High-temperature-resistant and high-fermentation-activity lactic acid bacteria and application thereof
By promoting the fermentation of highland barley rice wine with a multi-strain mixed fermentation agent, the problems of lack of aroma and unstable quality of rice wine caused by single-strain fermentation have been solved, and the production of highland barley rice wine with rich flavor and refreshing taste has been achieved.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- CHENGDU UNIV
- Filing Date
- 2024-07-19
- Publication Date
- 2026-05-22
Smart Images
Figure CN118652818B_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of fermentation and brewing technology, specifically to a highland barley rice wine fermentation agent, its preparation method, and its application. Background Technology
[0002] Barley wine, a representative of traditional brewed wines, boasts a long history and profound cultural heritage. It is made primarily from barley and glutinous rice, through a brewing process involving steaming, saccharification, and fermentation. It has a mellow and fragrant taste with a unique flavor. It is believed to have benefits such as invigorating qi and blood, promoting blood circulation, strengthening immunity, and aiding digestion.
[0003] Fermentation promotes the digestibility, nutritional value, and flavor formation of rice wine. The taste of rice wine is mainly composed of sugars, organic acids, amino acids, and polypeptides. Its aroma primarily originates from the raw materials and fermentation process, while the final flavor and quality are determined by a combination of factors, including the raw materials, production techniques, and fermentation agents. Currently, research on rice wine mainly focuses on its raw materials, yeast starter, and fermentation processes.
[0004] Yellow rice wine is typically brewed using wheat koji. Wheat koji, a saccharifying agent, is made from raw wheat and inoculated microorganisms. Fermentation with a single strain may result in yellow rice wine lacking aroma, failing to produce the multiple flavors required for certain types of yellow rice wine, such as ester and alcohol aromas, and also has disadvantages such as long fermentation time and unstable quality. Summary of the Invention
[0005] The purpose of this application is to provide a highland barley rice wine fermentation agent, its preparation method and application. The fermentation agent is prepared by mixing multiple strains of bacteria and is used for highland barley rice wine fermentation. It can effectively promote the rice wine fermentation process, improve brewing efficiency, and at the same time improve the flavor and quality of the highland barley rice wine product.
[0006] The technical solution of this application is as follows:
[0007] On one hand, embodiments of this application provide a highland barley rice wine fermentation agent, which is prepared from a suspension of microorganisms including the following types:
[0008] Mold suspension, yeast suspension, non-Saccharomyces cerevisiae suspension, and lactic acid bacteria suspension;
[0009] The aforementioned mold is Rhizopus arrhizus G01, with the accession number CGMCCNO.40839.
[0010] The yeast strain mentioned above is Saccharomyces cerevisiae XDN2, with the preservation number CGMCC NO.30936.
[0011] The aforementioned non-Saccharomyces cerevisiae is Pichiakudriavzevii XDB1, with the preservation number CGMCC NO.30935;
[0012] The lactic acid bacteria mentioned above is Lactobacillus plantarum, with the Latin name Lactobacillus plantarum and the accession number CGMCC NO.1.3919.
[0013] Furthermore, in some embodiments of this application, the weight ratio of the above-mentioned fermenting agent to the mold suspension, yeast suspension, non-Saccharomyces cerevisiae suspension and lactic acid bacteria suspension is (3-5):(1-3):(0.5-2):(0.5-2).
[0014] Furthermore, in some embodiments of this application, the weight ratio of the mold suspension, yeast suspension, non-Saccharomyces cerevisiae suspension and lactic acid bacteria suspension in the above-mentioned fermentation agent is 4:1:1:2.
[0015] This application also provides a method for preparing a highland barley rice wine fermentation agent, which includes the following steps:
[0016] Preparation of mold suspension: Rhizopus spp. was inoculated onto PDA solid medium and cultured in a constant temperature incubator. The spores were then washed with sterile water to prepare a spore suspension, thus obtaining the mold suspension.
[0017] Preparation of yeast suspension: Saccharomyces cerevisiae was cultured in liquid form. A loopful was picked onto PDA liquid medium and cultured in a constant temperature shaker. After centrifugation to remove the supernatant, the cells were washed with PBS buffer and then sterile water was added to prepare the yeast suspension.
