Bacillus subtilis BS-TS1 and application thereof in preparation of distiller's yeast and wine products

Abstract

The invention provides bacillus subtilis BS-TS1 and application thereof in preparation of distiller's yeast and wine products, and belongs to the technical field of wine brewing microorganisms. The preservation number of the bacillus subtilis BS-TS1 provided by the invention is CGMCC (China General Microbiological Culture Collection Center) No. 32285. The bacillus subtilis BS-TS1 provided by the invention has the advantages of high penicillium resistance and high yield of aroma substances. The yeast for making hard liquor prepared by using the bacillus subtilis BS-TS1 has a remarkable penicillium resistance characteristic, and is not easily polluted in the making process of the yeast for making hard liquor. The vinasse juice prepared from the yeast prepared from the bacillus subtilis BS-TS1 is rich in volatile aroma substances, the content of acetoin reaches 150.944 mg / L, the content of pyrazine reaches 301.353 mg / L, and the content of 2, 3-butanediol reaches 21.319 mg / L; in addition, other aroma components such as ethyl butyrate, phenethyl alcohol and the like are also detected.

Description

Technical Field

[0001] The invention belongs to the technical field of wine-making microorganisms, and particularly relates to a Bacillus subtilis BS-TS1 and an application thereof in preparing wine yeast and wine products. Background Art

[0002] There are five types of koji: Daqu, Xiaoqu, Wheat Qu, Red Qu and Wheat Qu. Daqu is made from wheat and is produced using natural microorganisms and specific processes for liquor fermentation. The microbial community inside it plays a vital role in the liquor fermentation process.

[0003] Bacillus subtilis is a type of Gram-positive bacteria that is long and rod-shaped and contains spores. When encountering adverse environments such as nutrient deficiency and drought, the organism will form spores. In recent years, with the continuous deepening of research on the microbial flora in Daqu, the complex process of Daqu in formation and flavor construction has become increasingly clear, and the key role played by Bacillus subtilis has gradually been revealed. Under conditions of high temperature, high salt, and low oxygen, it can produce a variety of biologically active metabolites, such as antibiotics, enzymes, and antibacterial peptides. These metabolites show great application potential in inhibiting the growth of pathogenic microorganisms, promoting plant growth, and enhancing plant disease resistance.

[0004] The aroma substances in liquor can be mainly divided into esters, alcohols and aldehydes. The main aroma substances produced by Bacillus subtilis include volatile fatty acids, such as acetic acid, propionic acid and butyric acid, which usually have sour and fermented aroma. In addition, alcohols such as ethanol and isopentanol show the characteristics of fruity and wine aroma; esters such as ethyl acetate and ethyl butyrate often have fruity aroma; ketones such as ethyl ketone have sweet aroma; and aldehydes such as acetaldehyde usually emit a fresh aroma. The production of these aroma substances is closely related to the metabolic activity of Bacillus subtilis and is widely used in food fermentation, spice production and biotechnology.

[0005] Bacillus strains may be affected by various factors during production and application, such as temperature, pH value, humidity, etc., which may directly or indirectly weaken the stability of the strains, resulting in a decrease in their antibacterial properties. Therefore, how to ensure the stability and activity of the strains is an important issue in current research. Summary of the invention

[0006] In view of this, the object of the present invention is to provide a Bacillus subtilis that is highly resistant to Penicillium and produces a high amount of aroma substances.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The invention provides a Bacillus subtilis BS-TS1. The Bacillus subtilis BS-TS1 is classified and named as Bacillus subtilis, and is preserved in the General Microbiological Center of China Microorganism Culture Collection Administration, with a preservation number of CGMCC No.32285.

[0009] The present invention also provides a method for fermenting the Bacillus subtilis BS-TS1, wherein the Bacillus subtilis BS-TS1 is inoculated into a fermentation substrate, and fermented at 30-40° C. for 48-96 hours; the fermentation speed is 120-200 r / min, and the pH of the fermentation substrate is 6.5-8.

