A preparation method and use of Bacillus mohaivi

Abstract

The present invention discloses a preparation method of Bacillus mohaievii and its application, which solves the defect that the cultivation method in the prior art cannot significantly increase the number of live bacteria while reducing the production cost of the strain. The present invention comprises: inoculating the strain of Bacillus mohaievii on an inorganic salt culture medium for activation to obtain a live bacteria count of 9.0×10 8 CFU / ml or more activated bacterial liquid; the activated bacterial liquid is accessed to the liquid culture medium according to the inoculation amount of 2% 6%, and the seed bacterial liquid is obtained after the shaking table is cultured for 15 24h; the seed bacterial liquid is accessed to the liquid culture medium of more than 10L according to the inoculation amount of 2% 6% for cultivation, and continuous ventilation is carried out during the cultivation process; the preparation method of the liquid culture medium is: 4 6 weight parts of yeast extract, 1 3 weight parts of KH2PO4, 12 18 weight parts of sucrose molasses, 1 3 weight parts of MgSO4 7H2O, 7 8 weight parts of ammonium chloride, 2 4 weight parts of sodium nitrate, a total of 1000 weight parts are configured with water, pH is adjusted to 6.5 7.2, and sterilization is obtained. The present invention has the advantages of lower cost and higher number of viable bacteria in the same culture time.

Description

Technical Field

[0001] The invention relates to the field of strain preparation, and in particular to a method for preparing Bacillus mohaievii. Background Art

[0002] With the rapid development of economy and technology and the continuous acceleration of urbanization, the amount of kitchen waste in urban garbage is increasing continuously. In the treatment of kitchen waste, there are problems such as insufficient treatment, long treatment time and easy generation of foul-smelling gas. In recent years, a kitchen waste treatment method has emerged at home and abroad - kitchen waste liquefaction degradation machine. Its principle is to use microbial agents to intermittently stir the materials so that the kitchen waste is continuously degraded and liquefied during the fermentation and degradation process, and finally produces a degradation liquid with very little residue. Therefore, it is urgent to prepare a composite bacterial preparation that can effectively and quickly degrade kitchen waste and reduce the odor of garbage.

[0003] Chinese document CN105816897A discloses an organic waste deodorant containing microbial strains and a preparation method thereof, specifically disclosing a fermentation broth containing Bacillus mohaiwei and Bacillus pumilus as active ingredients in the organic waste deodorant, and specifically disclosing that the fermentation culture medium is: 0.50-1.00% ammonium nitrate, 0.20-0.40% sodium chloride, 0.15-0.25% dipotassium hydrogen phosphate, 0.015-0.025% magnesium sulfate, 0.015-0.025% ferrous sulfate heptahydrate, 0.05-0.15% yeast extract, 1.50%-2.50% sucrose, 95.7-97.0% distilled water, and the pH value of the fermentation broth is adjusted to 7.0-7.5. When the fermentation broth is prepared, the culture medium can only be used for small-scale preparation in the laboratory. When directly applied to large-scale production, the number of viable bacteria never reaches 10 9 cuf / ml or above, resulting in low production efficiency and high production cost. Summary of the invention

[0004] Therefore, the technical problem to be solved by the present invention is to overcome the defect that the culture method in the prior art cannot significantly increase the number of viable bacteria while reducing the production cost of the strain, thereby providing a culture method with lower cost and higher number of viable bacteria in the same culture time, and the number of viable bacteria can even reach 10 9 A method for preparing Bacillus mohaiwei with a concentration of cuf / ml or more.

[0005] A method for preparing Bacillus mohaiwei, comprising:

[0006] Activation step: The strain of Bacillus mohaiwei was inoculated on the inorganic salt medium for activation to obtain a live bacterial count of 9.0×10 8 Activated bacterial solution with more than cfu / ml;

[0007] Seed culture: inoculate the activated bacterial solution into the liquid culture medium at an inoculation rate of 2%-6%, and culture on a shaking table for 15-24 hours to obtain the seed bacterial solution;

[0008] Fermentation culture: Inoculate the seed bacterial liquid into more than 10L of liquid culture medium at an inoculation rate of 2%-6% for culture, and continue to ventilate during the culture process;

[0009] The preparation method of the liquid culture medium is:

[0010] 4-6 parts by weight of yeast extract, 1-3 parts by weight of KH2PO4, 12-18 parts by weight of sucrose molasses, 1-3 parts by weight of MgSO4·7H2O, 7-8 parts by weight of ammonium chloride, and 2-4 parts by weight of sodium nitrate are prepared with water to a total of 1000 parts by weight, the pH is adjusted to 6.5-7.2, and the liquid culture medium is prepared after sterilization.