[0018] Preparation of non-Saccharomyces cerevisiae suspension: The yeast cells were cultured in liquid medium, a loopful was picked and placed on PDA liquid medium, cultured in a constant temperature shaker, centrifuged to remove the supernatant, washed with PBS buffer, and then sterile water was added to prepare the yeast suspension.
[0019] Preparation of lactic acid bacteria suspension: Lactobacillus plantarum was cultured in MRS liquid medium, then the supernatant was removed by centrifugation, the cells were washed with PBS buffer, and then sterile water was added to prepare lactic acid bacteria suspension.
[0020] Fermentation agent preparation: The prepared mold suspension, yeast suspension, non-Saccharomyces yeast suspension and lactic acid bacteria suspension are mixed in proportion to prepare the finished fermentation agent.
[0021] Furthermore, in some embodiments of this application, in the above-mentioned mold suspension preparation step, the mold is cultured in a constant temperature incubator at 30°C for 72 hours; the concentration of the above-mentioned mold suspension is 10.6 CFU / mL.
[0022] Furthermore, in some embodiments of this application, in the above-mentioned yeast suspension preparation step, the yeast is cultured in a constant temperature shaker at 30°C and 150 r / min for 48 h, and then centrifuged at 8000 rpm for 10 min to remove the supernatant; the concentration of the above-mentioned yeast suspension is 10. 6 CFU / mL.
[0023] Furthermore, in some embodiments of this application, in the above-mentioned non-Saccharomyces cerevisiae suspension preparation step, the culture is carried out at 30°C and 150 r / min in a constant temperature shaker for 48 h, and then centrifuged at 8000 rpm for 10 min to remove the supernatant; the concentration of the above-mentioned non-Saccharomyces cerevisiae suspension is 10. 6 CFU / mL.
[0024] Furthermore, in some embodiments of this application, in the above-mentioned lactic acid bacteria suspension preparation step, *Lactobacillus plantarum* is cultured in MRS liquid medium at 37°C for 48 hours; the concentration of the above-mentioned lactic acid bacteria suspension is 10... 6 CFU / mL.
[0025] On the other hand, this application also provides a barley rice wine, which is prepared by fermentation using the above-mentioned barley rice wine fermentation agent.
[0026] This application also provides a method for preparing the above-mentioned highland barley rice wine, which includes the following steps:
[0027] Barley and glutinous rice are soaked in distilled water, steamed, and then spread out to cool. The barley rice wine fermentation agent mentioned above is then added, mixed, and transferred to a wine tank. Sterile water is added for fermentation, and barley rice wine is obtained after fermentation.
[0028] Compared with the prior art, the embodiments of this application have at least the following advantages or beneficial effects:
[0029] To address the above aspects, this application provides a highland barley rice wine fermentation agent, its preparation method, and its application. Specifically, it provides a highland barley rice wine fermentation agent and highland barley rice wine prepared by fermentation using the agent. The fermentation agent is prepared by mixing mold suspension, yeast suspension, non-brewing yeast suspension, and lactic acid bacteria suspension in a certain proportion. By fermenting highland barley rice wine with multiple strains, more beneficial microorganisms can be introduced, promoting the fermentation process of the rice wine. This improves the quality of the rice wine from multiple aspects such as taste, flavor, aroma, and flavor substances, ultimately resulting in a highland barley rice wine product with a moderate sweet and sour taste, a refreshing taste, and rich flavor substances. Attached Figure Description
[0030] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.
[0031] Figure 1 This is a diagram showing the detection results of the electronic tongue in Experiment Example 2 of this application. Detailed Implementation
[0032] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.
[0033] It should be noted that the term "comprising," or any other variation thereof, is intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.
[0034] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other.
[0035] The features and performance of this application will be further described in detail below with reference to embodiments.
[0036] Example 1
[0037] This embodiment provides a barley rice wine fermentation agent, which is prepared from a suspension of microorganisms including the following types:
[0038] Mold suspension, yeast suspension, non-Saccharomyces cerevisiae suspension, and lactic acid bacteria suspension;
[0039] The mold mentioned above is Rhizopus oligosporus;
[0040] The yeast strain mentioned above is *Saccharomyces cerevisiae*.
[0041] The aforementioned non-Saccharomyces cerevisiae is *Pichia kudrica*.
[0042] The lactic acid bacteria mentioned above are *Lactobacillus plantarum*.
