CN115322920B - Lactobacillus plantarum and application thereof in yellow wine brewing - Google Patents
Lactobacillus plantarum and application thereof in yellow wine brewing Download PDFInfo
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Abstract
Lactobacillus plantarum F25-4 and application thereof in yellow wine brewing comprise the following steps: inoculating lactobacillus plantarum F25-4 inclined plane strain to an activated liquid culture medium, performing stationary culture at 36-38 ℃ for 30-40h, then inoculating the obtained culture to a first seed culture medium, performing stationary culture at 36-38 ℃ for 20-28h, then inoculating the obtained culture to a second seed culture medium, performing stationary culture at 36-38 ℃ for 20-28h, and obtaining a seed culture; step 2: adding seed culture into a rice soaking tank filled with rice and water, mixing, stirring, and soaking rice. Adding the produced rice milk into fermented mash for fermentation, filtering, decocting and storing after fermentation to obtain yellow wine liquor. The lactobacillus plantarum F25-4 can reduce acidity values of rice and mash, inhibit mixed bacteria, improve cooking characteristics of soaked rice, reduce steam consumption during rice steaming, and utilize rice slurry in a recycling way to improve flavor of yellow wine.
Description
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 a traditional cultural heritage of Chinese nationality, has long history, is derived from China, and is called as ancient wine for brewing with beer and wine in the world. The yellow wine is a brewed wine which is prepared by taking rice, wheat and the like as main raw materials, soaking rice, steaming, adding yeast, saccharifying, fermenting, squeezing, filtering, decocting, storing and blending. In the traditional yellow wine brewing process, rice needs to be soaked for a period of time, and rice milk water is a byproduct in the rice soaking process of yellow wine production.
Soaking rice is a key step of Chinese yellow wine fermentation, and can enable rice to absorb water so as to be convenient for cooking, and meanwhile bacteria in rice slurry water produce acid to acidify rice, so that the pH value of fermentation liquor is reduced, and the safe fermentation of yellow wine is ensured. Soaking is also a fermentation process in which metabolites produced by bacteria can improve the texture and organoleptic properties of rice. Enzymes and organic acids produced by 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 stage with the highest biogenic amine yield in the yellow wine production.
Rice soaking produces a large amount of rice slurry, which is the main waste in yellow wine production and also high-concentration organic wastewater. The rice milk contains rich amino acids, proteins, starch, saccharides, fat, vitamins, minerals, trace elements, etc., and a large number of microorganisms. The Biochemical Oxygen Demand (BOD) and Chemical Oxygen Demand (COD) contents of the rice slurry water are high, and the direct discharge causes serious water pollution to the local environment. At present, two main types of recycling of rice milk water exist, namely, the rice milk water is recycled by acidification and circulation rice soaking, but the acidity of the rice milk water in the process is increased along with the circulation times, so that yellow wine peracid can be caused. Secondly, the rice milk is added into the fermentation process after being subjected to enzymolysis treatment of protease and amylase, but the filtering speed in the squeezing stage is reduced due to the fact that the rice milk contains various bacteria and metabolites thereof, and the production efficiency is affected.
In the rice soaking process, microorganisms are propagated in a large quantity and produce acid, so that an acidic environment is provided for the subsequent fermentation process, and the growth of mixed bacteria is inhibited. With the increase of the rice soaking time, the total acid content of the rice milk is in an ascending trend. In the traditional rice soaking process, the variety of microorganisms is various, and the acidogenic bacteria contained in the traditional rice soaking process can improve the acidity of rice milk, but the acidity rising rate is slower. The rice soaking time in winter generally needs about 10-14d (the rice can be soaked in water within 1-2 days, and the long-time soaking is to acidify the rice to ensure the normal fermentation), and the time is long. However, rice soaking at higher temperature (25-40deg.C) often causes odor of rice slurry, and reduces yellow wine quality.
In the natural rice soaking process, rice slurry contains a plurality of microorganisms, wherein some microorganisms generate amino acid decarboxylase, so that the biogenic amine content in the rice slurry is higher, and researches show that the more biogenic amine is generated in the rice soaking link, the higher the biogenic amine content in the final yellow wine is. Biogenic amines generated in the rice soaking process are mainly putrescine, cadaverine, histamine and tyramine, wherein histamine has the largest toxicity, and intake of foods containing histamine (> 100 mg) can cause serious food poisoning, thereby causing symptoms such as migraine, headache, dizziness and hypertension; tyramine has obvious dilating and contracting effects on blood vessels and muscles, and burst hypertension can be caused by excessive tyramine content.
The proper safe acid-producing strain is used for soaking rice, so that the rice soaking time can be shortened, the biogenic amine content of the product can be reduced, and the rice soaking link is not influenced by seasons.
