Marine-derived flavor base and method for preparing the same

Abstract

This invention provides a method for preparing a seafood seasoning base and the method thereof. The preparation method includes: enzymatically hydrolyzing seafood raw materials to prepare an enzymatic hydrolysate; performing multi-stage isoelectric point separation of umami amino acids and umami peptides in the enzymatic hydrolysate to prepare seasoning base A and supernatant A; subjecting supernatant A to aroma-enhancing treatment to obtain seasoning base B; and mixing seasoning base A and seasoning base B to obtain the seafood seasoning base. First, the seafood raw materials are enzymatically hydrolyzed using a composite enzyme, which can increase the content of umami peptides and umami amino acids in the hydrolysate. Then, based on the difference in isoelectric points between umami peptides, glutamic acid, and aspartic acid and other types of amino acids and peptides, multi-stage isoelectric point separation of umami amino acids and umami peptides is performed in the enzymatic hydrolysate to separate umami amino acids and umami peptides from the enzymatic hydrolysate, thereby increasing the content of umami amino acids and umami peptides in the final seafood seasoning base and enhancing the umami flavor of the seafood seasoning base.

Description

Technical Field

[0001] This invention relates to the field of seasoning preparation technology, and in particular to a seafood seasoning base and its preparation method. Background Technology

[0002] With the upgrading of residents' consumption, there is an increasing preference for natural and highly umami seasonings; the demand for umami-type seasonings made from natural raw materials is becoming more prominent, so it is necessary to develop a natural, highly umami-rich seafood seasoning base.

[0003] In related technologies, the preparation methods of umami seasoning base materials mainly include enzymatic hydrolysis and cooking. The enzymatic hydrolysate prepared by conventional enzymatic hydrolysis has a low content of umami amino acids and umami peptides. High-temperature cooking after enzymatic hydrolysis will also lead to the loss of umami amino acids and umami peptides. Therefore, the products prepared cannot meet the requirements of high umami. Summary of the Invention

[0004] Therefore, it is necessary to provide a seafood seasoning base and its preparation method to increase the content of umami amino acids and umami peptides in the seafood seasoning base and enhance the umami flavor of the seafood seasoning base.

[0005] The first aspect of the present invention provides a method for preparing a seafood seasoning base, comprising the following steps:

[0006] Seafood raw materials are subjected to enzymatic hydrolysis to prepare enzymatic hydrolysate;

[0007] The enzymatic hydrolysate is subjected to multi-stage isoelectric point separation of umami amino acids and umami peptides to prepare seasoning base A and supernatant A;

[0008] The supernatant A is subjected to aroma-enhancing treatment to obtain flavoring base B;

[0009] The seafood seasoning base is prepared by mixing the seasoning base A and the seasoning base B.

[0010] In some embodiments, the step of performing multi-stage isoelectric point separation of umami amino acids and umami peptides on the enzymatic hydrolysate to prepare seasoning base A and supernatant A includes:

[0011] Adjust the pH of the enzymatic hydrolysate to the first pH, add salt, and perform a first heat treatment to obtain heat treatment solution A. Separate the heat treatment solution A into a first precipitate and a supernatant B.

[0012] The pH value of the supernatant B is adjusted to a second pH, and a second heat treatment is performed to obtain heat treatment solution B. The heat treatment solution B is then separated into a second precipitate and supernatant A.

[0013] The first precipitate and the second precipitate are combined to obtain the flavoring base A;

[0014] The first pH and the second pH are different.

[0015] In some embodiments, the first pH is 4.3-4.7; and / or the second pH is 2.8-3.2.

[0016] In some embodiments, the mass of the salt accounts for 5%-10% of the total mass of the enzymatic hydrolysate and the salt.

[0017] In some embodiments, the first heat treatment includes: allowing the enzymatic hydrolysate to stand at 65°C-80°C for 15 min-30 min.

[0018] In some embodiments, the second heat treatment includes: allowing the supernatant B to stand at 65°C-80°C for 15 min-30 min.

[0019] In some embodiments, the pH values ​​of the enzymatic hydrolysate and the supernatant B are adjusted using acid, respectively;

[0020] Optionally, the acid includes citric acid monohydrate.

[0021] In some embodiments, a compound enzyme is used to enzymatically hydrolyze the marine raw materials;

[0022] Optionally, the enzymatic hydrolysis treatment includes at least one of the following conditions:

[0023] (1) The mass percentage of the compound enzyme to the mass of the seafood raw material is 0.15%-0.25%;

[0024] (2) The complex enzyme includes neutral protease, flavor protease and glutaminase;

[0025] Optionally, the mass ratio of the neutral protease, the flavor protease, and the glutaminase in the complex enzyme is 1:1:1;

[0026] Optionally, the enzyme activities of the neutral protease, the flavor protease, and the glutaminase are all 80,000 u / g to 130,000 u / g;

[0027] (3) The temperature of the enzymatic hydrolysis treatment is 50℃-60℃;

[0028] (4) The enzymatic hydrolysis time is 5.5h-6.5h.