[0010] The present invention also provides the use of the Bacillus subtilis BS-TS1 in preparing distiller's yeast.

[0011] Preferably, the distiller's yeast includes Daqu.

[0012] The present invention also provides a Daqu that is resistant to Penicillium and rich in aroma substances. The bacteria used for fermenting and preparing the Daqu include the above-mentioned Bacillus subtilis BS-TS1.

[0013] The invention also provides application of the Daqu in preparing wine products.

[0014] Preferably, the wine product includes white wine.

[0015] Preferably, the liquor comprises sorghum liquor.

[0016] The invention also provides a liquor brewed from the above-mentioned Daqu.

[0017] Beneficial effects of the present invention:

[0018] The Bacillus subtilis BS-TS1 provided by the present invention has the advantages of high resistance to Penicillium and high production of aroma substances. The Daqu prepared using the Bacillus subtilis BS-TS1 provided by the present invention has significant anti-Penicillium properties and is not easily contaminated during the production process of Daqu. The lees juice prepared from the Daqu prepared using the Bacillus subtilis BS-TS1 of the present invention is rich in volatile aroma substances, among which the content of acetoin reaches 150.944 mg / L, the content of pyrazine reaches 301.353 mg / L, and the content of 2,3-butanediol reaches 21.319 mg / L; in addition, other aroma components such as ethyl butyrate, propyl butyrate, and phenylethyl alcohol are also detected.

[0019] The Daqu prepared by using the Bacillus subtilis BS-TS1 of the present invention not only has the advantages of high resistance to Penicillium and high yield of aroma substances, but also has the advantage of stable performance. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The morphological identification results of Bacillus BS-TS1, where the left picture shows the colony morphology and the right picture shows the Gram-stained bacterial morphology;

[0021] Figure 2 The plate confrontation results of BS-TS1 strain, from left to right are the results of 24h, 48h and 72h cultivation respectively;

[0022] Figure 3 The agar column method inhibition results of Bacillus BS-TS1, the left picture is Penicillium + BS-TS1, the right picture is Penicillium;

[0023] Figure 4 The effect of different fermentation temperatures on the antibacterial activity of the supernatant of strain BS-TS1;

[0024] Figure 5 The effect of different initial pH values ​​on the antibacterial activity of the supernatant of strain BS-TS1;

[0025] Figure 6 The effect of different shaking speeds on the antibacterial effect of the supernatant of strain BS-TS1;

[0026] Figure 7 The effect of different fermentation time on the antibacterial activity of the supernatant of strain BS-TS1;

[0027] Figure 8 This is the GC-MS total ion current result of the fermentation liquid of Bacillus subtilis BS-TS1 strain;

[0028] Fig. 9 This is a picture of the production of Daqu after inoculation with Penicillium and BS-TS1, in which from left to right are inoculation with only Saccharomyces cerevisiae, inoculation with Saccharomyces cerevisiae + BS-TS1, inoculation with Saccharomyces cerevisiae + Penicillium + BS-TS1, and inoculation with Saccharomyces cerevisiae + Penicillium;

[0029] Fig.10 This is the GC-MS total ion current result of Saccharomyces cerevisiae fermentation broth;

[0030] Fig.11 This is the GC-MS total ion chromatogram result of Saccharomyces cerevisiae + BS-TS1 Daqu fermentation liquid;

[0031] Fig.12 This is the GC-MS total ion chromatogram result of commercial Daqu fermentation broth.

[0032] Collection Instructions

[0033] The Bacillus subtilis BS-TS1 of the present invention is deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration Committee, classified and named Bacillus subtilis, and the deposit address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, the deposit number is CGMCC No.32285, and the deposit date is October 21, 2024. DETAILED DESCRIPTION

[0034] The invention provides a Bacillus subtilis BS-TS1. The Bacillus subtilis BS-TS1 is classified and named as Bacillus subtilis, and is preserved in the General Microbiological Center of China Microorganism Culture Collection Administration, with a preservation number of CGMCC No.32285.