Disclosure of Invention
The invention aims to provide lactobacillus plantarum and application thereof in yellow wine brewing.
To achieve the above and other related objects, the present invention provides the following technical solutions: lactobacillus plantarum (Lactobacillus plantarum) F25-4 is preserved in China Center for Type Culture Collection (CCTCC) on 12 th month 9 of 2021, and the preservation number is CCTCC No. M20211574.
To achieve the above and other related objects, the present invention provides the following technical solutions: application of Lactobacillus plantarum (Lactobacillus plantarum) F25-4 in yellow wine brewing is provided.
The preferable technical scheme is as follows: comprises the following steps:
step 1: inoculating lactobacillus plantarum F25-4 inclined plane strain to an activation culture medium, performing stationary culture at 36-38 ℃ for 30-40h, then inoculating the obtained culture to a first seed culture medium according to 0.3-1% of inoculum size, performing stationary culture at 36-38 ℃ for 20-28h, then inoculating the obtained culture to a second seed culture medium according to 0.3-1% of inoculum size, and performing stationary culture at 36-38 ℃ for 20-28h to obtain a seed culture;
step 2: inoculating seed culture in 0.3-1% of seed culture in rice soaking tank filled with rice and water, mixing, stirring, and soaking rice.
The preferable technical scheme is as follows: the seed culture concentration is 1-9×10 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, 2g of glucose and sterilizing at 121 ℃ for 15min; the preparation method of the rice milk comprises the following steps: soaking semen oryzae Sativae at 40deg.C at a ratio of 1:1 for 1 hr, and sieving with 10 mesh sieve to obtain rice slurry.
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, 0.3g of yeast powder and 15min of sterilization at 121 ℃; the preparation method of the rice milk comprises the following steps: soaking semen oryzae Sativae at 40deg.C at a ratio of 1:1 for 1 hr, and sieving with 10 mesh sieve to obtain rice slurry.
The preferable technical scheme is as follows: the method also comprises the following steps: separating to obtain rice slurry and soaked rice after rice soaking, introducing steam of 0.1-0.2 MPa into soaked rice, controlling the temperature at 102-107 ℃, steaming for 12-17min, adding water to cool to 35-45 ℃, adding sterilized (sterilizing mode is boiling for 15 min) rice slurry after tank dropping, adding Angel active dry distiller's yeast, stirring uniformly, and starting fermentation; the fermentation period is 0-5 days, stirring is required every 4-5 hours, the temperature is controlled at 25-32 ℃, the fermentation period is 6-18 days, and the temperature is controlled at 18-25 ℃.
The preferable technical scheme is as follows: and adding Angel active dry yeast and Angel Shaoxing wine flavor brewing yeast.
The preferable technical scheme is as follows: activation medium: 0.5g of tryptone, 0.5g of yeast powder, 0.5g of beef extract, 0.2g of citric acid diamine, 0.25g of sodium chloride and 0.05g,Tween 80 0.1g,MnSO g of glucose 4 0.005g，MgSO 4 0.02g，CaCO 3 0.01g，K 2 HPO 4 0.2g,FeSO 4 0.004g, distilled water 100mL, pH5.3-5.5, and sterilized at 121 ℃ 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 acidity values of rice and mash, inhibit mixed bacteria and improve cooking characteristics of soaked rice.
2. The bacterial strain does not produce amino acid decarboxylase in metabolism, and the biogenic amine content of rice milk is low.
3. The strain and rice milk can be inoculated, so that the rice milk can be recycled, and the flavor of the yellow wine can be improved.
Drawings
FIG. 1 growth of strain F25-4 in amino acid decarboxylase assay medium
FIG. 2 Lactobacillus plantarum F25-4 gram staining microscopy (magnification 10X 100).
FIG. 3 growth of Lactobacillus plantarum F25-4 on sugar medium (glucose, galactose, lactose, fructose, cellobiose, maltose, sucrose, mannitol, xylose in order from left to right).
FIG. 4 effect of fermentation with rice milk soaked in Lactobacillus plantarum F25-4 on sensory evaluation of yellow wine.
Fig. 5 is a process flow diagram of the present invention.
Detailed Description
Further advantages and effects of the present invention will become apparent to those skilled in the art from the disclosure of the present invention, which is described by the following specific examples.
Please refer to fig. 1-5. It should be understood that the structures, proportions, sizes, etc. shown in the drawings are for illustration purposes only and should not be construed as limiting the invention to the extent that it can be practiced, since modifications, changes in the proportions, or otherwise, used in the practice of the invention, are not intended to be critical to the essential characteristics of the invention, but are intended to fall within the spirit and scope of the invention. Also, the terms such as "upper," "lower," "left," "right," "middle," and "a" and the like recited in the present specification are merely for descriptive purposes and are not intended to limit the scope of the invention, but are intended to provide relative positional changes or modifications without materially altering the technical context in which the invention may be practiced.
