Complex microbial inoculant for fermenting soybean meal and application of complex microbial inoculant

Abstract

The invention provides a complex microbial inoculant for fermenting soybean meal and application of the complex microbial inoculant. Strain in the complex microbial inoculant for fermenting soybean meal comprises lactic acid bacteria, bacillus and saccharomycetes; the lactic acid bacteria comprise lactobacillus plantarum and pediococcus acidilactici; the bacillus comprises bacillus coagulans and bacillus subtilis. The five strains of lactobacillus plantarum, pediococcus acidilactici, bacillus coagulans, bacillus subtilis and saccharomycetes are combined for use, so that macromolecular protein in a substrate can be effectively decomposed, antinutritional factors are reduced, and the flavor of soybean meal is improved; the fermented soybean meal has a synergistic effect in the aspects of increasing the viable count of the fermented soybean meal product, increasing the weight of fed animals and reducing the diarrhea rate of the fed animals. The complex microbial inoculant for fermenting the soybean meal is used for fermenting the soybean meal, is high in microbial proliferation speed, high in viable count in a final product, good in flavor and low in fermentation cost, and has higher economic application value.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial agents, and relates to a composite microbial agent for fermenting soybean meal and an application thereof, and specifically relates to a composite microbial agent for fermenting soybean meal and an application thereof, a fermented soybean meal and a preparation method thereof. Background Art

[0002] The lack of protein feed resources has become a major obstacle to the sustainable development of the livestock industry. In 2021, the amount of imported soybeans exceeded 100 million tons. Soybean meal has a good amino acid composition and meets the needs of farmed animals, but its protein digestibility has not met expectations. In addition to the fact that soybean meal contains a certain amount of anti-nutritional factors, there is also the problem that some large molecular proteins are not easily decomposed and used by livestock and poultry. The use of microbial fermentation can effectively reduce anti-nutritional factors and decompose large molecular proteins. However, relying solely on a single strain of bacteria, it is difficult to achieve multiple effects such as fast fermentation speed, small nutritional loss, and reduced anti-nutritional factors.

[0003] CN110651898A discloses a soybean meal fermentation method. The method comprises: firstly crushing soybean meal, corn, rapeseed cake and wheat bran, then mixing soybean meal powder, corn powder, rapeseed cake powder and wheat bran powder with Bacillus, lactic acid bacteria, brown sugar, calcium, phosphorus, sodium, potassium, copper, cobalt, iodine, zinc, selenium, fluorine and manganese; then adding water to adjust the water content of the material to 60%-70%; then putting it into a container such as a tank or a pool for fermentation. The soybean meal fermentation product prepared by the inventive method is easy to digest, fast to absorb, good in taste, effectively stimulates the proliferation of beneficial bacteria in the intestine, regulates the structure of the microecological flora in the body, and the protein dissolution ratio can reach 80%-90%.

[0004] CN105441363A discloses a strain, which relates to the field of microorganisms. The strain belongs to Bacillus subtilis and is deposited in CGMCC (General Microbiological Center of China Microbiological Culture Collection Administration Committee), with a deposit number of CGMCC No: 6440. The fermented soybean meal obtained by fermenting the strain has a high content of small peptides. The invention also provides a method for preparing fermented soybean meal using the above-mentioned strain and the fermented soybean meal obtained.

[0005] Currently, soybean meal is often used as animal feed after fermentation. However, there is a contradiction that the fermentation effect is poor when the amount of microorganisms added is small, while the cost is high when the amount of microorganisms added is large. Therefore, it is very meaningful to develop a low-cost and efficient composite bacterial agent for soybean meal fermentation. Summary of the invention

[0006] In view of the deficiencies in the prior art, the object of the present invention is to provide a composite bacterial agent for fermenting soybean meal and its application, and specifically to a composite bacterial agent for fermenting soybean meal and its application, a fermented soybean meal and its preparation method.

[0007] To achieve this object, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a composite bacterial agent for fermenting soybean meal, wherein the strains in the composite bacterial agent for fermenting soybean meal include lactic acid bacteria, bacillus and yeast; the lactic acid bacteria include plant lactobacillus and Pediococcus acidilactici; the bacillus include Bacillus coagulans and Bacillus subtilis.

[0009] The present invention creatively applies a combination of lactic acid bacteria, bacillus and yeast to prepare a composite bacterial agent for fermenting soybean meal, wherein the lactic acid bacteria include plant lactobacillus and Pediococcus acidilactici; and the bacillus include bacillus coagulans and bacillus subtilis.