[0029] In some embodiments, the aroma-generating process includes:

[0030] The supernatant A and white sugar were mixed and then subjected to a cooking process.

[0031] Optionally, the mass percentage of the white sugar in the supernatant A is 10%-20%;

[0032] Optionally, the temperature of the cooking treatment is 95℃-100℃;

[0033] Optionally, the cooking time is 1.5h-2.5h.

[0034] In some embodiments, the seafood ingredient includes oyster meat.

[0035] A second aspect of the present invention provides a seafood seasoning base, which is prepared using the method of the first aspect of the present invention.

[0036] The above-mentioned seafood seasoning base and its preparation method involve first using a complex enzyme composed of neutral protease, flavor protease, and glutaminase to enzymatically hydrolyze the seafood raw materials, which can increase the content of umami peptides and umami amino acids in the hydrolysate. Then, based on the difference in isoelectric points between umami peptides, glutamic acid, and aspartic acid and other types of amino acids and peptides, the hydrolysate is subjected to multi-stage isoelectric point separation of umami amino acids and umami peptides to separate them from the hydrolysate, avoiding their loss due to subsequent thermal reactions. This increases the content of umami amino acids and umami peptides in the final seafood seasoning base and enhances its umami flavor. Detailed Implementation

[0037] To facilitate understanding of the present invention, a more complete description will be given below with reference to relevant embodiments. Preferred embodiments of the present invention are shown below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that a thorough and complete understanding of the disclosure of the present invention will be achieved.

[0038] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0039] In this invention, the technical features described in an open-ended manner include both closed-ended technical solutions composed of the listed features and open-ended technical solutions that include the listed features.

[0040] In this invention, numerical ranges are involved. Unless otherwise specified, the numerical ranges are considered continuous and include the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe features or characteristics, the ranges may be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0041] This document only specifically discloses some numerical ranges. However, any lower limit can be combined with any upper limit to form an unspecified range; and any lower limit can be combined with other lower limits to form an unspecified range, just as any upper limit can be combined with any other upper limit to form an unspecified range. Furthermore, each individually disclosed point or single value can itself serve as a lower or upper limit and be combined with any other point or single value or with other lower or upper limits to form an unspecified range.

[0042] Unless otherwise specified, the temperature parameters in this invention can be either constant temperature processing or processing within a certain temperature range. The constant temperature processing allows temperature fluctuations within the precision range controlled by the instrument.

[0043] In the description of the invention, "multiple" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0044] Unless otherwise specified, all embodiments and optional embodiments of the present invention can be combined with each other to form new technical solutions. Unless otherwise specified, all technical features and optional technical features of the present invention can be combined with each other to form new technical solutions.

[0045] Unless otherwise specified, all steps of the present invention may be performed sequentially or randomly, but are preferably performed sequentially.

[0046] In related technologies, the preparation methods of umami seasoning bases mainly include enzymatic hydrolysis and cooking, but the resulting umami seasoning bases cannot meet the requirements for high umami flavor. Furthermore, in related technologies, the separation technology of umami substances mainly utilizes dextran gels and chromatographic columns to separate materials, but this technology is limited to the laboratory and cannot be applied to actual production.

[0047] To address the aforementioned issues, this invention, in preparing seafood seasoning base, first optimizes the enzymatic hydrolysis process to increase the content of umami peptides and umami amino acids in the hydrolysate. Then, based on the amphoteric electrolyte properties of umami amino acids and umami peptides, the difference in their isoelectric points is utilized to assist in salting out and heat treatment to disrupt the protein hydration layer, precipitating the target umami amino acids and umami peptides. This achieves the separation of umami amino acids and umami peptides from the hydrolysate, avoiding the destruction of umami substances by high-temperature heating, thereby increasing the content of umami amino acids and umami peptides in the seafood seasoning base and enhancing its umami flavor.

[0048] The first aspect of the present invention provides a method for preparing a seafood seasoning base, comprising the following steps:

[0049] Seafood raw materials are subjected to enzymatic hydrolysis to prepare enzymatic hydrolysate;

[0050] The enzymatic hydrolysate was subjected to multi-stage isoelectric point separation of umami amino acids and umami peptides to prepare seasoning base A and supernatant A.

[0051] The supernatant A was subjected to aroma-enhancing treatment to obtain flavoring base B;

[0052] Mix seasoning base A and seasoning base B to prepare seafood seasoning base.

