CN115322920A

CN115322920A - Lactobacillus plantarum and application thereof in yellow wine brewing

Info

Publication number: CN115322920A
Application number: CN202210275144.2A
Authority: CN
Inventors: 杨柳; 梅璐; 孙汉巨; 徐尚英
Original assignee: Hefei University of Technology
Current assignee: Hefei University of Technology
Priority date: 2022-03-21
Filing date: 2022-03-21
Publication date: 2022-11-11
Anticipated expiration: 2042-03-21
Also published as: CN115322920B

Abstract

A strain of Lactobacillus plantarum F25-4 and application thereof in yellow wine brewing comprise the following steps of 1: inoculating lactobacillus plantarum F25-4 slant strain to an activated liquid culture medium, performing static culture at 36-38 ℃ for 30-40h, then inoculating the obtained culture to a culture medium filled with a first seed culture medium, performing static culture at 36-38 ℃ for 20-28h, then inoculating the obtained culture to a second seed culture medium, and performing static culture at 36-38 ℃ for 20-28h to obtain a seed culture; step 2: adding the seed culture into a rice soaking tank filled with rice and water, mixing and stirring uniformly, and soaking rice. Adding the produced rice milk into the fermented mash for fermentation, and filtering, decocting and storing after the fermentation is finished to obtain the yellow wine liquid. The lactobacillus plantarum F25-4 can reduce acidity values of rice and mash, inhibit bacteria, improve cooking characteristics of soaked rice, reduce steam consumption during rice cooking, utilize rice milk as resources and improve flavor of yellow wine.

Description

Lactobacillus plantarum and application thereof in yellow wine brewing

Technical Field

The invention relates to Lactobacillus plantarum (Lactobacillus plantarum) and application thereof in yellow wine brewing, and belongs to the technical field of microbial fermentation.

Background

Yellow wine is the traditional cultural heritage of Chinese nationality, has a long history, is originated from China, and is called as ancient wine brewed in three parts of the world together with beer and wine. The yellow wine is brewed wine prepared by using rice, wheat and the like as main raw materials and through rice soaking, cooking, starter adding, saccharifying, fermenting, squeezing, filtering, wine decocting, storing and blending. In the traditional yellow wine brewing process, rice needs to be soaked for a period of time, and rice slurry is a byproduct in the rice soaking procedure in yellow wine production.

Soaking rice is a key step of Chinese yellow wine fermentation, can enable the rice to absorb water and is convenient to cook, and meanwhile, bacteria in rice slurry produce acid to acidify the rice, so that the pH value of fermentation liquor is reduced, and the safe fermentation of the yellow wine is ensured. Soaking is also a fermentation process in which metabolites produced by the bacteria can improve the texture and organoleptic properties of the rice. Enzymes and organic acids produced by the microorganisms during the soaking process hydrolyze the amorphous regions of the rice, changing the ratio of amylose to amylopectin, and the gelatinization properties of the rice. The rice soaking is also the highest biogenic amine yield stage in the yellow wine production.

The rice soaking can generate a large amount of rice milk water which is a main waste in the production of yellow wine and is also high-concentration organic wastewater. The rice milk is rich in amino acids, proteins, starch, saccharides, fat, vitamins, minerals, trace elements, etc., and a large amount of microorganisms. The rice milk has high Biochemical Oxygen Demand (BOD) and Chemical Oxygen Demand (COD) contents, and the direct discharge causes serious water pollution to local environment. At present, the rice milk water is recycled mainly in two ways, namely, the rice is soaked in an acidification cycle mode, the rice milk water is recycled, but the acidity of the rice milk water in the process is increased along with the cycle number, and yellow rice wine is over-acid. And secondly, the rice milk is subjected to protease and amylase enzymolysis treatment and then added into the fermentation process, but the rice milk contains various bacteria and metabolites thereof, so that the filtering speed in the squeezing stage is reduced, and the production efficiency is influenced.

The microorganisms propagate in a large amount and produce acid in the rice soaking process, so that an acid environment is provided for the subsequent fermentation process, and the growth of mixed bacteria is inhibited. As the rice soaking time is increased, the total acid content of the rice milk is in an increasing trend. The traditional rice soaking process has a plurality of microorganisms, acid-producing bacteria contained in the traditional rice soaking process can improve the acidity of rice milk, but the acidity rising speed is slower. The rice soaking time in winter is about 10-14 days generally (the rice can absorb water in 1-2 days during rice soaking, and the long-time soaking is to acidify the rice to ensure the normal fermentation), and the time consumption is long. Soaking rice at a higher temperature (such as 25-40 ℃) often causes the rice milk to smell, and the quality of the yellow wine is reduced.

