CN114437970A - Leavening agent for efficiently transforming sodium glutamate, fermented milk and preparation method of fermented milk - Google Patents

Leavening agent for efficiently transforming sodium glutamate, fermented milk and preparation method of fermented milk Download PDF

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CN114437970A
CN114437970A CN202210081082.1A CN202210081082A CN114437970A CN 114437970 A CN114437970 A CN 114437970A CN 202210081082 A CN202210081082 A CN 202210081082A CN 114437970 A CN114437970 A CN 114437970A
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sodium glutamate
milk
fermented milk
streptococcus thermophilus
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CN114437970B (en
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洪环
鄢明辉
王伟军
杨欢东
陈波
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Shanghai Xiran Biotechnology Co ltd
Zhejiang Inm Food Co ltd
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Zhejiang Inm Food Co ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
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    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
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    • A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
    • A23C9/1322—Inorganic compounds; Minerals, including organic salts thereof, oligo-elements; Amino-acids, peptides, protein-hydrolysates or derivatives; Nucleic acids or derivatives; Yeast extract or autolysate; Vitamins; Antibiotics; Bacteriocins
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/005—Amino acids other than alpha- or beta amino acids, e.g. gamma amino acids

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Abstract

The application relates to the field of fermented foods, and particularly discloses a leavening agent for efficiently converting sodium glutamate, fermented milk and a preparation method of the fermented milk. The leaven comprises the following components: sugar, fermentation strain and sodium glutamate, wherein the weight ratio is 5-10: 0.5-3: 0.1-0.5; the fermentation strain adopts streptococcus thermophilus, and the preservation number of the streptococcus thermophilus is CGMCC No. 23415; the fermented milk is prepared by adopting a leavening agent as a raw material. The preparation method of the fermented milk comprises the following steps: s1, preparing a mixed milk raw material: mixing and dissolving raw milk, sugar and sodium glutamate, and sterilizing to obtain a mixed milk raw material; s2, preparing fermented milk: inoculating the fermentation strain into the mixed milk raw material, uniformly stirring, and fermenting at 30-45 deg.C for 40-55h to obtain fermented milk; and S3, refrigerating and storing. The fermented milk prepared by the method has a large amount of gamma-aminobutyric acid, effectively reduces the residual amount of sodium glutamate, and has a positive improvement effect on human health.

