Method for producing bacillus velezensis by solid-state fermentation of distiller's grains

Abstract

The invention discloses a method for producing bacillus velezensis through solid-state fermentation of distiller's grains, and belongs to the technical field of fermentation engineering. The preparation method comprises the following steps: carrying out acid-base modification, enzymolysis and stacking fermentation treatment on white spirit vinasse, adding bran and jerusalem artichoke powder, carrying out steam explosion, and adding corn steep liquor dry powder, urea and other substances to prepare a white spirit vinasse culture medium; and carrying out amplification culture on the bacillus velezensis subjected to activation and seed culture by using a white spirit vinasse culture medium, drying and collecting the bacillus velezensis. Experimental results show that the bacillus velezensis can be efficiently produced by utilizing the fermentation method provided by the invention, and the spore number of the bacillus velezensis is as high as 9.2 * 10 < 10 > cfu / g. The technical method for producing bacillus velezensis through fermentation effectively solves the problem of resource utilization of white spirit vinasse and the problems of high cost and sewage in the liquid fermentation process, and is low in fermentation cost and suitable for industrial production.

Description

Technical Field

[0001] The present invention relates to the field of fermentation engineering, and particularly to a method for solid-state fermentation of Bacillus velezensis using distillers' grains. Background Art

[0002] Bacillus velezensis is a new species of Bacillus in taxonomy in recent years. It has a high genetic affinity with Bacillus amyloliquefaciens, can produce spore structures, has strong stress resistance, is safe and non-toxic to animals, and was included in the list of biological agents recognized and recommended for safety qualifications in the European Union in 2020. Bacillus velezensis can secrete a variety of bioactive substances, including bacteriocins that inhibit the growth of bacteria, fungi, viruses, and nematodes, enzymes for production, fermentation, and improving digestibility, hormones for promoting plant growth, etc. Currently, it is widely used in fields such as feed, medicine, textile, aquaculture, sewage treatment, and plant protection.

[0003] Distillers' grains are the solid residues remaining after fermentation using sorghum, wheat, corn, grains, etc. as raw materials in the liquor fermentation process. Distillers' grains contain nutrients that were not fully utilized during the fermentation process, have a very high water content, are not easy to store, are prone to spoilage and deterioration, and pollute the environment. China is a major liquor-producing country, with tens of millions of tons of distillers' grains produced annually. The comprehensive utilization of distillers' grains is of great significance for China's resource development and environmental protection. Currently, there have been many studies on the utilization of distillers' grains and certain achievements have been made, such as fermenting biological feed with distillers' grains, producing single-cell protein, fermenting to produce organic fertilizer, improving soil, making distillers' grain biscuits, distillers' grain fish, and distillers' grain brine, re-producing original liquor, producing methane from distillers' grain liquid, producing polyurethane materials and distillers' grain-based wood-plastic decorative boards from distillers' grains, aiming to improve the efficient utilization of distillers' grains and enhance their economic value.

[0004] The mainstream culture method of Bacillus velezensis is liquid fermentation culture, but there are problems such as a large amount of wastewater, easy environmental pollution, and high equipment requirements. Therefore, how to provide a new low-cost fermentation culture method to avoid the defects of the above liquid fermentation culture is a technical problem that needs to be solved urgently at present. Summary of the Invention

[0005] The purpose of the present invention is to provide a method for solid-state fermentation of Bacillus velezensis using distillers' grains to solve the problems existing in the above prior art. The purpose of the present invention is to provide a method for solid-state fermentation of Bacillus velezensis using distillers' grains to avoid or reduce the disadvantages of liquid fermentation, while efficiently utilizing distillers' grain resources and producing spores of Bacillus velezensis with high value in the agricultural field.