[0043] The weight ratio of mold suspension, yeast suspension, non-Saccharomyces cerevisiae suspension and lactic acid bacteria suspension was 4:2:1:1.
[0044] Its preparation method is as follows:
[0045] Preparation of mold suspension: Rhizopus spp. was inoculated onto PDA solid medium and cultured at 30°C for 72 hours. The spores were then washed with sterile water to prepare a spore suspension with a concentration of 10%. 6 CFU / mL mold suspension;
[0046] Preparation of yeast suspension: Saccharomyces cerevisiae was cultured in liquid form. A loopful was picked and placed on PDA liquid medium and cultured in a constant temperature shaker at 30℃ and 150 rpm for 48 h. Then, the suspension was centrifuged at 8000 rpm for 10 min, the supernatant was removed, and the cells were washed twice with PBS buffer. Sterile water was then added to prepare a 10% concentration suspension. 6 CFU / mL yeast suspension;
[0047] Preparation of non-Saccharomyces cerevisiae suspension: Pichia kudrica was cultured in liquid form. A loopful was picked and placed on PDA liquid medium and cultured in a constant temperature shaker at 30℃ and 150 rpm for 48 h. Then, the suspension was centrifuged at 8000 rpm for 10 min, the supernatant was removed, and the cells were washed twice with PBS buffer. Finally, sterile water was added to prepare a suspension with a concentration of 10... 6 CFU / mL non-Saccharomyces cerevisiae suspension;
[0048] Preparation of lactic acid bacteria suspension: *Lactobacillus plantarum* was cultured in MRS liquid medium at 37°C for 48 h, then centrifuged at 8000 rpm for 10 min, the supernatant was removed, and the cells were washed twice with PBS buffer. Sterile water was then added to prepare a suspension with a concentration of 10... 6 CFU / mL lactic acid bacteria suspension;
[0049] Fermentation agent preparation: The prepared mold suspension, yeast suspension, non-Saccharomyces yeast suspension and lactic acid bacteria suspension are mixed in proportion to prepare the finished fermentation agent.
[0050] In this embodiment, the Rhizopus septemlobus used is from existing preservable materials; the Lactobacillus plantarum used is from existing commercially available materials; the Saccharomyces cerevisiae and Pichia kudrica were self-preserved by the applicant and deposited at the China General Microbiological Culture Collection Center, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with a deposit date of June 14, 2024.
[0051] Example 2
[0052] This embodiment provides a highland barley rice wine fermentation agent, which is based on Example 1. The difference between Example 1 and Example 1 is that the weight ratio of mold suspension, yeast suspension, non-brewing yeast suspension and lactic acid bacteria suspension is 5:1:1:1.
[0053] Example 3
[0054] This embodiment provides a highland barley rice wine fermentation agent, which is based on Example 1. The difference between Example 1 and Example 1 is that the weight ratio of mold suspension, yeast suspension, non-brewing yeast suspension and lactic acid bacteria suspension is 4:1:1:2.
[0055] Example 4
[0056] This embodiment provides a highland barley rice wine fermentation agent, which is based on Example 1. The difference between Example 1 and Example 1 is that the weight ratio of mold suspension, yeast suspension, non-brewing yeast suspension and lactic acid bacteria suspension is 4.5:2.5:0.5:0.5.
[0057] Example 5
[0058] This embodiment provides a type of highland barley rice wine, which is prepared by fermentation using the highland barley rice wine fermentation agent provided in Example 1. The preparation method is as follows:
[0059] Soak 150g of highland barley and glutinous rice in distilled water at a ratio of 0.5:1 for 12 hours, then steam them on a 1200W induction cooker for 35 minutes until the grains are distinct. Spread the steamed highland barley and glutinous rice on a sterilized work surface to cool to 30℃~35℃. Then, inoculate the highland barley and glutinous rice with the fermentation agent provided in Example 1 and mix thoroughly. Transfer the mixture to a wine tank, add 100mL of sterile water, and ferment at 28℃ for 5 days, then at 15℃ for 20 days. The total inoculum amount is 5% of the volume of the rice wine. Finally, filter with gauze, clarify, and pasteurize at 75℃ for 10 minutes to obtain a highland barley rice wine sample.
