CN115746979B - Brewing method of fermented water chestnut wine - Google Patents
Brewing method of fermented water chestnut wine Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a brewing method of fermented water chestnut wine, which comprises the following steps: (1) selecting and crushing water chestnut; (2) gelatinization; (3) liquefying; (4) saccharification; (5) alcoholic fermentation: adding a proper amount of white granulated sugar to adjust the sugar degree to 15-20 degrees Bx; subsequently adding gamma-aminobutyric acid; inoculating Saccharomyces cerevisiae into saccharification liquid, and fermenting at the temperature of 28-30 ℃ for 7-10 d hours, wherein the fermentation liquid is subjected to cooling and heating treatment every 12 hours, and then fermentation is carried out at the primary fermentation temperature. The invention has simple process, utilizes the combination of temperature rise and temperature reduction stress and high-concentration gamma-aminobutyric acid to generate stress effect on yeast, improves the fermentation performance of the saccharomyces cerevisiae, and obtains the low-alcoholicity water chestnut wine with strong bouquet.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to a brewing method of fermented water chestnut wine.
Background
Water chestnut, also called loin chestnut, is a fruit of an annual aquatic weed plant of the family Trapa, genus Trapa. Water chestnut is commonly found in tropical, subtropical and temperate regions, wherein the water chestnut is mainly cultivated and distributed in Yangtze river basin in China. The water caltrop can be divided into water caltrop without corner, water caltrop with two corners and water caltrop with four corners according to the corner; the color can be classified into red diamond, purple diamond, black diamond, etc. The water chestnut contains various active substances and has the characteristics of resisting oxidation, resisting cancer, enhancing immunity and the like. The water chestnut peel is the exocarp of the water chestnut fruit, has slightly bitter and astringent taste and cool nature, and has the effects of clearing heat and detoxicating, treating sore, relieving diarrhea with astringents, and clearing damp-heat; the water chestnut has tender and sweet meat quality, and has the advantages of cooling, quenching thirst and having nutritive value comparable with other nuts. The water chestnut processing still stays in the primary stage, and the research on deep processing products is less. At present, the processed products of the water chestnut comprise water chestnut vermicelli, water chestnut fine dried noodles, water chestnut biscuits, water chestnut jelly, water chestnut yoghourt and the like. The water chestnut has higher starch content, and amylase can convert starch into monosaccharide and oligosaccharide, so the water chestnut can be used as a substrate for fermentation.
Through retrieval, CN201510006469.0 discloses a production method of water chestnut wine, which comprises the steps of sun-drying fresh water chestnuts after shell removal, and crushing to obtain crushed water chestnuts for later use; adding water, steaming, saccharifying, fermenting, distilling, and receiving when alcohol content of distilled liquor is higher than 53% vol. The process takes more than three years, is complex and has extremely high requirements on storage environment conditions.
Ai Mei consultation data shows that the market size of the Chinese low alcohol wine is kept in a high-speed growth state in 2017-2021, and the market size of the Chinese low alcohol wine is expected to break through 5000 hundred million yuan in 2022. The low alcohol degree wine (alcohol degree is below 20 degrees) is matched with the Z-generation consumption concept in the aspects of product form, brand marketing and the like, and becomes a new choice of wine. The financing amount of the China low alcohol wine industry in 2021 reaches tens of times, and the initial round is the main. Currently, the low alcohol wine industry is in the "hundred contend" stage. In the future, mode innovations and developments may be sustainable or will become the key to winning low wine brand angles from industry racetracks.
Therefore, the low-alcoholicity water chestnut wine with simple development process and rich wine aroma has wide market development prospect, and simultaneously provides theoretical and practical basis for developing more water chestnut products.
Disclosure of Invention
The invention aims to: aiming at the problems in the prior art, the invention provides the brewing method of the fermentation type water chestnut wine, which utilizes the combination of temperature rising and temperature reducing stress and high-concentration gamma-aminobutyric acid to generate stress effect on yeast in the fermentation process of the water chestnut wine, improves the fermentation performance of the yeast, simultaneously promotes the remarkable increase of the content of ester substances, improves the aroma quality of the wine, and obtains the low-alcoholicity water chestnut wine with strong aroma.