In the natural rice soaking process, the rice pulp water contains more types of microorganisms, and some microorganisms generate amino acid decarboxylase, so that the biogenic amine content in the rice pulp water is higher. Biogenic amines produced in the rice soaking process are mainly putrescine, cadaverine, histamine and tyramine, wherein the histamine has the largest toxicity, and the ingestion of foods containing the histamine (> 100 mg) can cause severe food poisoning and cause symptoms such as migraine, headache, dizziness and hypertension; tyramine has obvious vasodilation and contraction effects on blood vessels and muscles, and sudden hypertension is caused by overhigh content of tyramine.

The use of proper safe acid-producing strains for rice soaking can not only shorten the rice soaking time and reduce the biogenic amine content of the product, but also prevent the rice soaking link from being influenced by seasons.

Disclosure of Invention

The invention aims to provide lactobacillus plantarum and application thereof in yellow wine brewing.

In order to achieve the above objects and other related objects, the present invention provides the following technical solutions: a strain of Lactobacillus plantarum F25-4, which has been preserved in China Center for Type Culture Collection (CCTCC) at 12 months and 9 days in 2021 with the preservation number of CCTCC No. M20211574.

In order to achieve the above objects and other related objects, the present invention provides the following technical solutions: application of Lactobacillus plantarum F25-4 in yellow wine brewing.

The preferable technical scheme is as follows: comprises the following steps:

step 1: inoculating lactobacillus plantarum F25-4 slant strain to an activation culture medium, performing static culture at 36-38 ℃ for 30-40h, then inoculating the obtained culture with an inoculation amount of 0.3-1% to a culture medium containing a first seed, performing static culture at 36-38 ℃ for 20-28h, then inoculating the obtained culture with an inoculation amount of 0.3-1% to a second seed culture medium, and performing static culture at 36-38 ℃ for 20-28h to obtain a seed culture;

step 2: inoculating seed culture in a rice soaking tank filled with rice and water in an amount of 0.3-1%, mixing, and soaking rice.

The preferable technical scheme is as follows: the concentration of the seed culture is 1-9X 10 ⁸ CFU/mL。

The preferable technical scheme is as follows: in the step 2, the temperature is 35-45 ℃ and the time is 30-40h.

The preferable technical scheme is as follows: the raw material formula of the first seed culture medium comprises: 100mL of rice milk, 1g of polished round-grained rice, 0.5g of yeast powder and 2g of glucose, and sterilizing for 15min at 121 ℃; the preparation method of the rice milk water comprises the following steps: soaking the polished round-grained rice for 1h at 40 ℃ according to the material-water ratio of 1.

The preferable technical scheme is as follows: the raw material formula of the second seed culture medium comprises: 100mL of rice milk, 3g of polished round-grained rice, 1g of yellow wine lees and 0.3g of yeast powder, and sterilizing at 121 ℃ for 15min; the preparation method of the rice milk water comprises the following steps: soaking the polished round-grained rice for 1h at 40 ℃ according to the material-water ratio of 1.

The preferable technical scheme is as follows: also comprises the following steps: separating rice milk and soaked rice after soaking rice, introducing steam of 0.1-0.2 MPa into the soaked rice, controlling the temperature at 102-107 ℃, cooking for 12-17min, adding water for cooling to 35-45 ℃, adding sterilized rice milk (the sterilization mode is boiling for 15 min), adding Angel active dry distiller's yeast, stirring uniformly, and starting fermentation; the fermentation period is 0-5 days, stirring every 4-5 hr, and the temperature is controlled at 25-32 deg.C, and the fermentation period is 6-18 days, and the temperature is controlled at 18-25 deg.C.

The preferable technical scheme is as follows: adding Angel active dry yeast and Angel Shao flavor distiller's yeast.

The preferable technical scheme is as follows: activating a culture medium: tryptone 0.5g, yeast powder 0.5g, beef extract 0.5g, citric acid diamine 0.2g, sodium chloride 0.25g, glucose 0.05g, tween 80.1 g, mnSO ₄ 0.005g，MgSO ₄ 0.02g，CaCO ₃ 0.01g，K ₂ HPO ₄ 0.2g,FeSO ₄ 0.004g, 100mL distilled water, pH5.3-5.5, sterilizing at 121 deg.C for 20min.

Due to the application of the technical scheme, compared with the prior art, the invention has the advantages that:

1. the lactobacillus plantarum F25-4 can reduce the acidity value of rice and mash, inhibit mixed bacteria and improve the cooking characteristic of soaked rice.

2. The strain does not produce amino acid decarboxylase in metabolism, and the biogenic amine content of rice pulp water is low.

3. The inoculation of the strain and the rice milk can not only recycle the rice milk, but also improve the flavor of the yellow wine.

Drawings

FIG. 1 growth of Strain F25-4 in amino acid decarboxylase assay Medium

FIG. 2 gram staining micrograph of Lactobacillus plantarum F25-4 (magnification 10 x 100).

