A umami extract

Abstract

The application discloses a kind of umami extract, belong to food field.The preparation method of the umami extract with clam meat as raw material, after fermentation by specific strain, then the fermentation liquor is subjected to enzymolysis by specific composition of composite enzyme, filtration, irradiation treatment and freeze-drying is prepared.The step of fermentation before enzymolysis in the present application, through the synergistic effect of microorganism and composite enzyme can effectively avoid the generation of bitter taste, fishy smell and other undesirable flavors in extract, and the free amino acid content of the umami extract prepared is significantly increased, and the percentage of glutamic acid, aspartic acid and other umami amino acids in total free amino acid is also significantly improved.In addition, the present application also proves by experiment that adding amylose, sugar cane molasses and bonito powder in clam meat slurry during fermentation can further increase the content of free amino acid and the percentage of umami amino acid in umami extract, so that the flavor of the umami extract prepared is better.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of food additives, in particular to a kind of umami extract. BACKGROUND

[0002] Clam is a kind of marine product belonging to bivalve lamellariada, its meat is delicious, and it is praised as "the first fresh in the world" and "the king of hundred tastes". Clam has the characteristics of high protein, high trace element, high iron, high calcium and low fat, but some people are allergic to some proteins in clam, so they can not enjoy the deliciousness of clam,

[0003] It is well known that umami is usually caused by free amino acids, so to improve umami, the content of free amino acids needs to be improved. In order to improve umami, most of the current seafood flavor products adopt crushing, homogenizing and then enzymatic hydrolysis, but this method has low extraction efficiency, high loss and poor cost performance. With the increasing of biotechnology, researchers found that using fermentation technology to ferment animal protein, different strains have different metabolic pathways in different culture media, resulting in different secondary metabolites produced by fermentation. On the basis of decomposing substrates, fermentation products can be further endowed with more flavor characteristics, but strain fermentation can also lead to excessive secretion of biogenic amines and putrefactive odor due to incomplete utilization of substrates or too long fermentation time, so a method for extracting umami substances from clam is needed, which can fully utilize substrates, avoid the production of biogenic amines and produce more free amino acids. SUMMARY

[0004] The purpose of the present application is to provide a kind of umami extract, the preparation method of the umami extract takes clam meat as raw material, is fermented by specific strain, and then is enzymatically hydrolyzed, filtered, irradiated and freeze-dried by specific composite enzyme.The umami extract obtained by the preparation method of the present application has significantly increased content of free amino acids, and the percentage of umami amino acids such as glutamic acid and aspartic acid in total free amino acids is also significantly improved.

[0005] In order to achieve the above purpose, the technical scheme adopted by the present application is as follows:

[0006] The present application provides a kind of umami extract, which is characterized by comprising the following steps:

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

[0008] S2: use weissella hellenn to ferment the clam meat slurry, filter after fermentation is completed, to obtain fermentation liquor, wherein the fermentation time is 18-32h, and the fermentation temperature is 28-38℃;

[0009] S3: the fermentation liquor is subjected to ultrasonic enzymolysis using bromelain, chymotrypsin and flavor protease, and after the enzymolysis is completed, the filtrate is obtained by filtration, and the filtrate is freeze-dried to obtain a freeze-dried powder, the freeze-dried powder is subjected to irradiation sterilization and enzyme inactivation, and a umami extract is obtained, wherein the ultrasonic power is 135-175w, the enzymolysis pH is 5-7, the enzymolysis temperature is 50-65 DEG C, and the enzymolysis time is 6-12h;

[0010] the veillonella dispar CGMCC NO: 1.2513;

[0011] The mass ratio of the clam meat to the deionized sterile water is 1:13, the mass ratio of the clam meat slurry to the veillonella dispar is 10:0.05-0.07, the viable bacterial count of the veillonella dispar is 1x10 7 -1x10 9 The mass ratio of the clam meat slurry, the bromelain, the chymotrypsin and the flavor protease is 10:0.06-0.08:0.01-0.03:0.05-0.07.

