Fresh flavor extract

Abstract

The invention discloses a delicate flavor extract, and belongs to the field of food. The preparation method of the delicate flavor extract comprises the following steps: with clam meat as a raw material, fermenting the clam meat with a specific bacterial strain, and then carrying out enzymolysis, filtration, irradiation treatment and freeze-drying on the fermentation liquor with a compound enzyme with specific composition, thereby obtaining the delicate flavor extract. According to the preparation method disclosed by the invention, the step of firstly fermenting and then carrying out enzymolysis is adopted, so that bad flavors such as bitter taste and fishy smell in the extract can be effectively avoided through the synergistic effect of microorganisms and compound enzymes, and the content of free amino acids in the prepared fresh-taste extract is remarkably increased; and the percentage of flavor amino acids such as glutamic acid and aspartic acid in the total free amino acids is also obviously improved. In addition, experiments prove that the content of free amino acids in the umami extract and the percentage of umami amino acids can be further increased by adding amylose, cane molasses and skipjack powder into the clam meat slurry during fermentation, so that the prepared umami extract is better in flavor.

Description

Technical Field

[0001] The invention relates to the field of food additives, in particular to an umami extract. Background Art

[0002] Clams are a type of seafood belonging to the order Clams of the class Bivalvia. They are delicious and are known as "the most delicious food in the world" and "the king of all flavors". Clams are high in protein, trace elements, iron, calcium, and low in fat, but a small number of people are allergic to certain proteins in clams and cannot enjoy the deliciousness of clams.

[0003] As we all know, umami is usually caused by free amino acids, so if you want to improve the umami, you need to increase the content of free amino acids. In order to improve the umami taste, most of the current seafood flavor products use crushing, homogenization and then enzymatic hydrolysis, but this method has low extraction efficiency, high loss and poor cost performance. With the growing growth of biotechnology, researchers have found that using fermentation technology to ferment animal protein, different strains of bacteria have different metabolic pathways in different culture media, resulting in different secondary metabolites produced by fermentation. On the basis of decomposing the substrate, the fermentation product can be further given more flavor characteristics, but the fermentation of bacteria will also lead to excessive secretion of biogenic amines and produce a corrupt smell because the substrate cannot be fully utilized or the fermentation time is too long. Therefore, it is necessary to provide a method for extracting umami substances from clams that can fully utilize the substrate, avoid the production of biogenic amines, and produce more free amino acids. Summary of the invention

[0004] The object of the present invention is to provide an umami extract, wherein the preparation method of the umami extract is to use clam meat as a raw material, ferment it with a specific strain, and then enzymolyze, filter, irradiate and freeze-dry the fermentation liquid with a composite enzyme of a specific composition. The umami extract obtained by the preparation method of the present invention has a significantly increased free amino acid content, and the percentage of umami amino acids such as glutamic acid and aspartic acid in the total free amino acids is also significantly increased.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] The present invention provides an umami extract, characterized in that it comprises the following steps:

[0007] S1: Wash and remove impurities from fresh clam meat, add deionized sterile water, and homogenize to obtain clam meat slurry;

[0008] S2: fermenting the clam pulp with Heronweissella, filtering after fermentation to obtain a fermentation liquid, wherein the fermentation time is 18-32 hours and the fermentation temperature is 28-38°C;

[0009] S3: using bromelain, chymotrypsin and flavor protease to perform ultrasonic enzymolysis on the fermentation broth, filtering to obtain a filtrate after the enzymolysis is completed, freeze-drying the filtrate to obtain a freeze-dried powder, irradiating the freeze-dried powder for sterilization and enzyme inactivation to obtain an umami extract, wherein the ultrasonic power is 135-175w, the enzymolysis pH is 5-7, the enzymolysis temperature is 50-65°C, and the enzymolysis time is 6-12h;

[0010] The Herrenweissella CGMCC NO: 1.2513;

[0011] The mass ratio of clam meat to deionized sterile water is 1:13, the mass ratio of clam meat slurry to Herrenweissella is 10:0.05-0.07, and the viable count of Herrenweissella is 1×10 7 -1×10 9 CFU / g, the mass ratio of clam pulp, bromelain, chymotrypsin and flavor protease was 10: 0.06-0.08: 0.01-0.03: 0.05-0.07.

