CN115624121A - Black vinegar health-care beverage and preparation method thereof - Google Patents
Black vinegar health-care beverage and preparation method thereof Download PDFInfo
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- CN115624121A CN115624121A CN202211309291.3A CN202211309291A CN115624121A CN 115624121 A CN115624121 A CN 115624121A CN 202211309291 A CN202211309291 A CN 202211309291A CN 115624121 A CN115624121 A CN 115624121A
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- 238000002360 preparation method Methods 0.000 title claims description 23
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- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 18
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
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- LTVDFSLWFKLJDQ-UHFFFAOYSA-N α-tocopherolquinone Chemical compound CC(C)CCCC(C)CCCC(C)CCCC(C)(O)CCC1=C(C)C(=O)C(C)=C(C)C1=O LTVDFSLWFKLJDQ-UHFFFAOYSA-N 0.000 claims description 13
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- 230000000844 anti-bacterial effect Effects 0.000 description 2
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 2
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- CSPHGSFZFWKVDL-UHFFFAOYSA-M (3-chloro-2-hydroxypropyl)-trimethylazanium;chloride Chemical compound [Cl-].C[N+](C)(C)CC(O)CCl CSPHGSFZFWKVDL-UHFFFAOYSA-M 0.000 description 1
- OCZVHBZNPVABKX-UHFFFAOYSA-N 1,1-diphenyl-2-(2,4,6-trinitrophenyl)hydrazine;ethanol Chemical compound CCO.[O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1NN(C=1C=CC=CC=1)C1=CC=CC=C1 OCZVHBZNPVABKX-UHFFFAOYSA-N 0.000 description 1
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- YOBAEOGBNPPUQV-UHFFFAOYSA-N iron;trihydrate Chemical compound O.O.O.[Fe].[Fe] YOBAEOGBNPPUQV-UHFFFAOYSA-N 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
- C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
- C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12J—VINEGAR; PREPARATION OR PURIFICATION THEREOF
- C12J1/00—Vinegar; Preparation or purification thereof
- C12J1/04—Vinegar; Preparation or purification thereof from alcohol
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Nutrition Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Mycology (AREA)
- Wood Science & Technology (AREA)
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- Molecular Biology (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Non-Alcoholic Beverages (AREA)
Abstract
The invention discloses a method for preparing a black vinegar health-care beverage, which comprises the following steps: weighing cherries, tomatoes, medlar, cucumbers and dragon fruits, removing kernels and peels, washing, draining, mixing, crushing into juice, and sieving to obtain mixed pulp; II, sterilizing the mixed slurry and cooling to room temperature; adding zymophyte, fermenting, and filtering to obtain fermentation filtrate; III, adding xylitol and black vinegar into the fermentation filtrate, and uniformly mixing to obtain the black vinegar health-care beverage. The black vinegar health drink has unique flavor, rich nutrition and good oxidation resistance.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to a black vinegar health-care beverage and a preparation method thereof.
Background
The fruits and vegetables are rich in a large amount of nutrient substances required by human bodies, and the fruits and vegetables are processed by a fermentation process, so that the metabolism of saccharides, lipids, proteins and the like in the fruits and vegetables can be promoted, and the fruits and vegetables are converted into beneficial metabolites such as organic acids, oligosaccharides, active enzymes, oligopeptides, amino acids and the like, and are more beneficial to the health of human bodies. The natural fruit and vegetable fermented beverage is popular among people due to the higher nutrition and health care functions and the unique flavor.
The black vinegar is prepared by fermenting and brewing grains such as sorghum, rice and the like as raw materials, and is rich in a plurality of amino acids, mineral substances and other trace active ingredients. The edible black vinegar has effects of promoting digestion and absorption, enhancing liver function, preventing cardiovascular diseases, and accelerating fat and saccharide metabolism. Therefore, the fruit and vegetable fermented beverage and the black vinegar can be combined with the taste, flavor and nutritional value to prepare the compound beverage by taking the fruit and vegetable as raw materials and fermenting and processing the fruit and vegetable and the black vinegar.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a black vinegar health-care beverage and a preparation method thereof.
The technical scheme adopted by the invention is as follows:
a method for preparing a black vinegar health-care beverage comprises the following steps:
weighing cherries, tomatoes, medlar, cucumbers and dragon fruits, removing kernels and peels, washing, draining, mixing, crushing into juice, and sieving to obtain mixed pulp;
II, sterilizing the mixed slurry and cooling to room temperature; adding zymophyte, fermenting, and filtering to obtain fermentation filtrate;
III, adding xylitol and black vinegar into the fermentation filtrate, and uniformly mixing to obtain the black vinegar health-care beverage.
Preferably, the preparation method of the black vinegar health-care beverage comprises the following steps:
weighing cherry, tomato, medlar, cucumber and dragon fruit according to the mass ratio of (0.5-1.5) to (1-3) to (0.1-0.6) to (0.5-1.5) to (1-3), removing kernels and peels, washing with water, draining, mixing, crushing into juice, and sieving with a 50-100 mesh sieve to obtain mixed pulp;
II, sterilizing the mixed slurry at 110-120 ℃ for 10-20min, and cooling to room temperature; adding zymocyte, fermenting at 27-30 deg.C for 50-70 hr, and filtering to obtain fermented filtrate; the using amount of the zymophyte is 3-6wt.%; the fermentation bacteria are formed by mixing schizosaccharomyces pombe and bacillus subtilis according to the mass ratio of 2 (1-2);
III, adding 0.1-0.6 part by weight of xylitol and 25-35 parts by weight of black vinegar into 30-60 parts by weight of fermentation filtrate, and uniformly mixing to obtain the black vinegar health-care beverage.
Preferably, the black vinegar is prepared by the following method:
s1, crushing grains and sieving the grains with a 10-50-mesh sieve to obtain grain powder; the grain is a mixture of glutinous rice, sorghum, black beans, black rice, rye and black tartary buckwheat according to the mass ratio of (13-20) to (10-13) to (3-7) to (1-5) to (7-12) to (1-5);
s2, mixing the grain powder obtained in the step S1 with amylase and water at 38-41 ℃, stirring at 80-120rpm, and carrying out enzymolysis for 7-10h to obtain grain enzymolysis liquid; the mass ratio of the grain powder to the water is (0.7-1.3) to (4-6); the addition amount of the amylase in each milliliter of water is 480-520U;
s3, mixing dry yeast and (2-4) wt.% of sucrose aqueous solution at 30-35 ℃, and stirring at 240-360rpm for 0.5-2h to obtain yeast sugar solution; the concentration of the yeast in the yeast sugar solution is 10 6 CFU/mL;
S4, mixing the grain enzymolysis liquid obtained in the step S2, the yeast sugar liquid obtained in the step S3 and the grape fermentation product according to the mass ratio of (220-260) to (1-5) to (80-120), and stirring at the temperature of 28-33 ℃ and under the anaerobic condition at the rpm of 20-50 for 7-10 days to obtain pre-fermentation liquid;
s5, sterilizing the pre-fermentation liquid obtained in the step S4, adding acetobacter into the pre-fermentation liquid, and stirring the mixture for 8 to 12 days at a speed of 20 to 40rpm at a temperature of between 34 and 38 ℃ under the condition of introducing oxygen to obtain crude vinegar; the inoculation amount of the acetobacter is 10 6 CFU/mL, the oxygen introduction amount is 4-6m 3 /(h·m 3 );
And S6, sterilizing and filtering the crude vinegar obtained in the step S5 to obtain the black vinegar.
