阅读说明：本技术 一种黑木耳黄酒及其制备方法 (Black fungus yellow wine and preparation method thereof ) 是由 王谦 王晨 刘敏 于 2021-06-22 设计创作，主要内容包括：本发明公开了一种黑木耳黄酒,包括以下质量百分比原材料：以黍米量计,黑木耳发酵醪液20-25％,甜酒曲3.5-4.0％和酿酒酵母4-6％。本发明通过合理的添加成分和制备方法得到黑木耳黄酒,多糖含量可达28.24g/L,多酚含量可达292.15μg/mL,黄酮含量可达89.33μg/mL；总抗氧化能力(FRAP值)可达1.025mmol/L；对DPPH·、O-2-·、·OH的清除能力依次为81.65％、80.24％、73.50％,抗氧化能力强,保证了产品营养的丰富度与功能性。(The invention discloses black fungus yellow wine which comprises the following raw materials in percentage by mass: the fermented liquid of black fungus is 20-25%, sweet distiller's yeast is 3.5-4.0% and Saccharomyces cerevisiae is 4-6% calculated by broomcorn millet. The black fungus yellow wine is obtained by reasonable addition of components and a preparation method, the polysaccharide content can reach 28.24g/L, the polyphenol content can reach 292.15 mu g/mL, and the flavone content can reach 89.33 mu g/mL; the total oxidation resistance (FRAP value) can reach 1.025 mmol/L; for DPPH and O 2 The removal capacity of < - > 81.65%, 80.24% and 73.50% of OH is high in oxidation resistance, and the richness and functionality of the product nutrition are guaranteed.)

1. The black fungus yellow wine is characterized by comprising the following raw materials: millet, black fungus fermented mash, sweet wine yeast and saccharomyces cerevisiae.

2. The black fungus yellow wine according to claim 1, which is characterized by comprising the following raw materials in percentage by mass: the fermented liquid of black fungus is 20-25%, sweet distiller's yeast is 3.5-4.0% and Saccharomyces cerevisiae is 4-6% calculated by broomcorn millet.

3. The black fungus yellow wine according to claim 2, which is characterized by comprising the following raw materials in percentage by mass: the fermented liquor of black fungus is 25%, the sweet wine yeast is 3.5% and the saccharomyces cerevisiae is 5% in terms of the amount of husked millet.

4. The black fungus yellow wine according to claim 2, further comprising 8-10% of white granulated sugar and 0.1% of citric acid.

5. The black fungus yellow wine according to any one of claims 1 to 3, wherein the preparation method of the black fungus fermented mash comprises the following steps: inoculating the activated black fungus strain to a liquid fermentation culture medium in an inoculation amount of 10%, culturing at 28 deg.C and 160r/min for 5d, and breaking the wall of the fermentation liquid to obtain black fungus fermented liquor.

6. The black fungus yellow wine according to claim 5, wherein the liquid fermentation culture medium comprises the following raw materials in percentage by mass: 2.5 percent of glucose, 0.5 percent of corn flour, 0.5 percent of soybean meal, 0.1 percent of magnesium sulfate, 1.0 percent of yeast extract, 0.2 percent of potassium dihydrogen phosphate and the balance of water.

7. The preparation method of the black fungus yellow wine is characterized by comprising the following steps:

(1) weighing raw materials of husked millet, black fungus fermented liquor, sweet wine yeast and saccharomyces cerevisiae according to mass percentage for later use;

(2) removing impurities from semen Panici Miliacei, soaking in water, steaming, sprinkling rice, and cooling;

(3) adding the black fungus fermented liquor into the cooled husked millet, performing saccharification and fermentation at 28-32 ℃ for 2-3 days, adding the saccharomyces cerevisiae, performing alcoholic fermentation at 29-33 ℃ for 10-12 days, and finally filtering to obtain a filtrate;

(4) and seasoning and sterilizing the filtrate to obtain the black fungus yellow wine.

