Brussell bouquet saccharomycetes BB-0104 and application thereof in sun vinegar
By screening and utilizing the metabolic characteristics of Brussels BB-0104, the problem of Sichuan vinegar not rich enough and short shelf life is solved, and the flavor and quality of Sichuan vinegar not improved, providing a solid theoretical foundation.
Patent Information
- Application Number
- CN202411557859.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-04
- Publication Date
- 2024-12-27
AI Technical Summary
The unique flavor and quality of Sichuan vinegar are difficult to effectively improve, and the existing technology is difficult to screen out key flavor microorganisms, resulting in insufficient flavor and short shelf life.
Brussels yeast BB-0104 was screened through pure culture, fermentation test and high-throughput sequencing. Its metabolic properties in the fermentation medium were used to generate volatile flavor substances, improving the quality and flavor of Sichuan vinegar.
The volatile flavor substances produced by Brussels BB-0104 during the fermentation process are the key ingredients that constitute the unique flavor of Sichuan vinegar, which significantly improves the quality and flavor of Sichuan vinegar, and screens and flavors of Sichuan vinegar. Quality improvement provides a theoretical basis.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of microbial strain protection, and in particular relates to a strain of Brettanomyces brusselsii BB-0104 and application thereof in sun-dried vinegar. Background Art
[0002] Sichuan sun-dried vinegar is a type of Sichuan bran vinegar. Its solid-state fermentation has the characteristics of "three-side fermentation in the same tank", that is, the three stages of starch saccharification, alcohol fermentation, and acetic acid fermentation are carried out in the same fermentation tank. Typical representatives of Sichuan sun-dried vinegar include Zigong Taiyuanjing sun-dried vinegar and Yibin Sipo sun-dried vinegar. Compared with other types of vinegar, sun-dried vinegar has the following characteristics: (1) Raw material brewing. A large number of microorganisms in the brewing process are carried into the raw materials themselves and fermented while saccharifying. (2) The medicinal koji is unique. The koji is made from a variety of Chinese medicinal materials that are both medicinal and edible. It not only provides the crude enzymes required in the brewing process, but the Chinese medicinal materials in the medicinal koji also have a certain antibacterial effect, which enriches the taste while increasing the safety of the fermentation process.
[0003] In the brewing of Sichuan sun-dried vinegar, the key substances to the flavor of sun-dried vinegar may include acetaldehyde, 4-ethylguaiacol (4-ethyl-2-methoxyphenol) and other volatile aroma components. In the brewing process of traditional fermented foods, adding an appropriate amount of acetaldehyde can make the flavor fuller and the aroma more intense; in the brewing process, acetaldehyde is produced by the degradation of starch into glucose under the action of enzymes, and then enters the aldehyde acid pathway to be deaminated and decarboxylated by amino acids. It can synergize with other substances to reconcile the aroma in vinegar and make the flavor components richer. 4-Ethylguaiacol is a compound with a unique aroma. It can add clove and smoky flavors to vinegar, thereby enhancing the overall flavor of vinegar. 4-Ethylguaiacol has an inhibitory effect on certain bacteria, which helps to maintain the sanitary quality of vinegar and extend its shelf life to a certain extent. As a phenolic compound, 4-Ethylguaiacol has certain antioxidant properties. It can help scavenge free radicals in the body and reduce oxidative stress, thereby helping to protect cells from damage and delay the aging process. It also has antioxidant and anti-inflammatory effects, which can help enhance the body's immunity and improve the body's resistance to disease.
[0004] As a type of Sichuan bran vinegar, the flavor of Sichuan sun-dried vinegar comes from the rich microorganisms during fermentation. Therefore, obtaining the microorganisms that play a key role in the flavor of Sichuan sun-dried vinegar is crucial to further improving the quality and flavor of Sichuan sun-dried vinegar. Summary of the invention
[0005] In view of this, the object of the present invention is to provide a strain of Brettanomyces brusselsii BB-0104 and its application in sun-dried vinegar. The present invention utilizes the metabolic characteristics of microorganisms to screen out microorganisms that produce target flavor substances through pure culture, fermentation test and other methods. The microbial community is analyzed through high-throughput sequencing, metagenomics and other technologies to further screen out the key flavor microorganism Brettanomyces brusselsii BB-0104. The fermentation medium is fermented using the Brettanomyces brusselsii BB-0104 in the present invention. The results show that the volatile flavor substances produced by Brettanomyces brusselsii BB-0104 during the fermentation process are the key components of the unique flavor of Sichuan sun-dried vinegar, and can be used to improve the quality and flavor of Sichuan sun-dried vinegar.