[0010] Among them, Bacillus subtilis and Bacillus coagulans can secrete digestive enzymes such as protease and cellulase, and Lactobacillus plantarum and Pediococcus acidilactici can synthesize lactic acid. The combination of five strains of Lactobacillus plantarum, Pediococcus acidilactici, Bacillus coagulans, Bacillus subtilis and yeast can effectively decompose macromolecular proteins in the substrate, reduce anti-nutritional factors, improve the flavor of soybean meal, and have a synergistic effect in increasing the number of live bacteria in the fermented soybean meal product, improving the weight of animals when the fermented soybean meal product is fed to animals, and reducing the diarrhea rate of fed animals. The composite microbial agent for fermenting soybean meal of the present invention is used for soybean meal fermentation, and the microorganisms proliferate quickly, the number of live bacteria in the final product is high, the flavor is good, the fermentation cost is low, and it has greater economic application value.

[0011] Preferably, the ratio of the number of live bacteria of the lactic acid bacteria, the bacillus and the yeast is (2-10):(5-20):1; more preferably, the ratio of the number of live bacteria of the lactic acid bacteria, the bacillus and the yeast is (5-8):(10-15):1;

[0012] When the ratio of the number of live bacteria of lactic acid bacteria, bacillus and yeast is (2-10): (5-20): 1, the total number of live bacteria in the fermented soybean meal final product is high, the weight of animals increases rapidly and the diarrhea rate is low when the fermented soybean meal is used for animal feeding. When the ratio of the number of live bacteria of lactic acid bacteria, bacillus and yeast is further reduced to (5-8): (10-15): 1, the number of live bacteria in the fermented soybean meal is higher, and the effect of increasing weight gain and improving diarrhea when the fermented soybean meal is used for animal feeding is better.

[0013] The specific value in (2-10) can be 2, 3, 4, 5, 6, 7, 8, 9 or 10, etc.;

[0014] The specific value in (5-20) can be 5, 7, 9, 11, 13, 15, 17, 19 or 20, etc.;

[0015] The specific value in (10-15) can be 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5 or 15, etc.;

[0016] The specific values ​​in (5-8) can be 5, 5.5, 6, 6.5, 7, 7.5 or 8, etc.;

[0017] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0018] Preferably, the ratio of the viable counts of Lactobacillus plantarum to Pediococcus acidilactici is 1:(0.5-5);

[0019] The specific value in (0.5-5) can be selected from 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5, etc.;

[0020] Preferably, the ratio of the number of live bacteria of Bacillus coagulans to that of Bacillus subtilis is 1:(1-8);

[0021] The specific value in (1-8) can be 1, 2, 3, 4, 5, 6, 7 or 8, etc.;

[0022] Preferably, the total bacterial viability in the composite bacterial agent for fermenting soybean meal is not less than 1×10 11 CFU / g or 1×10 11 CFU / mL, for example 1×10 11 CFU / g (CFU / mL), 3×10 11 CFU / g (CFU / mL), 5×10 11 CFU / g (CFU / mL), 8×10 11 CFU / g (CFU / mL), 1×10 12 CFU / g (CFU / mL), 5×10 12 CFU / g (CFU / mL), 1×10 13 CFU / g (CFU / mL), etc.

[0023] Preferably, the lactic acid bacteria further include any one of Lactococcus lactis, Streptococcus lactis or Bifidobacterium lactis, or a combination of at least two of them. The combination of at least two includes a combination of Lactococcus lactis and Streptococcus lactis, a combination of Streptococcus lactis and Bifidobacterium lactis, or a combination of Lactococcus lactis and Bifidobacterium lactis, and the like. Any other combinations may be selected and will not be described in detail herein.

[0024] In a second aspect, the present invention provides a fermented soybean meal, wherein the fermented soybean meal is obtained by fermenting the composite bacterial agent for fermented soybean meal described in the first aspect.

[0025] The fermentation strains used in the fermented soybean meal involved in the present invention include the strains in the composite bacterial agent for fermenting soybean meal as described in the first aspect. Compared with conventional fermented soybean meal, the final product fermented soybean meal has a higher total live bacteria count and stronger bacterial activity. When used for preparing feed, the animals gain weight faster and have less diarrhea.

[0026] In a third aspect, the present invention provides a method for preparing the fermented soybean meal as described in the second aspect, the preparation method comprising the following steps:

[0027] (1) inoculating the fermented soybean meal composite bacterial agent into a composite bacterial agent activation culture medium, and culturing at 30-40° C. for 16-24 hours to obtain a composite bacterial solution;

[0028] (2) Mixing dry soybean meal and water, inoculating the composite bacterial solution, spreading the mixed solution on a fermentation bed, and performing anaerobically fermentation at 30-40° C. for 60-96 hours to obtain the fermented soybean meal.

[0029] The preparation method of the fermented soybean meal of the present invention comprises the activation culture of the composite bacterial agent for fermenting the soybean meal and the fermentation culture inoculating the activated culture solution into the soybean meal. The preparation method of the present invention has a simple process and is suitable for industrial scale-up.