[0006] To achieve the above purpose, the present invention provides the following solutions:

[0007] The present invention provides a method for preparing a white wine lees culture medium, comprising the following steps:

[0008] Take white wine lees, adjust the pH to 11 - 14, let stand for 3 - 5 days, adjust the pH to 5 - 7, add cellulase, inoculate Bacillus subtilis and Bacillus licheniformis, mix well, stack into a long strip with a height of 50 - 80 cm and a width of 1 - 1.5 m, and ferment for 3 - 5 days to obtain degraded white wine lees;

[0009] By weight, mix 70 - 85 parts of the degraded white wine lees, 5 - 10 parts of wheat bran, and 4 - 7 parts of Jerusalem artichoke powder evenly, perform steam explosion, and collect the exploded material;

[0010] By weight, mix 60 - 70 parts of the exploded material, 10 - 20 parts of water, 10 - 15 parts of corn steep liquor dry powder, 5 - 8 parts of glucose, 3 - 5 parts of urea, and 0.25 - 0.75 parts of calcium carbonate evenly, sterilize and cool to obtain the white wine lees culture medium.

[0011] Preferably, the addition amount of the cellulase is 5 - 10 IU of cellulase per gram of white wine lees.

[0012] Preferably, the ratio of the viable bacteria count of Bacillus subtilis to that of Bacillus licheniformis is 1:1, and the total inoculation amount is (1 - 2)×10 8 cfu per gram of white wine lees.

[0013] Preferably, it comprises the following steps:

[0014] Take white wine lees, adjust the pH to 11, let stand for 5 days, adjust the pH to 6.4, add 9 IU / g of cellulase, inoculate Bacillus subtilis and Bacillus licheniformis according to the ratio of viable bacteria count of 1:1, with a total inoculation amount of 1.3×10 8 cfu / g, mix well, stack into a long strip with a height of 70 cm and a width of 1.3 m, and ferment for 3 days to obtain degraded white wine lees;

[0015] By weight, mix 80 parts of the degraded white wine lees, 7 parts of wheat bran, and 4 parts of Jerusalem artichoke powder evenly, perform steam explosion at 1.8 MPa for 11 min, and collect the exploded material;

[0016] By weight, mix 68 parts of the exploded material, 13 parts of water, 12 parts of corn steep liquor dry powder, 6 parts of glucose, 3 parts of urea, and 0.5 part of calcium carbonate evenly, sterilize and cool to obtain the white wine lees culture medium.

[0017] The present invention also provides a white wine lees culture medium obtained according to the above - mentioned preparation method.

[0018] The present invention also provides an application of the above - mentioned white wine lees culture medium in the fermentation production of Bacillus velezensis.

[0019] The present invention also provides a method for fermentatively producing Bacillus velezensis, which includes the step of culturing Bacillus velezensis using the distillers' grains medium described above.

[0020] Preferably, it includes the following steps:

[0021] Activating and culturing Bacillus velezensis on a solid plate, picking colonies, inoculating them into a liquid seed medium in a shake flask for culture to obtain the seed liquid for the fermenter;

[0022] Inoculating the obtained seed liquid for the fermenter into a soybean molasses liquid medium for fermentative culture to obtain a seed liquid;

[0023] Inoculating the seed liquid into the distillers' grains medium described above, culturing until the spores completely fall off, and drying to obtain the Bacillus velezensis.

[0024] Preferably, the shake flask culture is carried out at a rotation speed of 180 - 220 rpm for 20 - 28 h;

[0025] The conditions for the fermentative culture are: rotation speed 300 - 400 rpm, aeration 30 - 50 L / min, and culture for 20 - 28 h.

[0026] Preferably, the volume - mass ratio of the seed liquid to the distillers' grains medium is (1 - 5) mL:200 g.

[0027] The present invention discloses the following technical effects:

[0028] The present invention uses fresh distillers' grains as the main fermentation raw material to prepare Bacillus velezensis. Utilizing the starch and protein contained in the distillers' grains, together with the carbon and nitrogen source organic substances such as bran, Jerusalem artichoke powder, corn steep liquor dry powder, glucose added additionally, and inorganic substances such as urea contained in or added additionally to various raw materials, through a reasonable ratio, the nutrient components of the strain are formulated to meet the nutritional requirements for the reproduction of the strain, and at the same time, the resources of the distillers' grains are fully utilized.