Preferably, the preparation method of the grape fermentation product comprises the following steps:
mixing botrytis cinerea, modified chitosan, hyaluronic acid and water at 15-22 ℃, and stirring at 180-300rpm for 10-40min to obtain a botrytis cinerea liquid; the concentration of the botrytis cinerea in the botrytis cinerea liquid is 10 6 -10 8 CFU/mL; the mass ratio of the modified chitosan to the hyaluronic acid to the water is (2-5) to (0.05-0.12) to (80-120);
mixing the botrytis cinerea liquid obtained in the step and grapes according to a mass ratio of (0.7-1.3) to (4-6), and fermenting for 3-5 days in a light-resistant environment at the temperature of 8-11 ℃ and the relative humidity of 58-67%;
and thirdly, placing the grapes obtained in the step II in a light-shielding environment with the temperature of 14-17 ℃ and the relative humidity of 38-46% for continuous fermentation for 9-12d to obtain the grape leavening.
The invasion and growth of botrytis cinerea can volatilize the moisture of the grape raw material, change the components of saccharides, organic acids, polyhydric alcohols, terpenes and the like in the grapes, and finally obtain the grape fermentation product with high sugar content, strong fragrance and unique fragrance and taste; the modified chitosan prepared by the method has a selective bacteriostatic action on the grape fermentation product, can inhibit the physiological activities of other bacteria except botrytis cinerea, so that the fermentation activity of the botrytis cinerea on grapes is not disturbed by the other bacteria, and organic acid, polyhydric alcohol and terpenes with higher content are obtained; due to the molecular size and the unique hydrophilic characteristic of the hyaluronic acid, the hyaluronic acid can play a certain role in inhibiting mixed bacteria and promoting the propagation of the botrytis cinerea while moisturizing the grapes and facilitating the rapid parasitic invasion of the botrytis cinerea into the grapes. The 2-hydroxyethyl trimethyl ammonium chloride and the 3-chloro-2-hydroxypropyl trimethyl ammonium chloride are adopted to modify the chitosan, so that the selective bacteriostasis of the chitosan in the process of preparing grape fermentation products by utilizing the botrytis cinerea can be enhanced, namely, the inhibition effect on the physiological process of the botrytis cinerea is not generated while inhibiting the growth and the reproduction of other bacteria, because the selected botrytis cinerea has stronger resistance to the modified chitosan prepared by the specific method of the invention due to the fact that the surface of the selected botrytis cinerea has glycoprotein with special spatial configuration, the modified chitosan can grow and reproduce in the environment with the application of the modified chitosan, and other bacteria, such as escherichia coli and staphylococcus aureus, are inhibited without the protection of the glycoprotein; the attraction range between 2-hydroxyethyl in 2-hydroxyethyl trimethyl ammonium chloride and 2-hydroxypropyl in 3-chloro-2-hydroxypropyl trimethyl ammonium chloride can further enhance the selective antibacterial action of the obtained modified chitosan, thereby achieving the synergistic effect.
Preferably, the preparation method of the modified chitosan comprises the following steps:
(1) mixing chitosan and acetic acid water solution at 29-32 deg.C, and stirring at 240-360rpm for 20-40min to obtain chitosan acetic acid water solution; the concentration of the acetic acid aqueous solution is 1.5-2.1wt.%; the mass ratio of the chitosan to the acetic acid aqueous solution is (1-1.4) to (98.6-99);
(2) adjusting the pH value of the chitosan acetic acid aqueous solution obtained in the step (1) to 8.8-9.1 by using a NaOH aqueous solution with the concentration of 0.1-1mol/L at the temperature of 29-32 ℃, and filtering to obtain a filter cake;
(3) mixing the filter cake obtained in the step (2) with n-butanol and NaOH aqueous solution at 29-32 ℃, and stirring at 240-360rpm for 10-40min; then, dripping ammonium salt water solution at the speed of 0.5-1.5mL/min, heating to 65-70 ℃ after dripping is finished, and stirring at 120-240rpm for reaction for 7-10h; filtering to obtain primary filter residue after the reaction is finished, soaking the primary filter residue in absolute ethyl alcohol, stirring at 540-660rpm for 0.5-2h, filtering again to obtain secondary filter residue, and drying the secondary filter residue for 0.5-2h at 68-72 ℃ under the condition of 97-99.6kPa to obtain the modified chitosan; the concentration of the NaOH aqueous solution is 0.1-1mol/L; the concentration of the aqueous ammonium salt solution is 8-11wt.%; the mass ratio of the filter cake, the normal butanol, the NaOH aqueous solution and the ammonium salt aqueous solution is (0.6-1.5) to (3-7) to (2-3.5) to (1.2-2.3); the mass ratio of the primary filter residue to the absolute ethyl alcohol is (0.5-1.2) to (2.8-4.3).
The ammonium salt is 2-hydroxyethyl trimethyl ammonium chloride and/or 3-chloro-2-hydroxypropyl trimethyl ammonium chloride; preferably, the ammonium salt is a mixture of 2-hydroxyethyl trimethyl ammonium chloride and 3-chloro-2-hydroxypropyl trimethyl ammonium chloride according to the mass ratio of (1-3) to (1-3).
The invention also provides a black vinegar health-care beverage which is prepared by adopting the method.
The invention has the beneficial effects that: the black vinegar health-care beverage provided by the invention has unique flavor, harmonious and mellow fruit flavor and grain flavor, and has rich nutritional value and oxidation resistance.
Detailed Description
The above summary of the present invention is described in further detail below with reference to specific embodiments, but it should not be understood that the scope of the above subject matter of the present invention is limited to the following examples.
Introduction of some of the raw materials in this application:
glutinous rice, purchased from the food and oil company, the breed: the grain-out rate of fragrant glutinous rice is more than or equal to 10 percent, the water content is less than or equal to 10 percent, the polished rice rate is more than or equal to 61 percent, the impurities are less than or equal to 1.0 percent, the product number is as follows: 001. sorghum, purchased from Fucheng county Fuxing agricultural science and technology Co., ltd, variety: ji brewing II, the water content is less than or equal to 14 percent.