[0053] It should be noted that the "first" and "second" in the above-mentioned "first pH", "second pH", "first precipitation", "second precipitation", "first heat treatment" and "second heat treatment" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor should they be construed as implicitly indicating the importance or quantity of the indicated technical features.

[0054] Multi-level isoelectric point separation of umami amino acids and umami peptides refers to separating umami amino acids and umami peptides from other amino acids and peptides based on the difference in their isoelectric points.

[0055] Understandably, in preparing seafood seasoning base, the seafood raw materials are first enzymatically hydrolyzed using a complex enzyme composed of neutral protease, flavor protease, and glutaminase, which can increase the content of umami peptides and umami amino acids in the hydrolysate. Then, based on the difference in isoelectric points between umami peptides, glutamic acid, and aspartic acid and other types of amino acids and peptides, the hydrolysate is subjected to multi-stage isoelectric point separation of umami amino acids and umami peptides to separate them from the hydrolysate, avoiding their loss due to participation in subsequent thermal reactions. This increases the content of umami amino acids and umami peptides in the final seafood seasoning base, thereby enhancing its umami flavor.

[0056] In addition, the supernatant A can be treated to remove fishy smell, enhance sweetness and seafood aroma, and achieve concentration. When mixed with the separated umami substances, a highly umami seasoning base can be prepared.

[0057] Furthermore, the aroma-enhancing treatment of the supernatant A not only improves the utilization rate of raw materials but also provides a nitrogen source for the product, reducing the generation of scraps and waste.

[0058] To prepare umami seasoning bases, they are typically based on seafood ingredients. In some embodiments, the seafood ingredients include oyster meat.

[0059] When preparing a umami seasoning base using seafood as raw material, an equal mass of water is added to the seafood raw material and the mixture is minced. After mincing, the seafood raw material is subjected to enzymatic hydrolysis. In some embodiments, a compound enzyme is used to perform enzymatic hydrolysis on the seafood raw material.

[0060] In some alternative embodiments, during enzymatic hydrolysis, the complex enzyme includes a neutral protease, a flavor protease, and a glutaminase. In the complex enzyme, the neutral protease acts as an endonuclease, hydrolyzing the protein into peptides, and assists the flavor protease and glutaminase in further hydrolyzing the peptides into small molecular weight umami peptides and umami amino acids, thereby increasing the content of umami peptides and umami amino acids in the hydrolysate.

[0061] In some optional embodiments, the mass ratio of neutral protease, flavor protease, and glutaminase in the complex enzyme is 1:1:1. When the mass ratio of the three enzymes in the complex enzyme is within the above range, the enzymatic hydrolysate has a fresh sensory flavor and no particularly obvious fishy or bitter taste.

[0062] In some optional embodiments, the enzyme activities of neutral protease, flavor protease, and glutaminase are 80,000 u / g to 130,000 u / g, respectively. When the enzyme activities of neutral protease, flavor protease, and glutaminase are within the above range, the protein recovery rate of the enzymatic hydrolysate can reach more than 80%, wherein the protein recovery rate (%) = total protein in the enzymatic hydrolysate supernatant / protein content of the raw material × 100%.

[0063] In some optional embodiments, the mass percentage of the complex enzyme relative to the mass of the seafood raw material is 0.15%-0.25%; for example, it can be, but is not limited to, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, or any range preceding two of the above values. When the mass percentage of the complex enzyme relative to the mass of the seafood raw material is within the above range, the degree of protein hydrolysis is higher, reaching over 40%, wherein the degree of hydrolysis (%) = total amino acid nitrogen in the enzymatic hydrolysis supernatant / total nitrogen content in the enzymatic hydrolysis supernatant × 100%.

[0064] In some alternative embodiments, the enzymatic hydrolysis temperature is 50°C-60°C; for example, it can be, but is not limited to, 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C, 60°C or any two of the above values.

[0065] In some alternative embodiments, the enzymatic hydrolysis time is 5.5h-6.5h; for example, it can be, but is not limited to, 5.5h, 5.6h, 5.7h, 5.8h, 5.9h, 6h, 6.1h, 6.2h, 6.3h, 6.4h, 6.5h or any range between two of the above values.

[0066] After enzymatic hydrolysis, an enzymatic hydrolysate is obtained. This hydrolysate is then centrifuged to separate the precipitate and the enzymatic hydrolysate. The precipitate is removed, and the hydrolysate is used for subsequent processing. To reduce the destruction of umami amino acids and umami peptides in the hydrolysate during subsequent heating and cooking, the hydrolysate can undergo multi-stage isoelectric point separation of umami amino acids and umami peptides. In some embodiments, the steps of multi-stage isoelectric point separation of umami amino acids and umami peptides in the hydrolysate to prepare seasoning base A and supernatant A include: adjusting the pH of the hydrolysate to a first pH, adding salt, and performing a first heat treatment to obtain heat-treated solution A; separating heat-treated solution A into a first precipitate and supernatant B; adjusting the pH of supernatant B to a second pH, and performing a second heat treatment to obtain heat-treated solution B; separating heat-treated solution B into a second precipitate and supernatant A; and combining the first and second precipitates to obtain seasoning base A; wherein the first and second pH are different.

