CN112625929A - Tannin-tolerant yeast and application thereof in fruit vinegar fermentation - Google Patents
Tannin-tolerant yeast and application thereof in fruit vinegar fermentation Download PDFInfo
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Abstract
The invention provides a tannin-tolerant yeast and application thereof in fermenting fruit vinegar, belonging to the technical field of fermentation. The fermentation method is simple to operate, the fermentation time is short, the problem of tannin inhibition in the fermentation process of the persimmon cake vinegar is solved, the persimmon cake vinegar with pleasant fragrance and pure sour taste is brewed from the persimmon cakes, the flavor and the health-care effect of the persimmons can be kept to the maximum extent, persimmon resources are fully utilized, the flower and color varieties of vinegar are enriched, the environment is protected, and the economic benefit and the social benefit are improved.
Description
Technical Field
The invention relates to a tannin-tolerant yeast and application thereof in fermenting fruit vinegar, belonging to the technical field of food processing.
Background
The fruit vinegar is a novel beverage or acidic condiment with unique flavor, which is prepared by taking fruits as main raw materials and performing alcoholic fermentation and acetic acid fermentation, is rich in various organic acids, vitamins, amino acids, trace elements and other nutrient substances, has the functions of both fruits and table vinegar, and can relieve fatigue, resist anxiety and promote intestinal metabolism.
The persimmon is sweet in taste and cold in nature, can clear heat, promote fluid production and moisten lung, contains protein, sugar, fat, pectin and various vitamins and mineral substances, and has high nutritional value and medicinal value. However, because fresh persimmons are too astringent to be eaten directly and are not easy to store and transport for a long distance, some treated soft persimmons can be eaten or processed into dried persimmons, so that the economic value of the persimmons is limited, a new application field of the persimmons needs to be developed, and the brewing of fruit vinegar is a good field.
Persimmon vinegar is brewed from persimmon as a fermentation raw material, and fresh persimmons contain a high content of soluble tannin and have a strong astringent effect on the mucous membrane of the mouth and tongue, thus having astringent taste. The tannin content is reduced after fresh persimmons are made into dried persimmon, but the dried persimmon still contains a large amount of tannin, which not only affects the flavor of dried persimmon vinegar, but also inhibits the growth and fermentation efficiency of yeasts.
Disclosure of Invention
The technical problem is as follows: in order to solve the technical problems, the invention provides a tannin-tolerant yeast strain and application thereof in fermenting fruit vinegar, in particular tannin-rich fruit vinegar such as persimmon cake vinegar. The fermentation method is simple to operate and short in fermentation time, solves the problem of tannin inhibition in the fermentation process of the persimmon cake vinegar, and can furthest retain the flavor and the health-care effect of the persimmons.
The technical scheme is as follows: in order to achieve the purpose, the invention adopts the technical scheme that:
the first purpose of the invention is to provide a tannin-tolerant yeast strain.
A tannin-tolerant yeast (Saccharomyces cerevisiae) deposited at the china general microbiological culture collection center on 12 months and 10 days in 2020 with the collection address: the microbial research institute of China academy of sciences No. 3, Xilu No.1, Beijing, Chaoyang, with the collection number of CGMCC NO. 21206; the collection name is Saccharomyces cerevisiae JN-Y1.
Furthermore, the invention also claims to express the coding gene of the tannin-tolerant yeast, and the nucleotide sequence of the gene is shown as SEQ ID NO. 1.
The second purpose of the invention is to provide the application of the tannin-tolerant yeast, in particular to the tannin-tolerant yeast added in the alcoholic fermentation stage of fruit vinegar to prepare the fruit vinegar product rich in tannin fruits.
Furthermore, the tannin-rich fruit comprises dried persimmon, and the fruit vinegar is dried persimmon vinegar. Of course, fruits rich in tannin, such as black dates, pomegranates, grapes and the like, are also suitable for the method disclosed by the invention.
The third object of the present invention is to provide a method for preparing fruit vinegar by using the tannin tolerant yeast of claim 1.
Further, the method is characterized in that the tannin-tolerant yeast is added in the alcohol fermentation stage of fruit vinegar, namely, the tannin-tolerant yeast is inoculated in the fruit enzymolysis liquid to be fermented for fermentation.
Furthermore, the method specifically comprises the step of adding the tannin-tolerant yeast in the alcohol fermentation stage of the dried persimmon vinegar, namely inoculating the tannin-tolerant yeast in the to-be-fermented dried persimmon enzymatic hydrolysate for fermentation.