[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:
[0007] The present invention provides a strain of Brussels Brettanomyces bruxellensis BB-0104, wherein the Brussels Brettanomyces BB-0104 was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration on July 26, 2024, with a deposit number of CGMCC NO.31477.
[0008] Preferably, the ITS sequence of the Brettanomyces brusselsii strain BB-0104 is as shown in SEQ ID NO.1.
[0009] The present invention provides a fermentation method of Brettanomyces brusselsii BB-0104, comprising the following steps:
[0010] The brussels yeast BB-0104 was inoculated into the fermentation medium for fermentation.
[0011] Preferably, the method for preparing the fermentation medium is: mixing rice with water, sequentially subjecting the mixture to gelatinization, liquefaction and saccharification to obtain a primary fermentation medium, adjusting the pH of the primary fermentation medium to 2 to 5.5 and the ethanol concentration to 1% to 12% to obtain a fermentation medium;
[0012] The mass ratio of rice to water is 1:3-10;
[0013] The gelatinization temperature is 90-100°C, and the gelatinization time is 20-40 minutes;
[0014] The liquefaction treatment uses a thermostable α-amylase and is liquefied at 80 to 100° C. for 30 to 90 minutes;
[0015] The liquefied product is cooled to 50-70° C., and saccharifying enzyme is added for saccharification treatment. The saccharification treatment time is 1-3 hours.
[0016] Preferably, the fermentation temperature is 20-44° C., and the fermentation time is 3-8 days.
[0017] The invention provides a fermentation liquid containing the Brettanomyces brusselsii BB-0104.
[0018] The present invention also provides application of the Brettanomyces brusselsii BB-0104 or the fermentation liquid in producing solar-cured vinegar.
[0019] Preferably, the sun-dried vinegar is Sichuan sun-dried vinegar.
[0020] Preferably, the live bacteria concentration of the Brussels yeast strain BB-0104 used in the production of sun-cured vinegar is 5.2×10 5 ~2×10 7 CFU / mL.
[0021] Preferably, the inoculation amount of the Brettanomyces brusselsii BB-0104 strain is 0.5% to 3%.
[0022] Compared with the prior art, the present invention has the following beneficial effects: the present invention utilizes the metabolic characteristics of microorganisms to screen out microorganisms that produce target flavor substances through pure culture, fermentation test and other methods. The microbial community is analyzed by high-throughput sequencing, metagenomics and other technologies, and the key flavor microorganism of Sichuan sun-dried vinegar is further screened out as Brussels Brettanomyces BB-0104. Through purification culture technology, the screened Brussels Brettanomyces BB-0104 is rejuvenated and expanded to obtain a sufficient number of microorganisms. According to the metabolic characteristics of the simulated strains and the production conditions of the target flavor substances, the optimal culture conditions and the optimal culture medium of Brussels Brettanomyces BB-0104 are obtained. The fermentation medium is fermented with Brussels Brettanomyces BB-0104, and the results show that the volatile flavor substances produced by Brussels Brettanomyces BB-0104 during the fermentation process are the key components of the unique flavor of Sichuan sun-dried vinegar, which can be used to improve the quality and flavor of Sichuan sun-dried vinegar, and provide a solid theoretical basis for strain screening and flavor quality improvement of Sichuan sun-dried vinegar. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Figure 1 This is the colony morphology of Brettanomyces brusselsii BB-0104;
[0024] Figure 2 This is a microscopic morphology of Brettanomyces brusselsii BB-0104;
[0025] Figure 3 This is the growth curve of Brettanomyces brusselsii BB-0104;
[0026] Figure 4 The effect of temperature on Brettanomyces brusselsii BB-0104;
[0027] Figure 5 The effect of pH on Brettanomyces brusselsii BB-0104;
[0028] Figure 6 This is the effect of ethanol concentration on Brettanomyces brusselsii BB-0104.
