CN114703112B - Halophilic quadruple coccus halophilic subspecies and application thereof - Google Patents

Halophilic quadruple coccus halophilic subspecies and application thereof Download PDF

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CN114703112B
CN114703112B CN202210627383.XA CN202210627383A CN114703112B CN 114703112 B CN114703112 B CN 114703112B CN 202210627383 A CN202210627383 A CN 202210627383A CN 114703112 B CN114703112 B CN 114703112B
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史晓萌
刘进昌
勇倩倩
刘玮洁
庞雪威
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Yantai Xinhe Enterprise Food Co ltd
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    • A—HUMAN NECESSITIES
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Abstract

The invention relates to the technical field of food fermentation, and particularly discloses halophilic tetrasaccharide halophilic subspecies and application thereof. Wherein, the halophilic tetrasaccharide halophilic subspecies is named as a strain SP101, and the preservation number is CGMCC No. 23611. By applying the technical scheme of the invention, the strain is inoculated in the fermentation process of the sauce and the soy sauce, the growth and metabolism of the amine-producing bacteria in the sauce mash can be inhibited without changing the original fermentation control process, the biogenic amine content of the sauce and the soy sauce is obviously reduced, and the bean sauce and the soy sauce are endowed with excellent flavor.

Description

Halophilic quadruple coccus halophilic subspecies and application thereof
Technical Field
The invention relates to the technical field of food fermentation, in particular to halophilic tetrasococcus halophilic subspecies and application thereof.
Background
With the increase of the importance of food safety, the content of biogenic amine in traditional fermented food in China and abroad is more and more concerned. Biogenic amines are a generic term for a class of biologically active low molecular weight nitrogen-containing organic compounds, mainly histamine, tyramine, putrescine, cadaverine, tryptamine, phenylethylamine, spermine and spermidine. Biogenic amines are present in various tissues of animals and plants, are indispensable components of bioactive cells, and play an important role in regulating the synthesis of nucleic acids and proteins and the stability of biological membranes. Adequate amounts of biogenic amines contribute to normal physiological functions of the human body, but excess amounts can cause adverse physiological reactions.
In food, biogenic amines are formed primarily by decarboxylation of amino acids by microorganisms and are therefore closely related to the type and number of microorganisms in the environment. The sauce and soy sauce are seasonings which are prepared by fermenting through starter propagation by utilizing the action of microorganisms, so that some microorganisms in the sauce and soy sauce can possibly utilize the metabolism of amino acid to generate biogenic amine. The biogenic amine content in the commercial soy sauce is generally 200-2000 mg/L, and the biogenic amine and the histamine are mainly used. Although the national standard does not provide the limit regulation of the biogenic amine in the sauce and the soy sauce, the biogenic amine is researched and recognized from the food safety point of view, and a solution for reducing the biogenic amine content is actively searched, so that the biogenic amine can be used for protecting the quality and safety of products.
There are currently mainly the following three approaches to reduce biogenic amines:
the first method is the method of adding compound. For example, the invention patent application (application No. CN 201910058780.8) discloses a method for controlling the biogenic amine content of soy sauce by adding potassium sorbate to koji. The invention patent application (application number CN 201910058795.4) discloses a method for controlling biogenic amine content of soy sauce by adopting a red bayberry leaf extract. The invention patent (grant publication No. CN 102771752B) discloses a method for reducing biogenic amine content of soy sauce by adding glycine and citric acid.
And secondly, reducing the fermentation temperature. The invention patent application (application number CN 201811239914.8) discloses a method for reducing biogenic amine content of soy sauce by regulating and controlling temperature. The fermentation temperature of the soy sauce mash is controlled to be 30 ℃ for 0-30 days, then the fermentation temperature is 18-20 ℃, the total amount of biogenic amine in the finally obtained soy sauce is below 100mg/L, and the total amount of biogenic amine is reduced by more than 40% compared with that of a blank group.