8. The preparation method of black fungus yellow wine according to claim 7, wherein the soaking time in the step (2) is 12 hours, and the amount of water added is 2 times of the mass of the husked millet.

9. The method for preparing black fungus yellow wine according to claim 7, wherein the cooling temperature in the step (2) is 20-25 ℃.

10. The method for preparing black fungus yellow wine according to claim 7, wherein white granulated sugar and citric acid are added in the seasoning in the step (4).

Technical Field

The invention belongs to the technical field of yellow wine brewing, and particularly relates to black fungus yellow wine and a preparation method thereof.

Background

Yellow wine is a traditional fermentation product in China and has unique taste. Modern researches show that the yellow wine is rich in nutrition and has strong physiological effects. Yellow wine is rich in protein, carbohydrate, vitamins, inorganic salts, bioactive peptides, polyphenol, trace elements and the like, and is called as 'liquid cake'. Wherein, the functional oligosaccharide with higher content is beneficial to the change of the gastrointestinal micro-ecological environment. Yellow wine as brewing wine is not distilled in the making process, so various nutrient active substances produced in the fermentation process can be retained, the active ingredients are far higher than other wines such as vitamins, glutathione and the like, and the vitamin C of part of the yellow wine can reach 105.9 mg/L. The yellow wine also contains a large amount of phenolic substances, has strong free radical scavenging capacity, and in addition, active peptides, Maillard reaction products, biogenic amines, mineral substances and the like contained in the yellow wine also have certain antioxidant effect. .

Under the guidance of the 'big health' industry and national policies, people have new requirements on drinking, the healthy and nutritional drinking idea is gradually formed, and the functionality is also the future development direction of the yellow wine. In recent years, in order to meet the improvement of drinking requirements of the vast people, functional yellow wine is increasingly researched, and the health-care effect of the yellow wine can be improved by adding Chinese herbal medicines with homology of medicine and food or food materials with higher nutritional value. The current research on functional factors of yellow rice wine is mainly divided into two categories: firstly, rhizoma polygonati, poria cocos, Chinese wolfberry, cordyceps sinensis, red yeast rice, radix puerariae, radix polygoni multiflori, mulberry and other Chinese herbal medicines for both eating and taking medicine; the other is food material with high active ingredient such as Concha Ostreae, Coicis semen, Aloe, acerola cherry, Fortunella margarita (lour.) Swingle, fructus crataegi, etc.

The edible fungi are the traditional 'homology of medicine and food' crops in China, utilize the functional components of the edible fungi, are combined with the brewing technology of yellow wine, and have the unique advantage. At present, researches mostly concentrate on mushrooms, tremella and the like, only small-scale experiments are carried out, the conventional researches are not developed in a conventional mode, and industrial production cannot be realized; meanwhile, the black fungus has a very fast development speed and becomes the second largest edible fungus variety of the Chinese yield, the yield and the planting scale can ensure the smooth implementation of industrial production, but at present, no relevant report about the development of black fungus yellow wine exists.

Therefore, the black fungus yellow wine and the preparation method thereof are provided, and the research of the yellow wine by using the black fungus not only ensures the richness and functionality of the nutrition of the product, but also can promote the sustainable development of the black fungus industry and the yellow wine industry.

Disclosure of Invention

In view of the above, the black fungus yellow wine and the preparation method thereof are provided by the invention, the black fungus yellow wine is obtained by reasonable addition of components and the preparation method, the polysaccharide content can reach 28.24g/L, the polyphenol content can reach 292.15 mu g/mL, and the flavone content can reach 89.33 mu g/mL; the total oxidation resistance (FRAP value) can reach 1.025 mmol/L; for DPPH and O₂The removal capacity of < - > 81.65%, 80.24% and 73.50% of OH is high in oxidation resistance, and the richness and functionality of the product nutrition are guaranteed.

In order to achieve the purpose, the invention adopts the following technical scheme:

the black fungus yellow wine comprises the following raw materials: millet, black fungus fermented mash, sweet wine yeast and saccharomyces cerevisiae.