[0029] Biological Deposit Description
[0030] Brussels Brettanomyces bruxellensis BB-0104, Latin name Brettanomyces bruxellensis, is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, deposit date: July 26, 2024, deposit number: CGMCC NO.31477. DETAILED DESCRIPTION
[0031] The present invention provides a strain of Brussels Brettanomyces bruxellensis BB-0104, wherein the Brussels Brettanomyces BB-0104 was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration on July 26, 2024, with a deposit number of CGMCC NO.31477.
[0032] In the present invention, the ITS sequence of the Brettanomyces brusselsii BB-0104 strain is shown in SEQ ID NO.1, which is as follows:
[0033] .
[0034] The present invention provides a fermentation method of Brettanomyces brusselsii BB-0104, comprising the following steps:
[0035] The brussels yeast BB-0104 was inoculated into the fermentation medium for fermentation.
[0036] In the present invention, Brettanomyces brusselsii BB-0104 is inoculated into a fermentation medium and fermented at 20-44°C for 3-8 days; the fermentation temperature is preferably 25-36°C, more preferably 28°C, and the fermentation time is preferably 4-7 days, more preferably 5 days; the preparation method of the fermentation medium is: rice and water are mixed, and primary fermentation medium is obtained by gelatinization, liquefaction and saccharification in sequence, and the pH of the primary fermentation medium is adjusted to 2-5.5, preferably 3-5, more preferably 4.5, and the ethanol concentration is 1%-12%, preferably 2%-6%, more preferably 4%, to obtain a fermentation medium. The mass ratio of rice to water is 1:3-10, preferably 1:4-7, and more preferably 1:5; the temperature of the gelatinization treatment is 90-100°C, and the time of the gelatinization treatment is 20-40min, preferably 25-35min, and more preferably 30min; the liquefaction treatment uses a high-temperature resistant α-amylase at 80-100°C, preferably 85-95°C, and more preferably 90°C, and the liquefaction treatment is 30-90min, preferably 45-75min, and more preferably 60min. -The final concentration of amylase in the culture medium system is 5-20U / g, preferably 7-15U / g, and more preferably 10U / g; the liquefied product is cooled to 50-70°C, preferably 55-65°C, and more preferably 60°C, and a saccharifying enzyme is added for saccharification treatment, wherein the final concentration of the saccharifying enzyme in the culture medium system is 100-300U / g, preferably 150-250U / g, and more preferably 200U / g, and the saccharification treatment time is 1-3h, preferably 1.5-2.5h, and more preferably 2h.
[0037] The invention provides a fermentation liquid containing the Brettanomyces brusselsii BB-0104.
[0038] In the present invention, the method for preparing the fermentation broth is: inoculating Brussels yeast BB-0104 into YPD culture medium, fermenting at 20-44°C for 4-52h, the fermentation temperature is preferably 25-36°C, more preferably 28°C, the fermentation time is preferably 12-36d, more preferably 24h, to obtain the fermentation broth.
[0039] The present invention also provides application of the Brettanomyces brusselsii BB-0104 or the fermentation liquid in producing solar-cured vinegar.
[0040] In the present invention, the sun-dried vinegar is Sichuan sun-dried vinegar; the live bacteria concentration of the Brussels yeast BB-0104 strain used in the production of sun-dried vinegar is 5.2×10 5 ~2×10 7CFU / mL, the inoculation amount is 0.5% to 3%.
[0041] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.
[0042] Preparation of yeast extract powder glucose liquid medium (YPD liquid): weigh 20 g glucose, 20 g peptone, and 10 g yeast extract powder, dissolve in 1 L water, and sterilize at 121°C for 15 min.
[0043] Preparation of yeast extract powder glucose solid medium (YPD solid): weigh 20 g glucose, 20 g peptone, 10 g yeast extract powder, and 20 g agar, dissolve in 1 L water, and sterilize at 121°C for 15 min.
[0044] Preparation of Wallerstein's modified nutrient medium (WLN-1): Weigh 10 g of glucose, 50 g of peptone, 3 g of malt extract powder, 3 g of yeast extract powder, 0.022 g of bromocresol green, 0.1 g of p-coumaric acid, and 20 g of agar, dissolve in 1 L of water, adjust the pH to 5.4, sterilize at 121°C for 15 min, cool to 60°C, and add 0.03 g of cycloheximide and 0.04 g of chloramphenicol.
[0045] The YPD liquid medium, YPD solid medium and WLN-1 medium used in the subsequent examples were all prepared according to the above method.
