CN114214152B - Method for preparing roxburgh rose black glutinous rice wine through semi-solid fermentation - Google Patents
Method for preparing roxburgh rose black glutinous rice wine through semi-solid fermentation Download PDFInfo
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
- C12G3/024—Preparation of other alcoholic beverages by fermentation of fruits other than botanical genus Vitis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
- C12G3/021—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn
- C12G3/022—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn of botanical genus Oryza, e.g. rice
Abstract
The invention discloses a method for preparing black glutinous rice with roxburgh rose by a semi-solid fermentation method. The invention comprises the following steps: screening fructus Rosae Normalis juice prepared from fructus Rosae Normalis; flushing the black glutinous rice with water, soaking, filtering to remove water, steaming, taking out the black glutinous rice, cooling with cool white rice to obtain steamed and cooled black glutinous rice; adding saccharifying enzyme into the steamed and cooled black glutinous rice, and standing to obtain saccharified black glutinous rice; adding water into fructus Rosae Normalis juice, adding activated yeast, adding into saccharified black glutinous rice, stirring, fermenting, adding mixed lactobacillus, stirring, fermenting, filtering to remove residues, and aging to obtain final product. The roxburgh rose black glutinous rice wine prepared by specific raw material proportions and technological parameters has the advantages of excellent sensory quality, rich nutrition, high vitamin C content, high SOD activity, strong antioxidation capability and the like.
Description
Technical Field
The invention belongs to the technical processing field of alcoholic beverage production, in particular to a method for preparing roxburgh rose black glutinous rice wine by a semi-solid fermentation method.
Background
The Rosa roxburghii, also called silk reeling flower, chinese first fruit, and Chun Gui, is a perennial fallen leaf small shrub of Rosa genus of Rosaceae family, widely distributed in warm temperate zone and subtropical zone, and mainly grown in southwest area in China, wherein the quality of wild Rosa roxburghii grown in Guizhou is the best. The roxburgh rose not only has ornamental value, medicinal value and economic value, but also has high nutritional value, and is rich in functional active ingredients such as vitamin C, superoxide dismutase (Superoxide dismutase, SOD), polysaccharide, flavonoid compounds, polyphenol, triterpene, organic acid, trace elements and the like. The existing researches show that the roxburgh rose preparation has the functions of regulating immunity, protecting against radiation, resisting mutation and canceration, resisting atherosclerosis, resisting oxidation injury, relieving harmful metals such as lead, and enhancing the immunity of organisms.
Black glutinous rice is also called purple glutinous rice, red glutinous rice, medicated rice, etc., and has anticancer, antiallergic, anti-obesity, and arteriosclerosis preventing effects. The black glutinous rice is rich in crude protein, lysine, vegetable fat, cellulose, various minerals and rich vitamins necessary for human body, and the protein, amino acid and fat of the black glutinous rice are higher than those of common white rice, and also contain a plurality of trace elements, so that the black glutinous rice is an ideal healthy nourishing rice. Anthocyanin is a special active ingredient in black glutinous rice, has multiple bioactive functions, and can resist cancer, inflammation, allergy, diabetes, obesity, etc. Because the black glutinous rice is rich in bioactive substances such as anthocyanin and the like and has low amylose content, the black glutinous rice is combined with brewing wine, the wine brewed by the black glutinous rice has high wine yield, mellow wine body and unique appearance, has various physiological activity functions and health care effects, can enrich the variety and flavor of black glutinous rice products, can increase the added value of the products, improves the product quality to meet the demands of consumers, and is further suitable for the development requirements of the brewing industry.
The fruit wine is prepared from various fruits by mechanical crushing, squeezing to obtain juice, fermenting or soaking, and the like, and is prepared by carefully blending and brewing, wherein the alcoholicity is generally 7-18% vol, and the fruit wine belongs to low-alcoholicity alcoholic beverage wine. The fruit wine is rich in certain polyphenol chemical substances, and the polyphenol chemical substances can remove free radicals, so that the fruit wine has good antioxidant and anti-aging effects when being drunk properly, and further can play a role in maintaining beauty and keeping young. The research shows that compared with fruits, vitamins in the fruit wine are easier to be absorbed by human bodies, and the fruit wine can resist aging, prevent arteriosclerosis, hypertension, cardiovascular and cerebrovascular diseases and the like and can regulate emotion after being drunk for a long time with proper amount.
The roxburgh rose fruit is sour and astringent in taste, so that most of roxburgh rose products such as roxburgh rose juice, roxburgh rose preserved fruit, roxburgh rose wine and the like are presented in the market, but the astringency of the roxburgh rose wine is still heavy.
Disclosure of Invention
The invention aims to provide a method for preparing roxburgh rose black glutinous rice wine by semi-solid fermentation. The roxburgh rose black glutinous rice wine prepared by specific raw material proportions and technological parameters has the advantages of excellent sensory quality, rich nutrition, good efficacy, high vitamin C content, high SOD activity, strong antioxidant capacity and the like.
The invention adopts the following technical scheme to realize the aim of the invention:
a method for preparing fructus Rosae Normalis black glutinous rice wine by semi-solid fermentation comprises the following steps:
(1) Preparing fructus Rosae Normalis juice: screening fructus Rosae Normalis, removing calyx and pedicel, washing, air drying, slicing, removing seeds, crushing, filtering, and sterilizing to obtain fructus Rosae Normalis juice;
(2) Treating black glutinous rice: flushing the black glutinous rice with water, soaking, filtering to remove water, steaming, taking out the black glutinous rice, cooling with cool white rice to obtain steamed and cooled black glutinous rice;
(3) Saccharification: adding saccharifying enzyme into the steamed and cooled black glutinous rice, and standing to obtain saccharified black glutinous rice;
(4) Fermentation: adding water into fructus Rosae Normalis juice, adding activated yeast, adding into saccharified black glutinous rice, stirring, fermenting, adding mixed lactobacillus, stirring, fermenting, filtering to remove residues, and aging to obtain final product.