[0067] When performing multi-stage isoelectric point separation of umami amino acids and umami peptides in the enzymatic hydrolysate, the pH of the system is adjusted based on the differences in isoelectric points between umami peptides, glutamic acid, and aspartic acid and other types of amino acids and peptides to neutralize the umami amino acids and umami peptides to reach their isoelectric points. The concentration of sodium chloride is then adjusted to induce salting out, and additional heating treatment is used to disrupt the outer hydration layer of the protein, causing the umami amino acids and umami peptides to aggregate and precipitate. Centrifugation is then performed to separate the umami amino acids and umami peptides from the enzymatic hydrolysate in advance.

[0068] In addition, heat treatment also has the functions of sterilization and enzyme inactivation, which can reduce the damage to umami substances caused by repeated heating.

[0069] After enzymatic hydrolysis, the hydrolysate is subjected to multi-stage isoelectric point separation of umami amino acids and umami peptides to separate them from the hydrolysate. First, the pH of the hydrolysate is adjusted to a first pH. In some embodiments, an acid is used to adjust the pH of the hydrolysate; optionally, the acid includes citric acid monohydrate.

[0070] In some embodiments, the first pH is 4.3-4.7; for example, it can be, but is not limited to, 4.3, 4.4, 4.5, 4.6, 4.7 or any two of the above values. By adjusting the pH of the enzymatic hydrolysate to the above range, the separation efficiency of umami amino acids and umami peptides can be improved.

[0071] After adjusting the pH of the enzymatic hydrolysate to the first pH, salt is added to salt out the hydrolysate. In some embodiments, the mass of salt accounts for 5%-10% of the total mass of the enzymatic hydrolysate and salt; for example, it can be, but is not limited to, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, or any range between two of the above values.

[0072] After adding salt to the enzymatic hydrolysate, a first heat treatment is performed. In some embodiments, the first heat treatment includes: allowing the enzymatic hydrolysate to stand at 65°C-80°C for 15-30 minutes.

[0073] The temperature of the first heat treatment is 65℃-80℃; for example, it can be, but is not limited to, 65℃, 66℃, 67℃, 68℃, 69℃, 70℃, 71℃, 72℃, 73℃, 74℃, 75℃, 76℃, 77℃, 78℃, 79℃, 80℃, or any two of the above values. The time of the first heat treatment is 15min-30min; for example, it can be, but is not limited to, 15min, 16min, 17min, 18min, 19min, 20min, 21min, 22min, 23min, 24min, 25min, 26min, 27min, 28min, 29min, 30min, or any two of the above values.

[0074] After the first heat treatment is completed, heat treatment solution A is obtained. Heat treatment solution A is centrifuged at 8000 rpm for 5 min-10 min. After centrifugation, the first precipitate and supernatant B are obtained. The first precipitate is reserved.

[0075] Adjust the pH of supernatant B to a second pH. In some embodiments, an acid is used to adjust the pH of supernatant B; optionally, the acid includes citric acid monohydrate.

[0076] In some embodiments, the second pH is 2.8-3.2; for example, it can be, but is not limited to, 2.8, 2.9, 3.0, 3.1, 3.2, or any two of the above values. By adjusting the pH of the supernatant B to the above range, umami amino acids and umami peptides can be further separated based on the first separation step, thereby improving the separation effect.

[0077] After adjusting the pH value of supernatant B, supernatant B is subjected to a second heat treatment. In some embodiments, the second heat treatment includes: allowing supernatant B to stand at 65°C-80°C for 15-30 minutes.

[0078] The temperature of the second heat treatment is 65℃-80℃; for example, it can be, but is not limited to, 65℃, 66℃, 67℃, 68℃, 69℃, 70℃, 71℃, 72℃, 73℃, 74℃, 75℃, 76℃, 77℃, 78℃, 79℃, 80℃, or any two of the above values. The time of the second heat treatment is 15min-30min; for example, it can be, but is not limited to, 15min, 16min, 17min, 18min, 19min, 20min, 21min, 22min, 23min, 24min, 25min, or any two of the above values.

[0079] After the second heat treatment, heat treatment solution B is obtained. Heat treatment solution B is centrifuged at 8000 rpm for 5-10 minutes to obtain a second precipitate and supernatant A. The first and second precipitates are combined to obtain seasoning base A, which is the separated umami substance.