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 fermenting fermentation mash of 1d, 2d, 3d, 4d, 5d, 7d, 11d, 15d and 20d with yellow wine at 40 ℃, separating soaked rice from rice slurry, steaming, adding water, cooling, dropping, adding activated Angel yeast (Shaoxing wine flavor) and Angel active dry yeast, sampling, and performing high throughput sequencing analysis to find that a large amount of lactic acid bacteria exist in the rice slurry water soaked for 1d and 2d and fermentation 1d, 2d and 3 d.
High throughput sequencing method: extracting genomic DNA of the obtained sample by SDS-CTAB method, centrifuging 5mL of the sample 12000/r/min for 10min, collecting precipitate, adding 0.5 mL dd H 2 Transferring the suspension O into a 2mL EP tube, adding 10 mu L of lysozyme (50 mg/mL), standing at 37 ℃ for 30min, adding 125 mu L of 10% SDS, immediately adding 5 mu L of proteinase K (20 mg/mL), mixing uniformly, carrying out water bath at 65 ℃ for 1h (mixing uniformly by upside down every 10 min), adding 700 mu L of CTAB buffer solution, mixing uniformly, and carrying out water bath at 65 ℃ for 1h;12 Centrifuging at 000r/min for 10min, and collecting supernatant. The extracted genomic DNA was sent to Zhejiang Valley health Co., ltd, and its 16S V3-V4 hypervariable region was amplified and sequenced. After data quality control, blast is used to compare the sequence of a representative operation classification unit (OTU) with the sequence of the 16S r RNA gene in NCBI to determine species information of the OUT sequence.
2. Separation, purification and screening of lactic acid bacteria
1ml of rice slurry water (rice: water=1:1.5) and fermented mash are respectively diluted to 10≡7, 1ml of 10≡4, 10≡5 and 10≡6 diluted solutions are respectively coated on MRS plates containing light calcium carbonate and are placed on the plates for culturing for 48 hours at 37 ℃, colonies forming transparent rings on the plates are selected, and further separation and purification are carried out. Inoculating the purified strain into MRS liquid culture medium, culturing at 37deg.C for 36 hr, and measuring lactic acid in the culture solution by high performance liquid chromatography (chromatographic conditions: PLATISIL ODS 250×4.6mm, mobile phase: 0.0075mol/L sulfuric acid solution, detection wavelength: 210nm, column temperature):30 ℃, flow rate: sample injection amount of 0.8 ml/min: 20. Mu.L) to obtain 28 lactic acid-producing strains. The physiological and biochemical tests of lactic acid bacteria and basic physiological and biochemical property tests of the lactic acid bacteria are respectively carried out on each strain, and 10 strains which are gram-positive, contact enzyme 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, 15 ℃ growth and no hemolysis (gamma hemolysis) are screened. Inoculating 10 strains of lactobacillus respectively 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 0.1g,MnSO) 4 0.005g， MgSO 4 0.02g，CaCO 3 0.01g，K 2 HPO 4 0.2g,FeSO 4 0.004g, bromocresol purple 0.006g, thiamine 0.001g, pyridoxal 5' -phosphate 0.005g, histidine 0.2g, phenylalanine 0.2g, tyrosine 0.2g, tryptophan 0.2g, ornithine hydrochloride 0.2g, lysine 0.2g, distilled water 100mL, pH5.3-5.5, 121 ℃ for 20 min), at 37 ℃ for 36h. If the color of the culture solution changes from yellow to purple, the bacteria can produce the biogenic amine, if the culture solution does not change color, the bacteria can not produce the biogenic amine, and the determination shows that 5 strains of lactic acid bacteria can not produce the biogenic amine. The growth of strain F25-4 in the amino acid decarboxylase assay medium is shown in FIG. 1.
1mL of MRS liquid culture of 5 strains of bacteria is respectively added into a material containing 40g of rice and 60g of water according to the same inoculation amount, the temperature is kept for 36 hours at 37 ℃, the titratable acid amount in the material is measured by sampling every 4 hours (titration method comprises the steps of taking 1% phenolphthalein as an indicator and titrating with 0.1mol/L NaOH standard solution, wherein the total acid (g/L) =M multiplied by V multiplied by 0.090 multiplied by 1000 according to lactic acid, wherein the molar concentration of the M-NaOH standard solution, the mol/L, the titration volume of the V-NaOH standard solution, the mass of the mL, the g) of 0.090-1mmoL lactic acid is selected, the strain F25-4 with the fastest acid production is obtained by screening, and rice slurry water inoculated with the bacteria has no peculiar smell and has faint scent of rice.