[0035] The Bacillus subtilis BS-TS1 provided by the present invention is screened from the Daqu of the applicant's laboratory. When the Bacillus subtilis BS-TS1 grows on an LB plate, it forms a light yellow, translucent circular colony with a smooth and sticky surface, a protruding colony, and neat edges. These morphological characteristics are consistent with the typical characteristics of Bacillus, and the Bacillus subtilis BS-TS1 provided by the present invention is a Gram-positive bacterium with high resistance to Penicillium and high production of aroma substances, and the aroma substances include acetoin, 2,3-butanediol and pyrazine.

[0036] The present invention also provides a method for fermenting the Bacillus subtilis BS-TS1, wherein the Bacillus subtilis BS-TS1 is inoculated into a fermentation substrate, and fermented at 30-40° C. for 48-96 hours; the fermentation speed is 120-200 r / min, and the pH of the fermentation substrate is 6.5-8.

[0037] In the present invention, the fermentation substrate preferably includes a culture medium or a raw material for preparing koji, and the culture medium preferably includes LB liquid culture medium and a modified LB liquid culture medium, and the modified LB liquid culture medium is composed of 10.0 g of peptone, 5.0 g of yeast extract, 10.0 g of NaCl, 20 g of corn steep liquor powder, 80 g of glucose and 1000 mL of distilled water.

[0038] In the present invention, the fermentation temperature is preferably 33-38°C; the fermentation time is preferably 50-80°C, more preferably 60-75°C; the fermentation speed is preferably 130-180 r / min; and the pH of the fermentation substrate is preferably 7.

[0039] The present invention also provides the use of the above-mentioned Bacillus subtilis BS-TS1 in preparing distiller's yeast. In the use of the present invention, the distiller's yeast preferably includes Daqu.

[0040] The present invention also provides a Daqu that is resistant to Penicillium and rich in aroma substances, and the bacteria used to ferment the Daqu include the above-mentioned Bacillus subtilis BS-TS1. The Daqu prepared by fermenting the Bacillus subtilis BS-TS1 of the present invention has the advantages of high resistance to Penicillium and other bacteria, is not easily contaminated during the fermentation process, and has stable fermentation characteristics.

[0041] The present invention also provides the use of the above-mentioned Daqu in preparing wine products. In the use of the present invention, the wine product preferably includes liquor, and the liquor preferably includes sorghum liquor.

[0042] The present invention also provides a liquor brewed from the above-mentioned Daqu. The liquor brewed from the Daqu of the present invention has no bad flavor (musty smell, fungal smell) and has strong characteristic aroma substances such as acetoin, 2,3-butanediol and pyrazine.

[0043] 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.

[0044] In the following embodiments, unless otherwise specified, all of them are conventional methods.

[0045] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0046] Example 1

[0047] 1.1 Isolation and purification of Bacillus subtilis

[0048] First, crush the Daqu stored in the laboratory and weigh 1g, add it to 9mL of sterilized 0.9% saline, mix it thoroughly, and place it in an 80℃ water bath for 20min to kill non-spore microorganisms. Then, add this stock solution to 90mL LB liquid culture medium and perform enrichment culture at 37℃ for 24h. After the enrichment culture is completed, take 1mL of the bacterial solution and add it to 9mL of sterile saline, and dilute it step by step by 10 times until it reaches 10. -6 g / mL dilution concentration. Take 10 - 4 g / mL, 10 -5 g / mL and 10 -6 100 μL of each sample with a dilution of 100 μg / mL was inoculated onto an LB solid culture medium plate, evenly spread, and then placed in a 37°C constant temperature incubator for 18 to 24 hours. According to the morphological characteristics of Bacillus, a single colony was picked out and purified by streaking until a single pure strain was obtained. The isolated bacteria were morphologically observed and Gram-stained. Finally, the Bacillus that were positive under microscopic examination were transferred to LB solid culture medium and stored on a slant for later use.