FIG. 3 growth of Lactobacillus plantarum F25-4 on a sugar medium (glucose, galactose, lactose, fructose, cellobiose, maltose, sucrose, mannitol, xylose, in order from left to right).

FIG. 4 shows the effect of adding rice milk soaked with Lactobacillus plantarum F25-4 during fermentation process on sensory evaluation of yellow wine.

FIG. 5 is a flow chart of the process of the present invention.

Detailed Description

The following description of the embodiments of the present invention is provided for illustrative purposes, and other advantages and effects of the present invention will become apparent to those skilled in the art from the present disclosure.

Please refer to fig. 1-5. It should be understood that the structures, ratios, sizes, and the like shown in the drawings and described in the specification are only used for matching with the disclosure of the specification, so as to be understood and read by those skilled in the art, and are not used to limit the conditions under which the present invention can be implemented, so that the present invention has no technical significance, and any structural modification, ratio relationship change, or size adjustment should still fall within the scope of the present invention without affecting the efficacy and the achievable purpose of the present invention. In addition, the terms "upper", "lower", "left", "right", "middle" and "one" used in the present specification are used for clarity of description, and are not intended to limit the scope of the present invention, and the relative relationship between the terms and the terms may be changed or adjusted without substantial change in the technical content.

Example 1: lactobacillus plantarum and application thereof in yellow wine brewing

1. Screening Process

1. Sample screening

For 1:1.5 soaking rice slurry water of 1d and 2d and yellow wine in rice wine aqueous solution at 40 ℃ to ferment fermented mash of 1d, 2d, 3d, 4d, 5d, 7d, 11d, 15d and 20d (separating soaked rice from rice slurry water, introducing steam for cooking, adding water for cooling, falling a tank, adding activated Angel yeast (Shaoxing flavor) and Angel active dry yeast for starting fermentation), sampling, and carrying out high-throughput sequencing analysis to find that a large amount of lactic acid bacteria exist in the rice slurry water of 1d and 2d and the fermented rice slurry water of 1d, 2d and 3 d.

High throughput sequencing method: extracting the genomic DNA of the sample by SDS-CTAB method, centrifuging 5mL sample at 12000 r/min for 10min, collecting precipitate, adding 0.5 mL ddH ₂ O suspension, transferring to 2mL EP tube, adding 10. Mu.L lysozyme (50 mg/mL), standing at 37 ℃ for 30min, adding 125. Mu.L 10% SDS, immediately adding 5. Mu.L proteinase K (20 mg/mL), mixing uniformly, then, bathing at 65 ℃ for 1h (mixing the uniform sample upside down every 10 min), adding 700. Mu.L CTAB buffer, mixing uniformly, then, bathing at 65 ℃ for 1h;12 Centrifuging at 000r/min for 10min, and collecting supernatant. The extracted genomic DNA was sent to Zhejiang millet health Co., ltd, for which 1The 6S V3-V4 hypervariable region was amplified and sequenced. After data quality control, a representative operation classification unit (OTU) sequence is compared with a 16S r RNA gene sequence in NCBI by using Blast to determine species information of an OUT sequence.

2. Separation, purification and screening of lactic acid bacteria

1ml of rice milk water (rice: water = 1.5) and fermented mash are respectively diluted to 10^7, 1ml of 10^4, 10^5 and 10^6 diluent are respectively coated on an MRS plate containing light calcium carbonate and placed at 37 ℃ for culturing for 48 hours, colonies forming transparent rings on the plate are selected and further separated and purified. The purified strains were inoculated in MRS liquid medium and cultured at 37 ℃ for 36 hours, and lactic acid in the culture was measured by high performance liquid chromatography (chromatography conditions: chromatographic column: PLATISISIL ODS 250X 4.6mm, mobile phase: 0.0075mol/L sulfuric acid solution, detection wavelength: 210nm, column temperature: 30 ℃, flow rate: 0.8ml/min, sample intake: 20. Mu.L) to obtain 28 strains producing lactic acid. The physiological and biochemical tests of lactic acid bacteria and the basic physiological and biochemical property tests of Bergey's Manual of identification of bacteria are carried out on each strain respectively, and 10 strains of gram-staining positive, catalase negative, cytochrome oxidase negative, nitrate reduction negative, hydrogen sulfide production negative, citrate negative, amylase hydrolysis negative, methyl red positive, casein hydrolysis negative, gelatin liquefaction weak positive, growth at 15 ℃ and insoluble blood (gamma hemolysis) are screened out. Inoculating 10 strains of lactobacillus into liquid decarboxylase culture medium containing tyrosine, lysine, tryptophan, ornithine, phenylalanine, and histidine (liquid decarboxylase culture medium: tryptone 0.5g, yeast powder 0.5g, beef extract 0.5g, citric acid diamine 0.2g, sodium chloride 0.25g, glucose 0.05g, tween 80.1 g, mnSO) ₄ 0.005g， MgSO ₄ 0.02g，CaCO ₃ 0.01g，K ₂ HPO ₄ 0.2g,FeSO ₄ 0.004g, 0.006g of bromocresol purple, 0.001g of thiamine, 0.005g of pyridoxal 5' -phosphate, 0.2g of histidine, 0.2g of phenylalanine, 0.2g of tyrosine, 0.2g of tryptophan, 0.2g of ornithine hydrochloride, 0.2g of lysine, 100mL of distilled water, pH5.3-5.5, 121 ℃,20 min), and culturing at 37 ℃ for 36h. If the color of the culture solution changes from yellow to purple, the bacterium produces biogenic amine, if the bacterium is culturedIf the liquid is not discolored, the bacterium does not produce biogenic amine, and 5 strains of lactic acid bacteria are found to produce biogenic amine through measurement. The growth of strain F25-4 in the amino acid decarboxylase assay medium is shown in FIG. 1.