The technical scheme is as follows: the invention is realized by the following technical scheme: a brewing method of fermented water chestnut wine comprises the following steps:
step one, removing shells of water chestnuts, crushing and sterilizing;
Step two, gelatinization: weighing the water chestnut sterilized in the first step, mixing with sterilized distilled water, heating at the constant temperature of 90-95 ℃ for 30-40 min, continuously stirring, gelatinizing and cooling;
step three, liquefying: adding alpha-amylase into the product obtained in the step two, liquefying for 1.5-2 hours at the constant temperature of 92-96 ℃, inactivating enzyme, and cooling;
step four, saccharification: adding saccharifying enzyme into the product obtained in the step three, saccharifying for 1.5-2 hours at the constant temperature of 60-65 ℃, inactivating enzyme, and cooling;
Step five, alcohol fermentation: adding gamma-aminobutyric acid and saccharomyces cerevisiae into the obtained product of the step four after adjusting the sugar degree, and fermenting for 7-10 d at the temperature of 28-30 ℃;
And (3) carrying out cooling and heating treatment on the mixture in fermentation every 11-13 hours, and circulating the cooling and heating treatment in the fermentation process until the fermentation is completed.
Preferably, in the fifth step, the mass of the gamma-aminobutyric acid is 40-45 times of the content of the gamma-aminobutyric acid in the water caltrop weighing step.
Preferably, in the fifth step, the temperature-reducing and temperature-raising treatment is to reduce the temperature of the mixture in the fermentation to 0 ℃ at a rate of 3-5 ℃/min based on the temperature of 28-30 ℃ and then immediately raise the temperature back to the temperature of 28-30 ℃ at a rate of 5-8 ℃/min.
Preferably, in the fifth step, the saccharomyces cerevisiae is saccharomyces cerevisiae AWRI350,350.
Preferably, the mass of the saccharomyces cerevisiae AWRI is 0.3-0.5% of the mass of the weighed water chestnut in the second step.
Preferably, in the second step, the ratio of the water caltrop to the sterilized distilled water is 1 (8-10) (g/mL).
Preferably, in the third step and/or the fourth step, the enzyme deactivation treatment is to deactivate the enzyme at the constant temperature of 90-100 ℃ for 15-20 min.
Preferably, in the third step, the alpha-amylase is added in an amount of 20-30U/g.
Preferably, in the fourth step, the adding amount of the saccharifying enzyme is 100-140U/g.
Preferably, in the fifth step, the sugar degree is 15-20 degrees Bx.
The beneficial effects are that: the invention utilizes the combination of temperature rise and temperature reduction and high concentration gamma-aminobutyric acid to generate stress effect on yeast, improves the fermentation performance of the yeast, simultaneously promotes the remarkable increase of the content of esters, improves the aroma quality of wine, and has the specific beneficial effects that the obtained low-alcoholicity water chestnut wine has strong wine aroma:
1. The content of ester substances is obviously increased by combining heating and cooling stress with high-concentration gamma-aminobutyric acid reverse stress, so that the aroma quality of the wine is improved.
The tricarboxylic acid cycle is the most efficient way for the body to oxidize sugar or other substances to obtain energy, and the intermediate products (e.g., citric acid) of the cycle are raw materials for synthesizing fats and the like; citric acid can be broken down into acetyl-coa, a key substrate for the fatty acid synthesis pathway. NAD + is widely present in all living cells as a coenzyme and a reducing factor necessary for maintaining cellular metabolism, and participates in oxidative metabolic reactions within the cells. NAD + is an important cofactor involved in glycolysis and TCA cycle, and increasing the NAD +/NADH ratio can promote the efficiency of the reactions of these pathways. Saccharomyces cerevisiae is able to respond to stress of various substances and changes in the environment in response to metabolism. When the thalli are stressed by the outside, the saccharomyces cerevisiae can resist external injury and maintain the activity of the saccharomyces cerevisiae by changing the cell structure, physiological and biochemical indexes and the like. As a stress reaction of plant tissues to external conditions, enrichment of gamma-aminobutyric acid in plants is mainly generated by external stress, including but not limited to oxygen concentration, calcium ion concentration, hydrogen ion concentration, temperature, pressure and the like. In the fermentation process for preparing the water chestnut wine, the saccharomyces cerevisiae improves the NAD +/NADH ratio under the condition of combining external heating and cooling stress with high-concentration gamma-aminobutyric acid reverse stress, so that the glycolytic efficiency and TCA cycle efficiency are improved, intermediate products such as citric acid and the like enter cytoplasm, and the expression quantity of genes such as EHT1 and EEB1 coding alcohol acyl transferase (key enzyme responsible for catalyzing the reaction of acyl coenzyme A and ethanol to generate corresponding fatty acid ester), alcohol acetyl transferase ATF1 and ATF2, fatty acid acyl coenzyme A synthetase genes (FAA 1 and FAT 1) and the like is improved, so that the content of acetate and ethyl ester generated by saccharomyces cerevisiae metabolism is improved in the fermentation process, the content of ester substances is improved while the ethanol yield is maintained, the flavor of wine is increased, and further higher grading value is obtained.