[0060] Example 6
[0061] This embodiment provides a barley rice wine, which is based on embodiment 5. The difference between this embodiment and embodiment 5 is that it is prepared by fermentation using the barley rice wine fermentation agent provided in embodiment 2.
[0062] Example 7
[0063] This embodiment provides a barley rice wine, which is based on embodiment 5. The difference between this embodiment and embodiment 5 is that it is prepared by fermentation using the barley rice wine fermentation agent provided in embodiment 3.
[0064] Example 8
[0065] This embodiment provides a barley rice wine, which is based on embodiment 5. The difference between this embodiment and embodiment 5 is that it is prepared by fermentation using the barley rice wine fermentation agent provided in embodiment 4.
[0066] Comparative Example 1
[0067] This comparative example provides a barley rice wine based on Example 5. The difference between Example 5 and Example 5 is that the barley rice wine fermentation agent provided in this application is not used; instead, commercially available koji is used for fermentation.
[0068] Experimental Example 1
[0069] The physicochemical properties and sensory evaluation of the highland barley rice wine provided in Examples 5-8 of this application and Comparative Example 1 are shown in Table 1.
[0070] Table 1
[0071]
[0072]
[0073] As can be seen from the comparison, the reducing sugar content of the product provided by Comparative Example 1 is significantly higher than that of the other groups, indicating that the sugar metabolism of the other groups is more thorough, and therefore the alcohol content is also higher than that of the CK group.
[0074] The difference in the proportion of starter microorganisms between Example 5 and Example 6 resulted in significantly lower reducing sugar levels, indicating improved fermentation efficiency. This reduction can be attributed to the synergistic metabolic activity of the microbial symbiont, which enhances sugar absorption and conversion.
[0075] Total acidity is crucial for the palatability and microbial stability of Shaoxing wine. Comparative Example 1 had excessively high total acidity, resulting in an unbalanced flavor, while Examples 5-8 maintained relatively stable total acidity. This reduction in acidity may be due to the introduction of lactic acid bacteria into the designed fermentation agent, which is influenced by specific metabolic pathways of these bacteria. These pathways may utilize acid in different ways, affecting the final acidity profile.
[0076] Overall, the barley rice wine of Example 7 has a moderate sweet and sour taste, moderate alcohol content, and the best taste, meeting the requirements of GB / T13662-2018 Rice Wine.
[0077] Experimental Example 2
[0078] The electronic tongue sensor, manufactured in France, consists of seven sensors: AHS-Sourcess, CTS-Saltiness, NMS-umami, ANS, SCS, PKS, and CPS. Ag / AgCl is selected as the reference electrode. 80 mL of wine is taken and placed in a special beaker for electronic tongue detection.
[0079] Samples from Comparative Example 1 and Examples 5-8 were placed in slots 4, 5, 6, 7, and 8 of the electronic tongue autosampler, respectively; slots 1, 2, and 3 contained 80 mL of distilled water. Before testing the samples, the sensor was cleaned for 120 seconds, followed by a 120-second test. Each sample was tested six times, and the stable values from the last five tests within the 100-120 second range were used as the results for analysis. The test results are as follows: Figure 1 As shown.
[0080] Figure 1 In China, the tastes are categorized as follows: AHS (sour), ANS (sweet), CTS (salty), NMS (umami), SCS (bitter), PKS (general), and CPS (general).
[0081] according to Figure 1 It can be seen that Comparative Example 1 is more intense in terms of sourness, sweetness, saltiness, and bitterness, followed by Comparative Example 7. However, Comparative Example 7 has a slightly stronger umami flavor than Comparative Example 1. Comparative Example 1 is significantly more bitter than the other four groups. The content and types of bitter compounds produced in Comparative Example 1 are significantly different from the other four groups, resulting in a less harmonious sensory experience. In addition, apart from the significant difference in saltiness intensity, the flavor profiles of Examples 5-8 via the electronic tongue are basically similar, while Example 7 exhibits better flavor harmony than the other combinations.
[0082] The results showed that the fermenting agent provided in Example 3 was the optimal fermenting agent for highland barley rice wine. The fermented highland barley rice wine (Example 7) had a moderate sweet and sour taste, a refreshing taste, a moderate alcohol content, and the highest sensory score.