[0080] The above process utilizes the characteristics of the umami amino acids and umami peptides in the enzymatic hydrolysate, taking advantage of their difference in isoelectric point, to assist in salting out and heat treatment to destroy the protein hydration layer, thereby precipitating the target umami amino acids and umami peptides. Then, centrifugation is performed to separate them, thus pre-separating the umami amino acids and umami peptides from the enzymatic hydrolysate and avoiding the destruction of umami substances by subsequent high-temperature heating.

[0081] The non-umami-rich supernatant A obtained from the separation is subjected to an aroma-enhancing treatment to achieve aroma enhancement and concentration, thereby preparing flavor base B. In some embodiments, the aroma-enhancing treatment includes: mixing supernatant A with white sugar and then subjecting it to a cooking process.

[0082] In some alternative embodiments, the mass percentage of granulated sugar in the supernatant A is 10%-20%; for example, it can be, but is not limited to, 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%, 19%, 19.5%, 20%, or any range between two of the above values. When the mass percentage of granulated sugar in the second supernatant is within the above range, it can enhance the concentration of the Maillard reaction substrate, improve the aroma of the supernatant, remove fishy odors, and increase sweetness.

[0083] As one possible implementation, the temperature of the cooking treatment is 95℃-100℃; for example, it can be, but is not limited to, 95℃, 95.5℃, 96℃, 96.5℃, 97℃, 97.5℃, 98℃, 98.5℃, 99℃, 99.5℃, 100℃ or any two of the above values.

[0084] In some optional embodiments, the cooking time is 1.5h-2.5h; for example, it can be, but is not limited to, 1.5h, 1.6h, 1.7h, 1.8h, 1.9h, 2h, 2.1h, 2.2h, 2.3h, 2.4h, 2.5h or any range between two of the above values.

[0085] By cooking the supernatant A and white sugar, the fishy smell can be removed, and the sweet aroma and characteristic seafood fragrance can be enhanced, while also achieving a concentration effect. Mixing this with the separated umami substances can prepare a highly umami-rich seafood seasoning base. Furthermore, the cooking process improves the utilization rate of raw materials, provides a nitrogen source for the product, and reduces the generation of scraps and waste.

[0086] When preparing seafood seasoning base in the future, seasoning base A and seasoning base B can be mixed in a certain proportion according to needs to obtain seafood seasoning base.

[0087] A second aspect of the present invention provides a seafood seasoning base, which is prepared using the method of the first aspect of the present invention.

[0088] The seafood seasoning base provided by this invention has a rich umami flavor and a weak fishy smell. The sensory score can be improved from 4.8 points to 7.8 points, the umami threshold can be reduced from 150mg / L to 15mg / L, the glutamic acid content can be increased by 2.4 times, the aspartic acid content can be increased by 3.4 times, and the amino acid nitrogen content can be increased by about 18%, thus meeting the demand for high umami.

[0089] The technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0090] I. Preparation of Seasoning Base

[0091] Example 1

[0092] 1. Enzymatic hydrolysis: Oyster meat was collected, mixed with water in the same proportion, and then pulped using a pulping machine. 0.2% (by weight of the oyster meat) of a complex protease was added for enzymatic hydrolysis at 55℃ for 6 hours. The complex protease consisted of a neutral protease, a flavor protease, and a glutaminase in a mass ratio of 1:1:1, with each enzyme having an activity of 100,000 U / g. The hydrolysis products were separated by centrifugation to obtain a clear hydrolysate, and the precipitate was removed.

[0093] 2. Multi-stage isoelectric point separation of umami amino acids and umami peptides: The pH of the enzymatic hydrolysate was adjusted to 4.3 using citric acid monohydrate, and salt was added to adjust the salt content of the hydrolysate to 5%. The hydrolysate was placed in an 80℃ water bath and kept at that temperature for 20 minutes for the first heat treatment, yielding heat-treated solution A. Heat-treated solution A was centrifuged at 8000 rpm for 10 minutes to obtain the first precipitate and supernatant B. The first precipitate was reserved. The pH of supernatant B was adjusted to 2.8 using citric acid monohydrate, and supernatant B was placed in an 80℃ water bath and kept at that temperature for 20 minutes for the second heat treatment, yielding heat-treated solution B. Heat-treated solution B was centrifuged at 8000 rpm for 10 minutes to obtain the second precipitate and supernatant A. The first and second precipitates were combined to obtain seasoning base A.

[0094] 3. Flavoring treatment: Add 10% of the weight of white sugar to the clear liquid A, and cook at 100℃ for 1.5 hours to achieve flavoring and concentration, and obtain flavoring base B.