2. The fermentation performance of the saccharomyces cerevisiae is improved by combining temperature rise and temperature reduction stress with high-concentration gamma-aminobutyric acid reverse stress.
The yeast needs to respond to the temperature change and the gamma-aminobutyric acid pressure in the environment through the combination of temperature rise and temperature reduction stress and high-concentration gamma-aminobutyric acid reverse stress, the fermentation performance of the saccharomyces cerevisiae is improved, the fermentation blood sugar reduction rate is improved, the residual sugar content in the fermented wine body is obviously reduced, and the alcoholicity is obviously improved.
Drawings
FIG. 1 is a graph showing the relative expression levels of ATF1, ATF2, EHT1, EEB1, FAT1 and FAA1 genes in samples of examples 1 to 6 and comparative examples 1 to 3 according to the present invention;
FIG. 2 is a graph showing the sensory scores of the samples of examples 1 to 6 and comparative examples 1 to 3 according to the present invention;
FIG. 3 is a theoretical schematic of the present invention;
FIG. 4 is a graph showing NAD +/NADH ratio of samples of examples 1 to 6 and comparative examples 1 to 3 in the present invention.
Detailed Description
The invention is further described in connection with the following detailed description, in order to make the technical means, the creation characteristics, the achievement of the purpose and the effect of the invention easy to understand.
Example 1
Taking fresh water chestnut without diseases and insect pests as a raw material, removing shells, crushing by a crusher, and sterilizing at 100 ℃ for 15 min; weighing 1000g of sterilized water chestnut (containing 150mg of gamma-aminobutyric acid), adding sterilized distilled water according to a feed liquid ratio of 1:9 (g/mL), stirring and heating at 92 ℃ in a constant-temperature water bath for 35min, and then cooling for later use; then adding 27U/g of alpha-amylase, liquefying for 1.7 hours at 94 ℃ in a constant-temperature water bath, inactivating enzyme in the constant-temperature water bath at 94 ℃ for 16min, and cooling to room temperature; then 120U/g of saccharifying enzyme is added, saccharifying is carried out for 1.3 hours at the temperature of 63 ℃ in a constant-temperature water bath, then the saccharifying enzyme is put into the constant-temperature water bath at 96 ℃ for enzyme deactivation for 18 minutes, and the saccharifying enzyme is cooled to room temperature; adding a proper amount of white granulated sugar into the saccharification liquid to adjust the sugar degree to 17 degrees Bx; 6g of gamma-aminobutyric acid was then added; inoculating Saccharomyces cerevisiae with the mass of 0.4% of sterilized and dehulled water chestnut into saccharified liquid, and fermenting at 29 ℃, wherein the temperature of the fermentation liquid is reduced to 0 ℃ at a speed of 4 ℃/min on the basis of the original fermentation temperature every 12 hours, and then the temperature is immediately increased to the original fermentation temperature at a speed of 6 ℃/min for continuous fermentation, wherein the fermentation period is 9d. Before fermentation, the content of gamma-aminobutyric acid in the saccharification liquid is 693.7mg/L, and after fermentation, the concentration of gamma-aminobutyric acid in the fermentation liquid is 1175.6mg/L.
Example 2
Taking fresh water chestnut without diseases and insect pests as a raw material, removing shells, crushing by a crusher, and sterilizing at 100 ℃ for 15 min; weighing 1000g of sterilized water chestnut (containing 150mg of gamma-aminobutyric acid), adding sterilized distilled water according to a feed liquid ratio of 1:9 (g/mL), stirring and heating at 92 ℃ in a constant-temperature water bath for 35min, and then cooling for later use; then adding 27U/g of alpha-amylase, liquefying for 1.7 hours at 94 ℃ in a constant-temperature water bath, inactivating enzyme in the constant-temperature water bath at 94 ℃ for 16min, and cooling to room temperature; then 120U/g of saccharifying enzyme is added, saccharifying is carried out for 1.3 hours at the temperature of 63 ℃ in a constant-temperature water bath, then the saccharifying enzyme is put into the constant-temperature water bath at 96 ℃ for enzyme deactivation for 18 minutes, and the saccharifying enzyme is cooled to room temperature; adding a proper amount of white granulated sugar into the saccharification liquid to adjust the sugar degree to 17 degrees Bx; 6.75g of gamma-aminobutyric acid was then added; inoculating Saccharomyces cerevisiae with the mass of 0.4% of sterilized and dehulled water chestnut into saccharified liquid, and fermenting at 29 ℃, wherein the temperature of the fermentation liquid is reduced to 0 ℃ at a speed of 4 ℃/min on the basis of the original fermentation temperature every 12 hours, and then the temperature is immediately increased to the original fermentation temperature at a speed of 6 ℃/min for continuous fermentation, wherein the fermentation period is 9d. Before fermentation, the content of gamma-aminobutyric acid in the saccharification liquid is 783.6mg/L, and after fermentation, the concentration of gamma-aminobutyric acid in the fermentation liquid is 1426.7mg/L.