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

[0013] Compared with the prior art, the umami extract provided by the present application has the following advantages: the umami extract is prepared by using clam meat as a raw material, the clam meat is washed, homogenized with water to obtain a clam meat slurry, the clam meat slurry is first subjected to fermentation by using veillonella dispar, and a fermentation liquor is obtained by filtration, and then the fermentation liquor is subjected to enzymolysis by using bromelain, chymotrypsin and flavor protease, and the umami extract is prepared by filtration and freeze-drying after the enzymolysis is completed, wherein the fermentation of the clam meat slurry can produce volatile acids such as acetic acid, propionic acid and butyric acid, ester substances, exocellular polysaccharides and lactic acid, and many components in the above-mentioned substances have unique flavor characteristics, and the use of veillonella dispar, bromelain, chymotrypsin and flavor protease in the fermentation and enzymolysis steps can make the proteins in the clam meat be fully utilized, produce more free amino acids, and effectively avoid the generation of bitter taste, fishy smell and other undesirable flavors in the extract, experiments show that the content of free amino acids 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 obviously improved. In addition, the experiments also prove that the addition of amylose, cane molasses and bonito powder in the clam meat slurry during fermentation can further increase the content of free amino acids and the percentage of umami amino acids in the umami extract, so that the umami extract prepared has better flavor. DETAILED DESCRIPTION

[0014] The embodiments described below are part of the embodiments of the present application, but not all the embodiments of the present application. The embodiments of the present application are used to illustrate the present application, but not to limit the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0015] The experimental methods described in the following examples are generally carried out under conventional conditions, or under the conditions recommended by the manufacturer, unless otherwise specified. The materials, reagents, etc. used, if not specified, can be obtained commercially.

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

[0017] The clams are purchased from Jiangzidao Group Co., Ltd.;

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

[0019] The Weissella helleri is purchased from China General Microbiological Culture Collection Center, with the number CGMCC NO: 1.2513;

[0020] The Weissella cibaria is purchased from China General Microbiological Culture Collection Center, with the number CGMCC NO: 1.2474;

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

[0022] The chymotrypsin is purchased from Wuhan Haorong Biological Technology Co., Ltd., CAS: 9004-07-3, enzyme activity: 20,000 U / g.

[0023] The flavourzyme is purchased from Chongqing Tianlun Biological Products Co., Ltd., CAS: 9001-92-7, enzyme activity: 500,000 U / g.

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

[0025] The trypsin is purchased from Nanning Dongheng Huadao Biological Technology Co., Ltd., CAS: 9002-07-7, enzyme activity: 20,000 U / g.

[0026] The amylose is purchased from Wuhan Huaxiang Kej Biological Technology Co., Ltd., CAS: 9005-82-7.

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

[0028] The bonito powder is purchased from Qingdao Rishengchang Food Ingredients Co., Ltd.

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

[0030] Example 1:

[0031] S1: Fresh clam meat was washed and impurities were removed, deionized sterile water was added, and homogenate was obtained to obtain clam meat slurry;

[0032] S2: The clam meat slurry was fermented using Weisella hellenn, and after fermentation was completed, a fermentation liquor was obtained by filtration, wherein the fermentation time was 25 h, and the fermentation temperature was 33 DEG C;

[0033] S3: The fermentation liquor was subjected to ultrasonic enzymolysis using bromelain, chymotrypsin and flavor protease, and after the enzymolysis was completed, a filtrate was obtained by filtration, the filtrate was freeze-dried to obtain a freeze-dried powder, the freeze-dried powder was subjected to irradiation sterilization and enzyme inactivation, and an umami extract was obtained, wherein the ultrasonic power was 150 w, the enzymolysis pH was 6, the enzymolysis temperature was 60 DEG C, and the enzymolysis time was 8 h;

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

[0035] The mass ratio of the clam meat to deionized sterile water was 1:13, the mass ratio of the clam meat slurry to Weisella hellenn was 10:0.06, and the viable bacterial count of the Weisella hellenn was 1x10 8 CFU / g, and the mass ratio of the clam meat slurry, bromelain, chymotrypsin and flavor protease was 10:0.07:0.02:0.06.

[0036] Example 2:

[0037] S1: Fresh clam meat was washed and impurities were removed, deionized sterile water was added, and homogenate was obtained to obtain clam meat slurry;

[0038] S2: The clam meat slurry was fermented using Weisella hellenn, and after fermentation was completed, a fermentation liquor was obtained by filtration, wherein the fermentation time was 18 h, and the fermentation temperature was 28 DEG C;

[0039] S3: The fermentation liquor was subjected to ultrasonic enzymolysis using bromelain, chymotrypsin and flavor protease, and after the enzymolysis was completed, a filtrate was obtained by filtration, the filtrate was freeze-dried to obtain a freeze-dried powder, the freeze-dried powder was subjected to irradiation sterilization and enzyme inactivation, and an umami extract was obtained, wherein the ultrasonic power was 175 w, the enzymolysis pH was 5, the enzymolysis temperature was 65 DEG C, and the enzymolysis time was 6 h;

[0040] The Weisella hellenn 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 the clam meat to the deionized sterile water is 1:13, the mass ratio of the clam meat slurry to the brochothrix is 10:0.05, the viable count of the brochothrix is 1x10 7 The mass ratio of the clam meat slurry, the bromelain, the chymotrypsin and the flavor protease is 10:0.06:0.03:0.05.