[0011] In the fermentation culture, the liquid culture medium is contained in a fermentation tank, and the culture conditions in the fermentation tank are: the fermentation tank speed is 50-100r / min, the initial pH is 6.0-7.0, and the temperature is 35°C-40°C. After the seed bacterial liquid is cultured in the fermentation tank for 15-24h, it is cooled to below 25°C.

[0012] The culture temperature is 35℃-40℃.

[0013] The preparation method of the inorganic salt culture medium is:

[0014] 4-6 parts by weight of yeast extract, 1-3 parts by weight of KH2PO4, 8-12 parts by weight of sucrose, 1-3 parts by weight of MgSO4·7H2O, 7-8 parts by weight of ammonium chloride, and 2-4 parts by weight of sodium nitrate are prepared with water to a total of 1000 parts by weight, the pH is adjusted to 6.5-7.2, and the mixture is sterilized.

[0015] The preparation method of the inorganic salt culture medium is:

[0016] Add 5 g yeast extract, 2 g KH2PO4, 10 g sucrose, 2 g MgSO4·7H2O, 7.5 g ammonium chloride, and 3 g sodium nitrate, dilute to 1 L with water, adjust the pH to 6.5-7.2, and sterilize.

[0017] The yeast extract in the liquid culture medium is industrial-grade yeast extract;

[0018] The sterilization temperature of the liquid culture medium is 113-118°C;

[0019] The preparation method of the liquid culture medium is as follows: 5g of yeast extract, 2g of KH2PO4, 15g of sucrose molasses, 2g of MgSO4·7H2O, 7.5g of ammonium chloride, and 3g of sodium nitrate are diluted to 1L with water, the pH is adjusted to 6.5-7.2, and the liquid culture medium is sterilized.

[0020] The use of the Bacillus mohaiwei prepared by the above preparation method in preparing a kitchen waste degrading agent.

[0021] The kitchen waste degrading agent is also compounded with Lactobacillus acidophilus, yeast, Lactobacillus plantarum and Bacillus pumilus.

[0022] The ratio of the number of live bacteria of Bacillus mohaiwei: Lactobacillus acidophilus: yeast: Lactobacillus plantarum: Bacillus pumilus is (3-5):(1-2):(2-3):(1-2):(3-5).

[0023] The amount of live bacteria of the kitchen waste degradation agent added to the kitchen waste is 1×10 5 ~1×10 7 cfu / kg.

[0024] The technical solution of the present invention has the following advantages:

[0025] 1. The preparation method of Bacillus mohaiwei provided by the present invention mainly adjusts the inoculation amount and the formula of the liquid culture medium, specifically: ferrous sulfate heptahydrate is omitted, sucrose is replaced by sucrose molasses, and the composition concentration of each substance is optimized. Among them, the present invention effectively reduces the amount of ferrous sulfate heptahydrate by optimizing the culture medium, and uses sucrose molasses to replace sucrose with higher cost, effectively reducing the production cost;

[0026] In addition, the present invention effectively increases the number of viable bacteria of the cultured Bacillus mohaiwei by adjusting the culture medium formula, and at the same time, combined with the setting of the inoculation amount of 2%-6%, effectively increases the number of viable bacteria of the cultured Bacillus mohaiwei to 10 9 cfu / ml and above, the effect is very significant.

[0027] 2. The present invention further optimizes the sterilization temperature of the liquid culture medium. By lowering the sterilization temperature, the influence of temperature on substances in the liquid culture medium can be reduced, and the number of live bacteria can be further increased, with a more significant effect.

[0028] 3. The present invention provides the use of the Bacillus mohaiwei, which can achieve the purpose of better degradation of kitchen waste and effectively reduce the odor of garbage after being compounded with Lactobacillus acidophilus, yeast, Lactobacillus plantarum and Bacillus pumilus in a ratio of (3-5):(1-2):(2-3):(1-2):(3-5). DETAILED DESCRIPTION

[0029] Example 1

[0030] A method for preparing Bacillus mohaiwei, comprising:

[0031] Activation step: The strain of Bacillus mohaiwei was inoculated on the inorganic salt medium for activation to obtain a live bacterial count of 9.0×10 8 cfu / ml or above activated bacterial liquid; the preparation method of the inorganic salt culture medium is: add 5g yeast extract, 2g KH2PO4, 10g sucrose, 2g MgSO4·7H2O, 7.5g ammonium chloride, 3g sodium nitrate, dilute to 1L with water, adjust pH to 7.0, and sterilize at 121°C for 15min.