And thirdly, a biological method. The bacterial strains which have been reported at present and have the effect of reducing biogenic amine or do not produce biogenic amine are mainly used for fermenting sausages, pickles, yellow wine and the like, and are less used for fermenting sauce and soy sauce. For example, the invention patent (publication No. CN 109182188B) discloses that when a Lactobacillus plantarum strain is used for preparing a fermented sausage by using a leavening agent, the obtained fermented sausage does not produce biogenic amine in the curing or fermentation process, and the concentration of tyramine in the mature sausage is obviously lower than that in the prior art. The invention patent (publication No. CN 109401998B) discloses a Lactobacillus mindendori strain which has good amine-reducing capability, can obviously reduce the content of biogenic amine in the rice soaking process, and reduces biogenic amine brought into a yellow wine fermentation system.
The method for controlling biogenic amine by adding the compound has secondary risk and can influence the quality of soy sauce, and the original flavor of the soy sauce is greatly influenced by changing the fermentation temperature. Biological methods are currently the main methods for reducing biogenic amines in food products. However, the microbial community of the sauce and the soy sauce has complex structure, the fermentation salinity is higher than that of other foods, the biogenic amine degradation reaction is an aerobic reaction under the action of amine oxidase, and the fermentation environment of the sauce mash is anaerobic. Therefore, the biogenic amine-degrading strain is difficult to be used in the actual production process of sauces and soy sauce. The screening of the bacterial strains which are suitable for the fermentation of sauce mash of sauce and soy sauce, do not produce biogenic amine and can inhibit the growth of biogenic amine-producing microorganisms has important significance for producing healthy and safe sauce and soy sauce.
Disclosure of Invention
The invention aims to provide halophilic tetrasococcus halophilus subspecies and application thereof, which can reduce the content of biogenic amine in sauce and soy sauce without adding other compounds.
In order to realize the aim, the invention provides a halophilic tetragenococcus halophilic subspecies, which is named as a strain SP101 and is preserved in China general microbiological culture collection center in 10 months and 15 days 2021, the preservation address is Beijing, China, the preservation number is CGMCC number 23611, and the strain is named as the halophilic tetragenococcus according to classificationTetragenococcus halophilus。
The invention also provides application of the halophilic tetragenococcus halophilic subspecies in brewing of soy sauce or brewing sauce.
Further, comprising: mixing the finished starter with saline water to prepare soy sauce mash, inoculating the strain SP101 to the soy sauce mash fermented for 0-15 days for soy sauce or brewing sauce production fermentation, wherein the inoculation amount is 10 6 ～10 7 One per gram of soy sauce mash.
Further, soy sauce production fermentation includes: the early fermentation temperature is 8-10 ℃, and the fermentation time is 1-1.5 months; the middle-stage fermentation temperature is 20-25 ℃, and the fermentation time is 2.5-3 months; the later-stage fermentation temperature is 30-32 ℃, the time is 1-2 months, and the total of the early-stage fermentation, the middle-stage fermentation and the later-stage fermentation is 6 months of closed fermentation; preferably, the concentration of the saline water used in the soy sauce mash for producing the soy sauce is 16-20%, and the finished koji and the saline water are mixed according to the weight ratio of 1: 1-1: 1.5.
Further, the brew production fermentation includes: the fermentation temperature is kept at 25-35 ℃, and the fermentation time is 70-100 days; preferably, the concentration of the saline water used in the sauce mash for producing the brewing sauce is 20.5% -23.5%; the addition amount of the saline water is 0.9-1.5 times of the weight of the koji-forming agent.
Further, before inoculation, the method also comprises a step of activating the strain, wherein the activation comprises the following steps: inoculating the strain SP101 into an MRS culture medium containing 10% -20% of salt, and culturing at the temperature of 30-35 ℃ under an anaerobic condition for 48-72 h.
Further, the koji for producing soy sauce is prepared by the following steps: the starter propagation of soy sauce comprises: mixing the pretreated defatted soybeans and fried wheat according to the ratio of 1: 1-1: 1.5, inoculating aspergillus oryzae, and making starter propagation at 28-40 ℃ for 40-42 hours to obtain soy sauce finished starter; preferably, the pretreatment of defatted soybeans comprises: cleaning the defatted soybeans, controlling the temperature to be 115-120 ℃, controlling the pressure to be 0.05-0.15 MPa, and cooking for 8-10 minutes; the pretreatment of the fried wheat comprises the following steps: controlling the temperature to be 300-400 ℃, controlling the pressure to be 0.01-0.05 MPa, controlling the water content of the fried wheat to be 0.5-1%, and finally crushing the wheat, wherein the particle size of the crushed wheat is 20-100 meshes.