Adding 1mL of MRS liquid culture of 5 strains of bacteria into a material containing 40g of rice and 60g of water respectively according to the same inoculation amount, preserving the temperature for 36 hours at 37 ℃, sampling every 4 hours to measure the titratable acid amount in the material (titration method: taking 1% phenolphthalein as an indicator and using 0.1mol/L NaOH standard solution to titrate the total acid (in terms of lactic acid, g/L) = M multiplied by V multiplied by 0.090 multiplied by 1000. The molar concentration and mol/L of the M-NaOH standard solution, the titration volume and mL of the V-NaOH standard solution and the mass and g of 0.090-1mmoL of lactic acid) and screening out the strain F25-4 with the fastest acid production, wherein the rice milk inoculated with the bacteria has no peculiar smell and has faint scent of rice.

3. Identification of strains

The result of microscopic observation after gram staining of the strain F25-4 is shown in figure 2, the cells are stained purple, the shape is rod-shaped, and no spore exists, so the strain is preliminarily identified as G + bacillus. On the basis of the completed basic physiological and biochemical experiments of the lactic acid bacteria, the lactic acid bacteria are preliminarily identified. Further determining the utilization capacity of the strain F25-4 to sugar, and finding that the strain is turbid after being cultured in a culture medium of glucose, galactose, lactose, fructose, cellobiose, maltose, sucrose, mannitol and xylose for 24 hours at 30 ℃, which indicates that the strain can be preliminarily identified as the lactobacillus plantarum by Bergey bacteria identification handbook by utilizing various sugars in the culture medium. Extracting the DNA of the strain, determining the 16srDNA sequence, comparing the sequence with NCBI database, and identifying the strain as lactobacillus plantarum by combining physiological and biochemical properties, wherein the result is consistent with the physiological and biochemical experiment result of the strain. Lactobacillus plantarum F25-4, which has been deposited in China Center for Type Culture Collection (CCTCC) at 12 months and 9 days 2021, with the deposit number being CCTCC No. M20211574, china center for type culture Collection Address: eight-way No. 299 in Wuchang area of Wuhan city, hubei province.

4. Application of lactobacillus plantarum in yellow wine brewing

4.1 amplification culture of Strain F25-4

Strain F25-4 slant strain No. 1 loop was inoculated into a 18 x 180 test tube containing 15mL of activated broth and incubated at 37 ℃ for 36h. The 15mL culture was inoculated into a 1L Erlenmeyer flask containing 800mL of seed medium 1 and incubated at 37 ℃ for 24 hours. The culture was inoculated into a fermenter containing 10L of the seed medium 2 and subjected to static culture at 37 ℃ for 24 hours to obtain a seed culture.

4.2 application of Strain F25-4 in Rice soaking

Adding the above 10L culture of strain F25-4 (culture concentration is 10^8 CFU/mL) into a rice soaking tank containing 1t of rice and 1.5t of water, mixing, stirring, and soaking rice at 40 deg.C for 36h.

4.3 application of Rice slurry inoculated with Strain F25-4 in yellow wine fermentation

Separating rice slurry from rice by a sieve plate after soaking the rice, introducing steam of 0.1-0.2 MPa into the soaked rice, controlling the temperature to be about 105 ℃, cooking for about 15min, adding water for cooling to be about 40 ℃, dropping the rice slurry into a tank, adding sterilized rice slurry, adding Angel active dry distiller's yeast activated by 2% glucose solution and distiller's yeast (Shaoxing wine flavor), stirring uniformly, and starting fermentation. Fermenting for 0-5 days, stirring every 4-5 hr, controlling temperature at 25-32 deg.C, fermenting for 6-18 days, and fermenting for 15-18 days at 18-25 deg.C.

The yellow wine brewed by adopting the rice soaking process has the following beneficial effects.