[0095] 4. Mix seasoning base A and seasoning base B in a mass ratio of 1:3 to obtain seafood seasoning base.

[0096] Example 2

[0097] 1. Enzymatic hydrolysis: Oyster meat was collected, mixed with water in the same proportion, and then pulped using a pulping machine. 0.2% (by weight of the oyster meat) of a complex protease was added for enzymatic hydrolysis at 55℃ for 6 hours. The complex protease consisted of a neutral protease, a flavor protease, and a glutaminase in a mass ratio of 1:1:1, with each enzyme having an activity of 100,000 U / g. The hydrolysis products were separated by centrifugation to obtain a clear hydrolysate, and the precipitate was removed.

[0098] 2. Multi-stage isoelectric point separation of umami amino acids and umami peptides: The pH of the enzymatic hydrolysate was adjusted to 4.5 using citric acid monohydrate, and salt was added to adjust the salt content of the hydrolysate to 8%. The hydrolysate was placed in a 75℃ water bath and kept at that temperature for 15 minutes for the first heat treatment, yielding heat-treated solution A. Heat-treated solution A was centrifuged at 8000 rpm for 5 minutes to obtain the first precipitate and supernatant B. The first precipitate was reserved. The pH of supernatant B was adjusted to 3.0 using citric acid monohydrate, and supernatant B was placed in a 75℃ water bath and kept at that temperature for 15 minutes for the second heat treatment, yielding heat-treated solution B. Heat-treated solution B was centrifuged at 8000 rpm for 5 minutes to obtain the second precipitate and supernatant A. The first and second precipitates were combined to obtain seasoning base A.

[0099] 3. Flavoring process: Add 15% of the weight of white sugar to the clear liquid A, and cook at 97°C for 2 hours to achieve flavoring and concentration, thus obtaining flavoring base B.

[0100] 4. Mix seasoning base A and seasoning base B in a mass ratio of 1:3 to obtain seafood seasoning base.

[0101] Example 3

[0102] 1. Enzymatic hydrolysis: Oyster meat was collected, mixed with water in the same proportion, and then pulped using a pulping machine. 0.2% (by weight of the oyster meat) of a complex protease was added for enzymatic hydrolysis at 55℃ for 6 hours. The complex protease consisted of a neutral protease, a flavor protease, and a glutaminase in a mass ratio of 1:1:1, with each enzyme having an activity of 100,000 U / g. The hydrolysis products were separated by centrifugation to obtain a clear hydrolysate, and the precipitate was removed.

[0103] 2. Multi-stage isoelectric point separation of umami amino acids and umami peptides: The pH of the enzymatic hydrolysate was adjusted to 4.7 using citric acid monohydrate, and salt was added to adjust the salt content of the hydrolysate to 10%. The hydrolysate was placed in a 65℃ water bath and kept at that temperature for 30 minutes for the first heat treatment, yielding heat-treated solution A. Heat-treated solution A was centrifuged at 8000 rpm for 8 minutes to obtain the first precipitate and supernatant B. The first precipitate was reserved. The pH of supernatant B was adjusted to 3.2 using citric acid monohydrate, and supernatant B was placed in a 65℃ water bath and kept at that temperature for 30 minutes for the second heat treatment, yielding heat-treated solution B. Heat-treated solution B was centrifuged at 8000 rpm for 8 minutes to obtain the second precipitate and supernatant A. The first and second precipitates were combined to obtain seasoning base A.

[0104] 3. Flavoring treatment: Add 20% of the weight of white sugar to the clear liquid A, and cook at 95℃ for 2.5 hours to achieve flavoring and concentration, and obtain flavoring base B.

[0105] 4. Mix seasoning base A and seasoning base B in a mass ratio of 1:3 to obtain seafood seasoning base.

[0106] Comparative Example 1

[0107] 1. Take oyster meat, mix it with water in the same proportion, and then blend it using a blender. Add 0.2% alkaline protease by weight of oyster meat, adjust the pH of the system to 8.0 using baking soda, and enzymatically hydrolyze at 55℃ for 6 hours, with an enzyme activity of 100,000 u / g. Centrifuge the enzymatic hydrolysis product to obtain a clear enzymatic hydrolysate, and remove the precipitate.

[0108] 2. The enzymatic hydrolysate is boiled at 100℃ for 15 minutes to sterilize and inactivate the enzyme, thus obtaining the seasoning base.

[0109] Comparative Example 2

[0110] The main difference between Comparative Example 2 and Example 1 is that only the enzymatic hydrolysate underwent single-stage isoelectric point separation of umami amino acids and umami peptides. Details are as follows:

[0111] 1. Take oyster meat, mix it with water in the same proportion, and then blend it using a blender. Add 0.2% (by weight of oyster meat) of a complex protease for enzymatic hydrolysis at 55℃ for 6 hours. The complex protease consists of neutral protease, flavor protease, and glutaminase in a mass ratio of 1:1:1, with each enzyme having an activity of 100,000 u / g. Centrifuge the hydrolysate to obtain a clear hydrolysate, and remove the precipitate.