Further, a koji for use in the production of a brew is prepared by the steps of: the brewing sauce koji making comprises: mixing the pretreated soybean and/or wheat flour according to the ratio of 1: 10-1: 1, inoculating aspergillus oryzae, and making yeast at 28-35 ℃ for 38-45 hours to obtain brewed sauce yeast; preferably, the pretreatment of the soybeans comprises: after the soybeans are cleaned, the soybeans are cooked for 8-10 minutes at the temperature of 98-120 ℃; pretreatment of wheat flour includes: and (3) steaming at the temperature of 98-120 ℃ for 20-30 s, and controlling the water content to be 22-24%.
The invention also provides the soy sauce. The soy sauce is obtained by brewing soy sauce with the halophilic tetrasaccharide halophilic subspecies, and has biogenic amine content below 100 mg/L.
The invention also provides a brewing sauce, which is obtained by brewing the brewing sauce by adopting the halophilic tetrasaccharide halophilic subspecies, and the biogenic amine content of the brewing sauce is below 100 mg/L.
By applying the technical scheme of the invention, the halophilic tetrasococcus halophilic subspecies is inoculated in the fermentation process of the sauce and the soy sauce, the growth and metabolism of amine-producing bacteria in soy sauce mash can be inhibited without changing the original fermentation control process, the biogenic amine content of the sauce and the soy sauce is obviously reduced, and the bean sauce and the soy sauce are endowed with excellent flavor.
Detailed Description
It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present invention will be described in detail with reference to examples.
The invention is realized by the following technical scheme:
according to a typical embodiment of the present invention, there is provided a strain of Tetragenococcus halophilus subspecies halophilusTetragenococcus halophilus subsp. HalophilusSP101, used in sauce and soy sauce production process to reduce biogenic amine content.
Halophilic Tetragenococcus halophilic subspeciesT. halophilus subsp. HalophilusSP101, isolated from soy sauce mash fermented for 2 months. The obtaining method comprises the following steps: using 10% salt-containing MRS culture medium, adopting gradient dilution method to make separation culture, selecting strain with good growth vigor to make primary screening, after the primary screening strain is enriched, respectively inoculating them into soy sauce mash fermented for 0 day according to 1% inoculum size, after the fermentation is completed, utilizing high-performance liquid phase method to detect biogenic amine content to make secondary screening so as to finally obtain the bacterial strains SP101 and SP102 capable of reducing biogenic amine, and the molecular biological identification result shows that two bacterial strains are respectively halophilic tetragenococcus halophilic subspeciesT. halophilus subsp. HalophilusWherein SP101 has better effect of reducing biogenic amine.
SP101 strain characteristics were studied as follows: inoculating the strain SP101 on an MRS plate containing 10% of salt, and carrying out anaerobic culture at 30 ℃ for 4 days, wherein the bacterial colony is milky white, round and convex, opaque and neat in edge; the thalli are observed to be spherical under a microscope, are arranged in pairs or in quadruples, and are gram-positive. The results of the amino acid decarboxylase reaction show that it has no tyrosine and histidine decarboxylase activity, i.e., produces no tyramine and histamine. The strain SP101 was inoculated at 2% inoculum size into MRS broth with 10%, 14%, 18%, 20%, 22%, 24% salt concentration, respectively, and cultured at 30 ℃ for 48 hours with standing, the OD of which was 600nm The values are all larger than 1, which shows that the strain SP101 can grow and propagate under the salt concentration of 10% -24%, and can rapidly adapt to the soy sauce mash environment after being inoculated to the soy sauce mash and grow and propagate.
According to a typical embodiment of the invention, the use of strain SP101 comprises: mixing the finished starter with saline water to prepare soy sauce mash, inoculating the strain SP101 to the soy sauce mash fermented for 0-15 days for soy sauce or brewing sauce production fermentation, wherein the inoculation amount is 10 6 ～10 7 One per gram of soy sauce mash.