Black beans purchased from Shanghai elder Shen Fang food Co., ltd., crude protein content of not less than 40.0%, crude fat of not less than 20.0%.
The black rice is purchased from Guangzhou Linmiao food Co., ltd, the clear rate is more than or equal to 95%, the impurity content is less than or equal to 1.0%, and the water content is less than or equal to 8.0%.
Rye is purchased from Kaikuan city Xiang Fu Ouan wide crop planting farmer professional cooperative, the impurity content is less than or equal to 1.0 percent, and the water content is less than or equal to 12.5 percent.
Black tartary buckwheat is purchased from Shenzhen Longgang region Qizhen trade business, the impurity is less than or equal to 1.0 percent, and the water content is less than or equal to 12.5 percent.
Amylase, CAS:9000-92-4, available from Bailingwei science and technology, product number: d1115, purity: 30U/mg.
Dry yeast, CAS:68876-77-7, available from Shandong He shield New materials, inc., grade: food grade, cargo number: yuhs-02.
Acetic acid bacillus: acetobacter aceti, number: CGMCC 1.1809, purchased from China general microbiological culture Collection center.
Sucrose, CAS:57-50-1, available from western chemical technology (Shandong) Inc., cat #: c10185-500g, specification: and GR.
Grape, available from qinhuang island xin mao agri-technologies ltd, variety: chardonnay, sugar content is more than or equal to 18%, average grain weight: 2.4g, soluble solids: 17.8 percent.
Botrytis cinerea: botrytis cinerea, no.: CGMCC 3.3790, purchased from China general microbiological culture Collection center.
Schizosaccharomyces pombe, accession number: CICC 1260, purchased from China center for culture Collection of Industrial microorganisms.
Bacillus subtilis, and Bacillus subtilis with the number: CICC 10260, purchased from China center for culture Collection of Industrial microorganisms.
Cherry, tomato, medlar, cucumber and dragon fruit are all sold in the market.
Hyaluronic acid, CAS:9004-61-9, available from chemical technology of west asia (Shandong) Inc., cat #: a22873-100g, molecular weight: 100 ten thousand.
Polyglutamic acid, CAS:25513-46-6, available from sienna herbst biotechnology limited, molecular weight: 70 ten thousand.
Chitosan, CAS:9012-76-4, available from west asia chemical technology (shandong) ltd, cat # v: e14854-500g, degree of deacetylation not less than 95.0%, molecular weight: 62 ten thousand.
Acetic acid, CAS:64-19-7, available from western chemistry technologies (Shandong) Inc., cat #: d10086-500ml, specification: and GR.
NaOH, CAS:1310-73-2, available from western chemical technology (Shandong) Inc., cat #: a12087-500g, specification: and AR.
N-butanol, CAS:71-36-3, available from Nanjing Chemicals, inc., cat #: C0690522223.
absolute ethanol, CAS:64-17-5, available from Nanjing Chemicals, inc., cat #: c0692035010, specification: EP.
2-hydroxyethyl trimethylammonium chloride, CAS:67-48-1, available from western chemistry technologies (Shandong) Inc., cat #: b14469-2.5kg, specification: and (5) AR.
3-chloro-2-hydroxypropyltrimethylammonium chloride, CAS:3327-22-8, available from western chemical technology, santa limited, cat #: a11158-500ml, specification: 60wt.% aqueous solution.
Example 1
The preparation method of the black vinegar health-care beverage comprises the following steps:
weighing cherries, tomatoes, medlar, cucumbers and dragon fruits according to a mass ratio of 1;
II, sterilizing the mixed slurry at 115 ℃ for 15min, and cooling to room temperature; adding zymophyte, fermenting at 29 deg.C for 60 hr, and filtering to obtain fermentation filtrate; the amount of the zymophyte is 5wt.%; the fermentation bacteria are formed by mixing schizosaccharomyces pombe and bacillus subtilis 2:1 in mass ratio;
III, adding 0.5 part by weight of xylitol and 30 parts by weight of black vinegar into 50 parts by weight of fermentation filtrate, and uniformly mixing to obtain the black vinegar health-care beverage.
The black vinegar is prepared by the following method:
s1, crushing grains and sieving the grains with a 30-mesh sieve to obtain grain powder; the cereal is a mixture of glutinous rice, sorghum, black beans, black rice, rye and black tartary buckwheat according to a mass ratio of 15;
s2, mixing the grain powder obtained in the step S1 with amylase and distilled water at 40 ℃, stirring at 100rpm, and performing enzymolysis for 8 hours to obtain grain enzymolysis liquid; the mass ratio of the grain powder to the distilled water is 1:5; the addition amount of the amylase in per milliliter of distilled water is 500U;
s3, mixing dry yeast and 3.5wt.% of sucrose aqueous solution at 32 ℃, and stirring at 300rpm for 1h to obtain yeast sugar solution; the concentration of the yeast in the yeast sugar solution is 10 6 CFU/mL;
S4, mixing the grain enzymolysis liquid obtained in the step S2, the yeast sugar liquid obtained in the step S3 and the grape fermentation product according to the mass ratio of 250 to 100, and stirring at 30rpm for 8d under the anaerobic condition at 29 ℃ to obtain a pre-fermentation liquid;
s5, sterilizing the pre-fermentation liquid obtained in the step S4, adding acetobacter into the pre-fermentation liquid, and stirring the mixture for 10 days at the speed of 30rpm at the temperature of 36 ℃ under the condition of introducing oxygen to obtain crude vinegar; the inoculation amount of the acetobacter is 10 6 CFU/mL, the oxygen introduction amount is 5m 3 /(h·m 3 );
S6, sterilizing and filtering the crude vinegar obtained in the step S5 to obtain the black vinegar.
The preparation method of the grape fermentation product comprises the following steps:
mixing botrytis cinerea, modified chitosan, hyaluronic acid and distilled water at 19 ℃, and stirring at 240rpm for 30min to obtain a botrytis cinerea liquid; the concentration of the botrytis cinerea in the botrytis cinerea liquid is 10 9 CFU/mL; the mass ratio of the modified chitosan to the hyaluronic acid to the distilled water is 0.05;
mixing the botrytis cinerea liquid obtained in the step with grapes in a mass ratio of 1:5, and fermenting for 4d in a light-resistant environment at the temperature of 10 ℃ and the relative humidity of 65%;
and thirdly, placing the grapes obtained in the step II in a light-shading environment with the temperature of 15 ℃ and the relative humidity of 45% for continuous fermentation for 10d to obtain the grape fermentation product.