[0083] Experimental Example 3
[0084] To further determine the impact of the fermentation agent on the quality of highland barley rice wine, the volatile flavor compounds in the highland barley rice wines provided in Examples 5-8 were compared. The test methods are as follows:
[0085] The volatile compounds in rice wine were quantified by gas chromatography-mass spectrometry (GC-MS, Agilent 7890) using peak area normalization.
[0086] GC conditions: Agilent 7890GC; Capillary column: DB-5 (60m×0.25mm×0.25μm, Agilent Technologies, USA); Injector temperature: 250℃; Temperature program: Initial temperature 50℃, hold for 2 min; 2℃ / min to 115℃, hold for 3 min; 4℃ / min to 200℃; 6℃ / min to 230℃, hold for 10 min; Carrier gas: He; Flow rate: 1 mL / min; Splitless.
[0087] MS conditions: Aglient 5977BMS; Ionization method: EI source; Ionization voltage: 70 eV; Multiplier voltage: 1871 eV; Mass spectrometry interface temperature: 280℃; Full ion (SCAN) and Selected ion (SIM) scan range: 50-550 amu.
[0088] The identification of volatile flavor compounds was performed by searching the NIST mass spectrometry library and combining relevant data. Substances with a matching degree greater than 85 were selected as valid volatile components. The detection results are shown in Table 2.
[0089] Table 2
[0090]
[0091]
[0092]
[0093]
[0094] The results in Table 2 show that the volatile compounds in rice wine fermented with different starter cultures consist of alcohols, esters, acids, ketones, alkanes, and other compounds, with a total of 77 volatile compounds detected.
[0095] Comparative Example 1 detected a total of 37 compounds, Example 5 detected a total of 27 compounds, Example 6 detected a total of 25 compounds, Example 7 detected a total of 39 compounds, and Example 8 detected a total of 33 compounds. Example 7 showed superior diversity in volatile flavor compounds compared to the other four examples.
[0096] Among these flavor compounds, it can be seen that the main source of aroma in highland barley rice wine is esters. Ethyl hexanoate, ethyl octanoate, ethyl decanoate, phenylethyl acetate, ethyl laurate, ethyl palmitate, ethyl linoleate, and ethyl 9-hexadecenoate are the main esters. Volatile compounds such as ethyl palmitate, phenylethanol, and phenylethyl acetate have relatively high contents in the five wine samples. Different esters have different aroma characteristics and are formed during fermentation through yeast metabolism and esterification reactions. They contribute aromas such as apple, banana, and rose, thus enhancing the complexity of the rice wine's aroma. In addition, some important compounds (such as alcohols, acids, and ketones) were also identified as major compounds. Although they only constitute a small portion, they are also important for the aroma of the wine.
[0097] From the overall perspective of flavor components, the content and quantity of flavor components in Example 7 are generally higher than those in the other four groups. This may be because the microbial activity in Example 7 is high, resulting in the metabolism of a large number of enzymes and the decomposition of more and more complex flavor substances.
[0098] Overall, the barley rice wine (Example 7) obtained by fermentation with the fermenting agent provided in Example 3 has a harmonious taste and complex flavor, which meets the requirements of GB / T 13662-2018 Rice Wine.
[0099] In summary, this application provides a highland barley rice wine fermentation agent, its preparation method, and its application. The fermentation agent is prepared by mixing mold suspension, yeast suspension, non-brewing yeast suspension, and lactic acid bacteria suspension in a certain proportion. By fermenting highland barley rice wine with multiple strains, more beneficial microorganisms can be introduced to promote the fermentation process of rice wine. This improves the quality of the rice wine from multiple aspects such as taste, flavor, aroma, and flavor substances, ultimately resulting in a highland barley rice wine product with a moderate sweet and sour taste, a refreshing taste, and rich flavor substances.
[0100] The embodiments described above are some, but not all, of the embodiments of this application. The detailed description of the embodiments of this application is not intended to limit the scope of the claimed application, but merely to illustrate selected embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application.