[0046] Example 1 Obtaining the Brettanomyces brusselsii BB-0104 strain
[0047] Take a sample of the mother liquor of sun-dried vinegar from Sichuan Taiyuanjing Vinegar Co., Ltd., weigh 30g of the mother liquor and add it into a triangular flask containing 270mL YPD liquid culture medium, shake and culture at a constant temperature of 28℃ for 24h. Dilute it in 10-fold gradient to obtain 10 -2 ~10 -6 The dilution solution was taken and the sample solution of different gradients was spread on YPD solid medium and WLN-1 medium respectively, and cultured at 28℃ for 5 days, and then streaked 5 times to obtain pure strains. The colony characteristics of Russell Brettanomyces BB-0104, such as colony morphology, size, color, texture, transparency, and edge flatness, were observed under a microscope; and the shape, size, budding, and whether there was fission phenomenon were recorded. The results are shown in Figure 1 and Figure 2 .
[0048] Depend on Figure 1 It can be seen that the colony of the Russell Brettanomyces BB-0104 strain is oval in shape, milky white, moist, shiny, spherical, with smooth edges, and smells like wine and barn, indicating that it is budding.
[0049] Example 2 Identification of Brettanomyces brusselsii BB-0104 strain
[0050] The BB-0104 strain of Brettanomyces brusselsii was identified by referring to the physiological and biochemical identification methods in the Manual of Characteristics and Identification of Yeast.
[0051] Molecular identification The DNA of Brettanomyces brusselsii BB-0104 was extracted using a fungal DNA extraction kit (purchased from Beijing Solebao Technology Co., Ltd.), and the ITS region was amplified by PCR using universal primers ITS1 and ITS4 (purchased from Beijing Solebao Technology Co., Ltd.). The PCR amplification system was 25 μL, including 1 μL of template, 1 μL of ITS1, 1 μL of ITS4, 10 μL of PremixTaq 2×, and 12 μL of ddH2O. The PCR conditions were as follows: pre-denaturation of DNA double strands at 95°C for 1 min; denaturation at 95°C for 30 s; annealing at 55°C for 30 s; extension at 72°C for 1 min, final extension at 72°C for 10 min, 30 cycles, and storage at 4°C. The PCR products were detected by agarose gel electrophoresis, and after confirming that the concentration and band size were correct, the PCR products were sent to a sequencing company (Chengdu Shenggong Bioengineering (Chengdu) Co., Ltd.) for sequencing.
[0052] The ITS sequence of Brettanomyces brusselsii BB-0104 is shown in SEQ ID NO.1:
[0053] .
[0054] The sequences were compared with the NCBI database by Blast, and the sequences with high similarity were selected to determine that the screening strain was Brettanomyces brusselsii.
[0055] Example 3 Optimal culture conditions for Brettanomyces brusselsii BB-0104 strain
[0056] One ring of the strain obtained in Example 1 was picked and inoculated into YPD liquid culture medium, and fermented and cultured for 24 h at 28° C. and a rotation speed of 180 r / min in a shaking incubator to obtain a fermentation liquid (seed liquid).
[0057] Dilution: Use a hemocytometer to count the number of viable bacteria in the seed solution and dilute the seed solution with sterile saline until the number of viable bacteria is 5.2×10 5 ~2×10 7 CFU / mL, and the diluted seed solution was obtained for subsequent inoculation.
[0058] 3.1 Growth curve of Brettanomyces brusselsii BB-0104 strain
[0059] The diluted seed liquid was inoculated into 100 mL YPD liquid medium at 1% of the volume of the medium, and cultured in a shaking incubator at 28°C and 180 r / min. The number of viable bacteria in the medium was detected every 4 hours to observe the growth of the Brussels yeast strain BB-0104. The results are shown in Figure 3 .
[0060] Depend on Figure 3 It can be seen that during the culture period of 0 to 12 hours, the number of Brussels yeast BB-0104 increased slowly, and the strain growth was in the lag phase; during the culture period of 12 to 28 hours, the number of Brussels yeast BB-0104 increased exponentially, the strain entered the logarithmic growth phase, and the exponential growth was fastest at 24 hours; during the period of 28 to 36 hours, the yeast number tended to stabilize and entered the stable phase; after 36 hours of culture, the yeast strain entered the decline phase.