The preparation method of the roxburgh rose black glutinous rice wine comprises the following steps of: screening fresh, mature, full, no olive, no plant diseases and insect pests, no damage, no rot and no rot spot, removing calyx and pedicel, washing, airing, slicing, removing seeds, putting into a beater for breaking, filtering, and performing UHT sterilization to obtain the original juice of the roxburgh rose.
The method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method comprises the following steps of: selecting black glutinous rice which is free from mildew, worm damage and plump, washing with water for 1-2 times, soaking for 20-24h, draining water, steaming for 20-30min, taking out the black glutinous rice, cooling to 20-25 ℃ with cool white rice, and obtaining steamed and cooled black glutinous rice.
The method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method comprises the following steps of: 9-11.
The method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method comprises the following steps of: 1.
according to the method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method, the adding amount of the saccharifying enzyme is 0.3-0.5% of the mass of the black glutinous rice, and the saccharifying condition is saccharification at 28-32 ℃ for 40-56h. The preferred source of saccharifying enzyme is 10 ten thousand U/mL activity from leaf Biotechnology Inc.
According to the method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method, the adding amount of the saccharifying enzyme is 0.3-0.5% of the mass of the black glutinous rice, and the saccharifying condition is that the saccharifying is carried out at 30 ℃ for 48 hours.
The method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method is characterized in that the yeast is special for grape wine and fruit wine, the activation temperature is 30-35 ℃, and the activation time is 20-30min.
The method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method comprises the steps that the yeast is special for grape wine and fruit wine, the activation temperature is 32 ℃, and the activation time is 25 minutes.
The method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method comprises the steps of fermenting at 26-34 ℃ for 48-72h.
The method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method is characterized in that the primary fermentation temperature is 30 ℃ and the fermentation time is 48-72h.
In the method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method, the mixed lactobacillus contains lactobacillus rhamnosus H3: lactobacillus paracasei SR101: lactobacillus fermentum GZSC-1 is 2-3:2-3:2-3.
The method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method comprises the step of adding water into the roxburgh rose raw juice, wherein the water accounting for 90-110% of the roxburgh rose raw juice is added.
The method for preparing the roxburgh rose black glutinous rice wine by the semi-solid fermentation method comprises the step of adding water into the roxburgh rose raw juice in an equal amount.
The method for preparing fructus Rosae Normalis black glutinous rice wine by semi-solid fermentation comprises adding mixed lactobacillus, stirring, activating with MRS broth culture medium at 37deg.C for 2-3d, counting, and counting thallus to 1×10 8 CFU/mL to obtain mixed lactobacillus activating solution; adding water into fructus Rosae Normalis juice, adding mixed lactobacillus activating solution 1-2% of total amount of fructus Rosae Normalis juice, centrifuging to collect thallus, and adding 3Washing out thallus with 0.85% physiological saline solution of 0-100% lactobacillus activation solution, adding, and stirring.
The semi-solid fermentation method for preparing the roxburgh rose black glutinous rice wine comprises the following steps of (4) fermenting at 26-34 ℃ for 10-20d.
The semi-solid fermentation method for preparing the roxburgh rose black glutinous rice wine is characterized in that the secondary fermentation temperature in the step (4) is 30 ℃ and the fermentation time is 10-20d.
The semi-solid fermentation method for preparing the roxburgh rose black glutinous rice wine is characterized in that the secondary fermentation temperature in the step (4) is 30 ℃ and the fermentation time is 10-20d.
The semi-solid fermentation method for preparing the roxburgh rose black glutinous rice wine is characterized in that the secondary fermentation temperature in the step (4) is 30 ℃ and the fermentation time is 10-20d.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention combines the roxburgh rose and the black glutinous rice, takes the roxburgh rose as a main raw material and the black glutinous rice as an auxiliary material, adds saccharifying enzyme for saccharification after the black glutinous rice is cured, can provide a carbon source, does not need to add extra carbon source substances, and can provide a semi-solid fermentation mode.
2. The saccharomycetes and the mixed lactobacillus are added, and a secondary fermentation process is adopted, so that the obtained roxburgh rose black glutinous rice wine product is dark red, transparent and uniform in color; has the fragrance of fructus Rosae Normalis and black glutinous rice, and is coordinated; the taste is fine, mellow and soft, and the sweet taste is achieved; the product in the tissue state has uniform texture and crystal clear; the style of the wine is complete, the taste of the wine is coordinated, and the flavor is rich.
3. The roxburgh rose black glutinous rice wine prepared by specific raw material proportions and technological parameters has the advantages of excellent sensory quality, rich nutrition, good efficacy, high vitamin C content, high SOD activity, strong antioxidant capacity and the like.
4. The black glutinous rice is added to obviously weaken the astringency of the roxburgh rose wine, and is a good brewing raw material, so that the roxburgh rose and the black glutinous rice are used as raw materials for fermentation, the combined synergistic effect of antioxidant substances such as roxburgh rose vitamin C, SOD, black glutinous rice anthocyanin and the like can be realized, and a finished product with dark red and bright color, soft taste and rich flavor is obtained, and the effects of resisting aging and maintaining beauty are achieved; and various lactic acid bacteria are added in the secondary fermentation to participate in the fermentation, so that the fermentation time can be effectively shortened, the flavor of the product is enhanced, and the taste is softer. The invention not only enriches the types of fruit wine products in China, but also opens up a new field of application of the roxburgh rose and the black glutinous rice, and has higher economic value and application prospect.
5. In the invention, the black glutinous rice is cooked to gelatinize starch in the black glutinous rice, so that the starch is easier to be converted into sugar substances. The effect of cooling after cooking is to increase the cooling rate and allow the cooked black glutinous rice to absorb part of the water.
6. In the invention, water which is 1 time of the amount of the original juice of the roxburgh rose is added, the dosage ratio of the original juice of the roxburgh rose to the black glutinous rice is 1:1, and through experiments, the taste of the roxburgh rose wine is best only under the proportion, if the roxburgh rose is more, the wine taste is astringent; if the black glutinous rice has more fructus Rosae Normalis flavor, the flavor is not good.