Further, the production process of the method for preparing fruit vinegar specifically comprises the following steps:
1) preparing an enzymolysis solution: mixing fruits such as dried persimmon with water at a mass ratio of 1:4-8, pulping, adding pectase and cellulase 0.1-0.3% of the fruit mass, and performing enzymolysis at 50-60 deg.C for 2-3 hr to obtain enzymatic hydrolysate;
2) and (3) alcohol fermentation stage: pumping the enzymolysis liquid into an alcohol fermentation tank, cooling to 30 ℃, inoculating the activated tannin-resistant yeast into the fermentation tank according to the inoculation amount of 2% -4%, keeping the temperature at 30 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and carrying out the alcohol fermentation stage for 78 hours in total, wherein the residual sugar (calculated by reducing sugar) in the obtained alcohol fermentation liquid is 0.3g/L, and the alcoholic strength is 5.7%; the activation steps are as follows: inoculating tannin-tolerant yeast preserved in a glycerol tube into 20mL YPD liquid culture medium, and standing and culturing in a constant-temperature incubator at 30 ℃ for 36 h;
3) acetic acid fermentation: carrying out centrifugal filtration on the alcohol fermentation liquor, centrifuging to remove saccharomycetes in the alcohol fermentation process, pumping into an acetic acid fermentation tank, controlling the temperature to be 30 ℃, inoculating acetic acid bacteria seed liquor according to 2-4% of inoculation quantity, stirring at 400rpm, carrying out an acetic acid fermentation stage for 72 hours, obtaining acetic acid fermentation liquor after 72 hours, wherein the acetic acid content in the acetic acid fermentation liquor is 4.8g/100mL, and completing acetic acid fermentation when the alcohol is exhausted and the acetic acid does not rise any more, thus obtaining the fruit vinegar fermentation liquor; the fruit vinegar fermentation liquor is the fruit vinegar and can be regarded as a fruit vinegar primary product, and in order to optimize the flavor of the fruit vinegar primary product, the lactobacillus fermentation liquor of the fruit is introduced to be blended with the fruit vinegar primary product, so that the fruit vinegar product with better flavor is obtained.
4) And (3) fermenting lactic acid bacteria: pumping the enzymolysis liquid into an alcohol fermentation tank, then cooling to 37 ℃, inoculating activated lactobacillus with the inoculum size of 2% -4% into the fermentation tank, keeping the temperature at 37 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and performing the alcohol fermentation stage for 48 hours in total to obtain lactobacillus fermentation liquid.
5) Mixing: and mixing the lactobacillus fermentation liquor and the fruit vinegar fermentation liquor in proportion to obtain the lactobacillus fruit vinegar.
Further, the acetic acid bacteria is Acetobacter aceti CICC 20441.
Further, the dried persimmon is prepared dried persimmon.
Further, mixing the lactobacillus fermentation liquor and the fruit vinegar fermentation liquor according to a volume ratio of 1: 5-8, preferably 1: 6.
The second purpose of the invention is to provide the fruit vinegar prepared by the production process, including the dried persimmon vinegar.
Has the advantages that: compared with the prior art, the invention has the following advantages: the fermentation method is simple to operate, the fermentation time is short, the problem of tannin inhibition in the fermentation process of preparing the persimmon vinegar from tannin fruits such as the persimmon cake is solved, the persimmon vinegar with pleasant fragrance and pure sour taste is brewed from the persimmon cake, the flavor and the health-care effect of the persimmon can be kept to the maximum extent, the persimmon resources are fully utilized, the flower and color varieties of the table vinegar are enriched, the environment is protected, and the economic benefit and the social benefit are improved.
Detailed Description
The present invention will be further described with reference to the following examples.
Examples
The present invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the specific material ratios, process conditions and results thereof described in the examples are illustrative only and should not be taken as limiting the invention as detailed in the claims.