3. Identification of strains
As a result of microscopic observation after gram staining of the strain F25-4, the cells were stained purple in shape and in a rod-like form without spores, and thus, the strain was preliminarily identified as G+ bacillus. Based on the basic physiological and biochemical experiment of the finished lactobacillus, the lactobacillus is primarily identified. Further, the sugar utilization capacity of the strain F25-4 was measured, and the strain was found to be a Lactobacillus plantarum strain which was initially identified by means of the Burjie's bacteria identification manual by utilizing various sugars in the culture medium, after the strain was cultured in a glucose, galactose, lactose, fructose, cellobiose, maltose, sucrose, mannitol, xylose medium at 30℃for 24 hours, and the culture medium was turbid. The bacterial DNA is extracted, the 16srDNA sequence is determined, and is input into NCBI database for comparison, and the bacterial is identified as lactobacillus plantarum by combining with physiological and biochemical properties, and is consistent with the physiological and biochemical experimental results of the bacterial strain. Lactobacillus plantarum (Lactobacillus plantarum) F25-4 was deposited at China Center for Type Culture Collection (CCTCC) with the accession number of CCTCC No. M20211574 at the year 2021, month 12 and 9, china center for type culture collection address: no. 299 is eight paths in Wuchang district of Wuhan, hubei province.
4. Application of lactobacillus plantarum in yellow wine brewing
4.1 expanded culture of Strain F25-4
Strain F25-4 slant strain 1 was inoculated in a loop in 18 x 180 tubes containing 15mL of activated liquid medium and incubated at 37 ℃ for 36h. The 15mL culture was inoculated into a 1L flask containing 800mL of seed culture medium 1, and the culture was allowed to stand at 37℃for 24 hours. The culture was inoculated into a fermenter containing 10L of seed medium 2, and subjected to stationary culture at 37℃for 24 hours to obtain a seed culture.
4.2 application of Strain F25-4 in rice soaking
The 10L culture of strain F25-4 (culture concentration 10≡8 CFU/mL) is added into a rice soaking tank filled with 1t of rice and 1.5t of water, mixed and stirred evenly, and soaked for 36h at 40 ℃.
4.3 application of Rice slurry inoculated with strain F25-4 in yellow wine fermentation
After the rice soaking is finished, separating rice pulp water from rice by a sieve plate, introducing steam of 0.1-0.2 MPa into the soaked rice, controlling the temperature to be about 105 ℃, steaming for about 15min, adding water for cooling to about 40 ℃, dropping into a tank, adding sterilized rice pulp water, adding Angel active dried distiller's yeast and distiller's yeast (Shaoxing wine flavor) activated by 2% glucose solution, stirring uniformly, and starting fermentation. The fermentation period is 0-5 days, stirring is required every 4-5 hours, the temperature is controlled at 25-32 ℃, the fermentation period is 6-18 days, the temperature is controlled at 18-25 ℃, and the total fermentation is carried out for 15-18 days.
The rice soaking process for brewing yellow wine has the following beneficial effects.
(1) The acidity values of rice and mash can be quickly reduced, the bacteria can be inhibited, the cooking characteristics of soaked rice can be improved, and the rice soaking time can be shortened;
(2) The strain does not produce amino acid decarboxylase in metabolism, and the biogenic amine content of rice milk 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 rice milk has an effect of improving the flavor of yellow rice wine.
Inoculating strain F25-4 to reduce acidity value of rice and mash, and reducing biogenic amine content of rice slurry.
The pH of the rice milk was about 3.50 at 36h and the titratable acid content was 3.79g/L. Wherein the water absorption rate is related to acidity, and the larger the acidity is, the higher the water absorption rate is. The biogenic amine content in the rice milk is also obviously reduced, and the histamine and tyramine content is reduced from 13mg/L (the rice milk without adding lactobacillus is used as a control) to 8.5mg/L. The gelatinization temperature of the soaked rice was reduced from 71.49 ℃ (control was non-inoculated rice slurry) to 45.51 ℃. The minimum cooking time of rice was reduced from 25min (control was sterile rice slurry) to 15min.
TABLE 1 Effect of Natural soaking and inoculation of Lactobacillus plantarum F25-4 soaking on the content of titratable acid in Rice slurry (g/L)
TABLE 2 influence of natural soaking and inoculation of Lactobacillus plantarum F25-4 soaking on Water absorption (%) of rice
| Soaking time | Natural soaking (control) | 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 Effect of Natural soaking and seed Strain F25-4 soaking on histamine and tyramine in Rice slurry Water
The rice-soaked cooking characteristics of the lactobacillus plantarum F25-4 bacteria liquid are analyzed as follows:
the rice soaked in different modes is steamed, the boiling time of the rice grains is recorded through a rice milling method, and the water absorption index, the expansion coefficient and the water solubility index reflecting the boiling characteristics of the rice grains are shown in the table 4, and the rice steaming time is obviously reduced after the rice is soaked by the strain F25-4, the water absorption index, the expansion coefficient and the water solubility index are obviously increased, the higher the water absorption speed of the rice grains at the same temperature in the steaming process is, the better the steaming is, 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 is higher in the same time.