[0030] The specific value of 30-40°C can be 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C or 40°C, etc.;

[0031] The specific value of 16-24h can be selected from 16h, 17h, 18h, 19h, 20h, 21h, 22h, 23h or 24h, etc.;

[0032] The specific value of 60-96h can be 60h, 65h, 70h, 75h, 80h, 85h, 90h, 95h or 96h, etc.;

[0033] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0034] Preferably, the culture medium for activating the composite bacterial agent in step (1) comprises a carbon source, a nitrogen source and an inorganic salt;

[0035] Preferably, the carbon source includes any one of brown sugar, glucose, stachyose, raffinose, corn starch, glycerol or maltose, or a combination of at least two of them. The combination of at least two includes a combination of brown sugar and glucose, a combination of glucose and stachyose, or a combination of raffinose and corn starch, etc. Any other combination can be selected and will not be described here one by one.

[0036] More preferably, the carbon source comprises brown sugar.

[0037] Preferably, the nitrogen source includes any one of soybean meal, corn slurry powder, ammonium chloride, urea, peptone or yeast powder, or a combination of at least two of them; the combination of at least two includes a combination of soybean meal and corn slurry powder, a combination of corn slurry powder and ammonium chloride, or a combination of ammonium chloride and soybean meal, and any other combination can be selected, which will not be described here one by one.

[0038] More preferably, the nitrogen source comprises soybean meal and corn steep liquor powder;

[0039] When the nitrogen source in the culture medium for activating the composite bacterial agent of the present invention includes soybean meal and corn steep liquor powder, the cell proliferation rate of the five strains in the composite bacterial agent is faster and the cell activity is better. After being used for subsequent fermentation culture, the total number of viable bacteria in the final product is higher.

[0040] The carbon source and nitrogen source used in the culture medium for activating the composite bacterial agent of the present invention are low-cost nutrient sources, but have good fermentation culture effect, that is, the economic cost of fermentation is low and the cost performance is high.

[0041] Preferably, the inorganic salt includes any one of dipotassium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, ammonium dihydrogen sulfate or ammonium sulfate, or a combination of at least two of them; the combination of at least two includes a combination of dipotassium hydrogen phosphate and potassium dihydrogen phosphate, a combination of potassium dihydrogen phosphate and magnesium sulfate, or a combination of magnesium sulfate and ammonium dihydrogen sulfate, and the like. Any other combination may be selected and will not be described in detail here.

[0042] More preferably, the inorganic salt comprises dipotassium hydrogen phosphate.

[0043] Preferably, the culture medium for activating the composite bacterial agent further comprises a microbial activator;

[0044] Preferably, the microbial activator includes any one or a combination of at least two of 5-aminolevulinic acid, folic acid, lipoic acid, biotin, pantothenic acid, vitamin A or vitamin C. The combination of at least two includes a combination of 5-aminolevulinic acid and folic acid, a combination of folic acid and lipoic acid, or a combination of lipoic acid and biotin, and any other combinations can be selected, which will not be described one by one here.

[0045] Preferably, the mass ratio of the carbon source, the nitrogen source and the inorganic salt is (5-20):(10-30):1;

[0046] Preferably, the mass ratio of the soybean meal to the corn syrup dry powder is (1-10):1.

[0047] The specific value in (5-20) can be 5, 7, 9, 11, 13, 15, 17, 19 or 20, etc.;

[0048] The specific value in (10-30) can be 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 or 30, etc.;

[0049] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0050] The specific value in (1-10) can be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, etc.;

[0051] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0052] Preferably, the water content in step (2) is 30-60 wt %; more preferably, the water content in step (2) is 40-45 wt %;

[0053] The specific value of 30-60wt% can be selected from 30wt%, 35wt%, 40wt%, 45wt%, 50wt%, 55wt% or 60wt%, etc.;

[0054] The specific value of 40-45wt% can be selected from 40wt%, 41wt%, 42wt%, 43wt%, 44wt% or 45wt%, etc.;

[0055] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0056] Preferably, after the inoculation in step (2), the number of viable bacteria in the mixed solution is 10 7 -10 9 CFU / g.

[0057] 10 7 -10 9 The specific value in CFU / g can be 10 7 CFU / g, 5×10 7 CFU / g, 10 8 CFU / g, 5×10 8 CFU / g or 10 9 CFU / g, etc.

[0058] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0059] In a fourth aspect, the present invention provides the use of the composite bacterial agent for fermenting soybean meal as described in the first aspect, the fermented soybean meal as described in the second aspect, or the preparation method of the fermented soybean meal as described in the third aspect in the production of soybean meal feed.

[0060] Compared with the prior art, the present invention has the following beneficial effects:

[0061] 1. The present invention creatively applies a combination of lactic acid bacteria, bacillus and yeast to prepare a composite bacterial agent for fermenting soybean meal, wherein the lactic acid bacteria include plant lactobacillus and Pediococcus acidilactici; the bacillus include Bacillus coagulans and Bacillus subtilis.