[0029] The technical method of the present invention for solid - state fermentation of distillers' grains to produce Bacillus velezensis effectively solves the problems of resource utilization of distillers' grains and the high cost and sewage problems in the liquid - state fermentation process. After solid - state fermentation culture and drying, the spore count of Bacillus velezensis can reach 9.2×10 10 cfu / g.

[0030] The present invention uses distillers' grains as the main fermentation raw material, resulting in low fermentation cost and being suitable for industrial production. Detailed Embodiments

[0031] The various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation on the present invention, but rather as a more detailed description of certain aspects, features, and implementation schemes of the present invention.

[0032] It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0033] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0034] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific embodiments of the present invention specification, which are obvious to those skilled in the art. Other embodiments obtained from the specification of the present invention are obvious to those skilled in the art. The specification and examples of the present invention are merely exemplary.

[0035] Regarding the use of "comprising", "including", "having", "containing", etc. in this article, they are all open-ended terms, meaning including but not limited to.

[0036] Unless otherwise specified, the "parts" mentioned in the present invention are all calculated by mass parts.

[0037] All strains used in the embodiments of the present invention were purchased from the China Center for Type Culture Collection. The deposit numbers of Bacillus subtilis are CCTCC WB 2008762, the deposit numbers of Bacillus licheniformis are CCTCC AB 93195, and the deposit numbers of Bacillus velezensis are CCTCC AB 2018356.

[0038] The preparation method of the soybean molasses liquid medium used in the present invention is as follows:

[0039] According to the mass - volume ratio, take soybean meal and water and mix them evenly at a ratio of 1 g:10 mL. Adjust the pH value to 3.5 with dilute sulfuric acid, stir for 1 h, and filter to obtain the supernatant. Add 2.5% soybean meal powder, 0.5% sodium chloride, and 0.3% disodium hydrogen phosphate to the supernatant, and adjust the pH value to 6.5 - 7.0 to obtain the soybean molasses liquid medium.

[0040] Example 1

[0041] I. Preparation of distiller's grains medium

[0042] 1. Mix fresh distiller's grains with sodium hydroxide and stir to adjust the pH value to 13. Place it in a fermentation tank for 3 days to obtain alkali - pretreated distiller's grains.

[0043] 2. Use dilute sulfuric acid to adjust the pH value of the alkali - pretreated distiller's grains to 5. Add 6 IU of cellulase per gram of alkali - pretreated distiller's grains, and inoculate Bacillus subtilis and Bacillus licheniformis according to the viable bacteria number ratio of 1:1. A total of 1×10 8 cfu are inoculated per gram of alkali - pretreated distiller's grains. Stir evenly, pile it into a long strip with a height of 80 cm and a width of 1.2 m, and ferment for 3 days to obtain degraded distiller's grains.

[0044] 3. By weight, mix 70 parts of degraded distiller's grains, 5 parts of wheat bran, and 4 parts of Jerusalem artichoke powder evenly, put them into a steam explosion device, carry out steam explosion under the condition of a pressure of 1.5 MPa, and maintain the pressure for 15 min to collect the exploded material.

[0045] 4. By weight, take 60 parts of the exploded material, 10 parts of water, 10 parts of corn steep liquor dry powder, 6 parts of glucose, 3 parts of urea, and 0.25 parts of calcium carbonate, mix them evenly, sterilize at 121℃ for 1 h, and cool to room temperature to obtain the distiller's grains medium.

[0046] II. Production of Bacillus velezensis by solid - state fermentation using distiller's grains

[0047] 1. Streak the cryopreserved Bacillus velezensis on an LB solid plate and culture it at 37℃ until the colonies of Bacillus velezensis grow clearly.