The preparation method of the modified chitosan comprises the following steps:
(1) mixing chitosan and acetic acid aqueous solution at 30 ℃, and stirring at 300rpm for 30min to obtain chitosan acetic acid aqueous solution; the concentration of the acetic acid aqueous solution is 2wt.%; the mass ratio of the chitosan to the acetic acid aqueous solution is 1.2;
(2) adjusting the pH value of the chitosan acetic acid aqueous solution obtained in the step (1) to 9.0 by using a NaOH aqueous solution with the concentration of 0.5mol/L at the temperature of 30 ℃, and filtering to obtain a filter cake;
(3) mixing the filter cake obtained in the step (2) with n-butanol and NaOH aqueous solution at 30 ℃ and stirring at 300rpm for 30min; then, dropping an ammonium salt aqueous solution at the speed of 1mL/min, heating to 68 ℃ after the dropping is finished, and stirring at 180rpm for reaction for 8 hours; filtering to obtain primary filter residue after the reaction is finished, soaking the primary filter residue in absolute ethyl alcohol, stirring the soaked primary filter residue at 600rpm for 1 hour, filtering the primary filter residue again to obtain secondary filter residue, and drying the secondary filter residue for 1 hour at 70 ℃ under the condition of 99kPa to obtain the modified chitosan; the concentration of the NaOH aqueous solution is 0.5mol/L; the concentration of the aqueous ammonium salt solution was 10wt.%; the mass ratio of the filter cake to the n-butanol to the NaOH aqueous solution to the ammonium salt aqueous solution is 1; the mass ratio of the primary filter residue to the absolute ethyl alcohol is 1:3; the ammonium salt is a mixture of 2-hydroxyethyl trimethyl ammonium chloride and 3-chloro-2-hydroxypropyl trimethyl ammonium chloride according to a mass ratio of 1:1.
Example 2
The preparation method of the black vinegar health-care beverage comprises the following steps:
weighing cherries, tomatoes, medlar, cucumbers and dragon fruits according to a mass ratio of 1;
II, sterilizing the mixed slurry at 115 ℃ for 15min, and cooling to room temperature; adding zymophyte, fermenting at 29 deg.C for 60 hr, and filtering to obtain fermentation filtrate; the amount of the zymophyte is 5wt.%; the fermentation bacteria are formed by mixing schizosaccharomyces pombe and bacillus subtilis 2:1 in mass ratio;
III, adding 0.5 part by weight of xylitol and 30 parts by weight of black vinegar into 50 parts by weight of fermented filtrate, and uniformly mixing to obtain the black vinegar health-care beverage.
The black vinegar is prepared by the following method:
s1, crushing grains and sieving the grains with a 30-mesh sieve to obtain grain powder; the cereal is a mixture of glutinous rice, sorghum, black beans, black rice, rye and black tartary buckwheat according to a mass ratio of 15;
s2, mixing the grain powder obtained in the step S1 with amylase and distilled water at 40 ℃, stirring at 100rpm, and carrying out enzymolysis for 8 hours to obtain grain enzymolysis liquid; the mass ratio of the grain powder to the distilled water is 1:5; the addition amount of the amylase in per milliliter of distilled water is 500U;
s3, mixing dry yeast and 3.5wt.% of sucrose aqueous solution at 32 ℃, and stirring at 300rpm for 1h to obtain yeast sugar solution; the concentration of the yeast in the yeast sugar solution is 10 6 CFU/mL;
S4, mixing the grain enzymolysis liquid obtained in the step S2, the yeast sugar liquid obtained in the step S3 and the grape fermentation product according to the mass ratio of 250 to 100, and stirring at 30rpm for 8d under the anaerobic condition at 29 ℃ to obtain a pre-fermentation liquid;
s5, sterilizing the pre-fermentation liquid obtained in the step S4, adding acetobacter into the pre-fermentation liquid, and stirring the mixture for 10 days at the speed of 30rpm at the temperature of 36 ℃ under the condition of introducing oxygen to obtain crude vinegar; the inoculation amount of the acetobacter is 10 6 CFU/mL, the oxygen introduction amount is 5m 3 /(h·m 3 );
And S6, sterilizing and filtering the crude vinegar obtained in the step S5 to obtain the black vinegar.
The preparation method of the grape fermentation product comprises the following steps:
mixing botrytis cinerea, modified chitosan, hyaluronic acid and distilled water at 19 ℃, and stirring at 240rpm for 30min to obtain a botrytis cinerea liquid; the concentration of the botrytis cinerea in the botrytis cinerea liquid is 10 8 CFU/mL; the mass ratio of the modified chitosan to the hyaluronic acid to the distilled water is 0.05;
mixing the botrytis cinerea liquid obtained in the step with grapes in a mass ratio of 1:5, and fermenting for 4d in a light-resistant environment at the temperature of 10 ℃ and the relative humidity of 65%;
and thirdly, placing the grapes obtained in the step II in a light-shading environment with the temperature of 15 ℃ and the relative humidity of 45% for continuous fermentation for 10d to obtain the grape fermentation product.
The preparation method of the modified chitosan comprises the following steps:
(1) mixing chitosan and acetic acid aqueous solution at 30 ℃, and stirring at 300rpm for 30min to obtain chitosan acetic acid aqueous solution; the concentration of the acetic acid aqueous solution is 2wt.%; the mass ratio of the chitosan to the acetic acid aqueous solution is 1.2;
(2) adjusting the pH value of the chitosan acetic acid aqueous solution obtained in the step (1) to 9.0 by using a NaOH aqueous solution with the concentration of 0.5mol/L at the temperature of 30 ℃, and filtering to obtain a filter cake;
(3) mixing the filter cake obtained in the step (2) with n-butanol and NaOH aqueous solution at 30 ℃ and stirring at 300rpm for 30min; then, dropping an ammonium salt aqueous solution at the speed of 1mL/min, heating to 68 ℃ after the dropping is finished, and stirring at 180rpm for reaction for 8 hours; filtering to obtain primary filter residue after the reaction is finished, soaking the primary filter residue in absolute ethyl alcohol, stirring the mixture for 1 hour at 600rpm, filtering again to obtain secondary filter residue, and drying the secondary filter residue for 1 hour at 70 ℃ under 99kPa to obtain the modified chitosan; the concentration of the NaOH aqueous solution is 0.5mol/L; the concentration of the aqueous ammonium salt solution was 10wt.%; the mass ratio of the filter cake to the n-butanol to the NaOH aqueous solution to the ammonium salt aqueous solution is (1); the mass ratio of the primary filter residue to the absolute ethyl alcohol is 1:3; the ammonium salt is 2-hydroxyethyl trimethyl ammonium chloride.
Example 3
The preparation method of the black vinegar health-care beverage comprises the following steps:
weighing cherries, tomatoes, medlar, cucumbers and dragon fruits according to a mass ratio of 1;
II, sterilizing the mixed slurry at 115 ℃ for 15min, and cooling to room temperature; adding zymophyte, fermenting at 29 deg.C for 60 hr, and filtering to obtain fermentation filtrate; the amount of the zymophyte is 5wt.%; the zymocyte is formed by mixing schizosaccharomyces pombe and bacillus subtilis in a mass ratio of 2:1;
III, adding 0.5 part by weight of xylitol and 30 parts by weight of black vinegar into 50 parts by weight of fermentation filtrate, and uniformly mixing to obtain the black vinegar health-care beverage.