[0049] 1.2 Screening of Bacillus subtilis that inhibits Penicillium

[0050] The plate confrontation method was used to preliminarily screen the Bacillus that inhibit the growth of Penicillium, and then the agar column method was used as a re-screening method, and the inhibition rate was calculated. The steps of the plate confrontation method are as follows: First, the Penicillium picked up with an inoculation loop was inoculated in the center of the PDA culture medium, and then the Bacillus was inoculated around the Penicillium. The growth of the two strains was observed every 24 hours to screen out the Bacillus subtilis that has an inhibitory effect on the growth of Penicillium. The steps of the agar column method are: add the fermentation supernatant of Bacillus subtilis to the PDA culture medium at a ratio of 1:10, use a hole puncher to take out the Penicillium cake and place it in the center of the PDA culture medium, and culture it at a constant temperature of 28°C. Using the cross method, the diameter of the bacteria was measured every 12 hours until the bacteria in the blank control culture dish were full. The calculation formula for the inhibition rate of Bacillus subtilis is as follows:

[0051]

[0052] The results showed that Bacillus subtilis BS-TS1 had the best antibacterial effect. The morphological identification results of Bacillus subtilis BS-TS1 are shown in Figure 1 When the strain grows on the LB plate, it forms light yellow, translucent round colonies with a smooth and sticky surface, protruding colonies and neat edges. These morphological characteristics are consistent with the typical characteristics of Bacillus, and the strain is a Gram-positive bacterium. The plate confrontation results of Bacillus subtilis BS-TS1 are shown in Figure 2 The results of the agar column method for the inhibition of Bacillus subtilis BS-TS1 are shown in Figure 3 .

[0053] Example 2

[0054] Optimization of Fermentation Conditions of Bacillus subtilis BS-TS1

[0055] 2.1 Temperature

[0056] The Bacillus subtilis BS-TS1 obtained in Example 1 was inoculated in a conical flask containing 50 mL of LB liquid culture medium at an inoculum volume of 1%, and the conical flask was placed on a shaker at 140 r / min and cultured for 48 h at five temperature gradients of 25°C, 30°C, 37°C, 40°C and 45°C. After the culture was completed, the fermentation broth of the strain was centrifuged at 10,000 r / min for 20 min, the supernatant was taken out and added to the PDA culture medium. The antibacterial rate was calculated according to the agar column method (same as in Example 1), and a bar graph was drawn. Each experimental condition was repeated three times, and the results are shown in FIG. Figure 4Under the culture conditions of 25℃, 30℃, 37℃, 40℃ and 45℃, the average inhibition rates of Bacillus subtilis BS-TS1 were 37.98%, 66.21%, 81.84%, 56.11% and 49.23%, respectively. Among them, when the culture temperature was 37℃, the inhibition rate of strain BS-TS1 against Penicillium was the highest.

[0057] 2.2pH

[0058] The Bacillus subtilis BS-TS1 obtained in Example 1 was inoculated with 1% volume in a conical flask containing 50 mL LB liquid culture medium, and the pH values ​​of these culture media were set to 5, 6, 7, 8 and 9. At 37°C, the conical flask was placed on a shaker and cultured at 140 r / min for 48 hours. After the culture was completed, the strain fermentation broth was centrifuged at 10,000 r / min for 20 minutes, and the supernatant was taken out and added to the PDA culture medium. The antibacterial rate was calculated according to the agar column method (same as in Example 1), and a bar graph was drawn to determine the optimal culture pH value. Each experimental condition was repeated three times. The results are shown in Figure 5 The average antibacterial rates of the fermentation supernatant of strain BS-TS1 were 37.98%, 49.23%, 85.94%, 64.00% and 56.11% at initial pH values ​​of 5, 6, 7, 8 and 9, respectively. Among them, the highest antibacterial rate against Penicillium was achieved at initial pH value of 7.