[0061] 3.2 Effect of temperature on Brettanomyces brusselsii BB-0104 strain
[0062] The diluted seed solution was inoculated into YPD liquid medium at an inoculation rate of 1% of the volume of the medium, and cultured at different temperature gradients: the temperature gradient was set at 28, 32, 36, 40, and 44°C, and three groups were set in parallel for each gradient. The OD600 of the bacterial suspension was measured. The results are shown in Figure 4 .
[0063] Depend on Figure 4 It can be seen that when the temperature is not higher than 40℃, the strain maintains a high activity, indicating that the Brussels yeast BB-0104 strain has good tolerance to high temperature. In the early stage of solid-state fermentation of Sichuan sun-dried vinegar, the microbial metabolism is active and the temperature fluctuates between 28 and 44℃. The Brussels yeast BB-0104 strain can be used to prepare Sichuan sun-dried vinegar.
[0064] 3.3 Effect of pH on Brettanomyces brusselsii BB-0104 strain
[0065] The diluted seed liquid was inoculated into YPD liquid medium at an inoculum volume of 1% of the volume of the medium, and cultured at different pH gradients: the pH gradient was set to 2, 2.5, 3, 3.5, 4, 4.5, and 5. Three groups of parallels were set for each gradient, and the OD600 value of the bacterial suspension was measured. The results are shown in Figure 5 .
[0066] Depend on Figure 5 It can be seen that the BB-0104 strain of Brussels yeast can hardly grow under the condition of pH 2, but can maintain good biological activity in an environment with a pH greater than 2.5, and the optimal growth condition is a weakly acidic environment with a pH of 4.5, indicating that the BB-0104 strain of Brussels yeast has strong acid resistance and good adaptability to the pH environment of Sichuan sun-dried vinegar.
[0067] 3.4 Effect of ethanol concentration on Brettanomyces brusselsii BB-0104 strain
[0068] The diluted seed solution was inoculated into YPD liquid medium at an inoculation rate of 1% of the volume of the medium, and cultured at different ethanol concentration gradients: the ethanol concentration gradient was set to 0, 2%, 4%, 6%, 8%, 10%, and 12%, and three groups were set in parallel for each gradient. The OD600 value of the bacterial suspension was measured. The results are shown in Figure 6 .
[0069] Depend on Figure 6 It can be seen that the BB-0104 strain of Brettanomyces brusselsii still showed good tolerance in an environment with an ethanol concentration of 12%. When the ethanol concentration was 4%, the biomass of the BB-0104 strain of Brettanomyces brusselsii reached the best.
[0070] From the above results, it can be seen that the optimal culture conditions of Brussels yeast BB-0104 strain are 28°C, pH 4.5, and ethanol concentration of 4%, and the biomass reaches the maximum after 32 hours of culture.
[0071] Effect example
[0072] Preparation of primary fermentation medium: Mix rice and water in a mass ratio of 1:5, heat the mixture to 100°C, and gelatinize it. After gelatinization for 30 minutes, when the temperature drops to 90°C, add high-temperature resistant α-amylase with a final concentration of 10U / g, and liquefy it for 1 hour. After liquefaction, cool it to 60°C, add saccharifying enzyme with a final concentration of 200U / g, saccharify it for 2 hours, adjust the sugar content to 13°Bx, and obtain the primary fermentation medium.
[0073] Preparation of fermentation medium: The pH of the primary fermentation medium was adjusted to 4.5, and the final ethanol concentration was adjusted to 4%, thereby obtaining the fermentation medium.
[0074] The concentration of 1×10 7 CFU / mL Brussels yeast BB-0104 seed liquid was inoculated into the primary fermentation medium and the fermentation medium at an inoculation amount of 2% of the culture medium volume, respectively, and cultured at 28°C for 5 days. The volatile flavor substances in the obtained products were detected. The test results are shown in Table 1.