The black vinegar is prepared by the following method:
s1, crushing grains and sieving the grains with a 30-mesh sieve to obtain grain powder; the cereal is a mixture of glutinous rice, sorghum, black beans, black rice, rye and black tartary buckwheat according to a mass ratio of 15;
s2, mixing the grain powder obtained in the step S1 with amylase and distilled water at 40 ℃, stirring at 100rpm, and carrying out enzymolysis for 8 hours to obtain grain enzymolysis liquid; the mass ratio of the grain powder to the distilled water is 1:5; the addition amount of the amylase in per milliliter of distilled water is 500U;
s3, mixing dry yeast and 3.5wt.% of sucrose aqueous solution at 32 ℃, and stirring at 300rpm for 1h to obtain yeast sugar solution; the concentration of the yeast in the yeast sugar solution is 10 6 CFU/mL;
S4, mixing the grain enzymolysis liquid obtained in the step S2, the yeast sugar liquid obtained in the step S3 and the grape fermentation product according to a mass ratio of 250 to 100, and stirring at 30rpm for 8d under an anaerobic condition at 29 ℃ to obtain a pre-fermentation liquid;
s5, sterilizing the pre-fermentation liquid obtained in the step S4, adding acetobacter into the pre-fermentation liquid, and stirring the mixture for 10 days at the speed of 30rpm at 36 ℃ under the condition of oxygen introduction to obtain crude vinegar; the inoculation amount of the acetobacter is 10 6 CFU/mL, the oxygen introduction amount is 5m 3 /(h·m 3 );
And S6, sterilizing and filtering the crude vinegar obtained in the step S5 to obtain the black vinegar.
The preparation method of the grape fermentation product comprises the following steps:
mixing botrytis cinerea, modified chitosan, hyaluronic acid and distilled water at 19 ℃, and stirring at 240rpm for 30min to obtain a botrytis cinerea liquid; the concentration of the botrytis cinerea in the botrytis cinerea liquid is 10 8 CFU/mL; the mass ratio of the modified chitosan to the hyaluronic acid to the distilled water is 0.05;
mixing the botrytis cinerea liquid obtained in the step and grapes in a mass ratio of 1:5, and fermenting for 4d in a light-resistant environment at the temperature of 10 ℃ and the relative humidity of 65%;
and thirdly, placing the grapes obtained in the step II in a light-shading environment with the temperature of 15 ℃ and the relative humidity of 45% for continuous fermentation for 10d to obtain the grape fermentation product.
The preparation method of the modified chitosan comprises the following steps:
(1) mixing chitosan and acetic acid aqueous solution at 30 ℃, and stirring at 300rpm for 30min to obtain chitosan acetic acid aqueous solution; the concentration of the acetic acid aqueous solution is 2wt.%; the mass ratio of the chitosan to the acetic acid aqueous solution is 1.2;
(2) adjusting the pH value of the chitosan acetic acid aqueous solution obtained in the step (1) to 9.0 by using a NaOH aqueous solution with the concentration of 0.5mol/L at the temperature of 30 ℃, and filtering to obtain a filter cake;
(3) mixing the filter cake obtained in the step (2) with n-butanol and NaOH aqueous solution at 30 ℃ and stirring at 300rpm for 30min; then, dropwise adding an ammonium salt aqueous solution at the speed of 1mL/min, heating to 68 ℃ after dropwise adding is finished, and stirring at 180rpm for reaction for 8 hours; filtering to obtain primary filter residue after the reaction is finished, soaking the primary filter residue in absolute ethyl alcohol, stirring the mixture for 1 hour at 600rpm, filtering again to obtain secondary filter residue, and drying the secondary filter residue for 1 hour at 70 ℃ under 99kPa to obtain the modified chitosan; the concentration of the NaOH aqueous solution is 0.5mol/L; the concentration of the aqueous ammonium salt solution was 10wt.%; the mass ratio of the filter cake to the n-butanol to the NaOH aqueous solution to the ammonium salt aqueous solution is 1; the mass ratio of the primary filter residue to the absolute ethyl alcohol is 1:3; the ammonium salt is 3-chloro-2-hydroxypropyl trimethyl ammonium chloride.
Example 4
The preparation method of the black vinegar health drink comprises the following steps:
weighing cherries, tomatoes, medlar, cucumbers and dragon fruits according to a mass ratio of 1;
II, sterilizing the mixed slurry at 115 ℃ for 15min, and cooling to room temperature; adding zymophyte, fermenting at 29 deg.C for 60 hr, and filtering to obtain fermentation filtrate; the using amount of the zymophyte is 5wt.%; the fermentation bacteria are formed by mixing schizosaccharomyces pombe and bacillus subtilis 2:1 in mass ratio;
III, adding 0.5 part by weight of xylitol and 30 parts by weight of black vinegar into 50 parts by weight of fermentation filtrate, and uniformly mixing to obtain the black vinegar health-care beverage.
The black vinegar is prepared by the following method:
s1, crushing grains and sieving the grains with a 30-mesh sieve to obtain grain powder; the cereal is a mixture of glutinous rice, sorghum, black beans, black rice, rye and black tartary buckwheat according to a mass ratio of 15;
s2, mixing the grain powder obtained in the step S1 with amylase and distilled water at 40 ℃, stirring at 100rpm, and carrying out enzymolysis for 8 hours to obtain grain enzymolysis liquid; the mass ratio of the grain powder to the distilled water is 1:5; the addition amount of the amylase in per milliliter of distilled water is 500U;
s3, mixing dry yeast and 3.5wt.% of sucrose aqueous solution at 32 ℃, and stirring at 300rpm for 1h to obtain yeast sugar solution; the concentration of the yeast in the yeast sugar solution is 10 6 CFU/mL;
S4, mixing the grain enzymolysis liquid obtained in the step S2, the yeast sugar liquid obtained in the step S3 and the grape fermentation product according to a mass ratio of 250 to 100, and stirring at 30rpm for 8d under an anaerobic condition at 29 ℃ to obtain a pre-fermentation liquid;
s5, sterilizing the pre-fermentation liquid obtained in the step S4, adding acetobacter into the pre-fermentation liquid, and stirring the mixture for 10 days at the speed of 30rpm at the temperature of 36 ℃ under the condition of introducing oxygen to obtain crude vinegar; the inoculation amount of the acetobacter is 10 6 CFU/mL, the oxygen introduction amount is 5m 3 /(h·m 3 );
S6, sterilizing and filtering the crude vinegar obtained in the step S5 to obtain the black vinegar.