[0059] 2.3 Shaker speed

[0060] The Bacillus subtilis BS-TS1 obtained in Example 1 was inoculated with 1% volume in a conical flask containing 50 mL LB liquid culture medium (pH 7), and different shaking speed conditions were set, namely 40 r / min, 80 r / min, 120 r / min, 160 r / min and 200 r / min, and cultured at 37°C for 48 hours. After the culture was completed, the strain fermentation liquid was centrifuged at 10000 r / min for 20 minutes, and the supernatant was taken out and added to the PDA culture medium. The antibacterial rate was calculated according to the agar column method (same as Example 1), and a bar graph was drawn to determine the optimal culture shaking speed. Each experimental condition was repeated three times. The results are shown in Figure 6 Under the conditions of 40r / min, 80r / min, 120r / min, 160r / min and 200r / min, the average antibacterial rates of the fermentation supernatant of strain BS-TS1 were 49.23%, 51.00%, 69.75%, 79.75% and 48.52%, respectively. Among them, when the shaking speed was 160r / min, its antibacterial rate against Penicillium was the highest, so 160r / min was selected as the optimal shaking speed.

[0061] 2.4 Time

[0062] The Bacillus subtilis BS-TS1 obtained in Example 1 was inoculated with 1% volume in a conical flask containing 50 mL LB liquid culture medium (pH 7). Under 37°C conditions, the conical flask was cultured in a shaker at 160 r / min for 24 hours, 48 ​​hours, 72 hours, 96 hours and 120 hours. After the culture was completed, the strain fermentation liquid was centrifuged at 10000 r / min for 20 minutes, and the supernatant was taken out and added to the PDA culture medium. The antibacterial rate was calculated according to the agar column method (same as Example 1), and a bar graph was drawn to determine the optimal culture time. Each experimental condition was repeated three times. The results are shown in Figure 7 After culturing for 24 hours, 48 ​​hours, 72 hours, 96 hours and 120 hours, the inhibition rates of strain BS-TS1 were 64.00%, 79.75%, 85.89%, 72.64% and 68.34% respectively. Among them, the inhibition rate of BS-TS1 strain against Penicillium was the highest when it was cultured for 72 hours.

[0063] From the above test data results, it can be determined that strain BS-TS1 has the best antibacterial properties under the following conditions: the culture temperature is set at 37°C, the pH value of the initial culture medium is adjusted to 7.0, the shaker speed is controlled at 160r / min, and the culture is continued for 72 hours.

[0064] Example 3

[0065] 3.1 Experimental Materials

[0066] Fermentation medium: modified LB liquid medium: peptone 10.0 g, yeast extract 5.0 g, NaCl 10.0 g, corn steep liquor dry powder 20 g, glucose 80 g, distilled water 1000 mL, pH 7.0. Sterilize by high-pressure steam at 121°C for 20 min.

[0067] 3.2 Experimental methods

[0068] Bacillus subtilis BS-TS1 was fermented under the optimal antibacterial effect conditions obtained in Example 2. Finally, the aroma substances in the fermentation broth were determined using GC-MS headspace solid phase microextraction technology to determine the aroma production characteristics of the late Daqu. The specific fermentation process is as follows:

[0069] First, the Bacillus subtilis BS-TS1 screened out in Example 1 was inoculated into the fermentation medium at an inoculum size of 10%, and fermented for 72 hours at a set temperature of 37° C. and a rotation speed of 160 r / min. Finally, the types and percentages of aroma substances were determined.