Description

Leaven for efficiently converting sodium glutamate, fermented milk and preparation method of fermented milk
Technical Field
The application relates to the field of fermented foods, in particular to a leavening agent for efficiently transforming sodium glutamate, fermented milk and a preparation method of the fermented milk.
Background
Gamma-aminobutyric acid (GABA), also known as aminobutyric acid, is an important inhibitory neurotransmitter in the central nervous system and has biological functions of enhancing brain activity, soothing the nerves, regulating hormone secretion, improving lipid metabolism, lowering blood pressure and the like. GABA, a small molecular weight nonprotein amino acid, has been confirmed to be safe for eating, to have good water solubility and thermal stability, and to be useful for the production of foods such as beverages. Research shows that the ingestion of a certain amount of GABA has the physiological effects of improving the sleep quality of organisms, reducing blood pressure and the like. In recent years, research and development of GABA-rich food become a hot spot of research at home and abroad.
Nowadays, GABA has been industrially produced by a method of microbial fermentation, such as fermentation using lactic acid bacteria having GABA-producing ability in the related art to obtain GABA-enriched foods. Most of lactic acid bacteria capable of producing gamma-aminobutyric acid in the related art use glutamic acid or salts thereof as a substrate, and the substrate is decarboxylated by glutamate decarboxylase to synthesize gamma-aminobutyric acid, so that more glutamic acid or glutamate needs to be added to a product to ensure the yield of gamma-aminobutyric acid when preparing related products.
The inventor researches and discovers that the residual quantity of sodium glutamate in the related technology is high, and the application of the sodium glutamate in the field of food can cause more sodium glutamate to be ingested by consumers, so that the sodium glutamate is not beneficial to human health.
Disclosure of Invention
In order to reduce the residual quantity of sodium glutamate, the application provides a leaven for efficiently converting sodium glutamate, fermented milk and a preparation method of the fermented milk.
In a first aspect, the application provides a leaven for efficiently transforming sodium glutamate, which adopts the following technical scheme: a leaven for efficiently transforming sodium glutamate is characterized in that: the leavening agent comprises the following components in parts by weight: sugar, fermentation strain and sodium glutamate, wherein the weight ratio is 5-10: 0.5-5: 0.1 to 1;
the fermentation strain adopts streptococcus thermophilus, and the preservation number of the streptococcus thermophilus is CGMCC No. 23415.
By adopting the technical scheme, the streptococcus thermophilus adopted by the application can be fermented to produce a large amount of gamma-aminobutyric acid, has the capacity of efficiently converting sodium glutamate, and can effectively solve the problem of high residual quantity of sodium glutamate.
The streptococcus thermophilus adopted by the application uses sodium glutamate as a substrate, and the sodium glutamate is decarboxylated through glutamate decarboxylase to synthesize gamma-aminobutyric acid, so that the yield of the gamma-aminobutyric acid and the conversion rate of the sodium glutamate are effectively improved.
And the sugar can provide energy required by the activity of the streptococcus thermophilus so as to ensure sufficient fermentation.
Preferably, the weight ratio of the sugar to the fermentation strain to the sodium glutamate is 7: 2: 0.2.
by adopting the technical scheme, the yield of the gamma-aminobutyric acid is the highest at the specific addition ratio, and the residual quantity of the sodium glutamate is the lowest, so that the method is beneficial to the health of a human body to the maximum extent.
Preferably, the fermentation strain is subjected to an activation treatment, and the activation treatment comprises the following steps:
s1, culture of strains: inoculating streptococcus thermophilus into an MC culture medium, culturing and preliminarily activating strains;
s2, activation of the strain: and selecting a single colony with better growth from S1, transferring the single colony to an MRS culture medium, and culturing to obtain activated streptococcus thermophilus.
By adopting the technical scheme, the streptococcus thermophilus is cultured by the MC culture medium and the single colony with better growth vigor, namely higher activity, is transferred to the MRS culture medium for culture, so that the streptococcus thermophilus is further activated. The streptococcus thermophilus is cultured through the MC culture medium firstly, so that strains with high activity can be screened conveniently, useless mixed bacteria are screened out, and further pure-strain activation culture is carried out on the strains through the MRS culture medium.
Preferably, S1, culture of the species: sterilizing MC culture medium, cooling, pouring into aseptic culture dish, preparing plate, extracting Streptococcus thermophilus, streaking on MC plate, culturing in incubator, and activating strain.
By adopting the technical scheme, the strain is cultured and purified by adopting the MC plate streaking mode, so that the strain is beneficial to screening useless mixed bacteria and is also convenient for screening the strain with higher activity.