[0045] Example 3:

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

[0047] S2: Ferment the clam meat slurry using brochothrix, and filter the fermented liquid after fermentation is completed, wherein the fermentation time is 32h, and the fermentation temperature is 38℃;

[0048] S3: Perform ultrasonic enzymolysis on the fermented liquid using bromelain, chymotrypsin and flavor protease, filter the filtrate after the enzymolysis is completed, and obtain a freeze-dried powder by freeze-drying the filtrate, and perform irradiation sterilization and enzyme inactivation on the freeze-dried powder to obtain a umami extract, wherein the ultrasonic power is 135w, the enzymolysis pH is 7, the enzymolysis temperature is 50℃, and the enzymolysis time is 12h;

[0049] The brochothrix is purchased from CGMCC NO: 1.2513;

[0050] The mass ratio of the clam meat to the deionized sterile water is 1:13, the mass ratio of the clam meat slurry to the brochothrix is 10:0.07, the viable count of the brochothrix is 1x10 9 The mass ratio of the clam meat slurry, the bromelain, the chymotrypsin and the flavor protease is 10:0.08:0.01:0.07.

[0051] Example 4:

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

[0053] S2: Ferment the clam meat slurry using brochothrix, and filter the fermented liquid after fermentation is completed, wherein the fermentation time is 25h, and the fermentation temperature is 33℃;

[0054] S3: the fermentation liquor is subjected to ultrasonic enzymolysis using bromelain, pepsin and flavor protease, the filtrate is obtained after the enzymolysis is completed, the freeze-dried powder is obtained by freeze-drying the filtrate, the freeze-dried powder is subjected to irradiation sterilization and enzyme inactivation, and the umami extract is obtained, wherein the ultrasonic power is 150w, the enzymolysis pH is 6, the enzymolysis temperature is 60℃, and the enzymolysis time is 8h;

[0055] The Lactobacillus helveticus is purchased from CGMCC NO: 1.2513.

[0056] The mass ratio of the clam meat to the deionized sterile water is 1:13, the mass ratio of the clam meat paste to the Lactobacillus helveticus is 10:0.06, the viable bacterial count of the Lactobacillus helveticus is 1x10 8 CFU / g, and the mass ratio of the clam meat paste, the bromelain, the pepsin and the flavor protease is 10:0.07:0.02:0.06.

[0057] The step S2 further adds amylopectin, cane molasses and bonito powder to the clam meat paste, and the mass ratio of the clam meat, the amylopectin, the cane molasses and the bonito powder is 1:0.7:0.3:0.9.

[0058] Example 5:

[0059] S1: fresh clam meat is washed and impurities are removed, deionized sterile water is added, and the clam meat paste is obtained by homogenization;

[0060] S2: the clam meat paste is subjected to fermentation using Lactobacillus helveticus, and the fermentation liquor is obtained after the fermentation is completed, wherein the fermentation time is 25h, and the fermentation temperature is 33℃.

[0061] S3: the fermentation liquor is subjected to ultrasonic enzymolysis using bromelain, pepsin and flavor protease, the filtrate is obtained after the enzymolysis is completed, the freeze-dried powder is obtained by freeze-drying the filtrate, the freeze-dried powder is subjected to irradiation sterilization and enzyme inactivation, and the umami extract is obtained, wherein the ultrasonic power is 150w, the enzymolysis pH is 6, the enzymolysis temperature is 60℃, and the enzymolysis time is 8h;

[0062] The Lactobacillus helveticus is purchased from CGMCC NO: 1.2513.

[0063] The mass ratio of the clam meat to the deionized sterile water is 1:13, the mass ratio of the clam meat paste to the Lactobacillus helveticus is 10:0.06, the viable bacterial count of the Lactobacillus helveticus is 1x10 8 CFU / g, and the mass ratio of the clam meat paste, the bromelain, the pepsin and the flavor protease is 10:0.07:0.02:0.06.