The preparation method of the grape fermentation product comprises the following steps:
mixing botrytis cinerea, chitosan, hyaluronic acid and distilled water at 19 ℃ and stirring at 240rpm for 30min to obtain a botrytis cinerea liquid; the concentration of the botrytis cinerea in the botrytis cinerea liquid is 10 8 CFU/mL; the mass ratio of the chitosan to the hyaluronic acid to the distilled water is 0.05;
mixing the botrytis cinerea liquid obtained in the step and grapes in a mass ratio of 1:5, and fermenting for 4d in a light-resistant environment at the temperature of 10 ℃ and the relative humidity of 65%;
and thirdly, placing the grapes obtained in the step II in a light-shading environment with the temperature of 15 ℃ and the relative humidity of 45% for continuous fermentation for 10d to obtain the grape fermentation product.
Example 5
The preparation method of the black vinegar health drink comprises the following steps:
i, weighing cherries, tomatoes, chinese wolfberries, cucumbers and dragon fruits according to a mass ratio of 1;
II, sterilizing the mixed slurry at 115 ℃ for 15min, and cooling to room temperature; adding zymophyte, fermenting at 29 deg.C for 60 hr, and filtering to obtain fermentation filtrate; the amount of the zymophyte is 5wt.%; the fermentation bacteria are formed by mixing schizosaccharomyces pombe and bacillus subtilis 2:1 in mass ratio;
III, adding 0.5 part by weight of xylitol and 30 parts by weight of black vinegar into 50 parts by weight of fermented filtrate, and uniformly mixing to obtain the black vinegar health-care beverage.
The black vinegar is prepared by the following method:
s1, crushing grains and sieving the grains with a 30-mesh sieve to obtain grain powder; the cereal is a mixture of glutinous rice, sorghum, black beans, black rice, rye and black tartary buckwheat according to a mass ratio of 15;
s2, mixing the grain powder obtained in the step S1 with amylase and distilled water at 40 ℃, stirring at 100rpm, and carrying out enzymolysis for 8 hours to obtain grain enzymolysis liquid; the mass ratio of the grain powder to the distilled water is 1:5; the addition amount of the amylase in per milliliter of distilled water is 500U;
s3, mixing dry yeast and 3.5wt.% of sucrose aqueous solution at 32 ℃ and stirring at 300rpm for 1h to obtain yeast sugar solution; the concentration of the yeast in the yeast sugar solution is 10 6 CFU/mL;
S4, mixing the grain enzymolysis liquid obtained in the step S2, the yeast sugar liquid obtained in the step S3 and the grape fermentation product according to the mass ratio of 250 to 100, and stirring at 30rpm for 8d under the anaerobic condition at 29 ℃ to obtain a pre-fermentation liquid;
s5, sterilizing the pre-fermentation liquid obtained in the step S4, adding acetobacter into the pre-fermentation liquid, and stirring the mixture for 10 days at the speed of 30rpm at the temperature of 36 ℃ under the condition of introducing oxygen to obtain crude vinegar; the inoculation amount of the acetobacter is 10 6 CFU/mL, the oxygen introduction amount is 5m 3 /(h·m 3 );
And S6, sterilizing and filtering the crude vinegar obtained in the step S5 to obtain the black vinegar.
The preparation method of the grape fermentation product comprises the following steps:
mixing botrytis cinerea, modified chitosan, polyglutamic acid and distilled water at 19 ℃, and stirring at 240rpm for 30min to obtain a botrytis cinerea bacterial liquid; the concentration of the botrytis cinerea in the botrytis cinerea liquid is 10 8 CFU/mL; the mass ratio of the modified chitosan to the hyaluronic acid to the distilled water is 0.05;
mixing the botrytis cinerea liquid obtained in the step and grapes in a mass ratio of 1:5, and fermenting for 4d in a light-resistant environment at the temperature of 10 ℃ and the relative humidity of 65%;
and thirdly, placing the grapes obtained in the step II in a light-shading environment with the temperature of 15 ℃ and the relative humidity of 45% for continuous fermentation for 10d to obtain the grape fermentation product.
The preparation method of the modified chitosan comprises the following steps:
(1) mixing chitosan and acetic acid aqueous solution at 30 ℃, and stirring at 300rpm for 30min to obtain chitosan acetic acid aqueous solution; the concentration of the acetic acid aqueous solution is 2wt.%; the mass ratio of the chitosan to the acetic acid aqueous solution is 1.2;
(2) adjusting the pH value of the chitosan acetic acid aqueous solution obtained in the step (1) to 9.0 by using a NaOH aqueous solution with the concentration of 0.5mol/L at the temperature of 30 ℃, and filtering to obtain a filter cake;
(3) mixing the filter cake obtained in the step (2) with n-butanol and NaOH aqueous solution at 30 ℃ and stirring at 300rpm for 30min; then, dropping an ammonium salt aqueous solution at the speed of 1mL/min, heating to 68 ℃ after the dropping is finished, and stirring at 180rpm for reaction for 8 hours; filtering to obtain primary filter residue after the reaction is finished, soaking the primary filter residue in absolute ethyl alcohol, stirring the mixture for 1 hour at 600rpm, filtering again to obtain secondary filter residue, and drying the secondary filter residue for 1 hour at 70 ℃ under 99kPa to obtain the modified chitosan; the concentration of the NaOH aqueous solution is 0.5mol/L; the concentration of the aqueous ammonium salt solution was 10wt.%; the mass ratio of the filter cake to the n-butanol to the NaOH aqueous solution to the ammonium salt aqueous solution is 1; the mass ratio of the primary filter residue to the absolute ethyl alcohol is 1:3; the ammonium salt is a mixture of 2-hydroxyethyl trimethyl ammonium chloride and 3-chloro-2-hydroxypropyl trimethyl ammonium chloride according to a mass ratio of 1:1.
Example 6
The preparation method of the black vinegar health-care beverage comprises the following steps:
weighing cherries, tomatoes, medlar, cucumbers and dragon fruits according to a mass ratio of 1;
II, sterilizing the mixed slurry at 115 ℃ for 15min, and cooling to room temperature; adding zymophyte, fermenting at 29 deg.C for 60 hr, and filtering to obtain fermentation filtrate; the amount of the zymophyte is 5wt.%; the fermentation bacteria are formed by mixing schizosaccharomyces pombe and bacillus subtilis 2:1 in mass ratio;
III, adding 0.5 part by weight of xylitol and 30 parts by weight of black vinegar into 50 parts by weight of the fermentation filtrate, and uniformly mixing to obtain the black vinegar health-care beverage.