[0048] 2. Pick a single colony of Bacillus velezensis with clear colonies from the LB solid plate and inoculate it into an LB liquid seed medium. Culture it on a shaker at 180 rpm at 37℃ for 20 h to obtain the seed liquid for the fermenter.

[0049] 3. Inoculate the seed liquid of Bacillus velezensis for the fermenter into the soybean molasses liquid medium. Culture it at 37℃ with a rotation speed of 300 rpm and an aeration condition of 50 L / min for 20 h to obtain the seed liquid of Bacillus velezensis in the solid - state fermentation medium, with a viable bacteria number of 7.3×10 9 cfu / mL;

[0050] 4. Inoculate the Bacillus velezensis seed solution into the solid-state medium of distillers' grains according to the volume ratio of 4 mL:200 g, and incubate at a constant temperature of 35 °C for 3 d until the spores completely fall off;

[0051] 5. Dry the solid material after fermentation to obtain the solid-state fermented Bacillus velezensis.

[0052] Inoculate the Bacillus velezensis prepared in this example into LB medium, and incubate at a constant temperature of 37 °C for 20 h. The total viable count is determined by the plate counting method to be 9.3×10 10 cfu / g, and the spore count is 9.1×10 10 cfu / g.

[0053] Example 2

[0054] I. Preparation of distillers' grains medium

[0055] 1. Mix fresh distillers' grains with sodium hydroxide and stir to adjust the pH value to 12. Place it in a fermentation tank for 5 days to obtain alkali-pretreated distillers' grains;

[0056] 2. Adjust the pH value of the alkali-pretreated distillers' grains to 6 with dilute sulfuric acid. Add 5 IU of cellulase per gram of alkali-pretreated distillers' grains, and inoculate Bacillus subtilis and Bacillus licheniformis according to the viable count ratio of 1:1. A total of 1.5×10 8 cfu / g is inoculated per gram of alkali-pretreated distillers' grains. Stir evenly, pile it into a long strip with a height of 60 cm and a width of 1 m, and ferment for 5 days to obtain degraded distillers' grains;

[0057] 3. By weight, mix 80 parts of degraded distillers' grains, 10 parts of wheat bran, and 7 parts of Jerusalem artichoke powder evenly, put them into a steam explosion device, and perform steam explosion at a pressure of 2 MPa for 10 min to collect the explosion products;

[0058] 4. By weight, take 70 parts of explosion products, 20 parts of water, 15 parts of corn steep liquor dry powder, 8 parts of glucose, 5 parts of urea, and 0.75 parts of calcium carbonate, mix them evenly, sterilize at 115 °C for 2 h, and cool to room temperature to obtain the distillers' grains medium.

[0059] II. Production of Bacillus velezensis by solid-state fermentation using distillers' grains

[0060] 1. Streak the cryopreserved Bacillus velezensis on an LB solid plate and culture it at 37 °C until the Bacillus velezensis colony grows clearly;

[0061] 2. Pick a single colony of Bacillus velezensis with clear colonies from the LB solid plate, inoculate it into the LB liquid seed medium, and culture it in a shaker at 180 rpm at 37 °C for 20 h to obtain the seed liquid for the fermenter;

[0062] 3. Inoculate the Bacillus velezensis seed liquid for the fermenter into the soybean molasses liquid medium, and culture it at 400 rpm and under an aeration condition of 30 L / min at 37 °C for 24 h to obtain the Bacillus velezensis seed liquid for the solid-state fermentation medium, with the viable cell count being 5.6×10 9 cfu / mL;

[0063] 4. Inoculate the Bacillus velezensis seed liquid into the distillers grains solid medium according to the volume-mass ratio of 5 mL:200 g, and culture it at a constant temperature of 33 °C for 5 d until the spores completely fall off;

[0064] 5. Dry the solid material after fermentation to obtain the solid-state fermented Bacillus velezensis.

[0065] Inoculate the Bacillus velezensis prepared in this example into the LB medium, and culture it at a constant temperature of 37 °C for 20 h. The total viable cell count was determined to be 8.9×10 10 cfu / g by the plate counting method, and the spore count was 8.8×10 10 cfu / g.