Example 3
Taking fresh water chestnut without diseases and insect pests as a raw material, removing shells, crushing by a crusher, and sterilizing at 100 ℃ for 15 min; weighing 1000g of sterilized water chestnut (containing 150mg of gamma-aminobutyric acid), adding sterilized distilled water according to a feed liquid ratio of 1:9 (g/mL), stirring and heating at 92 ℃ in a constant-temperature water bath for 35min, and then cooling for later use; then adding 27U/g of alpha-amylase, liquefying for 1.7 hours at 94 ℃ in a constant-temperature water bath, inactivating enzyme in the constant-temperature water bath at 94 ℃ for 16min, and cooling to room temperature; then 120U/g of saccharifying enzyme is added, saccharifying is carried out for 1.3 hours at the temperature of 63 ℃ in a constant-temperature water bath, then the saccharifying enzyme is put into the constant-temperature water bath at 96 ℃ for enzyme deactivation for 18 minutes, and the saccharifying enzyme is cooled to room temperature; adding a proper amount of white granulated sugar into the saccharification liquid to adjust the sugar degree to 17 degrees Bx; 6.30g of gamma-aminobutyric acid was then added; inoculating Saccharomyces cerevisiae with the mass of 0.4% of sterilized and dehulled water chestnut into saccharified liquid, and fermenting at 29 ℃, wherein the temperature of the fermentation liquid is reduced to 0 ℃ at a speed of 4 ℃/min on the basis of the original fermentation temperature every 12 hours, and then the temperature is immediately increased to the original fermentation temperature at a speed of 6 ℃/min for continuous fermentation, wherein the fermentation period is 9d. Before fermentation, the content of gamma-aminobutyric acid in the saccharification liquid is 746.7mg/L, and after fermentation, the concentration of gamma-aminobutyric acid in the fermentation liquid is 1296.2mg/L.
Example 4
Taking fresh water chestnut without diseases and insect pests as a raw material, removing shells, crushing by a crusher, and sterilizing at 100 ℃ for 15 min; weighing 1000g of sterilized water chestnut (containing 150mg of gamma-aminobutyric acid), adding sterilized distilled water according to a feed liquid ratio of 1:9 (g/mL), stirring and heating at 92 ℃ in a constant-temperature water bath for 35min, and then cooling for later use; then adding 27U/g of alpha-amylase, liquefying for 1.7 hours at 94 ℃ in a constant-temperature water bath, inactivating enzyme in the constant-temperature water bath at 94 ℃ for 16min, and cooling to room temperature; then 120U/g of saccharifying enzyme is added, saccharifying is carried out for 1.3 hours at the temperature of 63 ℃ in a constant-temperature water bath, then the saccharifying enzyme is put into the constant-temperature water bath at 96 ℃ for enzyme deactivation for 18 minutes, and the saccharifying enzyme is cooled to room temperature; adding a proper amount of white granulated sugar into the saccharification liquid to adjust the sugar degree to 17 degrees Bx; 6.30g of gamma-aminobutyric acid was then added; inoculating Saccharomyces cerevisiae with the mass of 0.4% of sterilized and dehulled water chestnut into saccharified liquid, and fermenting at 29 ℃, wherein the temperature of the fermentation liquid is reduced to 0 ℃ at a speed of 3 ℃/min on the basis of the original fermentation temperature every 12 hours, and then the temperature is immediately increased to the original fermentation temperature at a speed of 5 ℃/min for continuous fermentation, wherein the fermentation period is 9d. Before fermentation, the content of gamma-aminobutyric acid in the saccharification liquid is 751.9mg/L, and after fermentation, the concentration of gamma-aminobutyric acid in the fermentation liquid is 1283.7mg/L.