[0032] Seed culture: inoculate the activated bacterial liquid into the liquid culture medium at an inoculation rate of 3%, and culture the liquid culture medium under shaking at 37°C for 20 hours, with the shaking speed of 180 r / min; the preparation method of the liquid culture medium is as follows: add 5g of yeast extract, 2g of KH2PO4, 15g of sucrose molasses, 2g of MgSO4·7H2O, 7.5g of ammonium chloride, and 3g of sodium nitrate, dilute to 1L with water, adjust the pH to 7.0, and sterilize at 115°C for 15min.

[0033] Fermentation culture: The seed bacterial liquid is inoculated at an inoculation rate of 3% into a fermentation tank filled with 10L of liquid culture medium for culture. The fermentation tank speed is 80r / min, the initial pH is 7.0, and the temperature is 37°C. After the seed bacterial liquid is cultured in the fermentation tank for 18h, it is cooled to below 25°C. Aeration is continued during the culture process, and the ventilation ratio is 1:0.5-1:1. The ventilation ratio preferably used in this embodiment is 1:1, that is, the ventilation volume in the fermentation tank with 10L of liquid culture medium is 10L / min.

[0034] The live bacteria count of the fermentation broth after culture was 2.5×10 9 cfu / ml.

[0035] Example 2

[0036] A method for preparing Bacillus mohaiwei, comprising:

[0037] Activation step: The strain of Bacillus mohaiwei was inoculated on the inorganic salt medium for activation to obtain a live bacterial count of 9.0×10 8 cfu / ml or above activated bacterial liquid; the preparation method of the inorganic salt culture medium is: add 5g yeast extract, 2g KH2PO4, 10g sucrose, 2g MgSO4·7H2O, 7.5g ammonium chloride, 3g sodium nitrate, dilute to 1L with water, adjust pH to 7.0, and sterilize at 121°C for 15min.

[0038] Seed culture: inoculate the activated bacterial liquid into the liquid culture medium at an inoculation rate of 6%, and culture the liquid culture medium under shaking at 37°C for 20 hours, with the shaking speed of 180 r / min; the preparation method of the liquid culture medium is as follows: add 6g of yeast extract, 1g of KH2PO4, 12g of sucrose molasses, 1g of MgSO4·7H2O, 8g of ammonium chloride, and 4g of sodium nitrate, dilute to 1L with water, adjust the pH to 7.0, and sterilize at 113°C for 15min.

[0039] Fermentation culture: The seed bacterial liquid is inoculated at an inoculation rate of 6% into a fermentation tank filled with 10L of liquid culture medium for culture. The fermentation tank speed is 80r / min, the initial pH is 7.0, and the temperature is 37°C. After the seed bacterial liquid is cultured in the fermentation tank for 18h, it is cooled to below 25°C. Aeration is continued during the culture process, and the ventilation ratio is 1:0.5-1:1. The ventilation ratio preferably used in this embodiment is 1:0.5, that is, the ventilation volume in the fermentation tank with 10L of liquid culture medium is 5L / min.

[0040] The live bacteria count of the fermentation broth was 2.9×10 9 cfu / ml.

[0041] Example 3

[0042] A method for preparing Bacillus mohaiwei, comprising:

[0043] Activation step: The strain of Bacillus mohaiwei was inoculated on the inorganic salt medium for activation to obtain a live bacterial count of 9.0×10 8 cfu / ml or above activated bacterial liquid; the preparation method of the inorganic salt culture medium is: add 5g yeast extract, 2g KH2PO4, 10g sucrose, 2g MgSO4·7H2O, 7.5g ammonium chloride, 3g sodium nitrate, dilute to 1L with water, adjust pH to 7.0, and sterilize at 121°C for 15min.

[0044] Seed culture: inoculate the activated bacterial liquid into the liquid culture medium at an inoculation rate of 2%, and culture the liquid culture medium under a shaking table at 37°C for 20 hours, with the shaking table rotating speed at 180 r / min; the preparation method of the liquid culture medium is as follows: add 4g of yeast extract, 3g of KH2PO4, 18g of sucrose molasses, 1g of MgSO4·7H2O, 7g of ammonium chloride, and 2g of sodium nitrate, dilute to 1L with water, adjust the pH to 7.0, and sterilize at 118°C for 15min.