The parameters used in the examples are specifically as follows:
TABLE 1 emphasis parameters used for the examples and comparative examples
Example 1:
example 1.1:
1. preparing the persimmon cake enzymatic hydrolysate: mixing dried persimmon and water according to the mass ratio of 1:4, pulping, adding pectinase and cellulase which are 0.1% of the mass of the dried persimmon respectively, and performing enzymolysis at 50 ℃ for 2h to obtain a dried persimmon enzymolysis liquid;
2. and (3) alcohol fermentation stage: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 30 ℃, inoculating activated yeast with the inoculum size of 2% into the fermentation tank, keeping the temperature at 30 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and carrying out the alcohol fermentation stage for 78 hours in total, wherein the residual sugar (calculated by reducing sugar) is 0.3g/L, and the alcoholic strength is 5.7%;
3. acetic acid fermentation: carrying out centrifugal filtration on an alcohol fermentation liquid, centrifugally removing saccharomycetes in the alcohol fermentation process, pumping the alcohol fermentation liquid into an acetic acid fermentation tank, controlling the temperature to be 30 ℃, inoculating acetic acid bacteria seed liquid according to 4% of inoculation quantity, stirring at 400r/min, carrying out an acetic acid fermentation stage for 72 hours, obtaining an acetic acid fermentation liquid after 72 hours, wherein the acetic acid content in the acetic acid fermentation liquid is 4.8g/100mL, and completing acetic acid fermentation when alcohol is exhausted and acetic acid does not rise any more to obtain a fruit vinegar fermentation liquid, namely persimmon cake vinegar;
4. and (3) fermenting lactic acid bacteria: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 37 ℃, inoculating activated lactobacillus with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 37 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and performing the alcohol fermentation stage for 48 hours in total.
5. Mixing: mixing the persimmon cake lactobacillus fermentation liquor and the fruit vinegar fermentation liquor according to the proportion of 1:5 to obtain the lactobacillus persimmon cake vinegar.
Example 1.2:
1. preparing the persimmon cake enzymatic hydrolysate: mixing dried persimmon and water according to the mass ratio of 1:4, pulping, adding pectinase and cellulase accounting for 0.2% of the mass of the dried persimmon respectively, and performing enzymolysis at 55 ℃ for 2.5h to obtain a dried persimmon enzymolysis liquid;
2. and (3) alcohol fermentation stage: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 30 ℃, inoculating activated yeast with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 30 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and carrying out the alcohol fermentation stage for 78 hours in total, wherein the residual sugar (calculated by reducing sugar) is 0.3g/L, and the alcoholic strength is 5.7%;
3. acetic acid fermentation: carrying out centrifugal filtration on an alcohol fermentation liquid, centrifugally removing saccharomycetes in the alcohol fermentation process, pumping the alcohol fermentation liquid into an acetic acid fermentation tank, controlling the temperature to be 30 ℃, inoculating acetic acid bacteria seed liquid according to 4% of inoculation quantity, stirring at 400r/min, carrying out an acetic acid fermentation stage for 72 hours, obtaining an acetic acid fermentation liquid after 72 hours, wherein the acetic acid content in the acetic acid fermentation liquid is 4.8g/100mL, and completing acetic acid fermentation when alcohol is exhausted and acetic acid does not rise any more to obtain a fruit vinegar fermentation liquid, namely persimmon cake vinegar;
4. and (3) fermenting lactic acid bacteria: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 37 ℃, inoculating activated lactobacillus with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 37 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and performing the alcohol fermentation stage for 48 hours in total.
5. Mixing: mixing the persimmon cake lactobacillus fermentation liquor and the fruit vinegar fermentation liquor according to the proportion of 1:5 to obtain the lactobacillus persimmon cake vinegar.
Example 1.3:
1. preparing the persimmon cake enzymatic hydrolysate: mixing dried persimmon and water according to the mass ratio of 1:4, pulping, adding pectinase and cellulase accounting for 0.3% of the mass of the dried persimmon respectively, and performing enzymolysis at 60 ℃ for 3h to obtain a dried persimmon enzymolysis liquid;
2. and (3) alcohol fermentation stage: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 30 ℃, inoculating activated yeast with the inoculum size of 4% into the fermentation tank, keeping the temperature at 30 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and carrying out the alcohol fermentation stage for 78 hours in total, wherein the residual sugar (calculated by reducing sugar) is 0.3g/L, and the alcoholic strength is 5.7%;
3. acetic acid fermentation: carrying out centrifugal filtration on an alcohol fermentation liquid, centrifugally removing saccharomycetes in the alcohol fermentation process, pumping the alcohol fermentation liquid into an acetic acid fermentation tank, controlling the temperature to be 30 ℃, inoculating acetic acid bacteria seed liquid according to 4% of inoculation quantity, stirring at 400r/min, carrying out an acetic acid fermentation stage for 72 hours, obtaining an acetic acid fermentation liquid after 72 hours, wherein the acetic acid content in the acetic acid fermentation liquid is 4.8g/100mL, and completing acetic acid fermentation when alcohol is exhausted and acetic acid does not rise any more to obtain a fruit vinegar fermentation liquid, namely persimmon cake vinegar;
4. and (3) fermenting lactic acid bacteria: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 37 ℃, inoculating activated lactobacillus with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 37 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and performing the alcohol fermentation stage for 48 hours in total.