[0070] 3.3 Detection methods

[0071] Take 8 mL of fermentation broth for GC-MS headspace solid phase microextraction to determine the type and percentage of aroma substances. The determination method is:

[0072] Add 1.5 g NaCl to a 20 mL screw-cap bottle, take 8 mL of fermentation broth into the screw-cap bottle, and sonicate for 15 min. Add 10 μL of internal standard, sonicate again for 10 min, and perform GC-MS headspace solid phase microextraction.

[0073] The detection method is:

[0074] GC conditions: the chromatographic column is an RTX-WAX capillary column (30m×0.25μm×0.25mm); the carrier gas is high-purity helium; splitless injection, the injection port temperature is 250℃; the column temperature is programmed: the programmed temperature is 40℃ for 3min, increased to 130℃ at 2℃ / min, increased to 220℃ at 10℃ / min, and maintained for 4min.

[0075] MS conditions: chromatographic-mass spectrometry interface temperature 250°C, ion source temperature 230°C; ionization mode: electron impact source (EI), bombardment energy 70 eV.

[0076] 3.4 Test results

[0077] The GC-MS total ion chromatogram results of the fermentation broth of Bacillus subtilis BS-TS1 strain are shown in Figure 8 The results of the determination of the fermentation products of Bacillus subtilis BS-TS1 are shown in Table 1:

[0078] Table 1 Determination results of fermentation products of Bacillus subtilis BS-TS1

[0079]

[0080]

[0081] As can be seen from Table 1, pyrazine accounts for 64.23% of the aroma components, acetoin accounts for 9.83%, and 2,3-butanediol accounts for 17.81%. This shows that the Bacillus subtilis BS-TS1 of the present invention has the characteristics of efficient fermentation production of pyrazine, acetoin and 2,3-butanediol, which is of great significance for the subsequent research on the flavor of liquor.

[0082] Example 4

[0083] 4.1 Experimental Materials

[0084] 15kg sorghum; Saccharomyces cerevisiae; Bacillus subtilis BS-TS1; Penicillium.

[0085] 4.2 Experimental methods

[0086] 4.2.1 Production of Daqu

[0087] Select high-quality wheat and barley as raw materials. Crush the selected grains and grind them into coarse powder for subsequent saccharification and fermentation. Mix the coarse powder with water and cook it to gelatinize the starch for the subsequent saccharification process. Cool the cooked mixture to 37°C and then inoculate Aspergillus koji. This step is to help convert starch into sugar. Let Aspergillus koji take effect and convert starch into fermentable sugars. Divide the saccharified substances into two groups and put them into fermentation containers: Group A adds 20g of brewer's yeast; Group B adds 20g of brewer's yeast and 20g of Bacillus subtilis BS-TS1; Group C adds 20g of brewer's yeast, 20g of Bacillus subtilis BS-TS1 and 20g of Penicillium and stirs evenly; Group D adds 20g of brewer's yeast and 20g of Penicillium and stirs evenly. Natural fermentation was carried out for 14 days under the same conditions. After the fermentation is completed, the Daqu is dried or dried to reduce its moisture. After reaching a state suitable for long-term storage, the cross-section effect of the Daqu is immediately observed, such as Fig. 9 shown.

[0088] 4.2.2 Fermentation test

[0089] First, 15kg of sorghum with full grains was steamed, divided into 3 groups, and spread to 35℃, and then 500g of group a wine yeast, 500g of group b wine yeast and commercial Daqu were added respectively. After fully stirring and mixing, the grains were collected and piled into ceramic jars for aerobic accumulation for 3 days. During this period, the microorganisms in the lees fully grew and multiplied, generating various aroma substances. After 3 days, when there was a clear wine aroma and sweet taste, 2kg of white sugar was added to each group, and the opening of the ceramic jar was wrapped with plastic film and covered with a ceramic lid for 30 days of fermentation.

[0090] 4.3 Detection methods

[0091] 8 mL of each of the three groups of fermented lees juice was taken for GC-MS headspace solid phase microextraction to determine the types and percentages of aroma substances. The determination method was the same as in Example 3.