[0066] Example 3

[0067] I. Preparation of the distillers grains medium

[0068] 1. Mix fresh distillers grains with sodium hydroxide and stir to adjust the pH value to 14. Place it in the fermentation tank for 3 days to obtain the alkali-pretreated distillers grains;

[0069] 2. Use dilute sulfuric acid to adjust the pH value of the alkali-pretreated distillers grains to 7. Add 10 IU of cellulase per gram of the alkali-pretreated distillers grains. Inoculate Bacillus subtilis and Bacillus licheniformis according to the viable cell ratio of 1:1, and a total of 2×10 8 cfu / g is inoculated per gram of the alkali-pretreated distillers grains. Stir evenly, stack it into a long strip with a height of 50 cm and a width of 1.5 m, and ferment for 4 days to obtain the degraded distillers grains;

[0070] 3. By weight, mix 85 parts of the degraded distillers grains, 10 parts of wheat bran, and 5 parts of Jerusalem artichoke powder evenly, put them into a steam explosion device, and carry out steam explosion under the condition of a pressure of 1.7 MPa for 13 min, and collect the explosion products;

[0071] 4. Take 65 parts by weight of explosives, 15 parts of water, 13 parts of dry corn steep liquor, 5 parts of glucose, 4 parts of urea, and 0.6 part of calcium carbonate, mix them evenly, sterilize at 112 °C for 2.5 h, and cool to room temperature to obtain the distiller's grains medium.

[0072] II. Production of Bacillus velezensis by solid-state fermentation of distiller's grains

[0073] 1. Streak the cryopreserved Bacillus velezensis on an LB solid plate and culture it at 37 °C until the colonies of Bacillus velezensis grow clearly.

[0074] 2. Pick a single colony of Bacillus velezensis with clear colonies from the LB solid plate, inoculate it into the LB liquid seed medium, and culture it on a shaker at 180 rpm at 37 °C for 20 h to obtain the seed liquid for the fermenter.

[0075] 3. Inoculate the seed liquid of Bacillus velezensis for the fermenter into the soybean molasses liquid medium, culture it at 350 rpm at 37 °C under aeration conditions of 50 L / min for 28 h to obtain the seed liquid of Bacillus velezensis for the solid-state fermentation medium, and the viable cell count is 7.9×10 9 cfu / mL;

[0076] 4. Inoculate the seed liquid of Bacillus velezensis into the distiller's grains solid medium according to the volume-to-mass ratio of 3 mL:200 g, and culture it at a constant temperature of 37 °C for 3 d until the spores completely fall off.

[0077] 5. Dry the solid material after fermentation to obtain solid-state fermented Bacillus velezensis.

[0078] Inoculate the Bacillus velezensis prepared in this example into the LB medium, culture it at a constant temperature of 37 °C for 20 h, and use the plate counting method to determine that the total viable cell count is 8.6×10 10 cfu / g, and the spore count is 8.7×10 10 cfu / g.

[0079] Example 4

[0080] I. Preparation of the distiller's grains medium

[0081] 1. Mix fresh distiller's grains with sodium hydroxide and stir to adjust the pH value to 11, and place it in the fermentation tank for 5 days to obtain alkali-pretreated distiller's grains.

[0082] 2. Adjust the pH value of the alkali-pretreated distiller's grains to 6.4 with dilute sulfuric acid, add 9 IU of cellulase per gram of alkali-pretreated distiller's grains, inoculate Bacillus subtilis and Bacillus licheniformis according to the viable cell count ratio of 1:1, and inoculate a total of 1.3×10 per gram of alkali-pretreated distiller's grains 8cfu / g, stir evenly, pile up into a long strip with a height of 70 cm and a width of 1.3 m, and ferment for 3 days to obtain degraded distillers' grains;

[0083] 3. By weight, mix 80 parts of degraded distillers' grains, 7 parts of wheat bran, and 4 parts of Jerusalem artichoke powder evenly, put them into a steam explosion device, and perform steam explosion under the condition of a pressure of 1.8 MPa for 11 minutes, and collect the explosion products;

[0084] 4. By weight, take 68 parts of explosion products, 13 parts of water, 12 parts of corn steep liquor dry powder, 6 parts of glucose, 3 parts of urea, and 0.5 part of calcium carbonate, mix evenly, sterilize at 108 °C for 3 h, and cool to room temperature to obtain the distillers' grains culture medium.