[0112] 2. The pH of the enzymatic hydrolysate was adjusted to 4.3 using citric acid monohydrate, and salt was added to adjust the salt content of the enzymatic hydrolysate to 3%. The enzymatic hydrolysate was placed in a 50℃ water bath and kept at that temperature for 20 minutes for the first heat treatment to obtain heat treatment solution A. Heat treatment solution A was centrifuged at 8000 rpm for 10 minutes to obtain the first precipitate and supernatant B. The first precipitate was used as seasoning base A.

[0113] 3. Add 10% of the weight of white sugar to the clear liquid B, and cook at 100℃ for 1.5 hours to achieve aroma development and concentration, thus obtaining flavoring base B.

[0114] 4. Mix seasoning base A and seasoning base B in a mass ratio of 1:3 to obtain seafood seasoning base.

[0115] Comparative Example 3

[0116] The difference between Comparative Example 3 and Example 1 is that the enzymatic hydrolysate was not subjected to multi-stage isoelectric point separation of umami amino acids and umami peptides. Specifically:

[0117] 1. Take oyster meat, mix it with water in the same proportion, and then blend it using a blender. Add 0.2% (by weight of oyster meat) of a complex protease for enzymatic hydrolysis at 55℃ for 6 hours. The complex protease consists of neutral protease, flavor protease, and glutaminase in a mass ratio of 1:1:1, with each enzyme having an activity of 100,000 u / g. Centrifuge the hydrolysate to obtain a clear hydrolysate, and remove the precipitate.

[0118] 2. Add 10% of the weight of white sugar to the enzymatic hydrolysate and cook at 100℃ for 1.5 hours to achieve aroma development and concentration, thus obtaining the seafood seasoning base.

[0119] Comparative Example 4

[0120] Dissolve a 1% concentration of monosodium glutamate (MSG) in tap water.

[0121] II. Sensory Evaluation

[0122] Sensory evaluations were performed on the flavoring bases prepared in Examples 1-3 and Comparative Examples 1-4. The sensory evaluation methods are as follows:

[0123] A sensory evaluation panel of 20 people was selected to comprehensively evaluate the aroma and taste of the test samples. Each indicator was scored on a scale of 0 to 10 (0 being the worst and 10 being the best). The total score was 10. The final score for aroma and taste was calculated based on the weight of each indicator (30% or 70%). The scoring criteria are shown in Table 1.

[0124] Table 1. Sensory Evaluation Criteria

[0125] Score Aroma (30%) Taste (70%) 8-10 It has no fishy smell, and is characterized by a rich oyster aroma and a sweet fragrance. No bitterness, rich umami flavor 6-8 It has a milder fishy smell, a stronger characteristic oyster aroma, and a more pronounced sweetness. Slightly bitter, with a strong umami flavor 4-6 It has a fishy smell, and the characteristic oyster flavor and sweetness are average. It has a bitter taste and a mild umami flavor. 2-4 It has a distinct fishy smell, a weak oyster aroma, and a weak sweet aroma. The bitterness is pronounced, while the umami flavor is weak. 0-2 It has a strong fishy smell, lacks the characteristic oyster aroma, and has no sweetness. It has a strong bitter taste and a weak umami flavor.

[0126] III. Umami Threshold Determination

[0127] The sensory threshold of umami peptides was determined by the taste dilution analysis method. A series of dilutions ranging from 5 to 200 mg / L were prepared in increments of 5 mg / L. The dilutions of the sample were evaluated sequentially from low to high concentration. The concentration at which a dilution was just different from that of pure water was identified as the taste threshold of this substance. The result was the average of multiple tasters.

[0128] The sensory evaluation results and umami threshold measurement results of the above embodiments and comparative examples are shown in Table 2.

[0129] Table 2

[0130]

[0131] As can be seen from the results of Examples 1-4 and Comparative Example 1 in Table 2, compared with the conventional method for preparing oyster seasoning base, the seafood seasoning base prepared by the method of the present invention has a significant improvement in umami. Among them, compared with Comparative Example 1, Example 2 reduced the umami threshold from 150 mg / L to 15 mg / L, increased the glutamic acid content by 2.4 times, increased the aspartic acid content by 3.4 times, increased the amino acid nitrogen content by about 18%, and had an umami level approximately equal to that of a 7% concentration of monosodium glutamate solution. The sensory score increased from 4.8 points to 7.8 points.