[0012] Preferably, amylose, cane molasses and bonito meal are further added to the clam meat slurry in step S2, and the mass ratio of the clam meat, amylose, cane molasses and bonito meal is 1:0.6-0.8:0.2-0.4:0.8-1.

[0013] Compared with the prior art, the invention has the following beneficial effects: the invention provides an umami extract, which uses clam meat as a raw material, washes it, and adds water to homogenize it to obtain clam meat pulp, the clam meat pulp is first fermented by Herrenweissella, filtered to obtain a fermentation liquid, and then the fermentation liquid is enzymatically hydrolyzed by bromelain, chymotrypsin and flavor protease, and after the enzymatic hydrolysis is completed, the umami extract is obtained by filtering and freeze-drying, wherein the clam meat pulp can produce volatile acids and ester substances such as acetic acid, propionic acid, butyric acid, and extracellular polysaccharides and lactic acid after fermentation, and various components in the above substances have unique flavor characteristics, and in the fermentation and enzymatic hydrolysis steps, the combined use of Herrenweissella and bromelain, chymotrypsin and flavor protease can make full use of the protein in the clam meat, produce more free amino acids, and effectively avoid the generation of undesirable flavors such as bitterness and fishy smell in the extract, and experiments show that the free amino acid content in the prepared umami extract is significantly increased, and the percentage of umami amino acids such as glutamic acid and aspartic acid in the total free amino acids is also significantly increased. In addition, the present invention also proves through experiments that adding amylose, cane molasses and bonito powder to clam pulp during fermentation can further increase the content of free amino acids in the umami extract and the percentage of umami amino acids, so that the prepared umami extract has a better flavor. DETAILED DESCRIPTION

[0014] The embodiments described below are part of the embodiments of the present application, rather than all of the embodiments. The embodiments of the present application are used to illustrate the present invention, rather than to limit the present invention. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present application.

[0015] The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or conditions recommended by the manufacturers. The materials and reagents used can be obtained from commercial sources unless otherwise specified.

[0016] Some raw materials and their sources are as follows:

[0017] The clams were purchased from Zhangzidao Group Co., Ltd.

[0018] The water used in the preparation process was deionized sterile water;

[0019] The Herrenweissella was purchased from China General Microbiological Culture Collection Center, number: CGMCCNO:1.2513;

[0020] The Weissella cannella was purchased from China General Microbiological Culture Collection Center, number: CGMCCNO:1.2474;

[0021] The bromelain was purchased from Guangzhou Huayu Biotechnology Co., Ltd., CAS: 9001-00-7, enzyme activity: 100,000 U / g.

[0022] The chymotrypsin was purchased from Wuhan Haorong Biotechnology Co., Ltd., CAS: 9004-07-3, enzyme activity: 2000 U / g.

[0023] The flavor protease was purchased from Chongqing Tianrun Biological Products Co., Ltd., CAS: 9001-92-7, enzyme activity: 50,000 U / g.

[0024] The papain was purchased from Shaanxi Chenming Biotechnology Co., Ltd., CAS: 9001-73-4, enzyme activity: 100,000 U / g.

[0025] The trypsin was purchased from Nanning Dongheng Huadao Biotechnology Co., Ltd., CAS: 9002-07-7, enzyme activity: 2000 U / g.

[0026] The amylose was purchased from Wuhan Huaxiang Kejie Biotechnology Co., Ltd., CAS: 9005-82-7.

[0027] The sugarcane molasses was purchased from Shandong Ruihongze Chemical Technology Co., Ltd.

[0028] The bonito meal was purchased from Qingdao Rishengchang Food Ingredients Co., Ltd.