[0085] II. Production of Bacillus velezensis by solid-state fermentation of distillers' grains

[0086] 1. Streak the cryopreserved Bacillus velezensis on an LB solid plate and culture it at 37 °C until the Bacillus velezensis colonies grow clearly;

[0087] 2. Pick a single colony of Bacillus velezensis with clear colonies from the LB solid plate, inoculate it into an LB liquid seed medium, and culture it on a shaker at 180 rpm at 37 °C for 20 h to obtain the seed liquid for the fermenter;

[0088] 3. Inoculate the Bacillus velezensis seed liquid for the fermenter into a soybean molasses liquid medium, culture it at 37 °C at 370 rpm under the condition of aeration at 45 L / min for 28 h to obtain the Bacillus velezensis seed liquid for the solid-state fermentation medium, and the viable count is 6.4×10 9 cfu / mL;

[0089] 4. Inoculate the Bacillus velezensis seed liquid into the distillers' grains solid medium according to the volume-to-mass ratio of 3 mL:200 g, and culture it at a constant temperature of 30 °C for 5 d until the spores completely fall off;

[0090] 5. Dry the solid material after fermentation to obtain solid-state fermented Bacillus velezensis.

[0091] Inoculate the Bacillus velezensis prepared in this example into an LB medium, culture it at a constant temperature of 37 °C for 20 h, and use the plate counting method to determine that the total viable count is 9.5×10 10 cfu / g, and the spore count is 9.2×10 10 cfu / g.

[0092] Example 5

[0093] I. Preparation of solid-state fermentation medium

[0094] 1. Weigh 15 parts of rice husk, 10 parts of wheat bran, and 4 parts of Jerusalem artichoke powder by weight fraction, mix them evenly, put them into a steam explosion device, and perform steam explosion under the condition of a pressure of 1.9 MPa for 12 minutes while maintaining the pressure. Then collect the exploded materials.

[0095] 2. Weigh 60 parts of the exploded materials, 65 parts of water, 12 parts of corn steep liquor dry powder, 7 parts of glucose, 4 parts of urea, and 0.35 part of calcium carbonate by weight, mix them evenly, sterilize at 121 °C for 1 h, and cool to room temperature to obtain a solid fermentation medium without distillers' grains.

[0096] II. Production of Bacillus velezensis by solid fermentation without distillers' grains

[0097] 1. Streak the cryopreserved Bacillus velezensis on an LB solid plate and culture it at 37 °C until the colonies of Bacillus velezensis grow clearly.

[0098] 2. Pick a single colony of Bacillus velezensis with clear colonies from the LB solid plate and inoculate it into an LB liquid seed medium. Culture it on a shaker at 180 rpm at 37 °C for 20 h to obtain the seed liquid for the fermenter.

[0099] 3. Inoculate the seed liquid of Bacillus velezensis for the fermenter into a soybean molasses liquid medium and culture it at 320 rpm at 37 °C under the aeration condition of 40 L / min for 28 h to obtain the seed liquid of Bacillus velezensis for the solid fermentation medium, with the viable cell count being 6.1×10 9 cfu / mL.

[0100] 4. Inoculate the seed liquid of Bacillus velezensis into the solid fermentation medium without distillers' grains according to the volume-mass ratio of 3 mL:200 g, and culture it at a constant temperature of 30 °C for 4 d until the spores completely fall off.

[0101] 5. Dry the solid material after fermentation to obtain the solid-fermented Bacillus velezensis.