[0132] A comparison of the results from Comparative Example 3 and Example 1 shows that, prior to the steaming and flavoring process, multi-stage isoelectric point separation of umami amino acids and umami peptides in the enzymatic hydrolysate significantly reduces the umami threshold of the seasoning base, increases the content of glutamic acid, aspartic acid, and amino acid nitrogen in the seasoning base, and produces a seasoning base without fishy odor and with a strong umami flavor. This indicates that by performing multi-stage isoelectric point separation of umami amino acids and umami peptides in the enzymatic hydrolysate, umami amino acids and umami peptides can be pre-separated from the enzymatic hydrolysate, avoiding the destruction of umami substances by subsequent high-temperature heating, increasing the content of umami amino acids and umami peptides in the final seafood seasoning base, and enhancing the umami flavor of the seafood seasoning base.

[0133] Comparison of the results of Comparative Example 2 and Example 1 shows that, compared with single-stage isoelectric point separation of umami amino acids and umami peptides in the enzymatic hydrolysate alone, multi-stage isoelectric point separation of umami amino acids and umami peptides in the enzymatic hydrolysate before cooking and aroma enhancement significantly reduces the umami threshold of the seasoning base, significantly increases the content of glutamic acid, aspartic acid, and amino acid nitrogen in the seasoning base, and significantly reduces the fishy smell value and significantly enhances the umami flavor. This indicates that multi-stage isoelectric point separation of umami amino acids and umami peptides in the enzymatic hydrolysate is more efficient in separating these components.

[0134] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0135] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A method for preparing a seafood seasoning base, characterized in that, Includes the following steps: A compound enzyme is used to enzymatically hydrolyze marine raw materials to prepare a hydrolysate; the compound enzyme includes neutral protease, flavor protease and glutaminase; The pH of the enzymatic hydrolysate is adjusted to a first pH, salt is added, and a first heat treatment is performed to obtain heat treatment solution A. The heat treatment solution A is then separated into a first precipitate and a supernatant B. The first pH is 4.3-4.7, and the temperature of the first heat treatment is 65℃-80℃. The pH value of the supernatant B is adjusted to a second pH, and a second heat treatment is performed to obtain heat treatment solution B. The heat treatment solution B is then separated into a second precipitate and supernatant A. The second pH is 2.8-3.

2. The temperature of the second heat treatment is 65℃-80℃. The first precipitate and the second precipitate are combined to obtain the flavoring base A; The supernatant A is subjected to aroma-enhancing treatment to obtain flavoring base B; The aroma-enhancing process includes: mixing the supernatant A and white sugar and then steaming it; The seafood seasoning base is prepared by mixing the seasoning base A and the seasoning base B.

2. The method for preparing the seafood seasoning base as described in claim 1, characterized in that, The mass of the salt accounts for 5%-10% of the total mass of the enzymatic hydrolysate and the salt.

3. The method for preparing the seafood seasoning base as described in claim 1, characterized in that, The first heat treatment includes: allowing the enzymatic hydrolysate to stand at 65°C-80°C for 15-30 minutes; and / or The second heat treatment includes: allowing the supernatant B to stand at 65℃-80℃ for 15min-30min.

4. The method for preparing the seafood seasoning base as described in claim 1, characterized in that, The pH values ​​of the enzymatic hydrolysate and the supernatant B were adjusted using acid, respectively.

5. The method for preparing the seafood seasoning base as described in claim 4, characterized in that, The acid includes citric acid monohydrate.

6. The method for preparing the seafood seasoning base as described in claim 1, characterized in that, The enzymatic hydrolysis treatment includes at least one of the following conditions: (1) The mass percentage of the compound enzyme to the mass of the seafood raw material is 0.15%-0.25%; (2) The mass ratio of the neutral protease, the flavor protease and the glutaminase in the complex enzyme is 1:1:1; (3) The temperature of the enzymatic hydrolysis treatment is 50℃-60℃; (4) The enzymatic hydrolysis time is 5.5h-6.5h.

7. The method for preparing the seafood seasoning base as described in claim 6, characterized in that, The enzyme activities of the neutral protease, the flavor protease, and the glutaminase are all 80,000 u / g to 130,000 u / g.

8. The method for preparing the seafood seasoning base as described in any one of claims 1 to 7, characterized in that, The mass percentage of the white sugar in the supernatant A is 10%-20%.

9. The method for preparing the seafood seasoning base as described in any one of claims 1 to 7, characterized in that, The cooking process is carried out at a temperature of 95℃-100℃ for 1.5h-2.5h.

10. The method for preparing the seafood seasoning base as described in any one of claims 1 to 7, characterized in that, The seafood ingredients include oyster meat.

11. A seafood seasoning base, characterized in that, It is prepared by the method according to any one of claims 1 to 10.

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