[0064] The step S2 further adds amylose, sugarcane molasses and bonito powder in the surf clam meat slurry, and the mass ratio of the surf clam meat, the amylose, the sugarcane molasses and the bonito powder is 1:0.6:0.2:1.

[0065] Example 6:

[0066] S1: fresh surf clam meat is taken, cleaned and impurities are removed, deionized sterile water is added, and homogenization is performed to obtain surf clam meat slurry;

[0067] S2: the surf clam meat slurry is fermented by using weissella hellenica, and after fermentation is completed, filtration is performed to obtain fermentation liquor, wherein the fermentation time is 25 h, and the fermentation temperature is 33℃;

[0068] S3: the fermentation liquor is subjected to ultrasonic enzymolysis by using bromelain, pepsin and flavor protease, and after enzymolysis is completed, filtration is performed to obtain filtrate, the filtrate is freeze-dried to obtain freeze-dried powder, the freeze-dried powder is subjected to irradiation sterilization and enzyme inactivation, and a fresh taste extract is obtained, wherein the ultrasonic power is 150 w, the enzymolysis pH is 6, the enzymolysis temperature is 60℃, and the enzymolysis time is 8 h;

[0069] The weissella hellenica is purchased from CGMCC NO:1.2513;

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

[0071] The step S2 further adds amylose, sugarcane molasses and bonito powder in the surf clam meat slurry, and the mass ratio of the surf clam meat, the amylose, the sugarcane molasses and the bonito powder is 1:0.8:0.4:0.8.

[0072] Comparative Example 1:

[0073] The difference between Example 1 and Comparative Example 1 is that weissella kandiahii is used to replace weissella hellenica in Comparative Example 1, and the remaining steps are the same as those in Example 1.

[0074] Comparative Example 2:

[0075] The difference between Example 1 and Comparative Example 2 is that the mass ratio of the surf clam meat slurry, the bromelain, the pepsin and the flavor protease is 10:0.04:0.07:0.04 in Comparative Example 2, and the remaining steps are the same as those in Example 1.

[0076] Comparative Example 3:

[0077] Different from example 1, only chymotrypsin and flavourzyme were used for enzymatic hydrolysis in comparative example 3, and the reduced mass of the above two was proportionally distributed to chymotrypsin and flavourzyme, and the remaining steps were the same as example 1.

[0078] Comparative example 4:

[0079] Different from example 1, only bromelain and chymotrypsin were used for enzymatic hydrolysis in comparative example 4, and the reduced mass of the above two was proportionally distributed to bromelain and chymotrypsin, and the remaining steps were the same as example 1.

[0080] Comparative example 5:

[0081] Different from example 1, only bromelain and flavourzyme were used for enzymatic hydrolysis in comparative example 5, and the reduced mass of the above two was proportionally distributed to bromelain and flavourzyme, and the remaining steps were the same as example 1.

[0082] Comparative example 6:

[0083] Different from example 1, trypsin was used to replace chymotrypsin in comparative example 6, and the remaining steps were the same as example 1.

[0084] Comparative example 7:

[0085] Different from example 1, papain was used to replace bromelain in comparative example 7, and the remaining steps were the same as example 1.

[0086] Comparative example 8:

[0087] Different from example 4, the mass ratio of clam meat, amylose, sugar cane molasses and bonito powder in comparative example 8 was 1:0.3:0.9:0.7, and the remaining steps were the same as example 4.

[0088] Blank:

[0089] S1: Fresh clam meat was washed and impurities were removed, deionized sterile water was added, and homogenate was obtained to obtain clam meat slurry;

[0090] S2: The clam meat slurry was oscillated and extracted at 60℃, the extraction time was 4h, the liquid after extraction was filtered, and then freeze-dried and concentrated to obtain the aqueous extract of clam meat.

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

[0092] Test 1: Determination of total free amino acids

[0093] Test sample: Examples 1-6, comparative examples 1-8, blank.

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

[0095] The free amino acid test kit was purchased from Wuhan Aidi Biological Technology Co., Ltd., and the item number was 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 shows the determination results of the total free amino acid content.