Inoculating halophilic tetrasococcus halophilic subspecies in sauce and soy sauce fermentation processT. halophilus subsp. HalophilusSP101 (SP 101 in the deposit survival evidence, Tetragenococcus halophilus)Tetragenococcus halophilus) The strain can inhibit the growth and metabolism of amine-producing bacteria in soy sauce mash without changing the original fermentation control process, obviously reduce the content of biogenic amine in the sauce and the soy sauce, and endow the bean sauce and the soy sauce with excellent flavor. Preferably, the soy sauce production fermentation comprises: the early fermentation temperature is 8-10 ℃, and the fermentation time is 1.5 months; the middle-stage fermentation temperature is 20-25 ℃, and the fermentation time is 2.5-3 months; the later-stage fermentation temperature is 30-32 ℃, the time is 1-2 months, and the total of the early-stage fermentation, the middle-stage fermentation and the later-stage fermentation is 6 months of closed fermentation; preferably, the concentration of the saline water used in the soy sauce mash for producing soy sauce is 16-20%, and the finished koji and the saline water are mixed according to the weight ratio of 1: 1-1: 1.5. Preferably, the brew production fermentation comprises: the fermentation temperature is kept at 25-35 ℃, and the fermentation time is 70-100 days; preferably, the concentration of the saline water used in the sauce mash for producing the brewing sauce is 20.5% -23.5%; the addition amount of the brine is 0.9 to 1.5 times of the weight of the koji forming agent. The sauce or brewed sauce produced by the process has good flavor.
In order to improve the fermentation activity of the strain, the method also comprises the step of activating the strain before inoculation, wherein the activation comprises the following steps: the strain SP101 is inoculated in MRS broth (Beijing Luqiao, product number CM 187) containing 10% -20% of salt, and is subjected to static culture at the temperature of 30-35 ℃ for 48-72 hours.
According to a typical embodiment of the present invention, the koji for producing soy sauce is prepared by the steps of: the soy sauce starter propagation method comprises the following steps: mixing the pre-treated defatted soybeans and fried wheat according to a ratio of 1: 1-1: 1.5, inoculating Aspergillus oryzae (any Aspergillus oryzae conventionally used in the field), and making starter at 28-40 ℃ for 40-42 h to obtain soy sauce finished starter; preferably, the pretreatment of defatted soybeans comprises: cleaning the defatted soybeans, controlling the temperature to be 115-120 ℃, controlling the pressure to be 0.05-0.15 MPa, and cooking for 8-10 minutes; the pretreatment of the fried wheat comprises the following steps: controlling the temperature to be 300-400 ℃, controlling the pressure to be 0.01-0.05 MPa, controlling the water content of the fried wheat to be 0.5-1%, and finally crushing the wheat, wherein the particle size of the crushed wheat is 20-100 meshes. After pretreatment, the aspergillus oryzae can be fully contacted with the raw materials, so that the utilization capacity of the raw materials is improved.
According to a typical embodiment of the present invention, a koji for use in the production of a brew is prepared by the steps of: the sauce brewing koji comprises: mixing the pretreated soybean and/or wheat flour according to a ratio of 1: 10-1: 1, inoculating Aspergillus oryzae (any Aspergillus oryzae conventionally used in the field), and making starter at 28-35 deg.C for 38-45 h to obtain fermented soybean paste; preferably, the pretreatment of the soybeans comprises: after cleaning soybeans, controlling the temperature to be 98-120 ℃, and cooking for 8-10 minutes; pretreatment of wheat flour includes: and (3) steaming at the temperature of 98-120 ℃ for 20-30 s, and controlling the water content to be 22-24%. After pretreatment, the aspergillus oryzae can be fully contacted with the raw materials, so that the utilization capacity of the raw materials is improved.
According to still another aspect of the present invention, there is provided a soy sauce. The soy sauce is prepared from above halophilic Tetragenococcus halophilus subspecies in soy sauce brewing, and has biogenic amine content below 100 mg/L.
According to a further aspect of the present invention, there is provided a brew sauce prepared by use of the above mentioned halophilous tetrasaccharide subspecies in brewing a brew sauce having a biogenic amine content of less than 100 mg/L.
The following examples are provided to further illustrate the advantageous effects of the present invention.