7. The saccharification effect of the invention is to change starch in the black glutinous rice into micromolecular sugar for microbial fermentation, the added saccharifying enzyme amount is 0.3-0.5% of the mass of the black glutinous rice, and the saccharification condition is that saccharification is carried out at 28-32 ℃ for 40-56h, so that the complete saccharification can be achieved, and the invention is favorable for the subsequent fermentation and the technical effect of flavor formation. If the saccharification condition is exceeded, the problem of incomplete saccharification or acid change of the saccharification liquid is caused, and the subsequent fermentation is not facilitated.
8. In the invention, the yeast is dry yeast, and the activation is to restore the activity of the yeast in a dormant state, so that the yeast is the basis and the guarantee for realizing the alcoholic fermentation. The activation temperature according to the activation conditions defined in the present invention is 30-35℃and the activation time is 20-30min, and the advantage of performing the activation is that the recovery of the yeast activity is relatively complete, and too low an activation temperature results in slow or incomplete activation, and too high an activation temperature results in death during the activation of the yeast. If the activation time is too short, the activation is incomplete, and if the activation time is too long, the carbon source in the activation environment is reduced, and the number of yeasts is reduced, so that the activity of the yeasts in alcohol fermentation is affected; the yeast content is 0.1-0.14% of fructus Rosae Normalis juice. The temperature of the primary fermentation is 26-34 ℃ (optimal 30 ℃), the fermentation time is 48-72 hours, the fermentation effect can be very good, the mixed lactobacillus is just fermented by the fermented product, and the taste of the product is ensured to be very good.
9. Compared with the prior lactobacillus, the lactobacillus fermentum GZSC-1 adopted by the invention has good activity and strong adaptability, and the lactobacillus provided by the invention has good antioxidation characteristic, has the effect of reducing blood sugar and has the effect of regulating blood sugar level, so that the roxburgh rose black glutinous rice wine adopting the lactobacillus has corresponding efficacy.
10. The invention activates lactobacillus and then inoculates and ferments, the purpose is to recover the activity of lactobacillus, which is beneficial to the formation of fermentation and flavor, after activation, the lactobacillus is washed out by normal saline, because the bacterial liquid is centrifugated to obtain supernatant and thalli, the supernatant is mainly culture medium component and is not suitable for eating, so the supernatant is needed to be poured out, the thalli is washed out by normal saline, the lactobacillus suspension is obtained by dilution and dissolution of normal saline, the lactobacillus suspension can be directly used as inoculation and fermentation, after a great deal of experiments, the lactobacillus activation condition provided by the invention is that the mixed lactobacillus is activated for 2-3 days after being subjected to 37 ℃ in MRS broth culture medium, the viable bacteria count is carried out, and the number of the thalli reaches 1 multiplied by 10 8 CFU/mL to obtain mixed lactobacillus activating solution; adding water into fructus Rosae Normalis juice, adding mixed lactobacillus activating solution 1-2% of total amount of fructus Rosae Normalis juice, centrifuging to collect thallus, washing out thallus with 0.85% physiological saline of 30-100% lactobacillus activating solution, and adding to obtain the final product with optimal taste.
11. Lactobacillus rhamnosus H3, lactobacillus paracasei SR101 and lactobacillus fermentum GZSC-1 adopted in the invention are all bacteria obtained by the prior experiments and screening of the inventor, and are all preserved. The mixed bacteria of lactobacillus rhamnosus H3, lactobacillus paracasei SR101 and lactobacillus fermentum GZSC-1 are adopted for fermentation, the fermentation effect is good, and the total acid content, the lactic acid content, the total polyphenol content, the total flavone content and the SOD activity are all better than those of products obtained by fermenting the lactobacillus rhamnosus H3, the lactobacillus paracasei or the lactobacillus fermentum GZSC-1 singly, so that the synergistic effect can be achieved when the bacteria are matched for use. The 3 bacteria are applied to the fermentation of the roxburgh rose, have the effects of quick acid production, good flavor and retention or synergism of functional components, and can quickly assist the saccharomycetes to start the alcoholic fermentation, shorten the fermentation time and ensure that the nutrient substances are retained to the maximum, and the 3 bacteria contain lactobacillus rhamnosus H3 according to the proportion of bacteria: lactobacillus paracasei SR101: lactobacillus fermentum GZSC-1 is 2-3:2-3:2-3 (optimal 1:1:1) and the finished product obtained by fermentation has optimal taste and the nutrition and functional substances are retained to the maximum extent. If the 3 bacteria are not fermented according to the proportion defined by the invention, the problems of over-sour taste, single flavor and poor efficacy can occur, and the quality of the finished product is affected.
Drawings
FIG. 1 is a graph showing the comparison of Vc content change of added lactobacillus (example 1) and non-added lactobacillus (example 2) in the fermentation process of roxburgh rose black glutinous rice wine;
FIG. 2 is a graph showing the comparison of SOD activity change of added lactobacillus (example 1) and non-added lactobacillus (example 2) in the fermentation process of Rosa roxburghii black glutinous rice wine;
FIG. 3 is a graph showing the comparison of the reducing sugar content of added lactobacillus (example 1) and non-added lactobacillus (example 2) in the fermentation process of Rosa roxburghii black glutinous rice wine;
FIG. 4 is a graph showing the comparison of the change in alcoholicity of the added lactic acid bacteria (example 1) and the un-added lactic acid bacteria (example 2) in the fermentation process of the Rosa roxburghii black glutinous rice wine;
FIG. 5 is a process flow diagram of a Rosa roxburghii black glutinous rice wine;
FIG. 6 is a sample graph of Rosa roxburghii Heteroglutinous rice wine (left is Rosa roxburghii Heteroglutinous rice wine before centrifugation, right is Rosa roxburghii Heteroglutinous rice wine after centrifugation);
FIG. 7 is a graph showing a comparison of Rosa roxburghii black glutinous rice wine (left is Rosa roxburghii black glutinous rice wine fermented without adding lactobacillus, right is Rosa roxburghii black glutinous rice wine fermented with adding lactobacillus according to the present invention);
FIG. 8 is a graph showing the dynamic change of lactic acid content of the vinegar obtained by fermenting roxburgh rose with different lactobacillus strains;
FIG. 9 is a graph showing the dynamic change of total acid content of the fermented Rosa roxburghii fruit vinegar by different lactobacillus strains;
FIG. 10 is a graph showing the dynamic change of total polyphenol content of the fermented Rosa roxburghii fruit vinegar by different lactobacillus strains;
FIG. 11 is a graph showing the dynamic change of total flavone content of the fermented Rosa roxburghii fruit vinegar by different lactobacillus strains;
FIG. 12 is a graph showing dynamic changes in SOD activity of fermented Rosa roxburghii fruit vinegar by different lactobacillus strains;
Detailed Description
The present invention will be described in further detail with reference to specific examples. The following experimental methods are all conventional methods in the art unless otherwise specified, and the ingredients or materials used, if not otherwise specified, are all commercially available. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention.