TABLE 4 comparison of cooking characteristics of rice after Natural soaking and Lactobacillus plantarum F25-4 soaking
Differential Scanning Calorimetry (DSC) also reflects the gelatinisation properties of rice. Compared with natural soaking, the gelatinization temperature of rice flour is obviously reduced after 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 by rice after being soaked by adding bacteria is reduced.
TABLE 5 thermal Properties of Rice after Natural soaking and Lactobacillus plantarum F25-4 grafting soaking
The influence of lactobacillus plantarum 25-4 on the brewing quality of yellow wine is as follows.
Steaming rice soaked by strain 25-4 at 105deg.C for 15min, cooling with water, sterilizing the rest rice slurry, adding into fermentation process as feed water, adding distiller's yeast and yeast, fermenting, and adding alcohol content of 149.65kg (alcohol concentration 14.9% and volume 2875L) after 5 days, wherein total acid reaches maximum value of 3.9g/L in the fifth day. The content of residual total sugar (calculated by glucose) is 10mg/ml, the content of phenethyl alcohol in the wine base is increased, the content of higher alcohol is reduced, and the content of lipid is increased.
Table 6 effects of brewing Water on beer alcohol content (V/V, 25 ℃) with rice slurry inoculated with Lactobacillus plantarum F25-4.
TABLE 7 influence of soaking rice milk in Strain F25-4 on volatile higher alcohol flavours in yellow wine
Table 8 influence of Lactobacillus plantarum F25-4 added to soak rice milk on volatile lipid flavor in yellow wine.
Table 7 reflects the effect of the addition of Lactobacillus plantarum F25-4 soaked rice milk on higher alcohols in the flavor of yellow wine, wherein the higher alcohols content in the control group was found to be significantly reduced compared to the higher alcohols content in the group containing the scion rice milk, by 14.9% and 19.6% respectively, except for phenethyl alcohol, and the higher alcohols species were not increased. Wherein the phenethyl alcohol content is obviously increased by 137.1 percent compared with the control component, and the phenethyl alcohol beta-phenethyl alcohol is also called 2-phenethyl alcohol, is an aromatic higher alcohol with rose fragrance and honey fragrance, is an important high-boiling point fragrance component in alcoholic beverages, widely exists in various alcoholic beverages, and is an important aromatic substance in yellow wine. Isobutanol and isoamyl alcohol are the main reasons for leading up the yellow wine, after the inoculation rice milk is added, the content of the isobutanol is slightly increased, but the content of the isoamyl alcohol is reduced by 18.2%, and 2, 3-butanediol can bring the sweet taste to the yellow wine, so that the taste of the yellow wine is improved. Therefore, the addition of the bacteria-grafting rice milk can reduce the content of higher alcohol in the wine, increase the content of phenethyl alcohol and effectively reduce the upper head feeling generated by the 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 group added with the grafted rice milk is obviously higher than that of the control group, and the lipid content is respectively increased by 50 percent and 72.8 percent. The number of species is not reduced, and compared with a control group, the addition of the grafted rice milk water group is added with hexadecenoic acid ethyl ester, dibutyl adipate, dodecanoic acid ethyl ester, 9-octadecenoic acid ethyl ester, linoleic acid ethyl ester, 2-acrylic acid, 3- (4-methoxyphenyl) -2-ethylhexyl ester, and ethyl butyrate, ethyl octanoate, benzoic acid-2-ethylhexyl ester and isopropyl palmitate. Wherein, the content of isoamyl acetate is reduced by 59.3 percent, but the content of 2-ethyl phenyl acetate is increased by 4.14mg/L, and the ethyl caproate and the isoamyl acetate are reduced and the ethyl phenylacetate is increased after rice slurry water is added as feed water for fermentation.
Yellow wine sensory evaluation consisted of 10 experienced individuals in an evaluation panel (5 female, 5 male). The organoleptic characteristics of the different yellow wine samples were evaluated. In the first step, panelists first observe, taste and sniff the yellow wine to identify and record the sensory attributes contained. In the second step, the team members discuss and determine attributes, and then build the final descriptors. The color is characterized by 14 yellow wine descriptors such as appearance (redness and turbidity), aroma (bouquet, fruit aroma, flower aroma and cereal aroma), taste (sweet, sour, hot, sticky and bitter), taste (astringency, durability and saturation feeling) and the like. In the final step, 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 strong). 30mL of the prepared yellow wine samples were placed in the same opaque disposable plastic cup, subjected to good swirling treatment, and then the samples were evaluated for randomization. The assay was repeated three times per sample.