[0062] Among them, Bacillus subtilis and Bacillus coagulans can secrete digestive enzymes such as protease and cellulase, and Lactobacillus plantarum and Pediococcus acidilactici can synthesize lactic acid. The combination of five strains of Lactobacillus plantarum, Pediococcus acidilactici, Bacillus coagulans, Bacillus subtilis and yeast can effectively decompose macromolecular proteins in the substrate, reduce anti-nutritional factors, improve the flavor of soybean meal, and have a synergistic effect in increasing the number of live bacteria in the fermented soybean meal product, improving the weight of animals when raising animals, and reducing the diarrhea rate of animals. The composite bacterial agent for fermenting soybean meal of the present invention is used for soybean meal fermentation, and the microorganisms proliferate quickly, the number of live bacteria in the final product is high, the flavor is good, the fermentation cost is low, and it has greater economic application value.

[0063] 2. The fermentation strain used in the fermented soybean meal involved in the present invention is obtained by fermentation using the aforementioned composite bacterial agent. Compared with conventional fermented soybean meal, the final product fermented soybean meal has a higher total number of live bacteria and stronger bacterial activity. When used for preparing feed, the animals gain weight faster and have less diarrhea.

[0064] 3. The method for preparing fermented soybean meal of the present invention comprises the activation culture of the composite bacterial agent for fermenting soybean meal and the fermentation culture inoculating the activated culture solution into the soybean meal. The preparation method of the present invention has a simple process and is suitable for industrial scale-up. DETAILED DESCRIPTION

[0065] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0066] The corn syrup powder involved in the following implementation case is a product purchased from Wudi Bohao Biotechnology Co., Ltd.; Lactobacillus plantarum is a strain deposited in the General Microbiology Center of the China National Microbiological Culture Collection with a deposit number of CGMCC21766, and its classification name is Lactobacillus plantarum. The preservation time is January 29, 2021, and the preservation address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. It is made into bacterial powder before use; Pediococcus acidilactici is a strain deposited in the China Center for Type Culture Collection with a deposit number of CCTCC NO: M2023230, and its classification name is Pediococcus acidilactici. The preservation time is March 2, 2023, and the preservation address is Wuhan University, Wuhan, China. It is made into bacterial powder before use; Bacillus coagulans is a strain deposited by Anmaican Biotechnology Co., Ltd. in the General Microbiology Center of the China National Microbiological Culture Collection with a deposit number of CGMCC 21879 strain, which is classified and named Bacillus coagulans, was preserved on March 8, 2021, and the preservation address is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing. It was made into bacterial powder before use; Bacillus subtilis was purchased from Green World with a specification of 100 billion; yeast was purchased from Angel Yeast with a specification of 20 billion; Candida was purchased from Guangzhou Weiyuan Biotechnology Co., Ltd. with a specification of 20 billion. Soybean meal was 46 soybean meal purchased from Yihai Kerry; fish meal was purchased from Hebei Haikang Company; oil was purchased from Yihai Kerry; premix was purchased from Wuxi Zhengda Company with a model of 4% fattening pig premix.

[0067] The preparation method of bacterial powder is as follows: inoculate the desired strain into MRS liquid culture medium, culture at 37°C for 24 hours for activation, and activate twice continuously to obtain activation solution; inoculate the activation solution into MRS liquid culture medium at an inoculum amount of 4% (v / v), culture at 37°C for 24 hours to obtain bacterial solution; centrifuge the bacterial solution at 5000 rpm at 4°C for 5 minutes, filter to obtain bacterial bodies, mix with 2 times the mass of a protective agent (20% skim milk), and freeze-dry to obtain bacterial powder.

[0068] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0069] Example 1

[0070] This embodiment provides a fermented soybean meal, and the preparation method of the fermented soybean meal is as follows:

[0071] (1) Mix 8.5 parts of brown sugar, 15 parts of soybean meal, 2.5 parts of corn syrup powder and 1 part of dipotassium hydrogen phosphate (the mass ratio of carbon source, nitrogen source and inorganic salt is 8.5:17.5:1, and the mass ratio of soybean meal to corn syrup powder is 6:1), add water to make up to 100 parts, and sterilize at 115° C. for 15 minutes to obtain the culture medium for activating the composite bacterial agent.

[0072] (2) mixing five kinds of bacterial powders, namely Lactobacillus plantarum, Pediococcus acidilactici, Bacillus coagulans, Bacillus subtilis and yeast, to prepare the composite bacterial agent for fermenting soybean meal, wherein the ratio of live bacteria counts is as follows: lactic acid bacteria: Bacillus: yeast = 7.5:12.5:1; Lactobacillus plantarum: Pediococcus acidilactici = 1:2; Bacillus coagulans: Bacillus subtilis = 1:4.