Example 1. A method for preparing fructus Rosae Normalis black glutinous rice wine by semi-solid fermentation method.
(1) Preparing roxburgh rose juice: screening fresh, mature, full, no olive, no diseases and insect pests, no damage, no rot and no rot, removing calyx and pedicel after selection, washing, airing, slicing, removing seeds, putting into a beater for breaking, filtering to obtain roxburgh rose juice, and weighing 50g of roxburgh rose juice for later use;
(2) Treatment of black glutinous rice: selecting black glutinous rice which is free of mildew, worm damage and plump in grains, weighing 50g, washing with water for 1 time, and soaking for 24 hours; filtering to remove water, steaming in a pot for 30min, taking out, and cooling to 20-25deg.C;
(3) Saccharification: adding 0.1g of saccharifying enzyme which is the mass of the black glutinous rice into the steamed and cooled black glutinous rice, and uniformly stirring;
(4) The first stage of fermentation, namely adding 50mL of water into the roxburgh rose juice, adding 0.06g of wine and yeast special for fruit wine, activating for 2 hours, pouring into black glutinous rice added with saccharifying enzyme, and fermenting for 48 hours at 30 ℃;
(5) And the second stage of fermentation, namely adding 0.1mL of lactobacillus fermentum GZSC-1 lactobacillus after the first stage of fermentation is finished, fermenting for 10 days at 30 ℃, filtering to remove residues, and ageing at room temperature.
Example 2. Preparation of black glutinous rice wine of Rosa roxburghii in comparative example 1 without lactobacillus fermentation.
(1) Preparing roxburgh rose juice: screening fresh, mature, full, no olive, no diseases and insect pests, no damage, no rot and no rot, removing calyx and pedicel after selection, washing, airing, slicing, removing seeds, putting into a beater for breaking, filtering to obtain roxburgh rose juice, and weighing 50g of roxburgh rose juice for later use;
(2) Treatment of black glutinous rice: selecting black glutinous rice which is free of mildew, worm damage and plump in grains, weighing 50g, washing with water for 1 time, and soaking for 24 hours; filtering to remove water, steaming in a pot for 30min, taking out, and cooling to 20-25deg.C;
(3) Saccharification: adding 0.1g of saccharifying enzyme which is the mass of the black glutinous rice into the steamed and cooled black glutinous rice, and uniformly stirring for later use;
(4) Adding 50mL water into fructus Rosae Normalis juice, adding 0.06g of wine and yeast special for fruit wine, activating for 2 hr, adding into black glutinous rice with saccharifying enzyme, fermenting at 30deg.C for 12d, filtering to remove residue, and aging.
The products of example 1 and example 2 were measured to obtain physicochemical indexes of table 1.
Table 1 physicochemical index of black glutinous rice wine of roxburgh rose in example 1 and example 2
Example 3 the semi-solid fermentation process of preparing black glutinous rice wine with Rosa roxburghii comprises the following steps:
(1) Preparing fructus Rosae Normalis juice: screening fresh, mature, full, no olive, no plant diseases and insect pests, no damage, no rot and no rot spot, removing calyx and pedicel, washing, airing, slicing, removing seeds, putting into a beater for breaking, filtering, and performing UHT sterilization to obtain the original juice of the roxburgh rose;
(2) Treating black glutinous rice: selecting black glutinous rice which is free of mildew, worm damage and plump in grain, washing for 1 time with water, soaking for 24 hours, draining water, steaming for 30 minutes in a water-proof mode, taking out the black glutinous rice, cooling to 25 ℃ with cool white boiled rice, and obtaining steamed and cooled black glutinous rice; the dosage ratio of the original juice of the rosa roxburghii and the black glutinous rice is 9:11;
(3) Saccharification: adding saccharifying enzyme accounting for 0.3% of the mass of the black glutinous rice into the steamed and cooled black glutinous rice, stirring, standing and saccharifying for 56 hours at 32 ℃ to obtain saccharified black glutinous rice;
(4) Fermentation: adding water into fructus Rosae Normalis juice, adding fructus Rosae Normalis juice yeast at 35deg.C and fruit wine yeast for 30min, adding yeast 0.1% of fructus Rosae Normalis juice, adding into saccharified black glutinous rice, fermenting at 37deg.C for 72 hr, activating mixed lactobacillus with MRS broth culture medium at 37deg.C for 3d, and counting to obtain thallus with number of 1×10 8 CFU/mL to obtain mixed lactobacillus activating solution; adding mixed lactobacillus activating solution with the total amount of 2% of the total amount of the roxburgh rose juice into the roxburgh rose juice, centrifuging to collect thalli, washing out thalli with 0.85% physiological saline with the amount of 100% lactobacillus activating solution, and adding the thalli, wherein the bacterium containing proportion of the mixed lactobacillus is lactobacillus rhamnosus H3: lactobacillus paracasei SR101: lactobacillus fermentum GZSC-1 was 2:2:3, stirring uniformly, carrying out secondary fermentation for 20d at 34 ℃, filtering to remove residues, and ageing at room temperature in dark place to obtain a finished product.