5. Mixing: mixing the persimmon cake lactobacillus fermentation liquor and the fruit vinegar fermentation liquor according to the proportion of 1:5 to obtain the lactobacillus persimmon cake vinegar.
Example 2:
example 2.1:
1. preparing the persimmon cake enzymatic hydrolysate: mixing dried persimmon and water according to the mass ratio of 1:6, pulping, adding pectinase and cellulase accounting for 0.1% of the mass of the dried persimmon respectively, and performing enzymolysis at 55 ℃ for 2h to obtain a dried persimmon enzymolysis liquid;
2. and (3) alcohol fermentation stage: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 30 ℃, inoculating activated yeast with the inoculum size of 2% into the fermentation tank, keeping the temperature at 30 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and carrying out the alcohol fermentation stage for 78 hours in total, wherein the residual sugar (calculated by reducing sugar) is 0.3g/L, and the alcoholic strength is 5.7%;
3. acetic acid fermentation: carrying out centrifugal filtration on an alcohol fermentation liquid, centrifuging to remove saccharomycetes in the alcohol fermentation process, pumping into an acetic acid fermentation tank, controlling the temperature to be 30 ℃, inoculating acetic acid bacteria seed liquid according to 4% of inoculation quantity, stirring at 400rpm, carrying out an acetic acid fermentation stage for 72 hours, obtaining an acetic acid fermentation liquid after 72 hours, wherein the acetic acid content in the acetic acid fermentation liquid is 4.8g/100mL, and completing acetic acid fermentation when alcohol is exhausted and acetic acid does not rise any more, thus obtaining a fruit vinegar fermentation liquid, namely persimmon cake vinegar;
4. and (3) fermenting lactic acid bacteria: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 37 ℃, inoculating activated lactobacillus with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 37 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and performing the alcohol fermentation stage for 48 hours in total.
5. Mixing: mixing the persimmon cake lactobacillus fermentation liquor and the fruit vinegar fermentation liquor according to the proportion of 1:8 to obtain the lactobacillus persimmon cake vinegar.
Example 2.2:
1. preparing the persimmon cake enzymatic hydrolysate: mixing dried persimmon and water according to the mass ratio of 1:6, pulping, adding pectinase and cellulase accounting for 0.2% of the mass of the dried persimmon respectively, and performing enzymolysis at 50 ℃ for 3h to obtain a dried persimmon enzymolysis liquid;
2. and (3) alcohol fermentation stage: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 30 ℃, inoculating activated yeast with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 30 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and carrying out the alcohol fermentation stage for 78 hours in total, wherein the residual sugar (calculated by reducing sugar) is 0.3g/L, and the alcoholic strength is 5.7%;
3. acetic acid fermentation: carrying out centrifugal filtration on an alcohol fermentation liquid, centrifuging to remove saccharomycetes in the alcohol fermentation process, pumping into an acetic acid fermentation tank, controlling the temperature to be 30 ℃, inoculating acetic acid bacteria seed liquid according to 4% of inoculation quantity, stirring at 400rpm, carrying out an acetic acid fermentation stage for 72 hours, obtaining an acetic acid fermentation liquid after 72 hours, wherein the acetic acid content in the acetic acid fermentation liquid is 4.8g/100mL, and completing acetic acid fermentation when alcohol is exhausted and acetic acid does not rise any more, thus obtaining a fruit vinegar fermentation liquid, namely persimmon cake vinegar;
4. and (3) fermenting lactic acid bacteria: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 37 ℃, inoculating activated lactobacillus with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 37 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and performing the alcohol fermentation stage for 48 hours in total.
5. Mixing: mixing the persimmon cake lactobacillus fermentation liquor and the fruit vinegar fermentation liquor according to the proportion of 1:8 to obtain the lactobacillus persimmon cake vinegar.