In the following examples, koji for soy sauce was prepared by the following steps: the pretreatment of defatted soybeans comprises: after cleaning the defatted soybeans, controlling the temperature to be 115-120 ℃, controlling the pressure to be 0.05-0.15 MPa, and cooking for 9 minutes; the pretreatment of the fried wheat comprises the following steps: controlling the temperature to be 300-400 ℃, controlling the pressure to be 0.01-0.05 MPa, controlling the water content of the fried wheat to be 0.8%, and finally crushing the wheat until the particle size is 20-100 meshes. And (3) mixing the pretreated defatted soybeans and fried wheat according to a ratio of 1:1.5, inoculating aspergillus oryzae, and making koji at 28-40 ℃ for 42h to obtain the soy sauce koji.
The finished yeast of the brewing sauce is prepared by the following steps: the pretreatment of soybeans comprises: after cleaning soybeans, controlling the temperature to be 98-120 ℃, and cooking for 9 minutes; pretreatment of wheat flour includes: and (3) steaming at the temperature of 98-120 ℃ for 200s, and controlling the water content to be 22%. Mixing the pretreated soybean and/or wheat flour according to a ratio of 1:2, inoculating aspergillus oryzae, and making yeast at 28-35 ℃ for 38 hours to obtain the fermented soybean paste.
The strain activation comprises the following steps: the strain is inoculated in an MRS culture medium containing 10% of salt, and is cultured for 48 hours at the temperature of 30-35 ℃ under the anaerobic condition.
Example 1
Strain SP101 has effect of reducing soy sauce biogenic amine
Halophilic tetrasococcus halophilic subspeciesT. halophilus subsp. Halophilus After SP101 activation culture, the inoculum size was 10 6 Each gram of soy sauce mash is inoculated to soy sauce mash fermented for 0 day (the soy sauce mash is formed by mixing finished koji and 17% saline water according to the weight ratio of 1: 1.5), and used as an experimental group 1, and halophilic tetrasococcus halophilic subspeciesT. halophilus subsp.Halophilus After SP102 activation culture, the inoculum size was 10 6 Each gram of soy sauce mash is respectively inoculated to the same batch of soy sauce mash fermented for 0 day as an experimental group 2; the same batch of 0 day moromi without SP101 inoculation was used as a control. The experimental group and the control group are controlled according to the conventional fermentation process of the high-salt dilute soy sauce (the early-stage fermentation temperature is 8-10 ℃, the fermentation time is 1.5 months, the middle-stage fermentation temperature is 20-25 ℃, the fermentation time is 2.5 months, and the later-stage fermentation temperature is 30-32 ℃, and the fermentation time is 2 months). After fermenting for 6 months, squeezing the fermented product, extracting raw soy sauce, and pasteurizing to obtain the raw soy sauce. The content of the biogenic amine is detected by a high performance liquid method according to the determination of biogenic amine in GB 5009.208-2016 food safety national standard food, and the content of amino acid nitrogen is detected according to the determination of amino acid nitrogen in GB 5009.235-2016 food safety national standard food. Compared with the control group, the contents of tyramine and histamine in the experimental group 1 and the experimental group 2 are both obviously reduced, and the casein in the experimental group 1The contents of amine and histamine are lower, which shows that SP101 and SP102 both have the effect of reducing the biogenic amine content of the soy sauce, and the effect of SP101 on reducing biogenic amine is better. In addition, the content of amino acid nitrogen in the experimental group 1 is obviously higher than that in the control group, which shows that the content of amino acid nitrogen is increased while the content of soy sauce biogenic amine is reduced by inoculating SP 101. The specific test results are shown in Table 1.
Table 1: contents of biogenic amine and amino acid nitrogen in raw soy sauce juice
Figure DEST_PATH_IMAGE001
Remarking: ND means not detected, and the detection limit is 50 mg/kg.