(1) Can quickly reduce acidity values of rice and mash, inhibit bacteria, improve cooking characteristics of soaked rice, and shorten rice soaking time;

(2) The strain does not produce amino acid decarboxylase in metabolism, and the biogenic amine content of rice milk water is low;

(3) The rice milk inoculated with the strain is added into the fermentation process, so that the rice milk can be recycled, and the flavor of the yellow wine is improved.

Inoculating the strain F25-4 to reduce the acidity value of rice and mash and reduce the content of biogenic amine in rice slurry water.

The pH of the rice milk water at 36h is about 3.50, and the titratable acid content is 3.79g/L. The water absorption rate is related to acidity, and the higher the acidity, the higher the water absorption. The biogenic amine content in the rice milk is also obviously reduced, and the content of histamine and tyramine is reduced from 13mg/L (compared with the rice milk without lactobacillus) to 8.5mg/L. The gelatinization temperature of the soaked rice was reduced from 71.49 deg.C (as compared to non-inoculated rice milk) to 45.51 deg.C. The minimum cooking time of the rice is reduced from 25min (compared with non-inoculated rice milk) to 15min.

TABLE 1 influence of Natural soaking and Lactobacillus plantarum F25-4 inoculation on titratable acid (g/L) content of rice milk

TABLE 2 influence of Natural soaking and Lactobacillus plantarum F25-4 inoculation on Rice Water absorption (%)

Soaking time	Natural soaking (contrast)	Inoculating F25-4 for soaking
			1h	38.3	38.3
3h	38.8	38.9
			5h	38.9	38.6
7h	38.7	39.2
			9h	38.6	38.7
12h	38.0	38.3
			24h	38.4	38.6
36h	41.2	45.1
			48h	41.3	45.6

TABLE 3 influence of Natural soaking and inoculation of Strain F25-4 soaking on Histamine and tyramine in Rice plasma Water

The digestion characteristics of the soaked rice added with the lactobacillus plantarum F25-4 bacterial liquid are analyzed as follows:

the rice soaked in different modes is cooked, the cooking time of the rice grains and the water absorption index, the expansion coefficient and the water solubility index reflecting the cooking characteristics of the rice grains are recorded by a rice milling method, the results are shown in table 4, the rice cooking time is obviously reduced after the rice is soaked by inoculating the strain F25-4, the water absorption index, the expansion coefficient and the water solubility index are obviously increased, the water absorption index shows that the rice grains are more beneficial to cooking when the water absorption speed is higher at the same temperature in the cooking process, the water solubility is increased, the soluble substances in the rice grains are dissolved into the water solution, and the rice soaked by the lactobacillus plantarum F25-4 has higher cooking effect in the same time.

TABLE 4 comparison of cooking characteristics of rice after natural soaking and inoculation with Lactobacillus plantarum F25-4

Differential Scanning Calorimetry (DSC) also reflects the gelatinisation behaviour of rice. Compared with natural soaking, the gelatinization temperature of the rice flour is obviously reduced after the lactobacillus plantarum is added for soaking, and the initial temperature (To), the peak temperature (Tp) and the conclusion temperature (Tc) are respectively reduced by 17.9%, 11.6% and 4.2%. Indicating that the cooking time required for the rice after the bacteria adding and soaking is reduced.

TABLE 5 thermal Properties of naturally soaked and inoculated Lactobacillus plantarum F25-4 soaked rice

The effect of Lactobacillus plantarum 25-4 on the quality of yellow wine brewing is as follows.

Cooking rice soaked by strain 25-4 at 105 deg.C for 15min, adding water, cooling, sterilizing the rest rice slurry, adding into fermentation process, adding distiller's yeast and yeast, fermenting for 5 days until the alcohol content reaches 149.65kg (alcohol concentration is 14.9%, volume is 2875L), and the total acid reaches maximum 3.9g/L on the fifth day. The content of residual total sugar (calculated by glucose) is 10mg/ml, the content of phenethyl alcohol in the raw wine is increased, the content of higher alcohol is reduced, and the content of lipid is increased.

TABLE 6 influence of fermentation of mash alcohol content (V/V, 25 ℃) with rice slurry water inoculated with Lactobacillus plantarum F25-4 as brewing water.

TABLE 7 influence of Rice slurry soaked with addition of Strain F25-4 on volatile higher alcohol flavor substances in yellow wine

TABLE 8 influence of addition of Lactobacillus plantarum F25-4 soaked rice pulp water on volatile lipid flavor substances in yellow wine.

