 mL of the hydrolyzed protein solution is taken to make a volume of 50 mL, 1 mL of the diluent is taken into a test tube, and 1 mL of distilled water and 1 mL of ninhydrin colorimetric agent are added, mixed and placed in a boiling water bath for 15 min, and a blank experiment is performed at the same time; The standard curve regression equation is: y = 0.0229x-0.0077, R 2 =0.9994, the content of peptide bonds in the hydrolyzed protein solution (mmol / g) was calculated using the standard curve; The following formula is used to calculate the size of DH: In the formula: DH is the degree of hydrolysis (%), h is the number of peptide bonds cleaved per gram of protein after hydrolysis (mmol / g), h1 is the number of peptide bonds per gram of raw protein (mmol / g), and kelp and clam peptides are taken as 8.

0.

7. The method for extracting high-concentration seafood flavor raw materials from kelp and clams according to claim 1, characterized in that: In S6, the pre-freezing temperature is -20°C and the time is 4 hours, the freeze-drying temperature is -55°C and the time is 24 hours, and the vacuum degree is maintained at 5Pa.

Images