Example 4, the semi-solid fermentation method for preparing fructus Rosae Normalis black glutinous rice wine comprises the following steps:
(1) Preparing fructus Rosae Normalis juice: screening fresh, mature, full, no olive, no plant diseases and insect pests, no damage, no rot and no rot spot, removing calyx and pedicel, washing, airing, slicing, removing seeds, putting into a beater for breaking, filtering, and performing UHT sterilization to obtain the original juice of the roxburgh rose;
(2) Treating black glutinous rice: selecting black glutinous rice which is free of mildew, worm damage and plump in grains, washing for 2 times with water, soaking for 20 hours, draining water, steaming for 20 minutes in a water-proof mode, taking out the black glutinous rice, cooling to 20 ℃ with cool white boiled rice, and obtaining steamed and cooled black glutinous rice; the dosage ratio of the original juice of the roxburgh rose to the black glutinous rice is 11:9, a step of performing the process;
(3) Saccharification: adding saccharifying enzyme accounting for 0.5% of the mass of the black glutinous rice into the steamed and cooled black glutinous rice, stirring, standing and saccharifying for 40 hours at 28 ℃ to obtain saccharified black glutinous rice;
(4) Fermentation: activating yeast special for grape wine and fruit wine at 30deg.C for 20min, adding water into fructus Rosae Normalis juice, adding yeast special for grape wine and fruit wine, adding yeast 0.14% of fructus Rosae Normalis juice, and adding sugarFermenting black glutinous rice at 26deg.C for 48 hr, activating mixed lactobacillus with MRS broth culture medium at 37deg.C for 2d, and counting to obtain 1×10 thallus 8 CFU/mL to obtain mixed lactobacillus activating solution; adding mixed lactobacillus activating solution with the total amount of 1% of the total amount of the roxburgh rose juice into the roxburgh rose juice, centrifuging to collect thalli, washing out thalli with 0.85% physiological saline with the activating solution amount of 30% of lactobacillus, and adding the thalli, wherein the bacterium containing proportion of the mixed lactobacillus is lactobacillus rhamnosus H3: lactobacillus paracasei SR101: lactobacillus fermentum GZSC was 2:3:2, stirring uniformly, carrying out secondary fermentation at 26 ℃ for 10d, filtering to remove residues, and ageing at room temperature in dark place to obtain a finished product.
Example 5, the semi-solid fermentation method for preparing fructus Rosae Normalis black glutinous rice wine comprises the following steps:
(1) Preparing fructus Rosae Normalis juice: screening fresh, mature, full, no olive, no plant diseases and insect pests, no damage, no rot and no rot spot, removing calyx and pedicel, washing, airing, slicing, removing seeds, putting into a beater for breaking, filtering, and performing UHT sterilization to obtain the original juice of the roxburgh rose;
(2) Treating black glutinous rice: selecting black glutinous rice which is free of mildew, worm damage and plump in grains, washing for 2 times with water, soaking for 22 hours, draining water, steaming for 25 minutes in a water-proof mode, taking out the black glutinous rice, cooling to 22 ℃ with cool white boiled rice, and obtaining steamed and cooled black glutinous rice; the dosage ratio of the roxburgh rose juice to the black glutinous rice is 1:1, a step of;
(3) Saccharification: adding saccharifying enzyme accounting for 0.4% of the mass of the black glutinous rice into the steamed and cooled black glutinous rice, stirring, standing and saccharifying for 48 hours at 30 ℃ to obtain saccharified black glutinous rice;
(4) Fermentation: adding water into fructus Rosae Normalis juice, adding fructus Rosae Normalis juice yeast at 32deg.C and fruit wine yeast for 25min, adding yeast 0.12% of fructus Rosae Normalis juice, adding into saccharified black glutinous rice, fermenting at 30deg.C for 60 hr, activating mixed lactobacillus with MRS broth culture medium at 37deg.C for 3d, and counting to obtain thallus with number of 1×10 8 CFU/mL to obtain mixed lactobacillus activating solution; adding water into fructus Rosae Normalis juice, adding lactobacillus-activated solution 1.5% of total amount of fructus Rosae Normalis juice, centrifuging, and collectingCollecting thallus, washing out thallus with 0.85% physiological saline with 50% lactobacillus activation liquid amount, and adding, wherein the mixed lactobacillus contains lactobacillus rhamnosus H3: lactobacillus paracasei SR101: lactobacillus fermentum GZSC-1 was 1:1:1, stirring uniformly, carrying out secondary fermentation at 30 ℃ for 15 days, filtering to remove residues, and ageing at room temperature in dark place to obtain a finished product.
Experimental example one. Effect test of Lactobacillus fermentum GZSC-1
The lactobacillus fermentum GZSC-1 mentioned in the invention is a newly preserved bacterium, and the preservation complete information is as follows:
deposit unit code: CCTCC (cctccc)
Address: university of Chinese, wuhan and Wuhan
Whether or not to survive: is that
Preservation date: 2017 12, 28
Preservation number: CCTCC NO: M2017847
Classification naming: lactobacillus fermentum GZSC-1
Lactobacillus fermentum GZSC-1
1. The preservation and culture of the antioxidant lactobacillus fermentum
The strain of the antioxidant lactobacillus fermentum is isolated from fermented pickled Chinese cabbage self-made by villages and towns in Qing town stations in Guiyang city of Guizhou province.
(1) And (3) strain preservation: inoculating solid MRS culture medium test tube inclined plane, checking bacterial count to be 1×10 7 After CFU/ml, glycerol is added into the upper part of the test tube, the test tube is sealed, and the test tube is preserved for 3 to 6 months at the temperature of minus 20 ℃. The strain can be prepared into freeze-dried powder and preserved for 1-5 years at the temperature of 1-8 ℃.
(2) Activating strains: inoculating a loop of lactobacillus into MRS broth culture medium under aseptic condition, and culturing at 37 ℃ for 24-48 h.