Sensory evaluation is represented by a sensory radar chart, and the result is shown in figure 4, and the characteristics of bouquet, persistence, color, flower fragrance, acidity and the like of the yellow wine fermentation added with the grafted rice milk are obviously higher than those of the control group. The turbidity, cereal aroma, bitter taste, astringency, richness and pungency of the yellow wine of the control group are higher than those of the rice milk water group added with the inoculation, which is probably due to the combined effect of amino acid and flavor substances in the yellow wine, and more acids and bitter amino acid with higher specific gravity can influence the sour taste and bitter taste and flavor development quality of the yellow wine. Yellow wine of the group added with the grafted rice milk is slightly better than that of the control group.
Compared with the traditional yellow wine brewing, the yellow wine brewed by adding the rice slurry soaked by the lactobacillus plantarum F25-4 can quickly improve the acidity of the rice slurry, shorten the rice soaking time, inhibit the growth of miscellaneous bacteria, improve the stability of the rice soaking process, effectively reduce the mass concentration of biogenic amine in the rice slurry, and ensure that the microbial acid produced in the rice soaking process is attached to rice to provide an acidic environment for the fermentation of the yellow wine, so that the fermentation can be normally carried out. The lactobacillus plantarum F25-4 improves the cooking property after being soaked, reduces the steam consumption and reduces the energy consumption. The rice milk is added into the fermentation process, so that the content of phenethyl alcohol in the product is increased, the content of isoamyl alcohol is reduced, the 'top feeling' after drinking wine is reduced, and the recycling of the rice milk reduces the treatment capacity of wastewater and further reduces the cost.
Example 2: lactobacillus plantarum and application thereof in yellow wine brewing
Lactobacillus plantarum (Lactobacillus plantarum) F25-4 is preserved in China Center for Type Culture Collection (CCTCC) on 12 th month 9 of 2021, and the preservation number is CCTCC No. M20211574.
Application of Lactobacillus plantarum (Lactobacillus plantarum) F25-4 in yellow wine brewing is provided.
The use according to claim 2, characterized in that: comprises the following steps:
step 1: inoculating lactobacillus plantarum F25-4 inclined plane strain to an activation culture medium, performing stationary culture at 36 ℃ for 30 hours, then inoculating the obtained culture to a first seed culture medium, performing stationary culture at 36 ℃ for 20 hours, then inoculating the obtained culture to a second seed culture medium, performing stationary culture at 36 ℃ for 20 hours, and obtaining a seed culture;
step 2: adding seed culture into a rice soaking tank filled with rice and water, mixing, stirring, and soaking rice.
The seed culture had a concentration of 1X 10 8 CFU/mL, seed cultures were added to a rice-dipping tank containing rice and water at 0.3% inoculum size.
In 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, 2g of glucose and sterilizing at 121 ℃ for 15min; the preparation method of the rice milk comprises the following steps: soaking semen oryzae Sativae at 40deg.C at a ratio of 1:1 for 1 hr, and sieving with 10 mesh sieve to obtain rice slurry.
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, 0.3g of yeast powder and 15min of sterilization at 121 ℃; the preparation method of the rice milk comprises the following steps: soaking semen oryzae Sativae at 40deg.C at a ratio of 1:1 for 1 hr, and sieving with 10 mesh sieve to obtain rice slurry.
The method also comprises the following steps: after the rice soaking is completed, separating to obtain rice slurry water and soaked rice, then introducing 0.1MPa steam into the soaked rice, controlling the temperature to be 102 ℃, steaming and boiling for 12min, adding water to cool to 35 ℃, adding sterilized rice slurry water after dropping into a tank, adding activated Angel active dry yeast and Shaoxing wine flavor brewing yeast, and starting fermentation after stirring uniformly; the fermentation period is 0-5 days, stirring is required every 4 hours, the temperature is controlled at 25 ℃, the fermentation period is 6 days, and the temperature is controlled at 18 ℃.
Activation medium: 0.5g of tryptone, 0.5g of yeast powder, 0.5g of beef extract, 0.2g of citric acid diamine, 0.25g of sodium chloride and 0.05g,Tween 80 0.1g,MnSO g of glucose 4 0.005g，MgSO 4 0.02g，CaCO 3 0.01g，K 2 HPO 4 0.2g, FeSO 4 0.004g, distilled water 100mL, pH5.3-5.5, and sterilized at 121 ℃ for 20min.
Example 3: lactobacillus plantarum and application thereof in yellow wine brewing
Lactobacillus plantarum (Lactobacillus plantarum) F25-4 is preserved in China Center for Type Culture Collection (CCTCC) on 12 th month 9 of 2021, and the preservation number is CCTCC No. M20211574.