According to the invention, by reasonably matching the millet and the black fungus fermented mash, the obtained black fungus yellow wine has high contents of polysaccharide, polyphenol and flavone and stronger total oxidation resistance, and the richness and functionality of the product nutrition are ensured.

Preferably, the material comprises the following raw materials in percentage by mass: the fermented liquid of black fungus is 20-25%, sweet distiller's yeast is 3.5-4.0% and Saccharomyces cerevisiae is 4-6% calculated by broomcorn millet.

Preferably, the material comprises the following raw materials in percentage by mass: the fermented liquor of black fungus is 25%, the sweet wine yeast is 3.5% and the saccharomyces cerevisiae is 5% in terms of the amount of husked millet.

The black fungus yellow wine prepared according to the proportion has the pH value of 4.2, the alcoholic strength of 13.2 vol%, the total acid content of 4.78g/L, the total sugar (calculated by glucose) content of 52.46g/L, the amino nitrogen content of 0.67 g/L, the non-sugar solid content of 17.88g/L and the calcium oxide content of less than 1.0g/L, and meets the national yellow wine standard. Simultaneously, the antioxidant active ingredients in the black fungus yellow wine are measured, wherein the polysaccharide content is 28.24g/L, the polyphenol content is 292.15 mug/mL, and the flavone content is 89.33 mug/mL; the total antioxidant capacity (FRAP value) is 1.025 mmol/L; for DPPH and O₂The removal capacity of-OH and OH is 81.65%, 80.24% and 73.50% in sequence, and the oxidation resistance is stronger.

Preferably, 8-10% of white granulated sugar and 0.1% of citric acid are also included.

Preferably, the preparation method of the black fungus fermented mash comprises the following steps: inoculating the activated black fungus strain to a liquid fermentation culture medium in an inoculation amount of 10%, culturing at 28 deg.C and 160r/min for 5d, and breaking the wall of the fermentation liquid to obtain black fungus fermented liquor.

The fermented liquor of black fungus has nutrient components which are not inferior to those of fruit bodies, contains a large amount of active substances such as polysaccharide, polyphenol, flavone and the like, and has the content far higher than that of the fruit bodies, so that the yellow wine obtained by adding the fermented liquor of black fungus is superior to the yellow wine in the prior art in taste and nutrient components.

Preferably, the black fungus strain activation method comprises the following steps: taking out the black fungus strain preserved at the temperature of 4 ℃, placing the black fungus strain in a constant temperature environment of 25 ℃ for constant temperature culture and activation for 24 hours, respectively transferring the black fungus strain to PDA slant culture media in an ultra-clean workbench after activation is finished, placing the black fungus strain in a constant temperature incubator of 26 ℃ for dark culture for 5-6 days, and after hyphae grow over the PDA slant culture media, completing activation and carrying out the next step of experiment.

Preferably, the PDA slant culture medium comprises the following raw materials in percentage by mass: potato (peeled) 20.0%, glucose 2.0%, bran 2.0%, agar 2.0%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.1%, and water in balance, and has a natural pH.

Preferably, the liquid fermentation medium comprises the following raw materials in percentage by mass: 2.5 percent of glucose, 0.5 percent of corn flour, 0.5 percent of soybean meal, 0.1 percent of magnesium sulfate, 1.0 percent of yeast extract, 0.2 percent of potassium dihydrogen phosphate and the balance of water.

The preparation method of the black fungus yellow wine specifically comprises the following steps:

(1) weighing raw materials of husked millet, black fungus fermented liquor, sweet wine yeast and saccharomyces cerevisiae according to mass percentage for later use;

(2) removing impurities from semen Panici Miliacei, soaking in water, steaming, sprinkling rice, and cooling;

(3) adding the black fungus fermented liquor into the cooled husked millet, performing saccharification and fermentation at 28-32 ℃ for 2-3 days, adding the saccharomyces cerevisiae, performing alcoholic fermentation at 29-33 ℃ for 10-12 days, and finally filtering to obtain a filtrate;

(4) and seasoning and sterilizing the filtrate to obtain the black fungus yellow wine.