2. Comparison of antioxidant Capacity of extracellular secretions of bacterial suspensions of different species of lactic acid bacteria
(1) Inoculating lactobacillus paracasei SR101 lactobacillus to MRS broth culture medium under aseptic condition, culturing in an aseptic incubator at 37deg.C for 48 hr, centrifuging to collect thallus and supernatant, wherein the supernatant is SR101 extracellular secretion; adjusting the concentration of bacterial liquid to 10 with 0.85% physiological saline 9 CFU/mL is the SR101 bacterial suspension.
(2) Inoculating a loop of Lactobacillus casei H1 lactobacillus into MRS broth culture medium under aseptic condition, culturing in an aseptic incubator at 37deg.C for 24 hr, centrifuging to collect thallus and supernatant, wherein the supernatant is H1 extracellular secretion; adjusting the concentration of bacterial liquid to 10 with 0.85% physiological saline 9 CFU/mL is H1 bacterial suspension.
(3) Inoculating a loop of GZSC-1 lactobacillus into MRS broth culture medium under aseptic condition, culturing in an aseptic incubator at 37deg.C for 60 hr, centrifuging to collect thallus and supernatant, wherein the supernatant is GZSC-1 extracellular secretion; adjusting the concentration of bacterial liquid to 10 with 0.85% physiological saline 9 CFU/mL is GZSC-1 bacterial suspension.
The SR101 extracellular secretion, SR101 bacterial suspension, H1 extracellular secretion, H1 bacterial suspension, GZSC-1 extracellular secretion and GZSC-1 bacterial suspension were analyzed to obtain the data of tables 1 to 3.
TABLE 1 determination of DPPH radical scavenging by different lactic acid bacteria
| Sample name | DPPH radical scavenging (%) | Vc equivalent (mg mL) -1 ) |
| SR101 bacterial suspension | 67.00 | 0.057 |
| SR101 extracellular secretion | 91.14 | 0.097 |
| H1 bacterial suspension | 87.00 | 0.090 |
| H1 extracellular secretion | 92.27 | 0.098 |
| GZSC-1 bacterial suspension | 63.20 | 0.052 |
| gZSC-1 extracellular secretion | 95.45 | 0.104 |
TABLE 2 determination of ABTS radical scavenging by different lactic acid bacteria
| Sample name | ABTS radical clearance (%) | Vc equivalent (mg mL) -1 ) |
| SR101 bacterial suspension | 34.00 | 0.064 |
| SR101 extracellular secretion | 68.65 | 0.118 |
| H1 bacterial suspension | 35.80 | 0.066 |
| H1 extracellular secretion | 39.23 | 0.072 |
| GZSC-1 bacterial suspension | 11.66 | 0.029 |
| gZSC-1 extracellular secretion | 50.00 | 0.089 |
TABLE 3 determination of the free radical scavengers of hydroxyl groups by different lactic acid bacteria
| Sample name | Radical scavenging rate of hydroxyl (%) | Vc equivalent (mg mL) -1 ) |
| SR101 bacterial suspension | 98.96 | 0.545 |
| SR101 extracellular secretion | 98.29 | 0.499 |
| H1 bacterial suspension | 98.00 | 0.497 |
| H1 extracellular secretion | 67.74 | 0.308 |
| GZSC-1 bacterial suspension | 80.27 | 0.386 |
| gZSC-1 extracellular secretion | 70.21 | 0.323 |
The antioxidant capacity of the 3 strains of lactobacillus suspension and extracellular secretion is expressed by Vc equivalent, and the higher the Vc equivalent value is, the stronger the antioxidant capacity is. As can be seen from Table 1, the clearance of the extracellular secretion of Lactobacillus fermentum GZSC-1 to DPPH free radical is up to 95.45%, which is significantly higher than that of Lactobacillus casei H1 and Lactobacillus paracasei SR101; in addition, the extracellular secretion of the lactobacillus fermentum GZSC-1 has higher clearance rate of ABTS free radicals, which proves that the lactobacillus fermentum GZSC-1 has higher antioxidation property, in particular to the culture or fermentation product thereof.
3. In vitro hypoglycemic capability of 3 lactobacillus suspensions and extracellular secretion
The above SR101 extracellular secretion, SR101 bacterial suspension, H1 extracellular secretion, H1 bacterial suspension, GZSC-1 extracellular secretion and GZSC-1 bacterial suspension were analyzed to obtain the data of Table 4.
TABLE 4 measurement of blood glucose levels by different lactic acid bacteria
| Sample name | Alpha-glucosidase inhibition Rate (%) | Acarbose equivalent (mg/mL) -1 ) |
| SR101 bacterial suspension | 97.89±1.11 a | 2.45×10 -5 |
| SR101 extracellular secretion | 94.07±1.56 a | 2.3×10 -5 |
| H1 bacterial suspension | 68.77±1.34 c | 1.42×10 -5 |
| H1 extracellular secretion | 94.52±2.51 a | 2.37×10 -5 |
| GZSC-1 bacterial suspension | 98.27±4.55 d | 2.96×10 -5 |
| gZSC-1 extracellular secretion | 88.96±0.67 b | 2.16×10 -5 |
The inhibition capacity of the 3 strains of lactobacillus suspension and extracellular secretion for inhibiting alpha-glycosidase is expressed by acarbose equivalent, and the higher the acarbose equivalent value is, the stronger the antioxidation capacity is. As can be seen from table 4: in an in vitro experiment, the highest acarbose equivalent value of the lactobacillus fermentum GZSC-1 bacterial suspension indicates that the antioxidant capacity of the lactobacillus fermentum GZSC-1 bacterial suspension is higher. In addition, the GZSC-1 bacterial suspension also has better blood sugar reducing effect from the aspect of the alpha-glycosidase inhibition rate.
Experimental example two. Test of mixed bacteria effect
Example 1 preparation method of mixed bacteria compound fermentation roxburgh rose fruit vinegar.