[0073] (3) The fermentation compound bacterial agent was inoculated into the compound bacterial agent activation medium. The number of viable bacteria in the medium after inoculation was 10 7 CFU / g, and cultured at 37℃ for 20h to obtain the composite bacterial solution.

[0074] (4) Mix 40% water and 60% dry soybean meal, inoculate the composite bacterial solution, and the number of live bacteria in the mixed solution after inoculation is 10 8 CFU / g, and then the mixed solution was spread on a fermentation bed and cultured at 37°C for 72 hours for anaerobic fermentation to obtain the fermented soybean meal.

[0075] Example 2

[0076] This embodiment provides a fermented soybean meal, and the preparation method of the fermented soybean meal is as follows:

[0077] (1) Mix 5 parts of brown sugar, 27.3 parts of soybean meal, 2.7 parts of corn syrup powder and 1 part of dipotassium hydrogen phosphate (the mass ratio of carbon source, nitrogen source and inorganic salt is 5:30:1, and the mass ratio of soybean meal to corn syrup powder is 10:1), add water to make up to 100 parts, and sterilize at 115° C. for 15 minutes to obtain the culture medium for activating the composite bacterial agent.

[0078] (2) mixing five kinds of bacterial powders, namely Lactobacillus plantarum, Pediococcus acidilactici, Bacillus coagulans, Bacillus subtilis and yeast, to prepare the composite bacterial agent for fermenting soybean meal, wherein the ratio of live bacteria counts is as follows: lactic acid bacteria: Bacillus: yeast = 8:15:1; Lactobacillus plantarum: Pediococcus acidilactici = 1:5; Bacillus coagulans: Bacillus subtilis = 1:8.

[0079] (3) The fermentation compound bacterial agent was inoculated into the compound bacterial agent activation medium. The number of viable bacteria in the medium after inoculation was 10 7 CFU / g, and fermented at 40℃ for 16h to obtain the composite bacterial solution.

[0080] (4) Mix 45% water and 55% dry soybean meal, inoculate the composite bacterial solution, and the number of live bacteria in the mixed solution after inoculation is 10 9 CFU / g, and then the mixed solution was spread on a fermentation bed, and cultured at 40°C for 60 hours for anaerobic fermentation to obtain the fermented soybean meal.

[0081] Example 3

[0082] This embodiment provides a fermented soybean meal, and the preparation method of the fermented soybean meal is as follows:

[0083] (1) Mix 20 parts of brown sugar, 5 parts of soybean meal, 5 parts of corn syrup powder and 1 part of dipotassium hydrogen phosphate (the mass ratio of carbon source, nitrogen source and inorganic salt is 20:10:1, and the mass ratio of soybean meal to corn syrup powder is 1:1), add water to make up to 100 parts, and sterilize at 115° C. for 15 minutes to obtain the culture medium for activating the composite bacterial agent.

[0084] (2) mixing five kinds of bacterial powders, namely Lactobacillus plantarum, Pediococcus acidilactici, Bacillus coagulans, Bacillus subtilis and yeast, to prepare the composite bacterial agent for fermenting soybean meal, wherein the ratio of live bacteria counts is as follows: lactic acid bacteria: Bacillus: yeast = 5:10:1; Lactobacillus plantarum: Pediococcus acidilactici = 1:4; Bacillus coagulans: Bacillus subtilis = 1:1.

[0085] (3) The fermentation compound bacterial agent was inoculated into the compound bacterial agent activation medium. The number of viable bacteria in the medium after inoculation was 10 7 CFU / g, and cultured at 30℃ for 24h to obtain the composite bacterial solution.

[0086] (4) Mix 40% water and 60% dry soybean meal, inoculate the composite bacterial solution, and the number of live bacteria in the mixed solution after inoculation is 10 7 CFU / g, and then the mixed solution was spread on a fermentation bed and cultured at 30°C for 96 hours for anaerobic fermentation to obtain the fermented soybean meal.

[0087] Example 4

[0088] This embodiment provides a fermented soybean meal. The preparation method of the fermented soybean meal is different from that of Example 1 only in that the ratio of the number of live bacteria in the composite bacterial agent for the fermented soybean meal is: lactic acid bacteria: Bacillus: yeast = 7.5:12.5:1 is replaced by the ratio of the number of live bacteria: lactic acid bacteria: Bacillus: yeast = 3:18:1, the total number of bacteria remains unchanged, and the rest is the same as Example 1.

[0089] Example 5

[0090] This embodiment provides a fermented soybean meal. The preparation method of the fermented soybean meal is different from that of Example 1 only in that the ratio of the number of live bacteria in the composite bacterial agent for the fermented soybean meal is replaced by the ratio of the number of live bacteria: lactic acid bacteria: Bacillus: yeast = 7.5:12.5:1 to the ratio of the number of live bacteria: lactic acid bacteria: Bacillus: yeast = 9:8:1, the total number of bacteria remains unchanged, and the rest is the same as Example 1.