[0102] Inoculate the Bacillus velezensis prepared in this example into an LB medium and culture it at a constant temperature of 37 °C for 20 h. The total viable cell count measured by the plate counting method is 2.7×10 10 cfu / g, and the spore count is 2.4×10 10 cfu / g.

[0103] The above embodiments are only descriptions of the preferred embodiments of the present invention and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention should fall within the protection scope determined by the claims of the present invention.

Claims

1. A method for preparing a white wine lees culture medium, characterized in that: The following steps are involved: Take white wine lees, adjust the pH to 11-14, stand for 3-5 days, adjust the pH to 5-7, add cellulase, inoculate Bacillus subtilis and Bacillus licheniformis, mix well, pile into strips with a height of 50-80 cm and a width of 1-1.5 m, ferment for 3-5 days, and obtain degraded white wine lees; By weight, 70-85 parts of the degraded white wine lees, 5-10 parts of bran and 4-7 parts of Jerusalem artichoke powder are mixed evenly, steam-exploded, and the exploded materials are collected; By weight, 60-70 parts of the explosive material, 10-20 parts of water, 10-15 parts of corn syrup powder, 5-8 parts of glucose, 3-5 parts of urea and 0.25-0.75 parts of calcium carbonate are mixed evenly, sterilized and cooled to obtain the white wine lees culture medium.

2. The preparation method according to claim 1, characterized in that The added amount of the cellulase is 5-10 IU of cellulase per gram of white wine grains.

3. The preparation method according to claim 1, characterized in that: The ratio of the number of live bacteria of the Bacillus subtilis to that of the Bacillus licheniformis is 1:1, and the total inoculation amount is (1-2)×10 8 cfu.

4. The preparation method according to any one of claims 1 to 3, characterized in that: The following steps are involved: Take the white wine lees, adjust the pH to 11, let it stand for 5 days, adjust the pH to 6.4, add 9 IU / g of cellulase, and inoculate Bacillus subtilis and Bacillus licheniformis at a ratio of 1:

1. The total inoculation amount is 1.3×10 8 cfu / g, mixed, piled into strips 70 cm high and 1.3 m wide, fermented for 3 days, and obtained degraded liquor lees; By weight, 80 parts of the degraded white wine lees, 7 parts of bran and 4 parts of Jerusalem artichoke powder were mixed evenly, and steam exploded at 1.8 MPa for 11 minutes, and the exploded materials were collected; By weight, 68 parts of the explosive material, 13 parts of water, 12 parts of corn syrup powder, 6 parts of glucose, 3 parts of urea and 0.5 parts of calcium carbonate are mixed evenly, sterilized and cooled to obtain the white wine lees culture medium.

5. A white wine grains culture medium obtained according to the preparation method according to any one of claims 1 to 4.

6. Use of the white wine grains culture medium as claimed in claim 5 in fermentation production of Bacillus Velez.

7. A method for producing Bacillus Velez by fermentation, characterized in that: The method comprises the step of culturing the Bacillus Velez using the white wine grains culture medium according to claim 5.

8. The method according to claim 7, characterized in that The following steps are involved: Bacillus Velez is activated and cultured on a solid plate, and colonies are picked and inoculated into a liquid seed culture medium for shaking flask culture to obtain a seed solution for tanking; Inoculating the seed liquid in the upper tank into a soybean molasses liquid culture medium, fermenting and culturing to obtain a seed liquid; The seed liquid is inoculated into the white wine lees culture medium according to claim 5, cultured until the spores completely fall off, and dried to obtain the Bacillus Velezii.

9. The method according to claim 8, characterized in that The shake flask culture is carried out at a rotation speed of 180-220 rpm for 20-28 hours; The fermentation culture conditions are: rotation speed 300-400 rpm, ventilation 30-50 L / min, and culture 20-28 h.

10. The method according to claim 8, characterized in that The volume mass ratio of the seed liquid to the white wine lees culture medium is (1-5) mL:200 g.