(1) Preparing original juice of fructus Rosae Normalis: screening fresh, mature, full, no olive, no disease and pest damage, no rot and no rot, removing calyx and pedicel after selection, washing, airing, slicing, removing seeds, putting into a beater for breaking, filtering, performing UHT sterilization to obtain roxburgh rose juice, and filling into a fermentation tank for standby;
(2) Dilution: placing 40mL of roxburgh rose juice in a fermentation tank, and adding 160mL of water to obtain a roxburgh rose juice diluent for later use;
(3) Sugar degree adjustment: adding white granulated sugar to adjust the sugar degree of the roxburgh rose diluent to 16 DEG Brix;
(4) Alcohol fermentation, namely adding 0.04g of wine fruit wine active dry yeast into 4% glucose and roxburgh rose juice diluent, carrying out water bath for 30min at 32 ℃, oscillating once every 10min, and inoculating 8mL into the roxburgh rose diluent after activation; activating lactobacillus strain with MRS broth culture medium at 37deg.C for 2d, centrifuging 14mL of the strain to collect thallus, eluting with 0.85% physiological saline, and inoculating into diluted solution of fructus Rosae Normalis juice; fermenting in a 30 ℃ incubator for 5 days to obtain the Rosa roxburghii fruit wine for later use; the lactobacillus strain is used in the proportion of 1:1:1, lactobacillus rhamnosus H3, lactobacillus paracasei SR101 and lactobacillus fermentum GZSC-1, and the concentration of each thallus after activation reaches 1X 10 8 CFU/mL；
(5) Acetic acid fermentation: and (3) picking the acetic acid bacteria strain preserved in the glycerol into an acetic acid bacteria liquid culture medium subjected to high-temperature and high-pressure sterilization, culturing the acetic acid bacteria strain in a shaking incubator with the rotating speed of 150r/min at the temperature of 30 ℃ for 50 hours, taking 14mL of bacteria liquid, centrifuging to collect bacteria, eluting with 0.85% physiological saline, inoculating the bacteria liquid into the Rosa roxburghii fruit wine, fermenting the bacteria liquid culture medium in the shaking incubator with the rotating speed of 140r/min at the temperature of 32 ℃ for 8 days, and obtaining the mixed bacteria compound fermentation Rosa roxburghii fruit vinegar after the fermentation is finished.
Comparative example 1. A yeast fermented fructus Rosae Normalis fruit vinegar and its preparation method are provided.
The specific procedure and starting materials of this comparative example were essentially the same as in example 1, except that: only saccharomycetes and no lactic acid bacteria are added in the alcohol fermentation stage.
Comparative example 2. A yeast fermented fructus Rosae Normalis fruit vinegar and its preparation method are provided.
The specific procedure and starting materials of this comparative example were essentially the same as in example 1, except that: in the alcohol fermentation stage, except yeast is added, lactobacillus is only added with lactobacillus rhamnosus H3 (the dosage of lactobacillus is the same as that of the mixed bacteria in the embodiment 1).
Lactobacillus rhamnosus H3 has been deposited with the following information:
deposit unit code: CCTCC (cctccc)
Address: university of Chinese, wuhan and Wuhan
Whether or not to survive: is that
Preservation date: 2016, 9, 26 days
Preservation number: CCTCC NO: M2016525
Classification naming: lactobacillus rhamnosus H3
Lactobacillus rhamnosus H3
Comparative example 3. A yeast fermented fructus Rosae Normalis fruit vinegar and its preparation method are provided.
The specific procedure and starting materials of this comparative example were essentially the same as in example 1, except that: in the alcohol fermentation stage, except yeast, only lactobacillus paracasei SR101 is added into lactobacillus.
Lactobacillus paracasei SR101 has been deposited with the following information:
deposit unit code: CCTCC (cctccc)
Address: university of Chinese, wuhan and Wuhan
Whether or not to survive: is that
Preservation date: 2016, 9, 26 days
Preservation number: CCTCC NO: M2016527
Classification naming: lactobacillus paracasei SR101
Lactobacillus paracasei SR101
Comparative example 4. A yeast fermented fructus Rosae Normalis fruit vinegar and its preparation method are provided.
The specific procedure and starting materials of this comparative example were essentially the same as in example 1, except that: in the alcohol fermentation stage, except yeast, only lactobacillus fermentum GZSC-1 is added into lactobacillus.
The products obtained in example 1 and comparative example were analyzed to obtain the results in Table 1.
TABLE 1 physicochemical index of original vinegar of Rosa roxburghii in example 1 and comparative example 1
According to the dynamic changes of a plurality of physical and chemical indexes (shown in figures 8-12) in the fermentation process of the embodiment 1 and the comparative embodiment 1, the total polyphenol and the total flavone are in a descending trend in the whole fermentation period, the descending of the alcohol fermentation stage is faster, the acetic acid fermentation later period has a stabilizing trend, the total polyphenol and the total flavone content of a lactic acid bacteria sample which is not added are lower than the content of the lactic acid bacteria sample, and the total acid content, the total flavone content and the SOD activity of the original vinegar of the Rosa roxburghii after the fermentation are higher than those of the original vinegar of the Rosa roxburghii without lactic acid bacteria, and the lactic acid bacteria can quickly start the fermentation, so that the fermentation is facilitated.
As is clear from the dynamic changes of the fermentation physicochemical indexes (as shown in FIGS. 8 to 12) of comparative example 1 and comparative examples 2 to 4, the total acid content, lactic acid content, total polyphenol content, total flavone content and SOD activity of the Rosa roxburghii raw vinegar of example 1 after fermentation by the mixed bacteria of Lactobacillus rhamnosus H3, lactobacillus paracasei SR101 and Lactobacillus fermentum GZSC-1 in example 1 are all higher than those of the Rosa roxburghii raw vinegar obtained by the comparative examples of Lactobacillus rhamnosus H3 or Lactobacillus paracasei or Lactobacillus fermentum GZSC-1 alone. The method shows that the lactobacillus rhamnosus H3, the lactobacillus paracasei SR101 and the lactobacillus fermentum GZSC-1 are mixed for fermentation, so that a better fermentation effect than that of a single strain is obtained.
The foregoing is only a preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art, who is within the scope of the present invention, should make equivalent substitutions or modifications according to the technical scheme of the present invention and the inventive concept thereof, and should be covered by the scope of the present invention.