Preferably, the soaking time in the step (2) is 12 hours, and the water addition amount is 2 times of the mass of the husked millet.

Preferably, the cooling temperature in step (2) is 20-25 ℃.

Preferably, white granulated sugar and citric acid are added to the seasoning in the step (4).

According to the technical scheme, compared with the prior art, the invention has the following beneficial effects: the black fungus yellow wine is obtained by reasonable addition of components and a preparation method, the polysaccharide content can reach 28.24g/L, the polyphenol content can reach 292.15 mu g/mL, and the flavone content can reach 89.33 mu g/mL; the total oxidation resistance (FRAP value) can reach 1.025 mmol/L; for DPPH and O₂The removal capacity of < - > 81.65%, 80.24% and 73.50% of OH is high in oxidation resistance, and the richness and functionality of the product nutrition are guaranteed.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.

FIG. 1 is a graph showing sensory results of the addition amount of the koji according to the present invention;

FIG. 2 is a graph showing sensory results of the addition of Saccharomyces cerevisiae according to the present invention;

FIG. 3 is a graph showing sensory results of the addition amount of fermented liquor of Auricularia auricula;

FIG. 4 is a sensory graph of the amount of white granulated sugar added in the present invention;

FIG. 5 is a plot of the response surface versus contour between the AB of the present invention;

FIG. 6 is a plot of the response surface versus contour between ACs of the present invention;

FIG. 7 is a plot of response surface versus contour between ADs of the present invention;

FIG. 8 is a plot of response surface versus contour between the BC of the present invention;

FIG. 9 is a plot of response surface versus contour between BDs of the present invention;

FIG. 10 is a plot of the response surface versus contour between CDs of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

A preparation method of black fungus yellow wine specifically comprises the following steps:

(1) weighing raw materials of husked millet, and then weighing 25% of black fungus fermented mash, 3.5% of sweet distiller's yeast, 5% of saccharomyces cerevisiae, 9% of white granulated sugar and 0.1% of citric acid for later use according to the mass of the husked millet; the preparation method of the black fungus fermented liquor comprises the following steps: inoculating the activated black fungus strain to a liquid fermentation culture medium in an inoculation amount of 10%, culturing at 28 ℃ and 160r/min for 5d, and performing wall breaking treatment on fermentation liquor to obtain black fungus fermentation mash, wherein the liquid fermentation culture medium comprises the following raw materials in percentage by mass: 2.5% of glucose, 0.5% of corn flour, 0.5% of soybean meal, 0.1% of magnesium sulfate, 1.0% of yeast extract, 0.2% of potassium dihydrogen phosphate and the balance of water;

(2) removing impurities from semen Panici Miliacei, soaking in 2 times of water for 12 hr, steaming, spraying rice, and cooling to room temperature of 25 deg.C;

(3) adding Auricularia auricula fermented liquor into cooled semen Panici Miliacei, performing diastatic fermentation at 30 deg.C for 2d, adding Saccharomyces cerevisiae, performing alcoholic fermentation at 30 deg.C for 10d, and filtering to obtain filtrate;

(4) adding white granulated sugar and citric acid into the filtrate, mixing uniformly, and sterilizing to obtain the black fungus yellow wine.