[0029] The preparation method of the umami extract is specifically as follows:

[0030] Embodiment 1:

[0031] S1: Wash and remove impurities from fresh clam meat, add deionized sterile water, and homogenize to obtain clam meat slurry;

[0032] S2: fermenting clam pulp with Heronweissella, filtering to obtain fermentation liquid after fermentation, wherein the fermentation time is 25 hours and the fermentation temperature is 33°C;

[0033] S3: using bromelain, chymotrypsin and flavor protease to perform ultrasonic enzymolysis on the fermentation broth, filtering to obtain a filtrate after the enzymolysis is completed, freeze-drying the filtrate to obtain a freeze-dried powder, and irradiating the freeze-dried powder for sterilization and enzyme inactivation to obtain an umami extract, wherein the ultrasonic power is 150w, the enzymolysis pH is 6, the enzymolysis temperature is 60°C, and the enzymolysis time is 8h;

[0034] The Herrenweissella was purchased from CGMCC NO: 1.2513;

[0035] The mass ratio of clam meat to deionized sterile water is 1:13, the mass ratio of clam meat slurry to Herrenweissella is 10:0.06, and the viable count of Herrenweissella is 1×10 8 CFU / g, the mass ratio of clam pulp, bromelain, chymotrypsin and flavor protease was 10:0.07:0.02:0.06.

[0036] Embodiment 2:

[0037] S1: Wash and remove impurities from fresh clam meat, add deionized sterile water, and homogenize to obtain clam meat slurry;

[0038] S2: fermenting clam pulp with Heronweissella, filtering to obtain fermentation liquid after fermentation, wherein the fermentation time is 18 hours and the fermentation temperature is 28°C;

[0039] S3: using bromelain, chymotrypsin and flavor protease to perform ultrasonic enzymolysis on the fermentation broth, filtering to obtain a filtrate after the enzymolysis is completed, freeze-drying the filtrate to obtain a freeze-dried powder, and irradiating the freeze-dried powder for sterilization and enzyme inactivation to obtain an umami extract, wherein the ultrasonic power is 175w, the enzymolysis pH is 5, the enzymolysis temperature is 65℃, and the enzymolysis time is 6h;

[0040] The Herrenweissella was purchased from CGMCC NO: 1.2513;

[0041] The CAS number of the bromelain is: 9001-00-7;

[0042] The CAS number of the chymotrypsin is: 9004-07-3;

[0043] The CAS number of the flavor protease is: 9001-92-7;

[0044] The mass ratio of clam meat to deionized sterile water is 1:13, the mass ratio of clam meat slurry to Herrenweissella is 10:0.05, and the viable count of Herrenweissella is 1×10 7 CFU / g, the mass ratio of clam pulp, bromelain, chymotrypsin and flavor protease was 10:0.06:0.03:0.05.

[0045] Embodiment 3:

[0046] S1: Wash and remove impurities from fresh clam meat, add deionized sterile water, and homogenize to obtain clam meat slurry;

[0047] S2: fermenting clam pulp with Heronweissella, filtering to obtain fermentation liquid after fermentation, wherein the fermentation time is 32 hours and the fermentation temperature is 38°C;

[0048] S3: using bromelain, chymotrypsin and flavor protease to perform ultrasonic enzymolysis on the fermentation broth, filtering to obtain a filtrate after the enzymolysis is completed, freeze-drying the filtrate to obtain a freeze-dried powder, and irradiating the freeze-dried powder for sterilization and enzyme inactivation to obtain an umami extract, wherein the ultrasonic power is 135w, the enzymolysis pH is 7, the enzymolysis temperature is 50°C, and the enzymolysis time is 12h;

[0049] The Herrenweissella was purchased from CGMCC NO: 1.2513;

[0050] The mass ratio of clam meat to deionized sterile water is 1:13, the mass ratio of clam meat slurry to Herrenweissella is 10:0.07, and the viable count of Herrenweissella is 1×10 9 CFU / g, the mass ratio of clam pulp, bromelain, chymotrypsin and flavor protease was 10:0.08:0.01:0.07.