Sequence listing
<110> university of Guizhou
<120> method for preparing fructus Rosae Normalis black glutinous rice wine by semi-solid fermentation method
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1538
<212> DNA
<213> Lactobacillus fermentum (Lactobacillus fermentum)
<400> 1
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aaggtacccc caccgacttt gggtgttaca aactctcatg gtgtgacggg cggtgtgtac 120
aaggcccggg aacgtattca ccgcggcatg ctgatccgcg attactagcg attccgactt 180
cgtgcagggc gagttgcagc ctgcagtccg aactgagaac ggttttaaga gatttgcttg 240
ccctcgcgag ttcgcgactc gttgtaccgt ccattgtagc acgtgtgtag cccaggtcat 300
aaggggcatg atgatctgac gtcgtcccca ccttcctccg gtttgtcacc ggcagtctca 360
ctagagtgcc caacttaatg ctggcaacta gtaacaaggg ttgcgctcgt tgcgggactt 420
aacccaacat ctcacgacac gagctgacga cgaccatgca ccacctgtca ttgcgttccc 480
gaaggaaacg ccctatctct agggttggcg caagatgtca agacctggta aggttcttcg 540
cgtagcttcg aattaaacca catgctccac cgcttgtgcg ggcccccgtc aattcctttg 600
agtttcaacc ttgcggtcgt actccccagg cggagtgctt aatgcgttag ctccggcact 660
gaagggcgga aaccctccaa cacctagcac tcatcgttta cggcatggac taccagggta 720
tctaatcctg ttcgctaccc atgctttcga gtctcagcgt cagttgcaga ccaggtagcc 780
gccttcgcca ctggtgttct tccatatatc tacgcattcc accgctacac atggagttcc 840
actaccctct tctgcactca agttatccag tttccgatgc acttctccgg ttaagccgaa 900
ggctttcaca tcagacttag aaaaccgcct gcactctctt tacgcccaat aaatccggat 960
aacgcttgcc acctacgtat taccgcggct gctggcacgt agttagccgt gactttctgg 1020
ttaaataccg tcaacgtatg aacagttact ctcatacgtg ttcttcttta acaacagagc 1080
tttacgagcc gaaacccttc ttcactcacg cggtgttgct ccatcaggct tgcgcccatt 1140
gtggaagatt ccctactgct gcctcccgta ggagtatggg ccgtgtctca gtcccattgt 1200
ggccgatcag tctctcaact cggctatgca tcatcgcctt ggtaggccat taccccacca 1260
acaagctaat gcaccgcagg tccatccaga agtgatagcg agaagccatc ttttaagcgt 1320
tgttcatgcg aacaacgttg ttatgcggta ttagcatctg tttccaaatg ttgtcccccg 1380
cttctgggca ggttacctac gtgttactca cccgtccgcc actcgttggc gaccaaaatc 1440
aatcaggtgc aagcaccatc aatcaattgg gccaacgcgt tcgacttgca tgtattaggc 1500
acaccgccag cgttcatcct gagccaggtt caaactct 1538
Claims (3)
1. A method for preparing roxburgh rose black glutinous rice wine by a semi-solid fermentation method is characterized in that: the method comprises the following steps:
(1) Preparing fructus Rosae Normalis juice: screening fructus Rosae Normalis, removing calyx and pedicel, washing, air drying, slicing, removing seeds, crushing, filtering, and sterilizing to obtain fructus Rosae Normalis juice;
(2) Treating black glutinous rice: selecting black glutinous rice which is free from mildew, worm damage and plump in grain, washing with water for 1-2 times, soaking for 20-24h, draining water, steaming for 20-30min in a water-proof mode, taking out the black glutinous rice, cooling to 20-25 ℃ with cool and white rice to obtain steamed and cooled black glutinous rice;
(3) Saccharification: adding saccharifying enzyme into the steamed and cooled black glutinous rice, wherein the adding amount of the saccharifying enzyme is 0.3-0.5% of the mass of the black glutinous rice, and saccharifying at 28-32 ℃ for 40-56h to obtain saccharified black glutinous rice;
(4) Fermentation: adding water into fructus Rosae Normalis juice, and adding activated yeast, wherein the yeast is special for wine and fruit wine, the activation temperature is 30-35deg.C, and the activation time is 20-30min; the yeast is added in an amount of 0.1-0.14% of the original juice of fructus Rosae Normalis, and is poured into saccharified black glutinous rice, and is stirred uniformly, and is subjected to primary fermentation for 48-72 hours at 26-34 ℃, then mixed lactobacillus is added, and is stirred uniformly, wherein the proportion of lactobacillus rhamnosus H3 in the mixed lactobacillus is: lactobacillus paracasei SR101: lactobacillus fermentum GZSC-1 is 2-3:2-3:2-3, performing secondary fermentation at 26-34 ℃ for 10-20d, filtering to remove residues, and aging to obtain a finished product;
the usage ratio of the roxburgh rose juice to the black glutinous rice is 9-11:9-11.
2. The method for preparing the black glutinous rice wine with roxburgh rose by the semi-solid fermentation method according to claim 1, which is characterized in that: the water is added into the original juice of fructus Rosae Normalis, and the amount of water is 90-110% of that of original juice of fructus Rosae Normalis.
3. The method for preparing the black glutinous rice wine with roxburgh rose by the semi-solid fermentation method according to claim 1, which is characterized in that: the mixed lactobacillus is added and stirred uniformly, the mixed lactobacillus is activated for 2-3 days at 37 ℃ through MRS broth culture medium, and then viable count is carried out, and the number of the thalli reaches 1 multiplied by 10 8 CFU/mL to obtain mixed lactobacillus activating solution; adding water into fructus Rosae Normalis juice, adding mixed lactobacillus activating solution 1-2% of total amount of fructus Rosae Normalis juice, centrifuging to collect thallus, washing thallus with 0.85% physiological saline of 30-100% lactobacillus activating solution, adding, and stirring.
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