Preferably, S2, activation of the species: and extracting single colonies with better growth potential on the MC plate in the S1, transferring the single colonies into an MRS culture medium, and putting the single colonies into a constant-temperature incubator with the temperature of 30-45 ℃ for culture and activation for 20-50h to obtain activated streptococcus thermophilus.
By adopting the technical scheme, a proper growth environment is provided for the streptococcus thermophilus by carrying out constant-temperature culture in the MRS culture medium so as to fully activate the streptococcus thermophilus and enable the strains to achieve the optimal activity.
In a second aspect, the present application provides a fermented milk, which adopts the following technical scheme:
a fermented milk characterized by: the fermentation agent for highly efficiently converting sodium glutamate as claimed in any one of claims 1 to 3 is used as a raw material.
By adopting the leavening agent as a raw material, the prepared fermented milk has the advantages of high content of gamma-aminobutyric acid and low residual quantity of sodium glutamate.
Preferably, the raw materials comprise 90-90.8% of raw milk and 9.2-10% of leaven by mass percentage.
Preferably, the raw materials comprise 90.8% of raw milk, 7% of sugar, 2% of fermentation strain and 0.2% of sodium glutamate by mass percentage.
By adopting the technical scheme, the content of the gamma-aminobutyric acid and the residual quantity of the sodium glutamate can reach optimal balance values.
In a third aspect, the present application provides a method for preparing fermented milk, which adopts the following technical scheme:
a preparation method of fermented milk comprises the following steps:
s1, preparing a mixed milk raw material: mixing and dissolving raw milk, sugar and sodium glutamate, and sterilizing to obtain a mixed milk raw material;
s2, preparing fermented milk: inoculating the fermentation strain into the mixed milk raw material, uniformly stirring, and fermenting at 30-45 deg.C for 40-55h to obtain fermented milk;
and S3, refrigerating and storing.
In summary, the present application has the following beneficial effects:
1. the fermented milk prepared by the method has a large amount of gamma-aminobutyric acid, effectively reduces the residual amount of sodium glutamate, and has a positive improvement effect on human health.
2. Activating and purifying the fermentation strain on an MC (methyl cellulose) plate, and then activating the fermentation strain by using an MRS (MRS) liquid culture medium to ensure that the strain achieves the optimal activity, so that the fermentation is more sufficient, and the fermentation effect is improved.
Detailed Description
The culture medium source is as follows:
MC medium, purchased from Beijing Luqiao technology GmbH;
MRS broth, purchased from Tokyo Luqiao technology corporation.
Examples
Example 1
The raw materials of the fermented milk rich in gamma-aminobutyric acid and low-residual sodium glutamate comprise, by mass, 90.8% of raw milk and 9.8% of a leavening agent, wherein the leavening agent specifically comprises 7% of white granulated sugar, 2% of a fermentation strain and 0.2% of sodium glutamate.
The preparation method of the fermented milk comprises the following steps:
s1, preparing a mixed milk raw material: mixing and dissolving raw milk, sugar and sodium glutamate, and sterilizing at 105 deg.C for 15min to obtain mixed milk raw material;
s2, preparing fermented milk: cooling the mixed milk raw material to 38-40 ℃, inoculating a fermentation strain into the mixed milk raw material, uniformly stirring, and fermenting at 39 ℃ for 48h to obtain fermented milk;
and S3, cooling to 4 ℃ and refrigerating for storage.
The fermentation strain is obtained by separating and screening streptococcus thermophilus with the preservation number of CGMCC No.23415 from traditional fermented yak yoghourt homemade by herdsmen in Ruegeng county in Alba Sichuan. Classified and named as Streptococcus thermophilus. The strain is currently preserved in China general microbiological culture Collection center, China Beijing, with the preservation date of 2021, 9 months and 15 days.
The streptococcus thermophilus is subjected to activation treatment, and the activation treatment comprises the following steps:
s1, purifying and culturing strains: weighing 76.1g of MC culture medium, dissolving in 1L of water, autoclaving at 121 deg.C for 15min, cooling to 50 deg.C, and pouring into sterile culture dish to prepare MC plate;
repeatedly burning the inoculating loop on an alcohol burner, taking the streptococcus thermophilus stored in the glycerin pipe in S1, marking on an MC plate, and culturing in a constant-temperature incubator at 37 ℃ for 48h to activate strains;
repeatedly burning the inoculating loop on an alcohol lamp, selecting a streptococcus thermophilus single bacterial colony growing better on the MC flat plate, streaking the streptococcus thermophilus single bacterial colony on a new MC flat plate, and culturing the streptococcus thermophilus single bacterial colony in a constant-temperature incubator at 37 ℃ for 48h to separate and purify strains;
s2, activation of the strain: weighing 55.2g of MRS broth, dissolving in 1L of water, shaking, subpackaging into test tubes according to 5mL standard, and autoclaving at 121 ℃ for 15min for later use;