[0051] Embodiment 4:

[0052] S1: Wash and remove impurities from fresh clam meat, add deionized sterile water, and homogenize to obtain clam meat slurry;

[0053] S2: fermenting clam pulp with Heronweissella, filtering to obtain fermentation liquid after fermentation, wherein the fermentation time is 25 hours and the fermentation temperature is 33°C;

[0054] S3: using bromelain, chymotrypsin and flavor protease to perform ultrasonic enzymolysis on the fermentation broth, filtering to obtain a filtrate after the enzymolysis is completed, freeze-drying the filtrate to obtain a freeze-dried powder, and irradiating the freeze-dried powder for sterilization and enzyme inactivation to obtain an umami extract, wherein the ultrasonic power is 150w, the enzymolysis pH is 6, the enzymolysis temperature is 60°C, and the enzymolysis time is 8h;

[0055] The Herrenweissella was purchased from CGMCC NO: 1.2513;

[0056] The mass ratio of clam meat to deionized sterile water is 1:13, the mass ratio of clam meat slurry to Herrenweissella is 10:0.06, and the viable count of Herrenweissella is 1×10 8 CFU / g, the mass ratio of clam pulp, bromelain, chymotrypsin and flavor protease was 10:0.07:0.02:0.06.

[0057] In step S2, amylose, cane molasses and bonito powder are further added to the clam meat slurry, and the mass ratio of the clam meat, amylose, cane molasses and bonito powder is 1:0.7:0.3:0.9.

[0058] Embodiment 5:

[0059] S1: Wash and remove impurities from fresh clam meat, add deionized sterile water, and homogenize to obtain clam meat slurry;

[0060] S2: fermenting clam pulp with Heronweissella, filtering to obtain fermentation liquid after fermentation, wherein the fermentation time is 25 hours and the fermentation temperature is 33°C;

[0061] S3: using bromelain, chymotrypsin and flavor protease to perform ultrasonic enzymolysis on the fermentation broth, filtering to obtain a filtrate after the enzymolysis is completed, freeze-drying the filtrate to obtain a freeze-dried powder, and irradiating the freeze-dried powder for sterilization and enzyme inactivation to obtain an umami extract, wherein the ultrasonic power is 150w, the enzymolysis pH is 6, the enzymolysis temperature is 60°C, and the enzymolysis time is 8h;

[0062] The Herrenweissella was purchased from CGMCC NO: 1.2513;

[0063] The mass ratio of clam meat to deionized sterile water is 1:13, the mass ratio of clam meat slurry to Herrenweissella is 10:0.06, and the viable count of Herrenweissella is 1×10 8 CFU / g, the mass ratio of clam pulp, bromelain, chymotrypsin and flavor protease was 10:0.07:0.02:0.06.

[0064] In step S2, amylose, sugarcane molasses and bonito powder are further added to the clam meat slurry, and the mass ratio of the clam meat, amylose, sugarcane molasses and bonito powder is 1:0.6:0.2:1.

[0065] Embodiment 6:

[0066] S1: Wash and remove impurities from fresh clam meat, add deionized sterile water, and homogenize to obtain clam meat slurry;

[0067] S2: fermenting clam pulp with Heronweissella, filtering after fermentation to obtain fermentation liquid, wherein the fermentation time is 25 hours and the fermentation temperature is 33°C;

[0068] S3: using bromelain, chymotrypsin and flavor protease to perform ultrasonic enzymolysis on the fermentation broth, filtering to obtain a filtrate after the enzymolysis is completed, freeze-drying the filtrate to obtain a freeze-dried powder, and irradiating the freeze-dried powder for sterilization and enzyme inactivation to obtain an umami extract, wherein the ultrasonic power is 150w, the enzymolysis pH is 6, the enzymolysis temperature is 60°C, and the enzymolysis time is 8h;

[0069] The Herrenweissella was purchased from CGMCC NO: 1.2513;

[0070] The mass ratio of clam meat to deionized sterile water is 1:13, the mass ratio of clam meat slurry to Herrenweissella is 10:0.06, and the viable count of Herrenweissella is 1×10 8 CFU / g, the mass ratio of clam pulp, bromelain, chymotrypsin and flavor protease was 10:0.07:0.02:0.06.