Example 2
Inhibition effect of strain SP101 on amine-producing bacteria in soy sauce mash
Taking high-salt dilute soy sauce mash fermented for 0 day (the same soy sauce mash of example 1), and separating Aminogen (halophilic Tetracoccus halophilus subspecies) in laboratoryT. halophilus subsp. Halophilus SP 133) and halophilic Tetragenococcus halophilus subspeciesT. halophilus subsp. Halophilus Respectively performing activated culture on SP101, inoculating Aminogen with the inoculum size of 10 4 ～10 5 Each gram of soy sauce mash is inoculated with SP101, the inoculation amount is 10 6 Each gram of moromi as experimental group 1; inoculating Aminogen with the inoculum size of 10 4 ～10 5 Each gram of soy sauce mash is inoculated with SP102, the inoculation amount is 10 6 Pieces/g of moromi as experimental group 2; inoculating amine-producing bacteria only with the inoculation amount of 10 4 ～10 5 The same batch of fermented 0-day moromi per gram of moromi was used as a control group 1, and the same batch of fermented 0-day moromi without inoculated strain was used as a control group 2. The experimental group and the control group are controlled according to the conventional normal fermentation process of the high-salt dilute soy sauce mash (the early-stage fermentation temperature is 8-10 ℃, the fermentation time is 1.5 months, the middle-stage fermentation temperature is 20-25 ℃, the fermentation time is 2.5 months, and the later-stage fermentation temperature is 30-32 ℃, and the fermentation time is 2 months). After fermentation for 6 months, squeezing the fermented product, extracting raw soy sauce, and pasteurizing to obtain the raw soy sauce. Squeezing the sauce mash,Sterilizing to obtain raw juice, detecting the content of biogenic amine by high performance liquid chromatography with reference to 'determination of biogenic amine in GB 5009.208-2016 food safety national standard food', and detecting the content of amino acid nitrogen with reference to 'determination of amino acid nitrogen in GB 5009.235-2016 food safety national standard food'. The content of tyramine in the control group 1 is obviously improved compared with the control group 2 because of the inoculation of the amine-producing bacteria. Compared with the control group 1, the contents of tyramine and histamine in the sauce mash of the experimental group 1 and the experimental group 2 are both obviously reduced, which shows that both SP101 and SP102 can generate certain inhibition effect on the amine-producing bacteria, wherein the contents of tyramine and histamine in the experimental group 1 are lower than those in the control group 2, which shows that the inhibition effect on the amine-producing bacteria is stronger, and the effect of reducing the content of biogenic amine is better. Since SP101 inhibits the growth and metabolism of amine-producing bacteria and reduces decarboxylation of amino acid to produce biogenic amine, the content of amino acid nitrogen in the fermented soy sauce raw juice is higher. The specific test results are shown in Table 2.
Table 2: contents of biogenic amine and amino acid nitrogen in raw soy sauce juice
Figure DEST_PATH_IMAGE002
Remarking: ND means not detected, and the detection limit is 50 mg/kg.
Example 3
Strain SP101 has effect of reducing biogenic amine of fermented sauce
Collecting fermented soybean paste sauce mash (the sauce mash is prepared by mixing finished koji and 22% saline water at a weight ratio of 1: 1), and adding halophilic Tetracoccus tetragenococcus halophilus subspeciesT. halophilus subsp. HalophilusAfter SP101 activation culture, the inoculum size was 10 6 The sauce mash per gram is respectively inoculated to sauce mash fermented for 0 day as an experimental group 1; halophilic tetrasococcus halophilic subspeciesT. halophilus subsp. HalophilusAfter SP102 activation culture, the inoculum size was 10 6 Each gram of moromi is respectively inoculated to moromi fermented for 0 day as an experimental group 2; the control group was soy sauce mash without SP101 inoculation. The experimental group and the control group are controlled according to the normal fermentation process of the broad bean paste (the fermentation temperature is maintained at 25-30 ℃, and the fermentation time is 90 days). Fermentation ofAfter the end, the raw bean paste is pasteurized to obtain raw bean paste, and the biogenic amine content of the raw bean paste is detected by a high performance liquid chromatography with reference to 'determination of biogenic amine in GB 5009.208-2016 food safety national standard food', and the content of amino acid nitrogen is detected by reference to 'determination of amino acid nitrogen in GB 5009.235-2016 food safety national standard food'. Compared with the control group, the content of tyramine and histamine in the experimental group 1 and the experimental group 2 is obviously reduced, which shows that the SP101 and the SP102 have the function of reducing the content of biogenic amine in fermented sauce. Among them, the lower contents of tyramine and histamine in experimental group 1 indicate that SP101 has a better effect of reducing biogenic amine. The specific test results are shown in Table 3.