Example 5
Taking fresh water chestnut without diseases and insect pests as a raw material, removing shells, crushing by a crusher, and sterilizing at 100 ℃ for 15 min; weighing 1000g of sterilized water chestnut (containing 150mg of gamma-aminobutyric acid), adding sterilized distilled water according to a feed liquid ratio of 1:9 (g/mL), stirring and heating at 92 ℃ in a constant-temperature water bath for 35min, and then cooling for later use; then adding 27U/g of alpha-amylase, liquefying for 1.7 hours at 94 ℃ in a constant-temperature water bath, inactivating enzyme in the constant-temperature water bath at 94 ℃ for 16min, and cooling to room temperature; then 120U/g of saccharifying enzyme is added, saccharifying is carried out for 1.3 hours at the temperature of 63 ℃ in a constant-temperature water bath, then the saccharifying enzyme is put into the constant-temperature water bath at 96 ℃ for enzyme deactivation for 18 minutes, and the saccharifying enzyme is cooled to room temperature; adding a proper amount of white granulated sugar into the saccharification liquid to adjust the sugar degree to 17 degrees Bx; 6.30g of gamma-aminobutyric acid was then added; inoculating Saccharomyces cerevisiae with the mass of 0.4% of sterilized and dehulled water chestnut into saccharified liquid, and fermenting at 29 ℃, wherein the temperature of the fermentation liquid is reduced to 0 ℃ at a speed of 5 ℃/min on the basis of the original fermentation temperature every 12 hours, and then the temperature is immediately increased to the original fermentation temperature at a speed of 8 ℃/min for continuous fermentation, wherein the fermentation period is 9d. Before fermentation, the content of gamma-aminobutyric acid in the saccharification liquid is 735.1mg/L, and after fermentation, the concentration of gamma-aminobutyric acid in the fermentation liquid is 1326.7mg/L.
Example 6
Taking fresh water chestnut without diseases and insect pests as a raw material, removing shells, crushing by a crusher, and sterilizing at 100 ℃ for 15 min; weighing 1000g of sterilized water chestnut (containing 150mg of gamma-aminobutyric acid), adding sterilized distilled water according to a feed liquid ratio of 1:9 (g/mL), stirring and heating at 92 ℃ in a constant-temperature water bath for 35min, and then cooling for later use; then adding 27U/g of alpha-amylase, liquefying for 1.7 hours at 94 ℃ in a constant-temperature water bath, inactivating enzyme in the constant-temperature water bath at 94 ℃ for 16min, and cooling to room temperature; then 120U/g of saccharifying enzyme is added, saccharifying is carried out for 1.3 hours at the temperature of 63 ℃ in a constant-temperature water bath, then the saccharifying enzyme is put into the constant-temperature water bath at 96 ℃ for enzyme deactivation for 18 minutes, and the saccharifying enzyme is cooled to room temperature; adding a proper amount of white granulated sugar into the saccharification liquid to adjust the sugar degree to 17 degrees Bx; 6.30g of gamma-aminobutyric acid was then added; inoculating Saccharomyces cerevisiae with the mass of 0.4% of sterilized and dehulled water chestnut into saccharified liquid, and fermenting at 29 ℃, wherein the temperature of the fermentation liquid is reduced to 0 ℃ at a speed of 4 ℃/min on the basis of the original fermentation temperature every 12 hours, and then the temperature is immediately increased to the original fermentation temperature at a speed of 7 ℃/min for continuous fermentation, wherein the fermentation period is 9d. Before fermentation, the content of gamma-aminobutyric acid in the saccharification liquid is 753.7mg/L, and after fermentation, the concentration of gamma-aminobutyric acid in the fermentation liquid is 1315.9mg/L.