[0071] In step S2, amylose, cane molasses and bonito powder are further added to the clam meat slurry, and the mass ratio of the clam meat, amylose, cane molasses and bonito powder is 1:0.8:0.4:0.8.

[0072] Comparative Example 1:

[0073] The difference compared with Example 1 is that Weissella cannella is used to replace Weissella herrenii in Comparative Example 1, and the remaining steps are the same as in Example 1.

[0074] Comparative Example 2:

[0075] The difference compared with Example 1 is that in Comparative Example 2, the mass ratio of clam pulp, bromelain, chymotrypsin and flavor protease is 10:0.04:0.07:0.04, and the remaining steps are the same as in Example 1.

[0076] Comparative Example 3:

[0077] The difference from Example 1 is that in Comparative Example 3, only chymotrypsin and flavor protease are used for enzymatic hydrolysis, and the reduced mass of the above two is distributed to chymotrypsin and flavor protease in proportion, and the remaining steps are the same as in Example 1.

[0078] Comparative Example 4:

[0079] The difference from Example 1 is that in Comparative Example 4, only bromelain and chymotrypsin are used for enzymolysis, and the reduced mass of the above two is proportionally distributed to bromelain and chymotrypsin, and the remaining steps are the same as in Example 1.

[0080] Comparative Example 5:

[0081] The difference from Example 1 is that in Comparative Example 5, only bromelain and flavor protease are used for enzymatic hydrolysis, and the reduced mass of the above two is allocated to bromelain and flavor protease in proportion, and the remaining steps are the same as in Example 1.

[0082] Comparative Example 6:

[0083] The difference compared with Example 1 is that trypsin is used to replace chymotrypsin in Comparative Example 6, and the remaining steps are the same as in Example 1.

[0084] Comparative Example 7:

[0085] The difference compared with Example 1 is that papain is used to replace bromelain in Comparative Example 7, and the remaining steps are the same as those in Example 1.

[0086] Comparative Example 8:

[0087] The difference compared with Example 4 is that the mass ratio of clam meat, amylose, sugarcane molasses and bonito meal in Comparative Example 8 is 1:0.3:0.9:0.7, and the remaining steps are the same as Example 4.

[0088] blank:

[0089] S1: Wash and remove impurities from fresh clam meat, add deionized sterile water, and homogenize to obtain clam meat slurry;

[0090] S2: The clam slurry is shaken and extracted at 60° C. for 4 hours, the extracted liquid is filtered, and then freeze-dried and concentrated to obtain a clam water extract.

[0091] The mass ratio of clam meat to deionized sterile water was 1:13; extraction was performed once with an oscillation frequency of 80 rpm.

[0092] Test 1: Total free amino acid determination

[0093] Test samples: Examples 1-6, Comparative Examples 1-8, and blank.

[0094] Test method: refer to the kit instructions.

[0095] The free amino acid detection kit was purchased from Wuhan Aidikang Biotechnology Co., Ltd., item number: AD9255.

[0096] Calculation method: Total free amino acid content = m (测定出总游离氨基酸量) / M (蛤蜊肉质量)

[0097] Table 1 Total free amino acid content determination results

[0098]

[0099]

[0100] Note: All data are expressed as mean ± standard deviation SD (repeated times n = 3); "*" indicates significant difference compared with the blank group.

[0101] The experimental results of Example 1 and Comparative Examples 1-7 show that the preparation process involving specific bacterial fermentation and complex enzyme hydrolysis of specific composition provided by the present invention can significantly increase the content of total free amino acids in the umami extract. By comparing Example 4 with Example 1, it can be seen that adding amylose, cane molasses and bonito meal to clam pulp can effectively further increase the total free amino acid content in the umami extract. By comparing Example 4 with Comparative Example 8, it can be seen that the addition ratio of amylose, cane molasses and bonito meal is also very important. The experiment shows that the total free amino acid content in the umami extract can be effectively increased within the specified range of the present invention.