Table 7 reflects the effect of the addition of the rice milk soaked with lactobacillus plantarum F25-4 on the higher alcohols in the flavor of yellow wine, wherein it was found that the content of the higher alcohols in the control group was significantly reduced compared to the content of the higher alcohols in the rice milk containing inoculated bacteria, the total amount was reduced by 14.9% and 19.6%, respectively, except for phenethyl alcohol, and the types of the higher alcohols were not increased. The phenethyl alcohol content is obviously increased and is respectively increased by 137.1 percent compared with a contrast component, and the phenethyl alcohol beta-phenethyl alcohol is also called as 2-phenethyl alcohol, is aromatic high-grade alcohol with rose fragrance and honey fragrance, is an important high-boiling-point aroma component in alcoholic beverages, is widely present in various alcoholic beverages, and is an important aromatic substance in yellow wine. Isobutanol and isoamylol are the main reasons for yellow wine to cause the first infection, the content of isobutanol is slightly increased after the inoculation rice milk water is added, but the content of isoamylol is reduced by 18.2 percent, and 2, 3-butanediol can bring sweet taste to the yellow wine and increase the taste of the yellow wine. Therefore, the addition of the inoculation rice milk can reduce the content of higher alcohol in the wine, increase the content of phenethyl alcohol and effectively reduce the top feeling of yellow wine.

The lipid in yellow wine is one of the main sources of yellow wine aroma. As shown in Table 8, the lipid content of the inoculated rice milk-containing water group was significantly higher than that of the control group, and increased by 50% and 72.8%, respectively. The species number is not reduced, and compared with a control group, the group added with the inoculation rice milk water increases ethyl hexadecanoate, dibutyl adipate, ethyl dodecanoate, 9-octadecenoic acid ethyl ester, linoleic acid ethyl ester, 2-acrylic acid and 3- (4-methoxyphenyl) -2-ethylhexyl ester, and reduces ethyl butyrate, ethyl octanoate, benzoic acid-2-ethylhexyl ester and isopropyl palmitate. Wherein the isoamyl acetate is reduced by 59.3 percent, but the 2-ethyl phenyl acetate is increased by 4.14mg/L, and after the rice milk water is added as supplemented water to ferment, the ethyl caproate and the isoamyl acetate are reduced, the ethyl phenylacetate is increased, and the like.

Sensory evaluation of yellow wine a panel (5 women, 5 men) was composed of 10 experienced persons. The sensory characteristics of different yellow wine samples were evaluated. In the first step, the panelists first observe, taste and smell the yellow wine to identify and record the sensory attributes contained. In the second step, the panelists discuss and determine the attributes, and then build the final descriptors. 14 yellow rice wine descriptors such as appearance (redness and turbidity), aroma (bouquet, fruity, flowery odour and grain aroma), taste (sweet, sour, spicy, sticky and bitter), and taste (astringent, lasting and saturated feeling) are used. In the last step, the panelists express their judgment by quantifying each sensory descriptor. The intensity scale was from 0 to 5 (0: none, 1: very weak, 2: normal, 3: mild, 4: strong, 5: most intense). 30mL samples of the prepared yellow wine were placed in the same opaque disposable plastic cup and subjected to a good vortex treatment before the samples were evaluated for randomization. Each sample was assayed in triplicate.

Sensory evaluation is represented by sensory radar chart, the result is shown in figure 4, and the characteristics of bouquet, persistence, color, fragrance, acidity and the like of the yellow wine fermentation added with the inoculated rice milk water group are obviously higher than those of the control group. The turbidity, the grain aroma, the bitter taste, the astringency, the richness and the piquancy of the yellow wine in the control group are higher than those of the group added with the inoculated rice milk water, which is probably caused by the combined action of amino acid and flavor substances in the wine, and the sour taste and the bitter taste quality of the yellow wine can be influenced by more acid and bitter amino acid with larger specific gravity. The yellow wine of the group added with the inoculated rice milk is slightly better than that of the control group.

Compared with the traditional yellow wine brewing, the yellow wine added with the rice milk water soaked by the lactobacillus plantarum F25-4 can quickly improve the acidity of the rice milk water, shorten the rice soaking time, inhibit the growth of mixed bacteria, improve the stability in the rice soaking process, effectively reduce the quality concentration of biogenic amine in the rice milk water, and can provide an acidic environment for yellow wine fermentation by attaching microorganisms to rice during the rice soaking process, thereby ensuring the normal fermentation. After the lactobacillus plantarum F25-4 is soaked, the cooking characteristic is improved, the steam consumption is reduced, and the energy consumption is reduced. The rice milk is added into the fermentation process, so that the content of the phenethyl alcohol in the product is increased, the content of isoamyl alcohol is reduced, the 'top feeling' of drinking is reduced, the resource utilization of the rice milk reduces the treatment capacity of wastewater, and the cost is further reduced.

Example 2: lactobacillus plantarum and application thereof in yellow wine brewing

A strain of Lactobacillus plantarum F25-4, which has been preserved in China Center for Type Culture Collection (CCTCC) at 12 months and 9 days in 2021 with the preservation number of CCTCC No. M20211574.