Example 2.3:
1. preparing the persimmon cake enzymatic hydrolysate: mixing dried persimmon and water according to the mass ratio of 1:6, pulping, adding pectinase and cellulase accounting for 0.3% of the weight of the dried persimmon respectively, and performing enzymolysis at 60 ℃ for 2.8h to obtain a dried persimmon enzymolysis liquid;
2. and (3) alcohol fermentation stage: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 30 ℃, inoculating activated yeast with the inoculum size of 4% into the fermentation tank, keeping the temperature at 30 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and carrying out the alcohol fermentation stage for 78 hours in total, wherein the residual sugar (calculated by reducing sugar) is 0.3g/L, and the alcoholic strength is 5.7%;
3. acetic acid fermentation: carrying out centrifugal filtration on an alcohol fermentation liquid, centrifuging to remove saccharomycetes in the alcohol fermentation process, pumping into an acetic acid fermentation tank, controlling the temperature to be 30 ℃, inoculating acetic acid bacteria seed liquid according to 4% of inoculation quantity, stirring at 400rpm, carrying out an acetic acid fermentation stage for 72 hours, obtaining an acetic acid fermentation liquid after 72 hours, wherein the acetic acid content in the acetic acid fermentation liquid is 4.8g/100mL, and completing acetic acid fermentation when alcohol is exhausted and acetic acid does not rise any more, thus obtaining a fruit vinegar fermentation liquid, namely persimmon cake vinegar;
4. and (3) fermenting lactic acid bacteria: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 37 ℃, inoculating activated lactobacillus with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 37 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and performing the alcohol fermentation stage for 48 hours in total.
5. Mixing: mixing the persimmon cake lactobacillus fermentation liquor and the fruit vinegar fermentation liquor according to the proportion of 1:8 to obtain the lactobacillus persimmon cake vinegar.
Example 3:
example 3.1:
1. preparing the persimmon cake enzymatic hydrolysate: mixing dried persimmon and water according to the mass ratio of 1:8, pulping, adding pectinase and cellulase accounting for 0.3% of the mass of the dried persimmon respectively, and performing enzymolysis at 55 ℃ for 3h to obtain a dried persimmon enzymolysis liquid;
2. and (3) alcohol fermentation stage: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 30 ℃, inoculating activated yeast with the inoculum size of 2% into the fermentation tank, keeping the temperature at 30 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and carrying out the alcohol fermentation stage for 78 hours in total, wherein the residual sugar (calculated by reducing sugar) is 0.3g/L, and the alcoholic strength is 5.7%;
3. acetic acid fermentation: carrying out centrifugal filtration on an alcohol fermentation liquid, centrifuging to remove saccharomycetes in the alcohol fermentation process, pumping into an acetic acid fermentation tank, controlling the temperature to be 30 ℃, inoculating acetic acid bacteria seed liquid according to 4% of inoculation quantity, stirring at 400rpm, carrying out an acetic acid fermentation stage for 72 hours, obtaining an acetic acid fermentation liquid after 72 hours, wherein the acetic acid content in the acetic acid fermentation liquid is 4.8g/100mL, and completing acetic acid fermentation when alcohol is exhausted and acetic acid does not rise any more, thus obtaining a fruit vinegar fermentation liquid, namely persimmon cake vinegar;
4. and (3) fermenting lactic acid bacteria: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 37 ℃, inoculating activated lactobacillus with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 37 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and performing the alcohol fermentation stage for 48 hours in total;
5. mixing: mixing the persimmon cake lactobacillus fermentation liquor and the fruit vinegar fermentation liquor according to the volume ratio of 1:6 to obtain the lactobacillus persimmon cake vinegar.
Example 3.2:
1. preparing the persimmon cake enzymatic hydrolysate: mixing dried persimmon and water according to the mass ratio of 1:8, pulping, adding pectinase and cellulase accounting for 0.3% of the mass of the dried persimmon respectively, and performing enzymolysis at 55 ℃ for 3h to obtain a dried persimmon enzymolysis liquid;
2. and (3) alcohol fermentation stage: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 30 ℃, inoculating activated yeast with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 30 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and carrying out the alcohol fermentation stage for 78 hours in total, wherein the residual sugar (calculated by reducing sugar) is 0.3g/L, and the alcoholic strength is 5.7%;
3. acetic acid fermentation: carrying out centrifugal filtration on an alcohol fermentation liquid, centrifuging to remove saccharomycetes in the alcohol fermentation process, pumping into an acetic acid fermentation tank, controlling the temperature to be 30 ℃, inoculating acetic acid bacteria seed liquid according to 4% of inoculation quantity, stirring at 400rpm, carrying out an acetic acid fermentation stage for 72 hours, obtaining an acetic acid fermentation liquid after 72 hours, wherein the acetic acid content in the acetic acid fermentation liquid is 4.8g/100mL, and completing acetic acid fermentation when alcohol is exhausted and acetic acid does not rise any more, thus obtaining a fruit vinegar fermentation liquid, namely persimmon cake vinegar;
4. and (3) fermenting lactic acid bacteria: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 37 ℃, inoculating activated lactobacillus with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 37 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and performing the alcohol fermentation stage for 48 hours in total;
5. mixing: mixing the persimmon cake lactobacillus fermentation liquor and the fruit vinegar fermentation liquor according to the volume ratio of 1:6 to obtain the lactobacillus persimmon cake vinegar.