The black vinegar is prepared by the following method:
s1, crushing grains and sieving the grains with a 30-mesh sieve to obtain grain powder; the cereal is a mixture of glutinous rice, sorghum, black beans, black rice, rye and black tartary buckwheat according to a mass ratio of 15;
s2, mixing the grain powder obtained in the step S1 with amylase and distilled water at 40 ℃, stirring at 100rpm, and carrying out enzymolysis for 8 hours to obtain grain enzymolysis liquid; the mass ratio of the grain powder to the distilled water is 1:5; the addition amount of the amylase in per milliliter of distilled water is 500U;
s3, mixing dry yeast and 3.5wt.% of sucrose aqueous solution at 32 ℃, and stirring at 300rpm for 1h to obtain yeast sugar solution; the concentration of the yeast in the yeast sugar solution is 10 6 CFU/mL;
S4, mixing the grain enzymolysis liquid obtained in the step S2 and the yeast sugar liquid obtained in the step S3 according to the mass ratio of 350 to 3, and stirring at 30rpm for 8d under the anaerobic condition at 29 ℃ to obtain a pre-fermentation liquid;
s5, sterilizing the pre-fermentation liquid obtained in the step S4, adding acetobacter into the pre-fermentation liquid, and stirring the mixture for 10 days at the speed of 30rpm at the temperature of 36 ℃ under the condition of introducing oxygen to obtain crude vinegar; of the acetobacterThe inoculation amount is 10 6 CFU/mL, the oxygen introduction amount is 5m 3 /(h·m 3 );
And S6, sterilizing and filtering the crude vinegar obtained in the step S5 to obtain the black vinegar.
Test example 1
Testing the content of amino acid in black vinegar: the content of gamma-aminobutyric acid in the black vinegar prepared in examples 1 to 6 was tested according to QB/T5197-2017 "high Performance liquid chromatography for determination of 12 free amino acids in wine"; the contents of glutamic acid, glycine and phenylalanine (flavor amino acids related to the flavor and taste of black vinegar) in the black vinegar prepared in examples 1 to 6 were tested according to GB 5009.124-2016, determination of amino acids in national food safety Standard food.
TABLE 1 the amino acid content of the black vinegar prepared in the examples
Example 1 (using a mixture of 2-hydroxyethyltrimethylammonium chloride, 3-chloro-2-hydroxypropyltrimethylammonium chloride in a mass ratio of 1:1 as the ammonium salt) had a flavor =9.86 minutes, a mouthfeel =9.81 minutes, γ -aminobutyric acid =432.1mg/100mL, glutamic acid =188.2mg/100mL, glycine =85.6mg/100mL, phenylalanine =80.4mg/100mL which is significantly better than examples 2-3 (using 2-hydroxyethyltrimethylammonium chloride or 3-chloro-2-hydroxypropyltrimethylammonium chloride as the ammonium salt), example 4 (using chitosan), example 5 (using polyglutamic acid), example 6 (using no grape fermentation) for a flavor = (5.62-9.23) minute, a flavor = (5.76-9.10) minute, γ -aminobutyric acid = (261.1.5-422.8) mg/100mL, 130.7-32 zxft 32 (130.32 mg-3732 mg) for phenylalanine 100mL, 100 mg/100mL, because this is 2-6 mg/100 mL: the 2-hydroxyethyl trimethyl ammonium chloride and the 3-chloro-2-hydroxypropyl trimethyl ammonium chloride are adopted to modify chitosan, so that the selective bacteriostasis of chitosan in preparing a grape fermentation product by utilizing botrytis cinerea can be enhanced, namely, the physiological process of the botrytis cinerea is not inhibited while the growth and the reproduction of foreign bacteria are inhibited, because the selected botrytis cinerea has stronger resistance to the modified chitosan prepared by the specific method of the invention due to glycoprotein with special spatial configuration on the surface, the modified chitosan can grow and reproduce in the environment applying the modified chitosan, and other foreign bacteria such as escherichia coli and staphylococcus aureus are inhibited without the protection of the glycoprotein; the attraction range between 2-hydroxyethyl in 2-hydroxyethyl trimethyl ammonium chloride and 2-hydroxypropyl in 3-chloro-2-hydroxypropyl trimethyl ammonium chloride can further enhance the selective bacteriostasis of the obtained modified chitosan, and simultaneously, the growth and the propagation of botrytis cinerea can not be inhibited, thereby achieving the synergistic effect. The invasion and growth of botrytis cinerea can volatilize moisture of grape raw materials, so that components such as saccharides, organic acids, polyhydric alcohols, terpenes and the like in grapes are changed, and finally, a grape fermentation product with high sugar content, strong fragrance and unique fragrance and taste is formed; the modified chitosan prepared by the method has a selective antibacterial effect, can inhibit physiological activities of sundry bacteria except botrytis cinerea, so that the fermentation activity of the botrytis cinerea on grapes is not disturbed by the sundry bacteria, organic acid, polyalcohol and terpenes with higher content are obtained, and when the substances are used in the subsequent fermentation process of black vinegar, the flavor of the finally obtained black vinegar can be improved, and the content of various amino acids in the black vinegar can be increased; due to the molecular size and the unique hydrophilic characteristic of the hyaluronic acid, the hyaluronic acid can play a certain role in inhibiting mixed bacteria and promoting the propagation of the botrytis cinerea while moisturizing the grapes and facilitating the rapid parasitic invasion of the botrytis cinerea into the grapes.
Test example 2
Stability testing of black vinegar health drink: the black vinegar health drink prepared in examples 1 to 6 was stored at 4 ℃ for 7 days. Accurately weighing 10g of black vinegar health drink, centrifuging at 5000rpm for 30min, collecting precipitate, weighing, and calculating precipitation rate as index for evaluating stability of black vinegar health drink. The precipitation rate% = mass of precipitate (g)/mass of black vinegar health drink (g) × 100%. Parallel to 5 groups, average.
Table 2 stability testing of black vinegar health beverages of the examples
Precipitation Rate (%) | |
Example 1 | 1.36 |
Example 2 | 1.58 |
Example 3 | 1.61 |
Example 4 | 2.10 |
Example 5 | 1.82 |
Example 6 | 2.73 |
Test example 3
And (3) testing the oxidation resistance of the black vinegar health-care beverage: respectively adding 2mL of the black vinegar health-care beverage prepared in the embodiment 1-3 into 18mL of deionized water, and uniformly mixing to obtain a sample solution to be detected; uniformly mixing 2.5mL of sample solution to be detected with 2.5mL of 0.1mM DPPH-ethanol solution; after being placed at room temperature in the dark for 30min, the absorbance A1 is measured at 517nm with deionized water as a reference. The blank is replaced by deionized water, and deionized water is used for replacing the sample solution to be detectedThe absorbance A0 was measured at 517nm as a reference. Dpph.clearance% = (1-a) 1 /A 0 ) X100%. Parallel 5 groups, take the average.
Table 3 stability test of black vinegar health drink of example
DPPH.Rate of removal (%) | |
Example 1 | 90.52 |
Example 2 | 88.67 |
Example 3 | 87.80 |
It should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and are not limited. Although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the scope of the present invention.