Application of Lactobacillus plantarum F25-4 in yellow wine brewing.

Use according to claim 2, characterized in that: comprises the following steps:

step 1: inoculating lactobacillus plantarum F25-4 slant strains to an activation culture medium, performing static culture at 36 ℃ for 30 hours, then inoculating the obtained culture to a culture medium containing a first seed culture medium, performing static culture at 36 ℃ for 20 hours, then inoculating the obtained culture to a second seed culture medium, and performing static culture at 36 ℃ for 20 hours to obtain a seed culture;

step 2: adding the seed culture into a rice soaking tank filled with rice and water, mixing, and soaking rice.

The concentration of the seed culture was 1X 10 ⁸ CFU/mL, the seed culture was added at 0.3% inoculum size to a rice-soaking tank containing rice and water.

In the step 2, the temperature is 35 ℃ and the time is 30h.

The raw material formula of the first seed culture medium comprises: 100mL of rice milk, 1g of polished round-grained rice, 0.5g of yeast powder and 2g of glucose, and sterilizing for 15min at 121 ℃; the preparation method of the rice milk water comprises the following steps: soaking the polished round-grained rice for 1h at 40 ℃ according to the material-water ratio of 1, and sieving the polished round-grained rice with a 10-mesh sieve to obtain rice milk.

The raw material formula of the second seed culture medium comprises: 100mL of rice milk, 3g of polished round-grained rice, 1g of yellow wine lees and 0.3g of yeast powder, and sterilizing at 121 ℃ for 15min; the preparation method of the rice milk water comprises the following steps: soaking the polished round-grained rice for 1h at 40 ℃ according to the material-water ratio of 1.

Also comprises the following steps: separating to obtain rice milk and soaked rice after soaking rice, introducing 0.1MPa steam into the soaked rice, controlling the temperature at 102 ℃, cooking for 12min, adding water to cool to 35 ℃, adding sterilized rice milk after dropping, adding activated Angel active dry yeast and Shaoxing rice wine flavor wine brewing yeast, stirring uniformly, and starting fermentation; the fermentation period is 0-5 days, the stirring is required to be carried out once every 4 hours, the temperature is controlled at 25 ℃, the fermentation period is 6 days, and the temperature is controlled at 18 ℃.

Activation of the medium: tryptone 0.5g, yeast powder 0.5g, beef extract 0.5g, citric acid diamine 0.2g, sodium chloride 0.25g, glucose 0.05g, tween 80.1g, mnSO ₄ 0.005g，MgSO ₄ 0.02g，CaCO ₃ 0.01g，K ₂ HPO ₄ 0.2g, FeSO ₄ 0.004g of distilled water, 100mL of distilled water, pH5.3-5.5, and sterilizing at 121 ℃ for 20min.

Example 3: lactobacillus plantarum and application thereof in yellow wine brewing

A Lactobacillus plantarum F25-4 has been preserved in China Center for Type Culture Collection (CCTCC) in 2021, 12 months and 9 days, with the preservation number of CCTCC No. M20211574.

Application of Lactobacillus plantarum F25-4 in yellow wine brewing.

The preferable technical scheme is as follows: comprises the following steps:

step 1: inoculating lactobacillus plantarum F25-4 slant strains to an activation culture medium, performing static culture at 38 ℃ for 40h, then inoculating the obtained culture to a culture medium containing a first seed according to the inoculation amount of 1%, performing static culture at 38 ℃ for 28h, then inoculating the obtained culture to a second seed culture medium according to the inoculation amount of 1%, and performing static culture at 38 ℃ for 28h to obtain a seed culture;

and 2, step: inoculating seed culture in a rice soaking tank filled with rice and water in an inoculation amount of 1%, mixing, and soaking rice.

The preferred embodiment is: the concentration of the seed culture was 9X 10 ⁸ CFU/mL。

The preferred embodiment is: in the step 2, the temperature is 45 ℃ and the time is 40h.

The preferred embodiment is: the raw material formula of the first seed culture medium comprises: 100mL of rice milk, 1g of polished round-grained rice, 0.5g of yeast powder and 2g of glucose, and sterilizing for 15min at 121 ℃; the preparation method of the rice milk water comprises the following steps: soaking the polished round-grained rice for 1h at 40 ℃ according to the material-water ratio of 1, and sieving the polished round-grained rice with a 10-mesh sieve to obtain rice milk.

The preferred embodiment is: the raw material formula of the second seed culture medium comprises: 100mL of rice milk, 3g of polished round-grained rice, 1g of yellow wine lees and 0.3g of yeast powder, and sterilizing at 121 ℃ for 15min; the preparation method of the rice milk water comprises the following steps: soaking the polished round-grained rice for 1h at 40 ℃ according to the material-water ratio of 1, and sieving the polished round-grained rice with a 10-mesh sieve to obtain rice milk.