Example 3.3:
1. preparing the persimmon cake enzymatic hydrolysate: mixing dried persimmon and water according to the mass ratio of 1:8, pulping, adding pectinase and cellulase accounting for 0.3% of the mass of the dried persimmon respectively, and performing enzymolysis at 55 ℃ for 3h to obtain a dried persimmon enzymolysis liquid;
2. and (3) alcohol fermentation stage: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 30 ℃, inoculating activated yeast with the inoculum size of 4% into the fermentation tank, keeping the temperature at 30 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and carrying out the alcohol fermentation stage for 78 hours in total, wherein the residual sugar (calculated by reducing sugar) is 0.3g/L, and the alcoholic strength is 5.7%;
3. acetic acid fermentation: carrying out centrifugal filtration on the alcoholic fermentation liquor, centrifuging to remove saccharomycetes in the alcoholic fermentation process, pumping into an acetic acid fermentation tank, controlling the temperature to be 30 ℃, inoculating acetic acid bacteria seed liquor according to 4% of inoculation quantity, stirring at 400rpm, carrying out an acetic acid fermentation stage for 72 hours, obtaining acetic acid fermentation liquor after 72 hours, wherein the acetic acid content in the acetic acid fermentation liquor is 4.8g/100mL, and completing acetic acid fermentation when the alcohol is exhausted and the acetic acid does not rise any more to obtain fruit vinegar fermentation liquor, namely obtaining persimmon cake vinegar;
4. and (3) fermenting lactic acid bacteria: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 37 ℃, inoculating activated lactobacillus with the inoculation amount of 3% into the fermentation tank, keeping the temperature at 37 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and performing the alcohol fermentation stage for 48 hours in total;
5. mixing: mixing the persimmon cake lactobacillus fermentation liquor and the fruit vinegar fermentation liquor according to the volume ratio of 1:6 to obtain the lactobacillus persimmon cake vinegar.
Example of blank control group:
1. preparing the persimmon cake enzymatic hydrolysate: mixing dried persimmon and water according to the mass ratio of 1:8, pulping, adding pectinase and cellulase accounting for 0.3% of the mass of the dried persimmon respectively, and performing enzymolysis at 55 ℃ for 3h to obtain a dried persimmon enzymolysis liquid;
2. and (3) alcohol fermentation stage: pumping the persimmon cake enzymatic hydrolysate into an alcohol fermentation tank, then cooling to 30 ℃, inoculating activated yeast with the inoculum size of 4% into the fermentation tank, keeping the temperature at 30 ℃, ventilating for 6 hours in the alcohol fermentation stage, stirring at 100rpm, then closing ventilation, slowly stirring at 50rpm, and carrying out the alcohol fermentation stage for 78 hours in total, wherein the residual sugar (calculated by reducing sugar) is 0.3g/L, and the alcoholic strength is 5.7%;
3. acetic acid fermentation: carrying out centrifugal filtration on the alcoholic fermentation liquor, centrifuging to remove saccharomycetes in the alcoholic fermentation process, pumping into an acetic acid fermentation tank, controlling the temperature to be 30 ℃, inoculating acetic acid bacteria seed liquor according to 4% of inoculation quantity, stirring at 400rpm, carrying out an acetic acid fermentation stage for 72 hours, obtaining acetic acid fermentation liquor after 72 hours, wherein the acetic acid content in the acetic acid fermentation liquor is 4.8g/100mL, and completing acetic acid fermentation when the alcohol is exhausted and the acetic acid does not rise any more, thus obtaining fruit vinegar fermentation liquor, namely the persimmon cake vinegar.
Example analysis of results:
the differences in the technical effects obtained by comparing sensory and physicochemical indices in the above examples were analyzed and are specifically shown in tables 1, 2 and 3.