[0045] Fermentation culture: The seed bacterial liquid is inoculated at an inoculation rate of 2% into a fermentation tank filled with 10L of liquid culture medium for culture. The fermentation tank speed is 80r / min, the initial pH is 7.0, and the temperature is 37°C. After the seed bacterial liquid is cultured in the fermentation tank for 18h, it is cooled to below 25°C. Aeration is continued during the culture process, and the ventilation ratio is 1:0.5-1:1. The ventilation ratio preferably used in this embodiment is 1:0.8, that is, the ventilation volume in the fermentation tank with 10L of liquid culture medium is 8L / min.

[0046] The live bacteria count of the fermentation broth was 1.5×10 9 cfu / ml.

[0047] Example 4

[0048] A method for preparing Bacillus mohaiwei, comprising:

[0049] Activation step: The strain of Bacillus mohaiwei was inoculated on the inorganic salt medium for activation to obtain a live bacterial count of 9.0×10 8 cfu / ml or above activated bacterial liquid; the preparation method of the inorganic salt culture medium is: add 6g of yeast extract, 1g of KH2PO4, 12g of sucrose, 2g of MgSO4·7H2O, 8g of ammonium chloride, and 3g of sodium nitrate, dilute to 1L with water, adjust the pH to 7.0, and sterilize at 121°C for 15min.

[0050] Seed culture: inoculate the activated bacterial liquid into the liquid culture medium at an inoculation rate of 3%, and culture the liquid culture medium on a shaking platform at 35°C for 16 hours, with the shaking speed of 200 r / min; the preparation method of the liquid culture medium is as follows: add 5g of yeast extract, 2g of KH2PO4, 15g of sucrose molasses, 2g of MgSO4·7H2O, 7.5g of ammonium chloride, and 3g of sodium nitrate, dilute to 1L with water, adjust the pH to 7.0, and sterilize at 115°C for 15min.

[0051] Fermentation culture: The seed bacterial liquid is inoculated at an inoculation rate of 3% into a fermentation tank filled with 10L of liquid culture medium for culture. The fermentation tank speed is 50r / min, the initial pH is 6.5, and the temperature is 35°C. After the seed bacterial liquid is cultured in the fermentation tank for 18h, it is cooled to below 25°C. Aeration is continued during the culture process, and the ventilation ratio is 1:0.5-1:1. The ventilation ratio preferably used in this embodiment is 1:0.5, that is, the ventilation volume in the fermentation tank with 10L of liquid culture medium is 5L / min.

[0052] The live bacteria count of the fermentation broth was 2.0×10 9 cfu / ml.

[0053] Example 5

[0054] A method for preparing Bacillus mohaiwei, comprising:

[0055] Activation step: The strain of Bacillus mohaiwei was inoculated on the inorganic salt medium for activation to obtain a live bacterial count of 9.0×10 8 cfu / ml or above activated bacterial liquid; the preparation method of the inorganic salt culture medium is: add 4g yeast extract, 3g KH2PO4, 8g sucrose, 1g MgSO4·7H2O, 7g ammonium chloride, 4g sodium nitrate, dilute to 1L with water, adjust pH to 7.2, and sterilize at 121°C for 15min.

[0056] Seed culture: inoculate the activated bacterial liquid into the liquid culture medium at an inoculation rate of 3%, and culture the liquid culture medium on a shaking platform at 35°C for 16 hours, with the shaking speed of 200 r / min; the preparation method of the liquid culture medium is as follows: add 5g of yeast extract, 2g of KH2PO4, 15g of sucrose molasses, 2g of MgSO4·7H2O, 7.5g of ammonium chloride, and 3g of sodium nitrate, dilute to 1L with water, adjust the pH to 7.0, and sterilize at 115°C for 15min.

[0057] Fermentation culture: The seed bacterial liquid is inoculated at an inoculation rate of 3% into a fermentation tank filled with 10L of liquid culture medium for culture. The fermentation tank speed is 80r / min, the initial pH is 6.0, and the temperature is 40°C. After the seed bacterial liquid is cultured in the fermentation tank for 15h, it is cooled to below 25°C. Aeration is continued during the culture process, and the ventilation ratio is 1:0.5-1:1. The ventilation ratio preferably used in this embodiment is 1:1, that is, the ventilation volume in the fermentation tank with 10L of liquid culture medium is 10L / min.