[0075] Detection of volatile flavor substances: The volatile flavor substances of the fermentation broth product were determined by headspace solid phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS). 5 mL of fermentation broth and 2 g of NaCl were placed in a headspace bottle, 10 μL of internal standard (methyl octanoate 0.8775 g / L) was added, and after preheating at 50°C for 30 min, the extraction needle was inserted into the headspace bottle for extraction for 30 min. After the extraction, the headspace fiber was inserted into the gas chromatography injection port and adsorbed at 230°C for 3 min. Each sample was repeated 3 times. Chromatographic (GC) and mass spectrometry (MS) conditions: starting temperature 35°C, maintained for 1 min, increased to 130°C at a rate of 5°C / min, maintained for 1 min, then increased to 180°C at 3°C / min, maintained for 1 min, and finally increased to 230°C at 8°C / min and maintained for 1 min. The ion source was 230°C, the MS quadrupole was 150°C, the solvent delay was 3 min, and the acquisition mode was full scan. The test results were processed through the NIST05 spectral library and manual search, and compounds with a matching degree ≥ 80% were selected for qualitative analysis; each component was semi-quantitatively analyzed using the internal standard method, and the mass concentration of the corresponding component was calculated and identified.
[0076] Table 1 Determination of volatile flavor substances in fermentation products of different culture media
[0077]
[0078] As shown in Table 1, 15 volatile flavor substances were detected in the fermentation broth of Brettanomyces brusselsii BB-0104, including 3 alcohols, 3 aldehydes, 2 acids, 5 esters and 2 phenols. By comparing the primary fermentation medium and the fermentation medium, the effect of fermentation of Brettanomyces brusselsii BB-0104 in different culture media on volatile flavor substances was explored. The results showed that the contents of phenylacetaldehyde, acetaldehyde, benzaldehyde and 4-ethyl-2-methoxyphenol were significantly increased in the fermentation medium. The volatile flavor substances produced by Brettanomyces brusselsii BB-0104 during the fermentation process mainly showed floral and fruity aroma and sauce aroma, among which compounds such as acetaldehyde, 4-ethyl-2-methoxyphenol and benzaldehyde were the key components of the unique flavor of Sichuan sun-dried vinegar. It can be seen that Brussels yeast BB-0104 can not only adapt to the specific fermentation environment of Sichuan sun-cured vinegar, but also plays a vital role in substrate transformation and characteristic flavor formation, providing a solid theoretical basis for the strain screening and flavor quality improvement of Sichuan sun-cured vinegar.
[0079] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.
Claims
1. A strain of Brettanomyces bruxellensis BB-0104, characterized in that: The Brussels yeast BB-0104 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on July 26, 2024, with the deposit number CGMCCNO.31477.
2. The Brettanomyces brusselsii BB-0104 according to claim 1, characterized in that The ITS sequence of the Brettanomyces brusselsii strain BB-0104 is shown in SEQ ID NO.
1.
3. The fermentation method of Brettanomyces brusselsii BB-0104 according to claim 1, characterized in that: The following steps are involved: The brussels yeast BB-0104 was inoculated into the fermentation medium for fermentation.
4. The fermentation method according to claim 3, characterized in that The method for preparing the fermentation medium comprises: mixing rice with water, sequentially subjecting the mixture to gelatinization, liquefaction and saccharification to obtain a primary fermentation medium, and adjusting the pH of the primary fermentation medium to 2 to 5.5 and the ethanol concentration to 1% to 12% to obtain a fermentation medium; The mass ratio of rice to water is 1:3-10; The gelatinization temperature is 90-100°C, and the gelatinization time is 20-40 minutes; The liquefaction treatment uses a thermostable α-amylase and is liquefied at 80 to 100° C. for 30 to 90 minutes; The liquefied product is cooled to 50-70° C., and saccharifying enzyme is added for saccharification treatment. The saccharification treatment time is 1-3 hours.
5. The fermentation method according to claim 3, characterized in that: The fermentation temperature is 20-44° C., and the fermentation time is 3-8 days. A fermentation broth comprising the Brettanomyces brusselsii BB-0104 according to claim 1.
7. Use of the Brettanomyces brusselsii BB-0104 according to claim 1 or 2 or the fermentation liquid according to claim 6 in producing solar-cured vinegar.
8. The use according to claim 7, characterized in that: The sun-dried vinegar is Sichuan sun-dried vinegar.
9. The use according to claim 7, characterized in that: The live bacteria concentration of the Brussels yeast strain BB-0104 used in the production of sun-cured vinegar is 5.2×10 5 ~2×10 7 CFU / mL.
10. The use according to claim 9, characterized in that: The inoculation amount of the Brussels yeast BB-0104 strain is 0.5% to 3%.