[0098]

[0099]

[0100] Note: The data are expressed as mean ± standard deviation SD (n = 3 times); "*" 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 provided by the present application involving fermentation of specific bacteria and enzyme hydrolysis of specific composite enzymes can significantly improve the content of total free amino acids in the umami extract. As can be seen from Comparative Example 4 and Example 1, the addition of amylose, sugarcane molasses and bonito powder in the clam meat slurry can effectively further improve the content of total free amino acids in the umami extract. As can be seen from Comparative Example 4 and Comparative Example 8, the addition ratio of amylose, sugarcane molasses and bonito powder is also important. The experiment shows that within the scope defined in the present application, the content of total free amino acids in the umami extract can be effectively improved.

[0102] Test 2: percentage of umami amino acids in total free amino acids

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

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

[0105] The glutamic acid test kit was purchased from Shanghai Enzyme-linked Biological Technology Co., Ltd., and the item number was ml077309.

[0106] The aspartic acid test kit was purchased from Shanghai Enzyme-linked Biological Technology Co., Ltd., and the item number was ml077312.

[0107] The alanine test kit was purchased from Shanghai Enzyme-linked Biological Technology Co., Ltd., and the item number was ml077312.

[0108] The glycine test kit was purchased from Shanghai Enzyme-linked Biological Technology Co., Ltd., and the item number was ml076512.

[0109] The results are shown in Table 2.

[0110] Test 3: umami evaluation

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

[0112] Concentration: 1 g / L;

[0113] Detection method: 30 trained sensory evaluation personnel were invited to evaluate the umami of each sample by scoring method (excellent, good, poor), and the total score was 10, 8-10 was excellent, 6-7.9 was good, and 5.9 or less was poor. The results are shown in Table 2.

[0114] Table 2: umami test and sensory evaluation

[0115]

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

[0117] According to the results of Example 1 and Comparative Example 2 in Table 2, the composition of the composite enzyme has a greater impact on the percentage of umami amino acids in the total free amino acids in the umami extract. The preparation process provided by the present application can significantly increase the percentage of umami amino acids in the total free amino acids in the umami extract. According to the results of Example 4 and Comparative Example 8, the addition ratio of amylose, sugarcane molasses and bonito powder in the clam meat slurry also affects the percentage of umami amino acids in the total free amino acids in the umami extract and the final umami evaluation results. Experiments show that the addition ratio of amylose, sugarcane molasses and bonito powder within the range defined in the present application can effectively increase the percentage of umami amino acids in the total free amino acids. And the sensory evaluation of Test 3 also supports the conclusion of Test 2.

[0118] The above-described embodiments are part of the embodiments of the present application, rather than all the embodiments. The detailed description of the embodiments of the present application is not intended to limit the scope of the claimed application, but only represents 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 creative labor fall within the scope of protection of the present application.

Claims

1. A umami extract characterized by, The preparation method of the umami extract comprises the following steps: S1: taking fresh clam meat, washing and removing impurities, adding deionized sterile water, and homogenizing to obtain clam meat slurry; S2: using Weisella hellenn to ferment the clam meat slurry, filtering the fermented liquid after fermentation is completed, wherein the fermentation time is 18-32 h, and the fermentation temperature is 28-38℃; S3: using bromelain, chymotrypsin and flavor protease for ultrasonic enzymolysis of the fermented liquid, filtering the filtrate after the enzymolysis is completed, freeze-drying the filtrate to obtain freeze-dried powder, irradiation sterilization and enzyme inactivation of the freeze-dried powder, and obtaining the umami extract, wherein the ultrasonic power is 135-175 w, the enzymolysis pH is 5-7, the enzymolysis temperature is 50-65℃, and the enzymolysis time is 6-12 h; The preservation number of the Weisella hellenn is CGMCC NO: 1.2513; The mass ratio of the surf clam meat to deionized sterile water is 1:13, the mass ratio of the surf clam meat slurry to the Weissella hellinckii is 10:0.05-0.07, the viable count of the Weissella hellinckii is 1×10 7 -1×10 9 CFU / g, and the mass ratio of the surf clam meat slurry, bromelain, chymotrypsin, and prolamin is 10:0.06-0.08:0.01-0.03:0.05-0.

07.

2. The umami extract according to claim 1, characterized by, The step S2 further adds amylose, cane molasses and bonito powder to the clam meat slurry, and the mass ratio of the clam meat slurry, the amylose, the cane molasses and the bonito powder is 1:0.6-0.8:0.2-0.4:0.8-1.

3. The use of the umami extract in claim 1 or 2 in the preparation of food or health products.

4. Use according to claim 3, characterized in that, The umami extract is used as a flavor, a flavoring agent or a taste enhancer.

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