Process optimization test

1. Single factor test of brewing process: under the brewing process, the addition amount of the black fungus fermented mash (10.0%, 15.0%, 20.0%, 25.0%, 30.0%, 35.0%), the addition amount of the sweet distiller's yeast (2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%), the addition amount of the saccharomyces cerevisiae (1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%), the addition amount of the white granulated sugar (2.0%, 4.0%, 6.0%, 8.0%, 10.0%, 12.0%) are taken as single factors, and the sensory score is taken as an evaluation basis to carry out the test, and the result is shown in figures 1-4;

wherein the sensory evaluation test refers to the national standard GB/T13662-2018 of yellow wine, the evaluation table is shown in Table 1,

TABLE 1 organoleptic evaluation table for Auricularia auricula-yellow wine

As can be seen from FIGS. 1-4, the optimum addition amount of the sweet wine yeast is 3.5%, the optimum addition amount of the saccharomyces cerevisiae is 5.0%, the optimum addition amount of the black fungus fermented liquor is 25.0%, and the optimum addition amount of the white granulated sugar is 8.0%.

2. Box-Behnken design test and verification

After the optimal response area is determined through a single-factor test, the interaction among all the influence factors is further optimized by adopting a response surface method, the optimal process route is determined, the result is shown in the graph of 5-10, and meanwhile, after the optimization is performed through a Box-Behnken design test, the optimized process route is verified, and the reliability and the accuracy of the process route are tested;

after determining the optimal response area through a single-factor test, further optimizing by adopting a 4-factor 3 horizontal Box-Behnken test, wherein the design table is shown in Table 2:

TABLE 2 Box-Behnken design test Table

Determining a BBD test scheme through Design-Expert 8.0.6 software, obtaining test results, and obtaining a multiple quadratic regression equation while response values vary from 82.97 to 95.22 as shown in Table 3:

Y＝94.75+0.39A+0.34B-O.48C+0.30D+0.023AB-0.15AC+0.37AD-0.18BC- 0.032BD-0.13CD-3.25A²-1.98B²-7.75C²+9.583E-003D²；

meanwhile, according to the variance of the software analysis result, see table 3, the P value of the model is less than 0.0001, which shows that the regression equation is remarkable and the fitting performance of the whole test area is good; the complex correlation coefficient R2 is 0.9933, which shows that the correlation is better; the correction decision coefficient AdjR2 is 0.9866, which shows that 98.66% of experimental data variation can be analyzed by the model, and the accuracy of the test is 13.99, which is more than 4.0, and is in a reasonable range;

the three-dimensional response surface plot and the contour plot may directly show the interaction between two factors, as shown in fig. 5-10, with the maximum value given by the software, Y-95.2855 (designated 95.29), corresponding to response values: the addition amount of the sweet wine yeast is 3.56%, the addition amount of the saccharomyces cerevisiae is 5.08%, the addition amount of the black fungus fermented mash is 24.92%, and the addition amount of the white granulated sugar is 8.96%, for convenience, the addition amount of the sweet wine yeast is recorded as 3.5%, the addition amount of the saccharomyces cerevisiae is recorded as 5.0%, the addition amount of the black fungus fermented mash is recorded as 25.0%, and the addition amount of the white granulated sugar is recorded as 9.0%.

TABLE 3 Box-Behnken design test protocol and results

TABLE 3 Box-Behnken design test regression analysis results

According to BBD tests, the addition amount of sweet distiller's yeast is 3.50%, the addition amount of black fungus fermented liquor is 25.0%, saccharification and fermentation are carried out for 2d at 30 ℃, the addition amount of saccharomyces cerevisiae is 5.0%, fermentation is carried out for 10d at 30 ℃, then seasoning is carried out by adjusting the contents of citric acid and white granulated sugar, the addition amounts are respectively 9.0% of white granulated sugar and 0.1% of citric acid, the results are shown in Table 4, and experience proves that the error between the verification test value and the expected value predicted by the model is 0.19%, and the requirement is met.