The preferred embodiment is: also comprises the following steps: separating rice milk water and soaked rice after soaking rice, introducing 0.2MPa steam into the soaked rice, controlling the temperature at 107 ℃, cooking for 17min, adding water for cooling to 45 ℃, dropping into a tank, adding sterilized rice milk water (the sterilization mode is boiling for 15 min), adding Angel active dry distiller's yeast, stirring uniformly, and starting fermentation; the fermentation period is 5 days, the stirring is required to be carried out once every 5 hours, the temperature is controlled at 32 ℃, the fermentation period is 18 days, and the temperature is controlled at 25 ℃.

The preferred embodiment is: adding Angel active dry yeast and Angel Shaoxing wine-flavored distiller's yeast.

The preferred embodiment is: activation of the medium: tryptone 0.5g, yeast powder 0.5g, beef extract 0.5g, citric acid diamine 0.2g, sodium chloride 0.25g, glucose 0.05g, tween 80.1g, mnSO ₄ 0.005g，MgSO ₄ 0.02g，CaCO ₃ 0.01g，K ₂ HPO ₄ 0.2g,FeSO ₄ 0.004g, 100mL distilled water, pH5.3-5.5, sterilizing at 121 deg.C for 20min.

16S rDNA gene sequence:

the foregoing is illustrative of the preferred embodiment of the present invention and is not to be construed as limiting thereof in any way, and any modifications or variations thereof that fall within the spirit of the invention are intended to be included within the scope thereof.

Claims

1. Lactobacillus plantarum (II)Lactobacillus plantarum) F25-4, which has been preserved in China Center for Type Culture Collection (CCTCC) at 12 months and 9 days in 2021 with the preservation number of CCTCC No. M20211574.

2. Lactobacillus plantarum (f)Lactobacillus plantarum) The application of F25-4 in yellow wine brewing.

3. Use according to claim 2, characterized in that: comprises the following steps:

step 1: inoculating lactobacillus plantarum F25-4 slant strain to an activation culture medium, performing static culture at 36-38 ℃ for 30-40h, then inoculating the obtained culture to a culture medium filled with a first seed culture medium, performing static culture at 36-38 ℃ for 20-28h, then inoculating the obtained culture to a second seed culture medium, and performing static culture at 36-38 ℃ for 20-28h to obtain a seed culture;

4. Use according to claim 2, characterized in that: the concentration of the seed culture is 1-9X 10 ⁸ CFU/mL, seed culture was added at an inoculum size of 0.3-1% to a rice-soaking tank containing rice and water.

5. Use according to claim 2, characterized in that: in the step 2, the temperature is 35-45 ℃ and the time is 30-40h.

6. Use according to claim 2, characterized in that: the raw material formula of the first seed culture medium comprises: 100mL of rice milk, 1g of polished round-grained rice, 0.5g of yeast powder and 2g of glucose, and sterilizing for 15min at 121 ℃; the preparation method of the rice milk water comprises the following steps: soaking the polished round-grained rice for 1h at 40 ℃ according to the material-water ratio of 1.

7. Use according to claim 2, characterized in that: the raw material formula of the second seed culture medium comprises: 100mL of rice milk, 3g of polished round-grained rice, 1g of yellow wine lees and 0.3g of yeast powder, and sterilizing at 121 ℃ for 15min; the preparation method of the rice milk water comprises the following steps: soaking the polished round-grained rice for 1h at 40 ℃ according to the material-water ratio of 1, and sieving the polished round-grained rice with a 10-mesh sieve to obtain rice milk.

8. Use according to claim 2, characterized in that: also comprises the following steps: separating rice pulp water and soaked rice after rice soaking, introducing steam of 0.1-0.2 MPa into the soaked rice, controlling the temperature at 102-107 ℃, cooking for 12-17min, adding water for cooling to 35-45 ℃, adding sterilized rice pulp water after the rice pulp water falls into a tank, adding activated Angel active dry yeast and Shaoxing wine flavor distiller's yeast, stirring uniformly, and starting fermentation; the fermentation period is 0-5 days, stirring every 4-5 hr, and the temperature is controlled at 25-32 deg.C, and the fermentation period is 6-18 days, and the temperature is controlled at 18-25 deg.C.

9. Use according to claim 2, characterized in that: activating a culture medium: tryptone 0.5g, yeast powder 0.5g, beef extract 0.5g, citric acid diamine 0.2g, sodium chloride 0.25g, glucose 0.05g, tween 80.1g, mnSO ₄ 0.005g，MgSO ₄ 0.02g，CaCO ₃ 0.01g，K ₂ HPO ₄ 0.2g, FeSO ₄ 0.004g of the extract, 100mL of distilled water, pH5.3-5.5, and sterilizing at 121 ℃ for 20min.