[0058] The live bacteria count of the fermentation broth was 2.3×10 9 cfu / ml.

[0059] Example 6

[0060] A method for preparing Bacillus mohaiwei, comprising:

[0061] Activation step: The strain of Bacillus mohaiwei was inoculated on the inorganic salt medium for activation to obtain a live bacterial count of 9.0×10 8 cfu / ml or above activated bacterial liquid; the preparation method of the inorganic salt culture medium is: add 5g yeast extract, 2g KH2PO4, 10g sucrose, 2g MgSO4·7H2O, 7.5g ammonium chloride, 3g sodium nitrate, dilute to 1L with water, adjust pH to 7.0, and sterilize at 121°C for 15min.

[0062] Seed culture: inoculate the activated bacterial liquid into the liquid culture medium at an inoculation rate of 3%, and culture the liquid culture medium under a shaking table at 37°C for 20 hours, with the shaking table rotating speed at 180 r / min; the preparation method of the liquid culture medium is as follows: add 5 g of yeast extract, 2 g of KH2PO4, 15 g of sucrose molasses, 2 g of MgSO4·7H2O, 7.5 g of ammonium chloride, and 3 g of sodium nitrate, dilute to 1 L with water, adjust the pH to 7.0, and sterilize at 121°C for 15 minutes.

[0063] Fermentation culture: The seed bacterial liquid is inoculated at an inoculation rate of 3% into a fermentation tank filled with 10L of liquid culture medium for culture. The fermentation tank speed is 80r / min, the initial pH is 7.0, and the temperature is 37°C. After the seed bacterial liquid is cultured in the fermentation tank for 18h, it is cooled to below 25°C. Aeration is continued during the culture process, and the ventilation ratio is 1:0.5-1:1. The ventilation ratio preferably used in this embodiment is 1:1, that is, the ventilation volume in the fermentation tank with 10L of liquid culture medium is 10L / min.

[0064] The live bacteria count of the fermentation broth was 1.3×10 9 cfu / ml.

[0065] Example 6

[0066] In this example, the bacterial solution of Bacillus mohaiwei prepared in Example 1 was used to prepare a kitchen waste degradation agent, specifically as follows:

[0067] 1000 ml of the bacterial solution prepared in Example 1 was taken, and Lactobacillus acidophilus, yeast, Lactobacillus plantarum and Bacillus pumilus were added to the bacterial solution according to the live bacteria ratio of Bacillus mohaiwei: Lactobacillus acidophilus: yeast: Lactobacillus plantarum: Bacillus pumilus = 4:1.5:2.5:1.5:4, and then 0.30 g of ethylenediaminetetraacetic acid, 5.02 g of diammonium hydrogen phosphate and 1.50 g of rhamnolipid were added to prepare a degradation agent.

[0068] The degradation agent prepared in this example and the No. 3 deodorant in the document CN105816897A were added to the kitchen waste at the same time, and the addition amount was: 1×10 6 cfu / kg, that is, the number of live bacteria added per kg of kitchen waste is 1×10 6 The degradation agents of cfu were then placed in two sealed boxes respectively and tested after being placed at room temperature for one week. The measurement results are shown in Table 1.

[0069] Example 7

[0070] The difference between this embodiment and embodiment 6 is only that the ratio of live bacteria in the degradation agent is different. In this embodiment, Bacillus mohaiwei:Lactobacillus acidophilus:Saccharomyces cerevisiae:Lactobacillus plantarum:Bacillus brevis=5:1:3:1:3.

[0071] Example 8

[0072] The difference between this embodiment and embodiment 6 is only that the ratio of live bacteria in the degradation agent is different. In this embodiment, Bacillus mohaiwei:Lactobacillus acidophilus:Saccharomyces cerevisiae:Lactobacillus plantarum:Bacillus brevis=3:2:2:2:5.

[0073] Table 1

[0074]

[0075] Comparative Example 1

[0076] A method for preparing Bacillus mohaiwei, comprising:

[0077] Activation step: The strain of Bacillus mohaiwei was inoculated on the inorganic salt medium for activation to obtain a live bacterial count of 9.0×10 8 cfu / ml or above activated bacterial liquid; the preparation method of the inorganic salt culture medium is: add 6g of yeast extract, 1g of KH2PO4, 12g of sucrose, 2g of MgSO4·7H2O, 8g of ammonium chloride, and 3g of sodium nitrate, dilute to 1L with water, adjust the pH to 7.0, and sterilize at 121°C for 15min.