[0091] Example 6

[0092] This embodiment provides a fermented soybean meal. The preparation method of the fermented soybean meal is different from that of Embodiment 1 only in that the culture medium for activating the composite bacterial agent does not contain soybean meal, and the mass fraction of the soybean meal is distributed to the corn steep liquor powder. The rest is the same as that of Embodiment 1.

[0093] Example 7

[0094] This embodiment provides a fermented soybean meal. The preparation method of the fermented soybean meal is different from that of Example 1 only in that the culture medium for activating the composite bacterial agent does not contain corn steep liquor powder, and the mass fraction of corn steep liquor powder is distributed to the soybean meal. The rest is the same as that of Example 1.

[0095] Comparative Example 1

[0096] This comparative example provides a fermented soybean meal. The preparation method of the fermented soybean meal is different from that of Example 1 only in that the composite bacterial agent for fermenting the soybean meal does not contain plant lactobacillus, the ratio of the live bacterial counts of other strains remains unchanged, the total bacterial count remains unchanged, and the remaining steps are the same as those of Example 1.

[0097] Comparative Example 2

[0098] This comparative example provides a fermented soybean meal. The preparation method of the fermented soybean meal is different from that of Example 1 only in that the composite bacterial agent for fermenting the soybean meal does not contain lactic acid bacteria, the total bacterial count remains unchanged, the ratio of the live bacterial counts of other strains remains unchanged, and the remaining steps are the same as those of Example 1.

[0099] Comparative Example 3

[0100] This comparative example provides a fermented soybean meal. The preparation method of the fermented soybean meal is different from that of Example 1 only in that the composite bacterial agent for fermenting the soybean meal does not contain Bacillus coagulans, the total bacterial count remains unchanged, the ratio of the live bacterial counts of other strains remains unchanged, and the remaining steps are the same as those of Example 1.

[0101] Comparative Example 4

[0102] This comparative example provides a fermented soybean meal. The preparation method of the fermented soybean meal is different from that of Example 1 only in that the composite bacterial agent for fermenting the soybean meal does not contain Bacillus subtilis, the total bacterial count remains unchanged, the ratio of the live bacterial counts of other strains remains unchanged, and the remaining steps are the same as those of Example 1.

[0103] Comparative Example 5

[0104] This comparative example provides a fermented soybean meal. The preparation method of the fermented soybean meal is different from that of Example 1 only in that the composite bacterial agent for fermenting the soybean meal does not contain yeast, the total bacterial count remains unchanged, the ratio of the live bacterial counts of other strains remains unchanged, and the remaining steps are the same as those of Example 1.

[0105] Comparative Example 6

[0106] This comparative example provides a fermented soybean meal. The preparation method of the fermented soybean meal is different from that of Example 1 only in that 1 part of Lactobacillus plantarum, 2 parts of Pediococcus acidilactici, 1 part of Bacillus coagulans, 4 parts of Bacillus subtilis, and 2 parts of yeast (the ratio of the number of live bacteria is: lactic acid bacteria: Bacillus: yeast = 7.5:12.5:1) in the composite bacterial agent for the fermented soybean meal is replaced by 3 parts of Lactobacillus plantarum, 5 parts of Bacillus subtilis, and 2 parts of Candida (the ratio of the number of live bacteria is: lactic acid bacteria: Bacillus: yeast = 7.5:12.5:1), the total bacterial count remains unchanged, and the rest is the same as Example 1.

[0107] Comparative Example 7

[0108] This comparative example provides a fermented soybean meal. The preparation method of the fermented soybean meal is different from that of Example 1 only in that step (3) is not included, and the fermented soybean meal is directly inoculated with the composite bacterial agent into a mixture of dry soybean meal and water. The rest is the same as that of Example 1.

[0109] Test Example 1

[0110] The performance indexes of the fermented soybean meal prepared in each embodiment and comparative example were evaluated, and the specific evaluation method was as follows:

[0111] The fermented soybean meal prepared in each embodiment or comparative example was evaluated for odor and pH performance. Ten assessors were asked to score each embodiment or comparative example, and the average value was calculated. The evaluation criteria are shown in Table 1. The total score is 10 points. The evaluation results are shown in Table 2.