Claims
1. A highland barley rice wine fermentation agent, characterized in that, It is prepared from a suspension of the following types of microorganisms: Mold suspension, yeast suspension, non-Saccharomyces cerevisiae suspension, and lactic acid bacteria suspension; The mold in question is *Rhizopus oligosporus*, scientifically known as *Rhizopus septemlobus*. Rhizopus arrhizus G01, with accession number CGMCCNO.40839; The yeast mentioned is *Saccharomyces cerevisiae*, scientifically known as *Saccharomyces cerevisiae*. Saccharomyces cerevisiae XDN2, accession number CGMCC NO.30936; The non-Saccharomyces cerevisiae strain is *Kudriazwibichthys*, scientifically known as *Kudriazwibichthys*. Pichia kudriavzevii XDB1, accession number CGMCC NO.30935; The lactic acid bacteria mentioned is *Lactobacillus plantarum*, scientifically known as Lactobacillus. Lactobacillus plantarum The accession number is CGMCCNO.1.3919; In the fermentation agent, the weight ratio of mold suspension, yeast suspension, non-Saccharomyces cerevisiae suspension and lactic acid bacteria suspension is 3-5:1-3:0.5-2:0.5-2.
2. The barley rice wine fermentation agent according to claim 1, characterized in that, The fermentation agent contains mold suspension, yeast suspension, non-Saccharomyces cerevisiae suspension and lactic acid bacteria suspension in a weight ratio of 4:1:1:
2.
3. A method for preparing a barley rice wine fermentation agent as described in any one of claims 1 to 2, characterized in that, It includes the following steps: Preparation of mold suspension: Rhizopus spp. was inoculated onto PDA solid medium and cultured in a constant temperature incubator. The spores were then washed with sterile water to prepare a spore suspension, thus obtaining the mold suspension. Preparation of yeast suspension: Saccharomyces cerevisiae was cultured in liquid form. A loopful was picked onto PDA liquid medium and cultured in a constant temperature shaker. After centrifugation to remove the supernatant, the cells were washed with PBS buffer and then sterile water was added to prepare the yeast suspension. Preparation of non-Saccharomyces cerevisiae suspension: The yeast cells were cultured in liquid medium, a loopful was picked and placed on PDA liquid medium, cultured in a constant temperature shaker, centrifuged to remove the supernatant, washed with PBS buffer, and then sterile water was added to prepare the yeast suspension. Preparation of lactic acid bacteria suspension: Lactobacillus plantarum was cultured in MRS liquid medium, then the supernatant was removed by centrifugation, the cells were washed with PBS buffer, and then sterile water was added to prepare lactic acid bacteria suspension. Fermentation agent preparation: The prepared mold suspension, yeast suspension, non-Saccharomyces yeast suspension and lactic acid bacteria suspension are mixed in proportion to prepare the finished fermentation agent.
4. The method for preparing the barley rice wine fermentation agent according to claim 3, characterized in that, In the preparation step of the mold suspension, the mold is cultured in a constant temperature incubator at 30°C for 72 hours; the concentration of the mold suspension is 10. 6 CFU / mL.
5. The method for preparing the barley rice wine fermentation agent according to claim 3, characterized in that, In the yeast suspension preparation step, the yeast cells were cultured in a constant temperature shaker at 30℃ and 150 r / min for 48 h, and then centrifuged at 8000 rpm for 10 min to remove the supernatant; the concentration of the yeast suspension was 10. 6 CFU / mL.
6. The method for preparing the barley rice wine fermentation agent according to claim 3, characterized in that, In the preparation step of the non-Saccharomyces cerevisiae suspension, the culture is carried out at 30℃ and 150 r / min in a constant temperature shaker for 48 h, followed by centrifugation at 8000 rpm for 10 min to remove the supernatant; the concentration of the non-Saccharomyces cerevisiae suspension is 10. 6 CFU / mL.
7. The method for preparing the barley rice wine fermentation agent according to claim 3, characterized in that, In the preparation step of the lactic acid bacteria suspension, *Lactobacillus plantarum* was cultured in MRS liquid medium at 37°C for 48 h; the concentration of the lactic acid bacteria suspension was 10. 6 CFU / mL.
8. A type of highland barley rice wine, characterized in that, It is prepared by fermentation using the barley rice wine fermentation agent as described in any one of claims 1 to 2.
9. A method for preparing highland barley rice wine as described in claim 8, characterized in that, It includes the following steps: Barley and glutinous rice are soaked in distilled water, steamed, and then spread out to cool. The barley rice wine fermentation agent is then added, mixed, and transferred to a wine tank. Sterile water is added for fermentation, and barley rice wine is obtained after fermentation.