[0078] Seed culture: inoculate the activated bacterial liquid into the liquid culture medium at an inoculation rate of 3%, and culture the liquid culture medium on a shaking platform at 35°C for 16 hours, with the shaking speed of 200 r / min; the preparation method of the liquid culture medium is as follows: add 5g of yeast extract, 2g of KH2PO4, 15g of sucrose molasses, 2g of MgSO4·7H2O, 7.5g of ammonium chloride, and 3g of sodium nitrate, dilute to 1L with water, adjust the pH to 7.0, and sterilize at 115°C for 15min.

[0079] Fermentation culture: The seed bacterial liquid is inoculated at an inoculation rate of 0.1% into a fermentation tank filled with 10L of liquid culture medium for culture. The fermentation tank speed is 50r / min, the initial pH is 6.5, and the temperature is 35°C. After the seed bacterial liquid is cultured in the fermentation tank for 18h, it is cooled to below 25°C. Aeration is continued during the culture process, and the ventilation ratio is 1:0.5-1:1. The ventilation ratio preferably used in this embodiment is 1:1, that is, the ventilation volume in the fermentation tank with 10L of liquid culture medium is 10L / min.

[0080] The live bacteria count of the fermentation broth was 8×10 8 cfu / ml.

[0081] Comparative Example 2

[0082] A method for preparing Bacillus mohaiwei, comprising:

[0083] Activation step: Place the strain of Bacillus mohaiwei in a slant test tube and place it in a constant temperature incubator at 32°C for 48 hours.

[0084] Seed culture: inoculate the activated bacteria into a potato culture medium, wherein the potato culture medium includes sucrose and potato juice, wherein the sucrose content is 20 g / L and the potato mass fraction in the potato juice is 20%. Inoculate the bacteria activated in the above activation step into the potato culture medium, and culture on a shaking table at 35° C. for 16 hours to obtain a seed bacterial solution, wherein the shaking table speed is 200 r / min.

[0085] Fermentation culture: The seed bacterial liquid is inoculated into a fermenter containing 10L liquid culture medium according to an inoculation amount of 0.1% for cultivation. The fermenter speed is 80r / min, the initial pH is 7.0, and the temperature is 37°C. After the seed bacterial liquid is cultured in the fermenter for 18h, it is cooled to below 25°C. During the culture process, ventilation is continued, and the ventilation ratio is 1:0.5-1:1. The ventilation ratio preferably used in this embodiment is 1:1, that is, the ventilation volume in the fermenter of 10L liquid culture medium is 10L / min. The preparation method of the liquid culture medium is: take 10.3g of ammonium nitrate, 4.1g of sodium chloride, 2.5g of dipotassium hydrogen phosphate, 0.2g of magnesium sulfate, 0.2g of ferrous sulfate heptahydrate, 1.5g of yeast extract, 25.7g of sucrose, and 1000ml of distilled water, mix them evenly to form a solution, and adjust the pH to 7.5, and then sterilize at 121°C for 30min.

[0086] The live bacteria count of the fermentation broth was 4×10 8 cfu / ml.

[0087] Comparative Example 3

[0088] A method for preparing Bacillus mohaiwei, comprising:

[0089] Activation step: Place the strain of Bacillus mohaiwei in a slant test tube and place it in a constant temperature incubator at 32°C for 48 hours.

[0090] Seed culture: inoculate the activated bacteria into a potato culture medium, wherein the potato culture medium includes sucrose and potato juice, wherein the sucrose content is 20 g / L and the potato mass fraction in the potato juice is 20%. Inoculate the bacteria activated in the above activation step into the potato culture medium, and culture on a shaking table at 35° C. for 16 hours to obtain a seed bacterial solution, wherein the shaking table speed is 200 r / min.