repeatedly burning the inoculating loop on an alcohol lamp, selecting a single streptococcus thermophilus colony growing better on an MC plate in S2, transferring the single streptococcus thermophilus colony to a test tube filled with MRS broth, and culturing in a constant-temperature incubator at 37 ℃ for 24 hours;
shaking culture solution in the test tube, sucking 100 μ L seed solution with sterile suction head, inoculating into new test tube containing MRS broth, and culturing in 37 deg.C incubator for 24h to obtain zymocyte for preparing fermented milk.
The 16S rDNA sequence of the streptococcus thermophilus is as follows:
CCTGGCTCAGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTAGAACGCTGAAGA GAGGAGCTTGCTCTTCTTGGATGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACCTGC CTTGTAGCGGGGGATAACTATTGGAAACGATAGCTAATACCGCATAACAATGGATGACAC ATGTCATTTATTTGAAAGGGGCAATTGCTCCACTACAAGATGGACCTGCGTTGTATTAGCT AGTAGGTGAGGTAATGGCTCACCTAGGCGACGATACATAGCCGACCTGAGAGGGTGATC GGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATC TTCGGCAATGGGGGCAACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGG ATCGTAAAGCTCTGTTGTAAGTCAAGAACGGGTGTGAGAGTGGAAAGTTCACACTGTG ACGGTAGCTTACCAGAAAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAG GTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTGATAAGTC TGAAGTTAAAGGCTGTGGCTCAACCATAGTTCGCTTTGGAAACTGTCAAACTTGAGTGC AGAAGGGGAGAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACA CCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCGTGGGG AGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGGTGTTG GATCCTTTCCGGGATTCAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTAC GACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCAT GTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCCGATGCTATTT CTAGAGATAGAAAGTTACTTCGGTACATCGGTGACAGGTGGTGCATGGTTGTCGTCAGC TCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCC ATCATTCAGTTGGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGAT GACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGTTGGTAC AACGAGTTGCGAGTCGGTGACGGCGAGCTAATCTCTTAAAGCCAATCTCAGTTCGGATT GTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCC GCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAA CACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCTAAGGTGGGACAGATGATT GGGGTGAAGTCGTAACAAGG。
example 2
The difference from the embodiment 1 is that the raw materials of the fermented milk comprise 90.6 percent of raw milk and 9.4 percent of leavening agent by mass percentage, and the leavening agent specifically comprises 7 percent of white granulated sugar, 2 percent of zymophyte and 0.4 percent of sodium glutamate.
Comparative example
Comparative example 1
The difference from the embodiment 1 is that the raw materials of the fermented milk comprise 90% of raw milk and 10% of a leavening agent by mass percentage, wherein the leavening agent specifically comprises 7% of white granulated sugar, 2% of a fermentation strain and 1% of sodium glutamate.
Comparative example 2
The difference from the embodiment 1 is that the raw materials of the fermented milk comprise 88.8% of raw milk and 11.2% of a leavening agent by mass percentage, wherein the leavening agent specifically comprises 7% of white granulated sugar, 4% of a fermentation strain and 0.2% of sodium glutamate.
Detection method
The content of gamma-aminobutyric acid in the sample is determined by referring to the method of QB _ T4587-2013 gamma-aminobutyric acid, and the content is determined by using a pre-column ortho-phthalaldehyde (OPA) derivatization HPLC method. The chromatographic conditions used were: acclaim TM 120C 18 column (4.6X 250mm, 5 μm), column temperature 40 deg.C, sample size 5 μ L, flow rate 1.0mL/min (gradient elution), DAD-3000RS (338nm) detector, detection results are shown in Table 1.
The content of glutamic acid was measured and the content of sodium glutamate was obtained by referring to the method described in GB 5009.124-2016 (measurement of amino acids in food safety State Standard food), and the results of the measurement are shown in Table 1.
TABLE 1 test results
Figure BDA0003485919680000051
It can be seen from the combination of the examples and the comparative examples and the combination of table 1 that the ratio in example 1 is the optimal ratio, which can effectively control the residual amount of sodium glutamate to be the lowest value while ensuring that the content of gamma-aminobutyric acid approaches the highest yield, and the residual amount of glutamine is significantly different from that in examples 2 and comparative examples 1-2.
According to the invention, raw milk, white granulated sugar and sodium glutamate are used as raw materials, the autonomously screened strain streptococcus thermophilus is used for natural fermentation to prepare the flavored fermented milk with low sodium glutamate and rich gamma-aminobutyric acid residues, the content of the gamma-aminobutyric acid reaches 1.8-1.9 mg/g, the effective dose can be reached by drinking 50-60g per day, and the sodium glutamate residues are below 0.3mg/g, so that the flavored fermented milk has positive promotion significance on human health, and simultaneously, the adverse effect on human body is effectively avoided.
The present embodiment is only for explaining the present application, and it is not limited to the present application, and those skilled in the art can make modifications of the present embodiment without inventive contribution as needed after reading the present specification, but all of them are protected by patent law within the scope of the claims of the present application.