Table 4 verifies test results and errors

Detection experiment

Detecting items

1. The product of the example 1 is subjected to content entry determination of alcoholic strength, total acid, amino acid nitrogen, total sugar, non-sugar and the like; wherein the content of the first and second substances,

and (3) measuring the alcoholic strength: with reference to GB 5009.225-2016 (national food safety Standard) -determination of ethanol concentration in wine, an alcometer method is adopted for determination;

determination of total acid: with reference to GB/T12456-;

determination of amino acid nitrogen: with reference to GB5009.235-2016 (national food safety Standard) -determination of amino acid nitrogen, an acidimetric method is adopted for determination;

and (3) measurement of other items: the determination of total sugar (calculated by glucose) refers to a 6.2.2 potassium ferrocyanide titration method in GB/T13662-; the determination of non-sugar solids is referenced to 6.3.1 gravimetric method; the determination of calcium oxide refers to a 6.6.2 potassium permanganate titration method; the pH is measured by reference to a 6.4 acidimeter method;

2. the active ingredients of the product of example 1 were tested, wherein,

the content of the total polysaccharide is measured by a phenol-sulfuric acid method;

determining the total phenol content by referring to a Folin-Ciocalteus method, determining a gallic acid standard curve, determining the OD value of the diluted wine sample to be determined, and calculating the total phenol content according to the curve;

measuring the content of total flavonoids by adopting an aluminum trichloride absorbance method, determining a rutin standard curve, measuring the OD value of the diluted wine sample to be measured, and calculating the content of the total flavonoids according to the curve;

the total oxidation resistance measured by the FRAP method adopts an iron ion reduction method, and the principle is as follows: under acidic conditions, Fe³⁺TPTZ complex (orange) + antioxidant → Fe²⁺-TPTZ (blue);

determination of radical scavenging ability: the determination of 3 kinds of free radicals takes 0.10mg/mL VC solution as positive control, and the VC equivalent antioxidant capacity of the wine sample to be determined, namely AEAC, is calculated, wherein the dibenzoyl bitter acyl free radical (DPPH. adopts a spectrophotometric method; superoxide anion radical (O)₂-. The) is measured by the tri-o-phenol autoxidation method; hydroxyl free radical (. OH) is measured by salicylic acid method;

finally, performing data statistical analysis by using Excel, SPSS 20.0 and origin8.0 software;

results

1 example 1 the detection results of various indexes of the product are shown in Table 5, and the detection results show that the indexes all meet the physicochemical requirements of yellow wine in 5.3.3 in the national standard GB/T13662-;

TABLE 5 yellow wine test items and results in example 1

2. Active ingredient determination:

the results of total polysaccharides, total polyphenols and total flavonoids are shown in table 6, and the addition of the fermented mash of black fungus to yellow wine brewing increased the active ingredients.

TABLE 6 measurement results of the components of Total polysaccharides, Total polyphenols and Total Flavonoids

Measurement of antioxidant ability

According to IC in Table 7₅₀The values of (a) and (b) indicate that the DPPH.removing ability of the 2 samples is, in order: VC solution at 0.1mg/mL, example 1; the conversion shows that the yellow wine in the unit volume (1mL) has the capability of removing free radicals which is equal to 0.0339mgVC solution respectively; (semi-inhibitory concentration, i.e., IC)₅₀And the concentration corresponding to the clearance rate of 50% is shown, and the concentration can be used for evaluating the clearance capacity of the wine sample to be tested on the free radicals, and the smaller the semi-inhibitory concentration is, the stronger the clearance capacity on the free radicals is, so that the antioxidant activity difference among the samples can be more accurately compared).

TABLE 7 DPPH.removing ability of wine sample to be tested

According to IC in Table 8₅₀Of 2 samples, it can be seen that₂The scavenging capacity of-is in order: example 1, 0.1mg/mL VC solution; the conversion shows that the yellow wine in the unit volume (1mL) has the capability of removing free radicals which is equal to 0.2648mgVC solution respectively;

TABLE 82 sample scavenging ability for O2- °

According to IC in Table 9₅₀The values of (a) and (b) can be seen, the OH scavenging capacity of the 2 samples is, in order: examples 1, 0.4mg/mL VC solution, and by conversion, the ability of example 1 to scavenge free radicals per unit volume (1mL) was found to correspond to 0.8505mg VC solution, respectively;

TABLE 92 sample cleaning ability for OH

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.