[0112] Table 1

[0113]

[0114] Table 2

[0115] Group Performance evaluation score Example 1 9.5 Example 2 8.5 Example 3 9 Example 4 7 Example 5 7.5 Example 6 9 Example 7 8.5 Comparative Example 1 5 Comparative Example 2 5 Comparative Example 3 6.5 Comparative Example 4 6.5 Comparative Example 5 5.5 Comparative Example 6 6 Comparative Example 7 8

[0116] It can be seen from the data in Table 2 that, compared with Example 1, the live count ratio of lactic acid bacteria: Bacillus: yeast in Example 4 and Example 5 is not within the preferred range of (5-8): (10-15): 1, and the performance evaluation score is lower than that in Example 1, indicating that when the live count of the three microorganisms is maintained within the preferred range of (5-8): (10-15): 1, the fermented soybean meal product obtained by fermentation has a fragrant smell, a strong fragrance, a low pH, and excellent product quality; compared with Example 1, the nitrogen source of the fermentation activation medium in Example 6 and Example 7 lacks one of soybean meal and corn syrup powder, respectively, and the performance evaluation score is lower than that in Example 1, indicating that when the nitrogen source selects a combination of soybean meal and corn syrup powder, the fermentation of the fermented soybean meal is better, and the obtained fermented soybean meal product has excellent performance in terms of smell and pH.

[0117] Compared with Example 1, Comparative Examples 1-5 lack one of Lactobacillus plantarum, Pediococcus acidilactici, Bacillus coagulans, Bacillus subtilis, and yeast, respectively, and their fermented soybean meal performance evaluation scores are lower than that of Example 1, indicating that the five microorganisms have a synergistic effect in improving the fermentation performance of fermented soybean meal products.

[0118] Compared with Example 1, Comparative Example 6 has different types of lactic acid bacteria, Bacillus and yeast, and its performance evaluation score is lower than that of Example 1, indicating that when the composite bacterial agent for fermenting soybean meal of the present invention specifically includes Lactobacillus plantarum, Pediococcus acidilactici, Bacillus coagulans, Bacillus subtilis and yeast, the fermented soybean meal product has excellent performance.

[0119] In Comparative Example 7, compared with Example 1, the composite bacterial agent for fermenting soybean meal was directly used for soybean meal fermentation without expansion culture, and its application cost was relatively high, and its performance evaluation score was also lower than that of Example 1, indicating that the composite bacterial agent for fermenting soybean meal was activated before fermentation culture, and the performance of the final product fermented soybean meal was better.

[0120] Test Example 2

[0121] The animal feeding effects of the fermented soybean meal prepared in the examples and comparative examples were evaluated, and the specific evaluation method was as follows:

[0122] (1) 160 Duchang growing and fattening pigs of similar weight and good health were selected, half male and half female, and divided into 16 groups, including 14 experimental groups corresponding to the embodiments or comparative examples, a blank group and a control group. The experimental group used the fermented soybean meal feed prepared in each embodiment or comparative example to feed the pigs, the blank group used the basic diet, and the control group used the feed containing commercially available fermented soybean meal, as shown in Table 3, and the pigs were fed for 30 consecutive days to monitor the overall weight growth, daily weight gain, daily feed intake, feed-to-meat ratio and diarrhea rate of the pigs. The results are shown in Table 4.

[0123] Table 3

[0124]

[0125] Table 4

[0126]

[0127]

[0128] It can be seen from the data in Table 4 that, compared with Example 1, the live count ratio of lactic acid bacteria: Bacillus: yeast in Example 4 and Example 5 is not within the preferred range of (5-8): (10-15): 1, and the feed-to-meat ratio and diarrhea rate are higher than those in Example 1, indicating that when the live count of the three microorganisms is maintained within the preferred range of (5-8): (10-15): 1, when the fermented soybean meal obtained by fermentation is used as pig feed, the growth efficiency of the pig is high and the diarrhea rate is low; compared with Example 1, the nitrogen source of the fermentation activated culture medium in Example 6 and Example 7 lacks one of soybean meal and corn syrup dry powder, respectively, and the feed-to-meat ratio is higher than that in Example 1, and the diarrhea rate is higher than that in Example 1, indicating that when the nitrogen source selects a combination of soybean meal and corn syrup dry powder, the fermentation performance of the fermented soybean meal is good, the live count is high, and when used for pig feeding, the growth efficiency of the pig is high and the diarrhea rate is low.

[0129] Compared with Example 1, Comparative Examples 1-5 lack one of Lactobacillus plantarum, Pediococcus acidilactici, Bacillus coagulans, Bacillus subtilis, and yeast, respectively, and their feed-to-meat ratio and diarrhea rate are higher than those of Example 1, indicating that the combined fermentation of the five microorganisms can synergistically enhance the effect, effectively reduce the diarrhea rate of farmed animals, and improve growth efficiency.

[0130] Compared with Example 1, the types of lactic acid bacteria, bacillus and yeast in Comparative Example 6 are different, and its feed-to-meat ratio and diarrhea rate are higher than those in Example 1, indicating that the composite bacterial agent for fermenting soybean meal of the present invention, compared with the bacterial agent in CN114403295A, has a high number of viable bacteria in the fermented soybean meal product, a low diarrhea rate for feeding animals, and improved growth efficiency.