Claims (6)
1. A method for preparing a black vinegar health-care beverage is characterized by comprising the following steps: the method comprises the following steps:
weighing cherries, tomatoes, medlar, cucumbers and dragon fruits, removing kernels and peels, washing, draining, mixing, crushing into juice, and sieving to obtain mixed pulp;
II, sterilizing the mixed slurry and cooling to room temperature; adding zymophyte, fermenting, and filtering to obtain fermentation filtrate;
III, adding xylitol and black vinegar into the fermentation filtrate, and uniformly mixing to obtain the black vinegar health-care beverage.
2. The method for preparing a black vinegar health drink according to claim 1, wherein the method comprises the following steps: the method comprises the following steps:
weighing cherries, tomatoes, medlar, cucumbers and dragon fruits according to the mass ratio of (0.5-1.5) to (1-3) to (0.1-0.6) to (0.5-1.5) to (1-3), removing kernels and peels, washing with water, draining, mixing, crushing into juice, and sieving with a 50-100-mesh sieve to obtain mixed pulp;
II, sterilizing the mixed slurry at 110-120 ℃ for 10-20min, and cooling to room temperature; adding zymophyte, fermenting at 27-30 deg.C for 50-70 hr, and filtering to obtain fermentation filtrate; the amount of the zymophyte is 3-6wt.%; the zymocyte is formed by mixing schizosaccharomyces pombe and bacillus subtilis according to the mass ratio of 2 (1-2);
III, adding 0.1-0.6 part by weight of xylitol and 25-35 parts by weight of black vinegar into 30-60 parts by weight of fermentation filtrate, and uniformly mixing to obtain the black vinegar health-care beverage.
3. The method for preparing a black vinegar health drink according to claim 1 or 2, wherein: the black vinegar is prepared by the following method:
s1, crushing grains and sieving the grains with a sieve of 10-50 meshes to obtain grain powder; the grain is a mixture of glutinous rice, sorghum, black beans, black rice, rye and black tartary buckwheat according to the mass ratio of (13-20) to (10-13) to (3-7) to (1-5) to (7-12) to (1-5);
s2, mixing the grain powder obtained in the step S1 with amylase and water at 38-41 ℃, stirring at 80-120rpm, and carrying out enzymolysis for 7-10h to obtain grain enzymolysis liquid; the mass ratio of the grain powder to the water is (0.7-1.3) to (4-6); the addition amount of the amylase in each milliliter of water is 480-520U;
s3, mixing dry yeast and (2-4) wt.% of sucrose aqueous solution at 30-35 ℃, and stirring at 240-360rpm for 0.5-2h to obtain yeast sugar solution; in the yeast sugar solutionThe concentration of yeast is 10 6 CFU/mL;
S4, mixing the grain enzymolysis liquid obtained in the step S2, the yeast sugar liquid obtained in the step S3 and the grape fermentation product according to the mass ratio of (220-260) to (1-5) to (80-120), and stirring at the temperature of 28-33 ℃ and under the anaerobic condition at the rpm of 20-50 for 7-10 days to obtain pre-fermentation liquid;
s5, sterilizing the pre-fermentation liquid obtained in the step S4, adding acetobacter into the pre-fermentation liquid, and stirring the mixture for 8 to 12 days at a speed of 20 to 40rpm at a temperature of between 34 and 38 ℃ under the condition of introducing oxygen to obtain crude vinegar; the inoculation amount of the acetobacter is 10 6 CFU/mL, the oxygen introduction amount is 4-6m 3 /(h·m 3 );
And S6, sterilizing and filtering the crude vinegar obtained in the step S5 to obtain the black vinegar.
4. The method for preparing a black vinegar health drink according to claim 3, wherein the method comprises the following steps: the preparation method of the grape fermentation product comprises the following steps:
mixing botrytis cinerea, modified chitosan, hyaluronic acid and water at 15-22 ℃, and stirring at 180-300rpm for 10-40min to obtain a botrytis cinerea liquid; the concentration of the botrytis cinerea in the botrytis cinerea liquid is 10 6 -10 8 CFU/mL; the mass ratio of the modified chitosan to the hyaluronic acid to the water is (2-5) to (0.05-0.12) to (80-120);
mixing the botrytis cinerea liquid obtained in the step and grapes according to a mass ratio of (0.7-1.3) to (4-6), and fermenting for 3-5 days in a light-resistant environment at the temperature of 8-11 ℃ and the relative humidity of 58-67%;
and thirdly, placing the grapes obtained in the step II in a light-shielding environment with the temperature of 14-17 ℃ and the relative humidity of 38-46% for continuous fermentation for 9-12d to obtain the grape leavening.
5. The method for preparing the black vinegar health drink according to claim 3, wherein the modified chitosan is prepared by the following steps:
(1) mixing chitosan and acetic acid aqueous solution at 29-32 deg.C, and stirring at 240-360rpm for 20-40min to obtain chitosan acetic acid aqueous solution; the concentration of the acetic acid aqueous solution is 1.5-2.1wt.%; the mass ratio of the chitosan to the acetic acid aqueous solution is (1-1.4) to (98.6-99);
(2) adjusting the pH value of the chitosan acetic acid aqueous solution obtained in the step (1) to 8.8-9.1 by using a NaOH aqueous solution with the concentration of 0.1-1mol/L at the temperature of 29-32 ℃, and filtering to obtain a filter cake;
(3) mixing the filter cake obtained in the step (2) with n-butanol and NaOH aqueous solution at 29-32 ℃, and stirring at 240-360rpm for 10-40min; then, dripping ammonium salt water solution at the speed of 0.5-1.5mL/min, heating to 65-70 ℃ after finishing dripping, and stirring at 120-240rpm for reaction for 7-10h; filtering to obtain primary filter residue after the reaction is finished, soaking the primary filter residue in absolute ethyl alcohol, stirring at 540-660rpm for 0.5-2h, filtering again to obtain secondary filter residue, and drying the secondary filter residue for 0.5-2h at 68-72 ℃ under the condition of 97-99.6kPa to obtain the modified chitosan; the concentration of the NaOH aqueous solution is 0.1-1mol/L; the concentration of the aqueous ammonium salt solution is 8-11wt.%; the mass ratio of the filter cake, the normal butanol, the NaOH aqueous solution and the ammonium salt aqueous solution is (0.6-1.5) to (3-7) to (2-3.5) to (1.2-2.3); the mass ratio of the primary filter residue to the absolute ethyl alcohol is (0.5-1.2) to (2.8-4.3); the ammonium salt is 2-hydroxyethyl trimethyl ammonium chloride and/or 3-chloro-2-hydroxypropyl trimethyl ammonium chloride.
6. A black vinegar health drink is characterized in that: made by the method of any one of claims 1-5.
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