Claims (9)

1. A leaven for efficiently transforming sodium glutamate is characterized in that: the leavening agent comprises the following components in parts by weight: sugar, fermentation strain and sodium glutamate, wherein the weight ratio is 5-10: 0.5-3: 0.1-0.5;
the fermentation strain adopts streptococcus thermophilus, and the preservation number of the streptococcus thermophilus is CGMCC No. 23415.
2. The leaven for highly efficiently transforming sodium glutamate according to claim 1, wherein: the weight ratio of the sugar to the fermentation strain to the sodium glutamate is 7: 2: 0.2.
3. the leaven for highly efficiently transforming sodium glutamate according to claim 1, wherein: the fermentation strain is subjected to activation treatment, and the activation treatment comprises the following steps:
s1, culture of strains: inoculating streptococcus thermophilus into an MC culture medium, culturing and preliminarily activating strains;
s2, activation of the strain: and selecting a single colony with better growth from S1, transferring the single colony to an MRS culture medium, and culturing to obtain activated streptococcus thermophilus.
4. The leaven for highly efficiently transforming sodium glutamate according to claim 3, wherein: s1, culture of strains: sterilizing MC culture medium, cooling, pouring into aseptic culture dish, preparing plate, extracting Streptococcus thermophilus, streaking on MC plate, culturing in incubator, and activating strain.
5. The leaven for highly efficiently transforming sodium glutamate according to claim 3, wherein: s2, activation of strains: and extracting single colonies with better growth potential on the MC plate in the S1, transferring the single colonies into an MRS culture medium, and putting the single colonies into a constant-temperature incubator with the temperature of 30-45 ℃ for culture and activation for 20-50h to obtain activated streptococcus thermophilus.
6. A fermented milk characterized by: the fermentation agent for highly efficiently converting sodium glutamate as claimed in any one of claims 1 to 3 is used as a raw material.
7. A fermented milk according to claim 6, characterized in that: the raw materials comprise 90-90.8% of raw milk and 9.2-10% of leaven by mass percent.
8. A fermented milk according to claim 7, characterized in that: the raw materials comprise 90.8 percent of raw milk, 7 percent of sugar, 2 percent of fermentation strain and 0.2 percent of sodium glutamate by mass percentage.
9. A method for producing fermented milk according to any one of claims 6 to 8, comprising the steps of:
s1, preparing a mixed milk raw material: mixing and dissolving raw milk, sugar and sodium glutamate, and sterilizing to obtain a mixed milk raw material;
s2, preparing fermented milk: inoculating the fermentation strain into the mixed milk raw material, uniformly stirring, and fermenting at 30-45 deg.C for 40-55h to obtain fermented milk;
and S3, refrigerating and storing.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104293699A (en) * 2014-09-17 2015-01-21 山东大学 Streptococcus thermophilus with synthetic capability of gamma-aminobutyric acid and glutathione and application of streptococcus thermophilus
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Publication number Priority date Publication date Assignee Title
CN104293699A (en) * 2014-09-17 2015-01-21 山东大学 Streptococcus thermophilus with synthetic capability of gamma-aminobutyric acid and glutathione and application of streptococcus thermophilus
CN110283855A (en) * 2019-06-20 2019-09-27 绿雪生物工程(深圳)有限公司 A method of it improving 210202 bacterial strain of streptococcus thermophilus and produces GABA ability
JP2021058093A (en) * 2019-10-03 2021-04-15 株式会社ミル総本社 Method of producing lactic acid fermented food
CN113272418A (en) * 2021-03-17 2021-08-17 微康益生菌(苏州)股份有限公司 Streptococcus thermophilus for producing gamma-aminobutyric acid and application thereof

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Title
LIN CHEN等: "γ-Aminobutyric acid-rich yogurt fermented by Streptococcus salivarius subsp. thermophiles fmb5 apprars to have anti-diabetic effect on streptozotocin-induced diabetic mice", 《JOURNAL OF FUNCTIONAL FOODS》 *
刘韩等: "高产γ-氨基丁酸乳酸菌的筛选及鉴定", 《中国奶牛》 *

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