[0131] In Comparative Example 7, compared with Example 1, the composite bacterial agent for fermenting soybean meal was directly used for soybean meal fermentation without expansion culture, and its feed-to-meat ratio and diarrhea rate were higher than those of Example 1, indicating that the composite bacterial agent for fermenting soybean meal was activated before fermentation culture, and the cell activity of the microorganisms was stronger during fermentation. When the obtained fermented soybean meal was used to feed animals, the diarrhea rate of the animals decreased and the growth efficiency was improved.

[0132] The control group used commercially available fermented soybean meal, and its feed-to-meat ratio and diarrhea rate were higher than those in Example 1, indicating that the fermented soybean meal prepared with the composite bacterial agent for fermented soybean meal of the present invention has better animal feeding effect.

[0133] The applicant declares that the present invention illustrates the method and process of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0134] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0135] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A composite bacterial agent for fermenting soybean meal, characterized in that: The strains in the composite bacterial agent for fermenting soybean meal include lactic acid bacteria, bacillus and yeast; the lactic acid bacteria include plant lactobacillus and Pediococcus acidilactici; the bacillus include bacillus coagulans and bacillus subtilis.

2. The composite bacterial agent for fermenting soybean meal according to claim 1, characterized in that The ratio of the number of live bacteria of the lactic acid bacteria, the bacillus and the yeast is (2-10):(5-20):1; more preferably, the ratio of the number of live bacteria of the lactic acid bacteria, the bacillus and the yeast is (5-8):(10-15):1; Preferably, the ratio of the viable counts of Lactobacillus plantarum to Pediococcus acidilactici is 1:(0.5-5); Preferably, the ratio of the number of live bacteria of Bacillus coagulans to that of Bacillus subtilis is 1:(1-8); Preferably, the total bacterial viability in the composite bacterial agent for fermenting soybean meal is not less than 1×10 11 CFU / g or 1×10 11 CFU / mL; Preferably, the lactic acid bacteria further include any one of Lactococcus lactis, Streptococcus lactis or Bifidobacterium lactis, or a combination of at least two thereof.

3. A fermented soybean meal, characterized in that: The fermented soybean meal is obtained by fermenting the composite bacterial agent according to claim 1 or 2.

4. The method for preparing fermented soybean meal according to claim 3, characterized in that: The preparation method comprises the following steps: (1) inoculating the fermented soybean meal composite bacterial agent into a composite bacterial agent activation culture medium, culturing at 30-40° C. for 16-24 hours to obtain a composite bacterial solution; (2) Mixing dry soybean meal and water, inoculating the composite bacterial solution, spreading the mixed solution on a fermentation bed, and performing anaerobically fermentation at 30-40° C. for 60-96 hours to obtain the fermented soybean meal.

5. The method for preparing fermented soybean meal according to claim 4, characterized in that: The culture medium for activating the composite bacterial agent in step (1) comprises a carbon source, a nitrogen source and an inorganic salt; Preferably, the carbon source comprises any one or a combination of at least two of brown sugar, glucose, stachyose, raffinose, corn starch, glycerol or maltose; more preferably, the carbon source comprises brown sugar; Preferably, the nitrogen source comprises any one or a combination of at least two of soybean meal, corn steep powder, ammonium chloride, urea, peptone or yeast powder; more preferably, the nitrogen source comprises soybean meal and corn steep powder; Preferably, the inorganic salt includes any one of dipotassium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, ammonium dihydrogen sulfate or ammonium sulfate, or a combination of at least two thereof; more preferably, the inorganic salt includes dipotassium hydrogen phosphate.

6. The method for preparing fermented soybean meal according to claim 4 or 5, characterized in that: The culture medium for activating the composite bacterial agent also includes a microbial activator; Preferably, the microbial activator comprises any one of 5-aminolevulinic acid, folic acid, lipoic acid, biotin, pantothenic acid, vitamin A or vitamin C, or a combination of at least two thereof.

7. The method for preparing fermented soybean meal according to claim 5, characterized in that: The mass ratio of the carbon source, the nitrogen source and the inorganic salt is (5-20):(10-30):1; Preferably, the mass ratio of the soybean meal to the corn syrup dry powder is (1-10):

1.

8. The method for preparing fermented soybean meal according to any one of claims 4 to 7, characterized in that: The water content in step (2) is 30-60 wt %; more preferably, the water content in step (2) is 40-45 wt %.

9. The method for preparing fermented soybean meal according to any one of claims 4 to 8, characterized in that: After the inoculation in step (2), the number of viable bacteria in the mixed solution is 10 7 -10 9 CFU / g.

10. Use of the composite bacterial agent for fermented soybean meal according to claim 1 or 2, the fermented soybean meal according to claim 3, or the preparation method of the fermented soybean meal according to any one of claims 4 to 9 in the production of soybean meal feed.

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