Application of Lactobacillus plantarum (Lactobacillus plantarum) F25-4 in yellow wine brewing is provided.
The preferable technical scheme is as follows: comprises the following steps:
step 1: inoculating lactobacillus plantarum F25-4 inclined plane strain to an activation culture medium, performing stationary culture at 38 ℃ for 40 hours, inoculating the obtained culture to a first seed culture medium according to an inoculum size of 1%, performing stationary culture at 38 ℃ for 28 hours, inoculating the obtained culture to a second seed culture medium according to an inoculum size of 1%, and performing stationary culture at 38 ℃ for 28 hours to obtain a seed culture;
step 2: inoculating seed culture in rice soaking tank containing rice and water at an inoculum size of 1%, mixing, stirring, and soaking rice.
The preferred embodiments are: the seed culture had a concentration of 9X 10 8 CFU/mL。
The preferred embodiments are: in step 2, the temperature is 45 ℃ and the time is 40h.
The preferred embodiments are: 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, 2g of glucose and sterilizing at 121 ℃ for 15min; the preparation method of the rice milk comprises the following steps: soaking semen oryzae Sativae at 40deg.C at a ratio of 1:1 for 1 hr, and sieving with 10 mesh sieve to obtain rice slurry.
The preferred embodiments are: 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, 0.3g of yeast powder and 15min of sterilization at 121 ℃; the preparation method of the rice milk comprises the following steps: soaking semen oryzae Sativae at 40deg.C at a ratio of 1:1 for 1 hr, and sieving with 10 mesh sieve to obtain rice slurry.
The preferred embodiments are: the method also comprises the following steps: after the rice soaking is completed, separating to obtain rice slurry and soaked rice, then introducing 0.2MPa steam into the soaked rice, controlling the temperature to be 107 ℃, steaming and boiling for 17min, adding water to cool to 45 ℃, adding sterilized (sterilizing mode is boiling for 15 min) rice slurry after dropping into a tank, adding Angel active dry distiller's yeast, and starting fermentation after stirring uniformly; the fermentation period is 5 days, stirring is required every 5 hours, the temperature is controlled at 32 ℃, the fermentation period is 18 days, and the temperature is controlled at 25 ℃.
The preferred embodiments are: and adding Angel active dry yeast and Angel Shaoxing wine flavor brewing yeast.
The preferred embodiments are: activation medium: 0.5g of tryptone, 0.5g of yeast powder, 0.5g of beef extract, 0.2g of citric acid diamine, 0.25g of sodium chloride and 0.05g,Tween 80 0.1g,MnSO g of glucose 4 0.005g，MgSO 4 0.02g，CaCO 3 0.01g，K 2 HPO 4 0.2g,FeSO 4 0.004g, distilled water 100mL, pH5.3-5.5, and sterilized at 121 ℃ for 20min.
16S rDNA gene sequence:
the foregoing description of the preferred embodiment of the invention is not intended to be limiting in any way, but rather, it is intended to cover all modifications or variations of the invention which fall within the spirit and scope of the invention.
SEQUENCE LISTING
<110> university of synthetic fertilizer industry
<120> Lactobacillus plantarum strain and application thereof in yellow wine brewing
<140> 2022102751442
<141> 2022-03-21
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1468
<212> DNA
<213> 16S rDNA Gene sequence
<400> 1
cgtacctgtg ctataatgca gtcgaacgaa ctctggtatt gattggtgct tgcatcatga 60
tttacatttg agtgagtggc gaactggtga gtaacacgtg ggaaacctgc ccagaagcgg 120
gggataacac ctggaaacag atgctaatac cgcataacaa cttggaccgc atggtccgag 180
cttgaaagat ggcttcggct atcacttttg gatggtcccg cggcgtatta gctagatggt 240
ggggtaacgg ctcaccatgg caatgatacg tagccgacct gagagggtaa tcggccacat 300
tgggactgag acacggccca aactcctacg ggaggcagca gtagggaatc ttccacaatg 360
gacgaaagtc tgatggagca acgccgcgtg agtgaagaag ggtttcggct cgtaaaactc 420
tgttgttaaa gaagaacata tctgagagta actgttcagg tattgacggt atttaaccag 480
aaagccacgg ctaactacgt gccagcagcc gcggtaatac gtaggtggca agcgttgtcc 540
ggatttattg ggcgtaaagc gagcgcaggc ggttttttaa gtctgatgtg aaagccttcg 600
gctcaaccga agaagtgcat cggaaactgg gaaacttgag tgcagaagag gacagtggaa 660
ctccatgtgt agcggtgaaa tgcgtagata tatggaagaa caccagtggc gaaggcggct 720