[0102] Experiment 2: Umami amino acids as a percentage of total free amino acids

[0103] Samples: Example 1, Example 4, Comparative Example 2, Comparative Example 8;

[0104] Test method: refer to the kit instructions;

[0105] Glutamate detection kit was purchased from Shanghai ELISA Biotechnology Co., Ltd., catalog number: ml077309;

[0106] Aspartate detection kit was purchased from Shanghai ELISA Biotechnology Co., Ltd., catalog number: ml077312;

[0107] Alanine detection kit was purchased from Shanghai ELISA Biotechnology Co., Ltd., catalog number: ml077312;

[0108] The glycine detection kit was purchased from Shanghai ELISA Biotechnology Co., Ltd., catalog number: ml076512.

[0109] The results are shown in Table 2.

[0110] Experiment 3: Umami taste evaluation

[0111] Samples: Example 1, Example 4, Comparative Example 2, Comparative Example 8;

[0112] Concentration: 1g / L;

[0113] Testing method: 30 trained sensory assessors were asked to rate the umami taste of each sample (excellent, good, poor) using a scoring method, where the total score was 10, with 8-10 being excellent; 6-7.9 being good; and 5.9 or below being poor. The results are shown in Table 2.

[0114] Table 2 Umami test and sensory evaluation

[0115]

[0116] Note: All data are expressed as mean ± standard deviation SD (repeated times n = 3).

[0117] According to the results of Example 1 and Comparative Example 2 in Table 2, the composition of the complex enzyme has a great influence on the percentage of umami amino acids in the umami extract to the total free amino acids. The preparation process provided by the present invention can significantly increase the percentage of umami amino acids in the umami extract to the total free amino acids. According to the results of Example 4 and Comparative Example 8, the addition ratio of amylose, cane molasses and bonito meal to the clam slurry will also affect the percentage of umami amino acids in the umami extract to the total free amino acids and the final umami evaluation result. The experiment shows that the addition ratio of amylose, cane molasses and bonito meal can effectively increase the percentage of umami amino acids in the total free amino acids within the specified range of the present invention; and the sensory evaluation of Experiment 3 also confirms the conclusion of Experiment 2.

[0118] The embodiments described above are part of the embodiments of the present application, rather than all of the embodiments. The detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents the selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present application.

Claims

1. A method for preparing an umami extract, characterized in that: The preparation method comprises the following steps: S1: Wash and remove impurities from fresh clam meat, add deionized sterile water, and homogenize to obtain clam meat slurry; S2: fermenting the clam pulp with Heronweissella, filtering to obtain the fermentation liquid after the fermentation, wherein the fermentation time is 18-32 hours and the fermentation temperature is 28-38°C; S3: using bromelain, chymotrypsin and flavor protease to perform ultrasonic enzymolysis on the fermentation broth, filtering to obtain a filtrate after the enzymolysis is completed, freeze-drying the filtrate to obtain a freeze-dried powder, irradiating the freeze-dried powder for sterilization and enzyme inactivation to obtain an umami extract, wherein the ultrasonic power is 135-175w, the enzymolysis pH is 5-7, the enzymolysis temperature is 50-65°C, and the enzymolysis time is 6-12h; The CGMCC NO of the Herrenweissella is 1.2513; The mass ratio of clam meat to deionized sterile water is 1:13, the mass ratio of clam meat slurry to Herrenweissella is 10:0.05-0.07, and the viable count of Herrenweissella is 1×10 7 -1×10 9 CFU / g, the mass ratio of clam pulp, bromelain, chymotrypsin and flavor protease was 10: 0.06-0.08: 0.01-0.03: 0.05-0.

07.

2. The preparation method according to claim 1, characterized in that: In step S2, amylose, cane molasses and bonito powder are further added to the clam meat slurry, and the mass ratio of the clam meat, amylose, cane molasses and bonito powder is 1:0.6-0.8:0.2-0.4:0.8-1.

3. An umami extract obtained according to the preparation method of claim 1 or 2.

4. Use of the umami extract according to claim 3 in preparing food or health products.

5. The use according to claim 4, characterized in that: The umami extract is used as an essence, flavoring agent or flavor enhancer.

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