TABLE 2 evaluation criteria for persimmon vinegar
TABLE 3 comparison of sensory indexes of persimmon vinegar produced in each example and comparative example
TABLE 4 comparison of physicochemical indices of persimmon vinegar produced in each example and comparative example
The product obtained by the control group is a primary product, namely the persimmon vinegar fermentation liquor, which is not blended by the lactobacillus fermentation liquor, and the comparison with the examples 1 to 3 shows that the properties of the primary product in all aspects are good, and particularly the physicochemical indexes are not greatly different; however, the sensory index, in particular the aroma and taste score, is relatively low, which also corresponds to our previous description of the impact of the tannin on the flavour. Therefore, the lactobacillus fermentation liquor is additionally added for blending, and the indexes of the blended fruit vinegar product in various aspects such as flavor and the like are greatly improved.
In addition, as can be seen from tables 2 and 3, the sensory score of the lactic acid bacteria dried persimmon vinegar in example 3 is higher than the sensory indexes of the lactic acid bacteria dried persimmon vinegar brewed in examples 1 and 2, especially two sensory indexes of flavor and taste, and the mixing ratio of the dried persimmon lactic acid bacteria fermentation liquid and the dried persimmon vinegar in example 3 is more suitable than that in examples 1 and 2; the lactic acid bacteria dried persimmon vinegar brewed in example 3 for external use has a higher total content of the bioactive substances such as non-volatile acids and polysaccharides than those in examples 1 and 2, and has a higher total ester content and a larger variety of volatile flavor substances. Example 3.3 in example 3 the lactic acid bacteria dried persimmon vinegar had a sensory score higher than that of examples 3.1 and 3.2, and the contents of the bioactive substances such as non-volatile acids and polysaccharides were higher than those of examples 3.1 and 3.2, and the total ester content was higher and the kinds of the volatile flavor substances were more abundant than those of example 3.3 and 3.3. From the above data, it is found that the lactic acid bacteria dried persimmon vinegar brewed with the yeast inoculum size of 4% and the mixing volume ratio of 1:6 is the best.
The above-mentioned embodiments are merely preferred embodiments for fully illustrating the present invention, and the scope of the present invention is not limited thereto. The equivalent substitution or change made by the technical personnel in the technical field on the basis of the invention is all within the protection scope of the invention. The protection scope of the invention is subject to the claims.
Sequence listing
<110> university of south of the Yangtze river
Hebei mingjiuchun Liquor Co.,Ltd.
<120> tannin-tolerant yeast and application thereof in fruit vinegar fermentation
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 437
<212> DNA
<213> tannin-tolerant yeast (Saccharomyces cerevisiae)
<400> 1
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ttttgaaaat ggattttttt gttttggcaa gagcatgaga gcttttactg ggcaagaaga 120
caagagatgg agagtccagc cgggcctgcg cttaagtgcg cggtcttgct aggcttgtaa 180
gtttctttct tgctattcca aacggtgaga gatttctgtg cttttgttat aggacaatta 240
aaaccgtttc aatacaacac actgtggagt tttcatatct ttgcaacttt ttctttgggc 300
attcgagcaa tcggggccca gaggtaacaa acacaaacaa ttttatttat tcattaaatt 360
tttgtcaaaa acaagaattt tcgtaactgg aaattttaaa atataaaaaa ctttcaacaa 420
cggatctctt ggttctc 437
Claims (10)
1. A tannin tolerant yeast (Saccharomyces cerevisiae) characterized by: the tannin-tolerant yeast is preserved in China general microbiological culture Collection center (CGMCC) at 12 months and 10 days in 2020, and the preservation address is as follows: the microbial research institute of China academy of sciences No. 3, Xilu No.1, Beijing, Chaoyang, with the collection number of CGMCC NO. 21206; the collection name is Saccharomyces cerevisiae JN-Y1.
2. The tannin-tolerant yeast gene according to claim 1, wherein: the nucleotide sequence of the gene is shown in SEQ ID NO. 1.
3. A fruit vinegar prepared by applying the tannin-tolerant yeast of claim 1, wherein the tannin-tolerant yeast is characterized in that: and inoculating the tannin-tolerant yeast into the fruit enzymolysis liquid to be fermented for fermentation.
4. The fruit vinegar of claim 3, wherein: the fruit vinegar is derived from tannin fruits including dried persimmon, and the fruit vinegar is tannin fruit vinegar including dried persimmon vinegar.