gtctggtctg taactgacgc tgaggctcga aagtatgggt agcaaacagg attagatacc 780
ctggtagtcc ataccgtaaa cgatgaatgc taagtgttgg agggtttccg cccttcagtg 840
ctgcagctaa cgcattaagc attccgcctg gggagtacgg ccgcaaggct gaaactcaaa 900
ggaattgacg ggggcccgca caagcggtgg agcatgtggt ttaattcgaa gctacgcgaa 960
gaaccttacc aggtcttgac atactatgca aatctaagag attagacgtt cccttcgggg 1020
acatggatac aggtggtgca tggttgtcgt cagctcgtgt cgtgagatgt tgggttaagt 1080
cccgcaacga gcgcaaccct tattatcagt tgccagcatt aagttgggca ctctggtgag 1140
actgccggtg acaaaccgga ggaaggtggg gatgacgtca aatcatcatg ccccttatga 1200
cctgggctac acacgtgcta caatggatgg tacaacgagt tgcgaactcg cgagagtaag 1260
ctaatctctt aaagccattc tcagttcgga ttgtaggctg caactcgcct acatgaagtc 1320
ggaatcgcta gtaatcgcgg atcagcatgc cgcggtgaat acgttcccgg gccttgtaca 1380
caccgcccgt cacaccatga gagtttgtaa cacccaaagt cggtggggta accttttagg 1440
aaccagccgc ctaaggtcac aggagatg 1468
Claims (9)
1. Lactobacillus plantarum strainLactobacillus plantarum) F25-4 is preserved in China Center for Type Culture Collection (CCTCC) at the 12 th month 9 of 2021, and the preservation number is CCTCC No. M20211574.
2. Lactobacillus plantarum strainLactobacillus plantarum) The application of F25-4 in yellow wine brewing is characterized in that: the lactobacillus plantarum is [ ]Lactobacillus plantarum) F25-4 is preserved in China Center for Type Culture Collection (CCTCC) at the 12 th month 9 of 2021, and the preservation number is CCTCC No. M20211574.
3. The use according to claim 2, characterized in that: comprises the following steps:
step 1: inoculating lactobacillus plantarum F25-4 inclined plane strain to an activation culture medium, performing stationary culture at 36-38 ℃ for 30-40h, then inoculating the obtained culture to a first seed culture medium, performing stationary culture at 36-38 ℃ for 20-28h, then inoculating the obtained culture to a second seed culture medium, performing stationary culture at 36-38 ℃ for 20-28h, and obtaining a seed culture;
step 2: adding seed culture into a rice soaking tank filled with rice and water, mixing, stirring, and soaking rice.
4. A use according to claim 3, characterized in that: the seed culture concentration is 1-9×10 8 CFU/mL, seed cultures were added to a rice-dipping tank containing rice and water at an inoculum size of 0.3-1%.
5. A use according to claim 3, characterized in that: in the step 2, the temperature is 35-45 ℃ and the time is 30-40h.
6. A use according to claim 3, 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, 2g of glucose and sterilizing at 121 ℃ for 15min; the preparation method of the rice milk comprises the following steps: soaking semen oryzae Sativae at 40deg.C at a ratio of 1:1 for 1 hr, and sieving with 10 mesh sieve to obtain rice slurry.
7. A use according to claim 3, 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, 0.3g of yeast powder and 15min of sterilization at 121 ℃; the preparation method of the rice milk comprises the following steps: soaking semen oryzae Sativae at 40deg.C at a ratio of 1:1 for 1 hr, and sieving with 10 mesh sieve to obtain rice slurry.
8. A use according to claim 3, characterized in that: the method also comprises the following steps: after the rice soaking is completed, separating to obtain rice slurry and soaked rice, then introducing steam of 0.1-0.2 MPa into the soaked rice, controlling the temperature to be 102-107 ℃, steaming for 12-17min, adding water to cool to 35-45 ℃, adding sterilized rice slurry after dropping into a tank, adding activated Angel active dry yeast and Shaoxing wine flavor brewing yeast, and starting fermentation after stirring uniformly; the fermentation period is 0-5 days, stirring is required every 4-5 hours, the temperature is controlled at 25-32 ℃, the fermentation period is 6-18 days, and the temperature is controlled at 18-25 ℃.
9. A use according to claim 3, characterized in that: activation medium: 0.5g of tryptone, 0.5g of yeast powder, 0.5g of beef extract, 0.2g of citric acid diamine, 0.25g of sodium chloride and 0.05g,Tween 80 0.1g,MnSO g of glucose 4 0.005g，MgSO 4 0.02g，CaCO 3 0.01g，K 2 HPO 4 0.2g, FeSO 4 0.004g, distilled water 100mL, pH5.3-5.5, and sterilization at 121 ℃ for 20min.
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