Comparative example 1
Taking fresh water chestnut without diseases and insect pests as a raw material, removing shells, crushing by a crusher, and sterilizing at 100 ℃ for 15 min; weighing 1000g of sterilized water chestnut (containing 150mg of gamma-aminobutyric acid), adding sterilized distilled water according to a feed liquid ratio of 1:9 (g/mL), stirring and heating at 92 ℃ in a constant-temperature water bath for 35min, and then cooling for later use; then adding 27U/g of alpha-amylase, liquefying for 1.7 hours at 94 ℃ in a constant-temperature water bath, inactivating enzyme in the constant-temperature water bath at 94 ℃ for 16min, and cooling to room temperature; then 120U/g of saccharifying enzyme is added, saccharifying is carried out for 1.3 hours at the temperature of 63 ℃ in a constant-temperature water bath, then the saccharifying enzyme is put into the constant-temperature water bath at 96 ℃ for enzyme deactivation for 18 minutes, and the saccharifying enzyme is cooled to room temperature; adding a proper amount of white granulated sugar into the saccharification liquid to adjust the sugar degree to 17 degrees Bx; inoculating Saccharomyces cerevisiae with the mass of 0.4% of sterilized and dehulled water chestnut into saccharified liquid, and fermenting at 29 ℃, wherein the temperature of the fermentation liquid is reduced to 0 ℃ at a speed of 4 ℃/min on the basis of the original fermentation temperature every 12 hours, and then the temperature is immediately increased to the original fermentation temperature at a speed of 6 ℃/min for continuous fermentation, wherein the fermentation period is 9d. Before fermentation, the content of gamma-aminobutyric acid in the saccharification liquid is 15.6mg/L, and after fermentation, the concentration of gamma-aminobutyric acid in the fermentation liquid is 16.4mg/L.
Comparative example 2
Taking fresh water chestnut without diseases and insect pests as a raw material, removing shells, crushing by a crusher, and sterilizing at 100 ℃ for 15 min; weighing 1000g of sterilized water chestnut (containing 150mg of gamma-aminobutyric acid), adding sterilized distilled water according to a feed liquid ratio of 1:9 (g/mL), stirring and heating at 92 ℃ in a constant-temperature water bath for 35min, and then cooling for later use; then adding 27U/g of alpha-amylase, liquefying for 1.7 hours at 94 ℃ in a constant-temperature water bath, inactivating enzyme in the constant-temperature water bath at 94 ℃ for 16min, and cooling to room temperature; then 120U/g of saccharifying enzyme is added, saccharifying is carried out for 1.3 hours at the temperature of 63 ℃ in a constant-temperature water bath, then the saccharifying enzyme is put into the constant-temperature water bath at 96 ℃ for enzyme deactivation for 18 minutes, and the saccharifying enzyme is cooled to room temperature; adding a proper amount of white granulated sugar into the saccharification liquid to adjust the sugar degree to 17 degrees Bx; 6.30g of gamma-aminobutyric acid was then added; inoculating 0.4% of sterilized and shelled water chestnut quality Saccharomyces cerevisiae into saccharified liquid, and fermenting at 29 deg.C for 9d. Before fermentation, the content of gamma-aminobutyric acid in the saccharification liquid is 729.7mg/L, and after fermentation, the concentration of gamma-aminobutyric acid in the fermentation liquid is 742.4mg/L.
Comparative example 3
Taking fresh water chestnut without diseases and insect pests as a raw material, removing shells, crushing by a crusher, and sterilizing at 100 ℃ for 15 min; weighing 1000g of sterilized water chestnut (containing 150mg of gamma-aminobutyric acid), adding sterilized distilled water according to a feed liquid ratio of 1:9 (g/mL), stirring and heating at 92 ℃ in a constant-temperature water bath for 35min, and then cooling for later use; then adding 27U/g of alpha-amylase, liquefying for 1.7 hours at 94 ℃ in a constant-temperature water bath, inactivating enzyme in the constant-temperature water bath at 94 ℃ for 16min, and cooling to room temperature; then 120U/g of saccharifying enzyme is added, saccharifying is carried out for 1.3 hours at the temperature of 63 ℃ in a constant-temperature water bath, then the saccharifying enzyme is put into the constant-temperature water bath at 96 ℃ for enzyme deactivation for 18 minutes, and the saccharifying enzyme is cooled to room temperature; adding a proper amount of white granulated sugar into the saccharification liquid to adjust the sugar degree to 17 degrees Bx; inoculating 0.4% of sterilized and shelled water chestnut quality Saccharomyces cerevisiae into saccharified liquid, and fermenting at 29 deg.C for 9d. Before fermentation, the content of gamma-aminobutyric acid in the saccharification liquid is 15.7mg/L, and after fermentation, the concentration of gamma-aminobutyric acid in the fermentation liquid is 16.5mg/L.