Table 3: the contents of biogenic amine and amino acid nitrogen in the raw juice of soy sauce
Figure DEST_PATH_IMAGE003
Remarking: ND means not detected, and the detection limit is 50 mg/kg.
Example 4
Application of strain SP101 in fermentation of sauce and soy sauce for influencing flavor of products
Sensory evaluation was performed on the soy sauce raw juice of example 1 and the raw sauce of example 3. 12 professional appraisers in the interior are selected, and appraisal is carried out on the body state, the color and luster, the fragrance, the sour taste, the sweet taste, the bitter taste, the salty taste, the delicate flavor, the sauce fragrance and the comprehensive taste, wherein the score is 0-5, and the higher the score is, the better the index is. The results are shown in tables 4 and 5.
The data in table 4 show that the experimental group 1, the experimental group 2 and the control group have no significant difference in a single sensory dimension, but the experimental group 1 is superior to the experimental group 2 and the control group in a comprehensive taste score. The application of the strain SP101 to soy sauce fermentation can enable the flavor of the soy sauce to be more harmonious and endow the soy sauce with good flavor.
The data in table 5 show that the sour and sweet scores of the experimental group 1 are significantly higher than those of the experimental group 2 and the control group, and the panelists feedback that the control group and the experimental group 2 are slightly sour, which results in the sour and sweet scores being affected, while the experimental group 1 is moderately sour and sweet. The possible reason is that the strain SP101 inhibits the growth and metabolism of part of acid-producing bacteria in the moromi while inhibiting the amine-producing bacteria in the moromi. And the comprehensive mouthfeel score also shows that the comprehensive mouthfeel of the experimental group 1 is the best. In conclusion, sensory score data indicate that strain SP101, when used in sauce fermentation, also imparts good flavour to the sauce.
Table 4: effect of Strain SP101 on Soy sauce flavor
Figure DEST_PATH_IMAGE004
Table 5: effect of Strain SP101 on sauce flavor
Figure DEST_PATH_IMAGE005
From the above description, it can be seen that the above-described embodiments of the present invention achieve the following technical effects:
the invention patent application (application number CN 202010259690.8) discloses a method for inhibiting biogenic amine accumulation of amine-producing bacteria by using hurdle technology, which adopts a mode of combining salinity and lactobacillus plantarum to form multiple inhibition on the amine-producing strains, thereby reducing the biogenic amine content. The invention patent (grant publication No. CN 110499271B) discloses that the compound inoculation of lactobacillus plantarum QR19 and zygosaccharomyces rouxii S96 is used for a soy sauce production process, so that the content of biogenic amine in soy sauce is reduced, and the SOD activity is improved. The invention patent (grant publication No. CN 109938243B) utilizes the combined fermentation of staphylococcus xylosus and tetragenococcus halophilus to control the nitrogen metabolism hazard in soy sauce.
Compared with the invention patent application (application No. CN 202010259690.8), the invention does not need to increase the salinity, and only passes halophilic subspecies of halophilic tetragenococcusT. halophilus subsp. HalophilusSP101 can achieve the effect of inhibiting the amine-producing strains, and the cost is lower because the salinity of the fermentation system does not need to be increased, and the influence on the flavor characteristics of the product is smaller because the salinity of the fermentation system is unchanged and the influence on microorganisms in the fermentation process is smaller.
Invention patent (grant publication No. CN 11049927)1B) The invention patent (No. CN 109938243B) combines two strains to reduce biogenic amine in soy sauce, the halophilic tetragenococcus halophilus subspecies used in the inventionT. halophilus subsp. HalophilusSP101 can tolerate the salt concentration of 10% -24%, can grow and reproduce under the anaerobic condition, has good adaptability in soy sauce mash, and can achieve the inhibition effect on amine-producing bacteria only by using the strain, thereby reducing the content of biogenic amine in soy sauce. Compared with inoculating two strains, the method has the advantages of lower cost of increasing the strains by using a single strain and more convenient control of the fermentation process. In addition, the strain can tolerate the salt concentration of 10% -24%, not only can be used for reducing the biogenic amine content of soy sauce, but also can be used for reducing the biogenic amine content of fermented sauce, and the application range of the strain is wider, so that the invention has remarkable advantages.