5. The method for preparing fruit vinegar according to claim 3 or 4, wherein the method comprises the following steps: the tannin-tolerant yeast is added in the alcohol fermentation stage of fruit vinegar, and the method specifically comprises the following steps:
1) preparing an enzymolysis liquid: mixing the fruits with water, pulping, adding pectinase and cellulase for enzymolysis to obtain an enzymatic hydrolysate;
2) alcohol fermentation: pumping the enzymolysis liquid into an alcohol fermentation tank, inoculating activated tolerant tannin yeast with the inoculation amount of 2-4% at 30 ℃, and then closing ventilation stirring after ventilation stirring to obtain alcohol fermentation liquid;
3) acetic acid fermentation: centrifuging alcohol fermentation liquor to remove the tannin-tolerant yeast, pumping into an acetic acid fermentation tank, inoculating acetic acid bacteria seed liquid according to the inoculation amount of 2% -4% at 30 ℃, stirring to obtain acetic acid fermentation liquor, and completing acetic acid fermentation when alcohol is exhausted and acetic acid does not rise any more to obtain fruit vinegar fermentation liquor, namely fruit vinegar.
6. The method for preparing fruit vinegar according to claim 5, wherein the method comprises the following steps: further comprising the steps of:
4) and (3) fermenting lactic acid bacteria: pumping the enzymolysis liquid into a fermentation tank, inoculating activated lactobacillus according to the inoculation amount of 2-4% at 37 ℃, and closing ventilation stirring after ventilation stirring to obtain lactobacillus fermentation liquid;
5) mixing: and mixing the lactobacillus fermentation liquor with the fruit vinegar fermentation liquor to obtain the lactobacillus fruit vinegar.
7. The method for preparing fruit vinegar according to claim 6, wherein the method comprises the following steps: in the step 5), mixing the lactobacillus fermentation liquor and the fruit vinegar fermentation liquor according to the volume ratio of 1: 5-8.
8. The method for preparing fruit vinegar according to claim 5 or 6, wherein the method comprises the following steps: the specific method of the step 1) comprises the following steps: mixing dried persimmon with water according to the mass ratio of 1:4-8, pulping, adding pectinase and cellulase accounting for 0.1-0.3% of the dried persimmon, and performing enzymolysis at 50-60 deg.C for 2-3h to obtain the enzymatic hydrolysate of dried persimmon.
9. The method for preparing fruit vinegar according to claim 5 or 6, wherein the method comprises the following steps: in the step 2), keeping the temperature at 37 ℃, ventilating for 6 hours, stirring at 100rpm, then closing ventilation, and stirring at 50rpm for 72 hours to obtain alcohol fermentation liquor; in the obtained alcoholic fermentation liquor, the residual sugar is 0.3g/L calculated by reducing sugar, and the alcoholic strength is 5.7%.
10. The method for preparing fruit vinegar according to claim 5 or 6, wherein the method comprises the following steps: in the step 3), the acetic acid bacteria are acetic acid bacillus CICC 20441, and are stirred at 400rpm for 72 hours to obtain acetic acid fermentation liquor; the acetic acid content in the obtained acetic acid fermentation liquor is 4.8g/100 mL.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104877923A (en) * | 2015-05-25 | 2015-09-02 | 大兴安岭百盛蓝莓科技开发有限公司 | Saccharomycetes used for improving quality of blueberry juice and application of saccharomycetes |
| CN106085799A (en) * | 2016-07-28 | 2016-11-09 | 广西壮族自治区农业科学院农产品加工研究所 | A kind of multi-cultur es co-immobilization fermentation Fructus Mori fruit vinegar and preparation method thereof |
| CN106635731A (en) * | 2016-12-21 | 2017-05-10 | 陕西时代酿造科技有限公司 | Preparation method for persimmon vinegar |
| CN108641883A (en) * | 2018-04-10 | 2018-10-12 | 湖北食为天药业股份有限公司 | A kind of persimmon vinegar and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104877923A (en) * | 2015-05-25 | 2015-09-02 | 大兴安岭百盛蓝莓科技开发有限公司 | Saccharomycetes used for improving quality of blueberry juice and application of saccharomycetes |
| CN106085799A (en) * | 2016-07-28 | 2016-11-09 | 广西壮族自治区农业科学院农产品加工研究所 | A kind of multi-cultur es co-immobilization fermentation Fructus Mori fruit vinegar and preparation method thereof |
| CN106635731A (en) * | 2016-12-21 | 2017-05-10 | 陕西时代酿造科技有限公司 | Preparation method for persimmon vinegar |
| CN108641883A (en) * | 2018-04-10 | 2018-10-12 | 湖北食为天药业股份有限公司 | A kind of persimmon vinegar and preparation method thereof |
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