The results of comparing the fermentation performance in the samples of examples 1 to 6 and comparative examples 1 to 3 are shown in Table 1:
TABLE 1 comparison of fermentation Performance of samples of different examples and comparative examples of the invention
| Alcohol content (v/v, 20 ℃ C.) | Residual sugar (g/L) | |
| Example 1 | 12.57±0.14b B | 3.34±0.05a A |
| Example 2 | 12.76±0.09b C | 3.88±0.03a C |
| Example 3 | 12.34±0.11b B | 3.73±0.06a B |
| Example 4 | 12.56±0.06b B | 3.69±0.06a B |
| Example 5 | 12.72±0.04b C | 3.77±0.04a B |
| Example 6 | 12.83±0.06b C | 3.41±0.03a A |
| Comparative example 1 | 10.13±0.21b A | 4.56±0.06a D |
| Comparative example 2 | 10.24±0.14b A | 4.47±0.13a D |
| Comparative example 3 | 10.16±0.15b A | 4.50±0.09a D |
The lower case letters of the same row differ significantly (p < 0.05); different capital letters in the same column represent significant differences (p < 0.05)
As can be seen from Table 1, the yeast needs to respond to the temperature change and the gamma-aminobutyric acid pressure in the environment by combining the temperature rising and reducing stress with the high-concentration gamma-aminobutyric acid reverse stress, and the fermentation performance and the fermentation blood sugar reducing rate of the Saccharomyces cerevisiae are improved in the process, so that compared with comparative examples 1 to 3, the residual sugar content in the wine after the fermentation of examples 1 to 6 is obviously reduced, and the alcoholic strength is obviously improved.
The types and contents of the ester components in the samples of examples 1 to 6 and comparative examples 1 to 3 were tested, and the results are shown in Table 2:
table 2 types and contents (mg/L) of ester components in fermented wine.
Different lower case letters in the same column represent significant differences (p < 0.05)
As can be seen from Table 2, during fermentation, saccharomyces cerevisiae is able to respond accordingly to stress of various substances and changes in the environment. When the thalli are stressed by the outside, the saccharomyces cerevisiae can resist external injury and maintain the activity of the saccharomyces cerevisiae by changing the cell structure, physiological and biochemical indexes and the like. As a stress reaction of plant tissues to external conditions, enrichment of gamma-aminobutyric acid in plants is mainly generated by external stress, including but not limited to oxygen concentration, calcium ion concentration, hydrogen ion concentration, temperature, pressure and the like. The Saccharomyces cerevisiae performs metabolic response under the stress of adding gamma-aminobutyric acid outside and increasing and decreasing the temperature of the outside environment, and the relative expression amounts of ATF1, ATF2, EHT1, EEB1, FAT1 and FAA1 genes are increased (figure 1), so that the contents of acetate and ethyl ester generated by metabolism of the Saccharomyces cerevisiae are increased in the fermentation process, the content of ester substances is increased while the yield of ethanol is maintained, and the flavor of wine is increased, so that compared with comparative examples 1-3, the contents of acetate and ethyl ester in wine bodies after fermentation of examples 1-6 are obviously higher.
Sensory evaluation in terms of color, aroma, taste and the like was performed on the water chestnut wines prepared in examples 1 to 6 and comparative examples 1 to 3, the sensory evaluation criteria are shown in table 3, and the sensory evaluation results are shown in fig. 2.
Table 3 sensory evaluation table
As can be seen from FIG. 2, the fermented wine of examples 1 to 6 had a higher aroma quality and a stronger aroma than those of comparative examples 1 to 3, and thus a higher sensory score was obtained.
The NAD +/NADH ratio in the samples of examples 1-6 and comparative examples 1-3 was analyzed and the results are shown in FIG. 4:
The tricarboxylic acid cycle is the most efficient way for the body to oxidize sugar or other substances to obtain energy, and the intermediate products (e.g., citric acid) of the cycle are raw materials for synthesizing fats and the like; citric acid can be broken down into acetyl-coa, a key substrate for the fatty acid synthesis pathway. NAD + is widely present in all living cells as a coenzyme and a reducing factor necessary for maintaining cellular metabolism, and participates in oxidative metabolic reactions within the cells. NAD + is an important cofactor involved in glycolysis and TCA cycle, and increasing the NAD +/NADH ratio can promote the efficiency of the reactions of these pathways. The improvement of NAD +/NADH ratio results in the improvement of glycolytic efficiency and TCA cycle efficiency, is favorable for intermediate products such as citric acid and the like to enter cytoplasm, promotes the increase of gene expression quantity such as EHT1 and EEB1 coding alcohol acyl transferase (key enzyme responsible for catalyzing the reaction of acyl coenzyme A and ethanol to generate corresponding fatty acid ester), alcohol acetyl transferase ATF1 and ATF2, fatty acid acyl coenzyme A synthetase genes (FAA 1, FAT 1) and the like, and accelerates the synthesis of ester substances (figure 3). Compared with the examples, the environmental temperature rise and reduction change in comparative example 1 and the external addition of gamma-aminobutyric acid in comparative example 2 have no significant influence on the fermentation performance of the fermentation strain (table 1), no significant increase in the content of the ester substances is promoted, the aroma quality of the wine is improved (table 2), and no significant influence on the sensory evaluation value of the fermented wine (fig. 2) is also achieved. As shown in FIG. 4, although various stress reactions were generated by fermentation strains by temperature increase and decrease or reverse increase of high concentration of gamma-aminobutyric acid, they did not significantly affect the NAD +/NADH ratio when they were reacted alone with temperature change in the environment or gamma-aminobutyric acid pressure, so that the comparative example did not significantly affect glycolysis and TCA cycle during fermentation as compared with the examples, which is shown in that they did not significantly affect fermentation performance and expression amounts of ATF1, ATF2, EHT1, EEB1, FAT1 and FAA1 genes, and further did not promote increase of ester content as compared with the examples, and thus flavor was not significantly changed.