In conclusion, the above patent applications and patents can achieve the effect of reducing biogenic amine in soy sauce by using the microorganism combination hurdle technology or using two strains of bacteria. Compared with the prior art, the invention only uses halophilous tetrasococcus halophilous subspeciesT. halophilus subsp. HalophilusSP101 can inhibit the amine-producing strain, reduce the content of biogenic amine, reduce decarboxylation of amino acid to generate biogenic amine, increase the content of amino acid nitrogen, and endow sauce and soy sauce with excellent flavor.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (12)

1. The halophilic tetragenococcus halophilic subspecies is named as a strain SP101, is preserved in China general microbiological culture Collection center in 2021 year 10 month 15 days, has the preservation address of Beijing, China and the preservation number of CGMCC number 23611, and is classified and named as the halophilic tetragenococcusTetragenococcus halophilus。
2. Use of the tetrasaccharide halophilus subspecies of claim 1 in soy sauce or brew brewing.
3. The use according to claim 2, comprising: mixing finished koji with saline water to prepare soy sauce mash, inoculating the strain SP101 to the soy sauce mash fermented for 0-15 days for soy sauce or brewing sauce production fermentation, wherein the inoculation amount is 10 6 ～10 7 One per gram of soy sauce mash.
4. Use according to claim 2, wherein the soy sauce production fermentation comprises: the early fermentation temperature is 8-10 ℃, and the fermentation time is 1-1.5 months; the middle-stage fermentation temperature is 20-25 ℃, and the fermentation time is 2.5-3 months; and the later-stage fermentation temperature is 30-32 ℃, the time is 1-2 months, and the early-stage fermentation, the middle-stage fermentation and the later-stage fermentation are totally sealed and fermented for 6 months.
5. The use according to claim 3, characterized in that the concentration of the brine used in the moromi for producing soy sauce is 16-20%, and the starter and the brine are mixed in a weight ratio of 1: 1-1: 1.5.
6. Use according to claim 2, characterized in that the brew production fermentation comprises: the fermentation temperature is kept at 25-35 ℃, and the fermentation time is 70-100 days.
7. Use according to claim 3, characterized in that in the moromi used for the production of the brew, the brine is used in a concentration of 20.5% to 23.5%; the addition amount of the brine is 0.9-1.5 times of the weight of the koji forming agent.
8. Use according to claim 2, characterized in that it further comprises, before inoculation, a step of activation of the strain, said activation comprising: inoculating the strain SP101 to an MRS culture medium containing 10% -20% of salt, and culturing at the temperature of 30-35 ℃ under an anaerobic condition for 48-72 hours.
9. Use according to claim 2, characterized in that the koji for producing soy sauce is prepared by the following steps: the soy sauce starter propagation method comprises the following steps: mixing the pretreated defatted soybeans and fried wheat according to a ratio of 1: 1-1: 1.5, inoculating aspergillus oryzae, and carrying out starter propagation at 28-40 ℃ for 40-42 h to obtain the soy sauce finished starter.
10. Use according to claim 9, wherein the pre-treatment of defatted soybeans comprises: after cleaning the defatted soybeans, controlling the temperature to be 115-120 ℃, controlling the pressure to be 0.05-0.15 MPa, and cooking for 8-10 minutes; the pretreatment of the fried wheat comprises the following steps: controlling the temperature to be 300-400 ℃, controlling the pressure to be 0.01-0.05 MPa, controlling the water content of the fried wheat to be 0.5-1%, and finally crushing the wheat, wherein the particle size of the crushed wheat is 20-100 meshes.
11. Use according to claim 2, characterized in that the koji used for the production of a brew is prepared by the following steps: the brewing sauce koji comprises: mixing the pretreated soybean and/or wheat flour according to the ratio of 1: 10-1: 1, inoculating aspergillus oryzae, and making yeast at 28-35 ℃ for 38-45 hours to obtain the fermented soy sauce.
12. Use according to claim 11, wherein the pre-treatment of the soybeans comprises: after the soybeans are cleaned, the soybeans are cooked for 8-10 minutes at the temperature of 98-120 ℃; the pretreatment of the wheat flour comprises the following steps: and (3) steaming at the temperature of 98-120 ℃ for 20-30 s, and controlling the water content to be 22-24%.
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