[0092] 4.4 The test results are shown in Tables 2 to 4, and Figure 10 to Figure 12 .

[0093] Table 2a Group Daqu fermentation product determination results

[0094] name Content (mg / L) EthylAcetate 5.56(±0.46) 1-Propanol,2-methyl-(isobutyl alcohol) 5.17(±0.72) 1-Butanol,3-methyl-,acetate(Isoamyl acetate) 10.45(±1.68) 1-Butanol,3-methyl-(isoamyl alcohol) 133.29(±4.66) Hexanoic acid, ethyl ester (ethyl hexanoate) 32.96(±1.97) Octanoic acid, ethyl ester (octanoic acid ethyl ester) 57.24(±3.41) Decanoic acid, ethyl ester (decanoic acid ethyl ester) 13.39(±1.05) Aceticacid,2-phenylethylester(Aceticacid,2-phenylethylester) 2.55(±0.31) Dodecanoic acid, ethyl ester (ethyl laurate) 1.08(±0.12) PhenylethylAlcohol 29.71(±1.87)

[0095] Table 3b Group Daqu fermentation product determination results

[0096] name Content (mg / L) 2,3-Butanedione 9.172(±2.34) Butanoic acid, 2-methyl-, ethylester (butanoic acid ethyl ester) 2.745(±0.65) 1-Propanol,2-methyl-(isobutyl alcohol) 5.717(±1.71) Acetoin 150.944(±8.69) Pyrazine,tetramethyl-(Pyrazine) 301.353(±12.74) 2,3-Butanediol,[R-(R*,R*)]-(2,3-Butanediol) 21.319(±2.41) Propanoic acid, 2-methyl-(ethyl isobutyrate) 17.671(±2.26) Butanoic acid, 2-methyl-(propyl butanoate) 31.257(±4.36) PhenylethylAlcohol 13.734(±2.08)

[0097] Table 4 Determination results of commercial Daqu fermentation products

[0098]

[0099]

[0100] It can be seen from the results in Tables 2 to 4 that the contents of aroma substances such as pyrazine, 2,3-butanediol, acetoin, propyl butyrate, 2,3-butanedione and butyl acetate in the lees juice obtained by fermentation of the Daqu prepared by the strain BS-TS1 of the present invention are higher than those after fermentation of the commercial Daqu and without adding the strain BS-TS1. Through the subsequent distillation processing, the distilled liquor has a great sauce-flavored smell, thereby enhancing the market competitiveness of the fragrant liquor prepared by the strain of the present invention.

[0101] 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 Bacillus subtilis BS-TS1, characterized in that The Bacillus subtilis BS-TS1 is named as Bacillus subtilis and is deposited in the General Microbiological Center of China Microbiological Culture Collection Administration with a deposit number of CGMCC No.32285.

2. A method for fermenting Bacillus subtilis BS-TS1 according to claim 1, characterized in that: The Bacillus subtilis BS-TS1 of claim 1 is inoculated into a fermentation substrate, and fermented at 30-40° C. for 48-96 hours; the fermentation speed is 120-200 r / min, and the pH of the fermentation substrate is 6.5-8.

3. Use of the Bacillus subtilis BS-TS1 described in claim 1 in preparing distiller's yeast.

4. The use according to claim 3, characterized in that: The distiller's yeast includes Daqu.

5. A Daqu that is resistant to Penicillium and rich in aroma substances, characterized in that: The bacteria used for fermenting and preparing Daqu include the Bacillus subtilis BS-TS1 described in claim 1.

6. Use of the Daqu described in claim 5 in preparing wine products.

7. The use according to claim 6, characterized in that: The wine products include white wine.

8. The use according to claim 7, characterized in that: The liquor includes sorghum liquor.

9. A liquor, characterized in that: Obtained by brewing the Daqu as claimed in claim 5.

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