The foregoing has shown and described the basic principles and main features of the present invention and the advantages of the present invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, and that the above embodiments and descriptions are merely illustrative of the principles of the present invention, and various changes and modifications may be made without departing from the spirit and scope of the invention, which is defined in the appended claims. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (9)
1. A brewing method of fermented water chestnut wine is characterized by comprising the following steps:
step one, removing shells of water chestnuts, crushing and sterilizing;
Step two, gelatinization: weighing the sterilized water caltrop in the first step, mixing with sterilized distilled water, heating at the constant temperature of 90-95 ℃ for 30-40 min, continuously stirring, gelatinizing and cooling;
Step three, liquefying: adding alpha-amylase into the product obtained in the step two, liquefying at the constant temperature of 92-96 ℃ for 1.5-2 h, inactivating enzyme, and cooling;
Step four, saccharification: adding saccharifying enzyme into the product obtained in the step three, saccharifying at the constant temperature of 60-65 ℃ for 1.5-2 h, inactivating enzyme, and cooling;
Step five, alcohol fermentation: adding gamma-aminobutyric acid and saccharomyces cerevisiae into the obtained product obtained in the step four after regulating the sugar degree, and fermenting at the temperature of 28-30 ℃ for 7-10 d; wherein, the mixture in fermentation is subjected to cooling treatment and heating treatment at a time every 11-13 h, and the cooling treatment and the heating treatment are circulated in the fermentation process until the fermentation is completed; the quality of the gamma-aminobutyric acid is 40-45 times of the content of the gamma-aminobutyric acid in the water caltrop weighed in the second step; the temperature-reducing and temperature-raising treatment is to firstly reduce the temperature of the mixture in the fermentation to 0 ℃ on the basis of 28-30 ℃ and then immediately raise the temperature back to 28-30 ℃.
2. The brewing method of the fermented water chestnut wine according to claim 1, which is characterized by comprising the following steps: and fifthly, the temperature-reducing and temperature-raising treatment is to firstly reduce the temperature of the mixture in fermentation to 0 ℃ at a speed of 3-5 ℃/min on the basis of 28-30 ℃, and then immediately raise the temperature back to 28-30 ℃ at a speed of 5-8 ℃/min.
3. The brewing method of the fermented water chestnut wine according to claim 1, which is characterized by comprising the following steps: in the fifth step, the Saccharomyces cerevisiae is Saccharomyces cerevisiae AWRI350,350.
4. A method for brewing fermented water chestnut wine according to claim 3, wherein: the mass of the saccharomyces cerevisiae AWRI is 0.3-0.5% of the mass of the water chestnut weighed in the second step.
5. The brewing method of the fermented water chestnut wine according to claim 1, which is characterized by comprising the following steps: in the second step, the ratio of the weighed water caltrop to the sterilized distilled water is 1 g:8-10 mL.
6. The brewing method of the fermented water chestnut wine according to claim 1, which is characterized by comprising the following steps: and step three and/or step four, the enzyme deactivation treatment is to deactivate the enzyme at the constant temperature of 90-100 ℃ for 15-20 min.
7. The brewing method of the fermented water chestnut wine according to claim 1, which is characterized by comprising the following steps: in the third step, the addition amount of the alpha-amylase is 20-30U/g.
8. The brewing method of the fermented water chestnut wine according to claim 1, which is characterized by comprising the following steps: in the fourth step, the adding amount of the saccharifying enzyme is 100-140U/g.
9. The brewing method of the fermented water chestnut wine according to claim 1, which is characterized by comprising the following steps: in the fifth step, the sugar degree is 15-20 degrees Bx.
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