[0091] Fermentation culture: The seed bacterial liquid is inoculated into a fermenter containing 10L liquid culture medium according to the inoculation amount of 3% for cultivation, the fermenter speed is 80r / min, the temperature is 37°C, the seed bacterial liquid is cultured in the fermenter for 18h, and then cooled to below 25°C. The ventilation is continued during the cultivation process, and the ventilation ratio is 1:0.5-1:1. The ventilation ratio preferably used in this embodiment is 1:1, that is, the ventilation volume in the fermenter of 10L liquid culture medium is 10L / min. The preparation method of the liquid culture medium is: take 10.3g of ammonium nitrate, 4.1g of sodium chloride, 2.5g of dipotassium hydrogen phosphate, 0.2g of magnesium sulfate, 0.2g of ferrous sulfate heptahydrate, 1.5g of yeast extract, 25.7g of sucrose, and 1000ml of distilled water, mix them evenly to form a solution, and adjust the pH to 7.5, and then sterilize at 121°C for 30min.

[0092] The live bacteria count of the fermentation broth was 9×10 8 cfu / ml.

[0093] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A method for preparing Bacillus mohaiwei, characterized in that: include: Activation step: The strain of Bacillus mohaiwei was inoculated on the inorganic salt medium for activation to obtain a live bacterial count of 9.0×10 8 Activated bacterial solution with more than cfu / ml; Seed culture: inoculate the activated bacterial solution into the liquid culture medium at an inoculation rate of 2%-6%, and culture on a shaking table for 15-24 hours to obtain the seed bacterial solution; Fermentation culture: Inoculate the seed bacterial liquid into more than 10L of liquid culture medium at an inoculation rate of 3%-6%, and continue to ventilate during the culture process; The preparation method of the liquid culture medium is: 4-6 parts by weight of yeast extract, 1-3 parts by weight of KH2PO4, 12-18 parts by weight of sucrose molasses, 1-3 parts by weight of MgSO4·7H2O, 7-8 parts by weight of ammonium chloride and 2-4 parts by weight of sodium nitrate are prepared with water to a total of 1000 parts by weight, the pH is adjusted to 6.5-7.2, and the liquid culture medium is prepared after sterilization at 113-118°C.

2. The preparation method according to claim 1, characterized in that: In the fermentation culture, the liquid culture medium is contained in a fermentation tank, and the culture conditions in the fermentation tank are: the fermentation tank speed is 50-100r / min, the initial pH is 6.0-7.0, and the temperature is 35°C-40°C. After the seed bacterial liquid is cultured in the fermentation tank for 15-24h, it is cooled to below 25°C.

3. The preparation method according to claim 1, characterized in that: During the seed cultivation process, the cultivation temperature is 35°C-40°C.

4. The preparation method according to any one of claims 1 to 3, characterized in that: The preparation method of the inorganic salt culture medium is: 4-6 parts by weight of yeast extract, 1-3 parts by weight of KH2PO4, 8-12 parts by weight of sucrose, 1-3 parts by weight of MgSO4·7H2O, 7-8 parts by weight of ammonium chloride, and 2-4 parts by weight of sodium nitrate are prepared with water to a total of 1000 parts by weight, the pH is adjusted to 6.5-7.2, and the mixture is sterilized.

5. The preparation method according to claim 4, characterized in that: The preparation method of the inorganic salt culture medium is: Add 5 g yeast extract, 2 g KH2PO4, 10 g sucrose, 2 g MgSO4·7H2O, 7.5 g ammonium chloride, and 3 g sodium nitrate, dilute to 1 L with water, adjust the pH to 6.5-7.2, and sterilize.

6. The preparation method according to any one of claims 1 to 3, characterized in that: The preparation method of the liquid culture medium is as follows: 5g of yeast extract, 2g of KH2PO4, 15g of sucrose molasses, 2g of MgSO4·7H2O, 7.5g of ammonium chloride, and 3g of sodium nitrate are diluted to 1L with water, the pH is adjusted to 6.5-7.2, and the liquid culture medium is sterilized.

7. Use of Bacillus mohaiwei prepared by the preparation method according to any one of claims 1 to 6 in preparing a kitchen waste degrading agent.

8. The use according to claim 7, characterized in that The kitchen waste degrading agent is also compounded with Lactobacillus acidophilus, yeast, Lactobacillus plantarum and Bacillus pumilus.

9. The use according to claim 8, characterized in that The ratio of the number of live bacteria of Bacillus mohaiwei:Lactobacillus acidophilus:Saccharomyces cerevisiae:Lactobacillus plantarum:Bacillus pumilus is (3-5):(1-2):(2-3):(1-2):(3-5).

10. The use according to claim 8 or 9, characterized in that The amount of live bacteria of the kitchen waste degradation agent added to the kitchen waste is 1×10 5 ~1×10 7 cfu / kg.

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