Process for fermenting peanut meal by complex bacteria and fermented product thereof

Abstract

The application discloses a peanut meal fermentation process by using composite bacteria and a fermentation product thereof, and belongs to the technical field of peanut meal fermentation. The process comprises the following steps: fermenting peanut meal mixture by using composite bacteria; the composite bacteria are bacillus velezensis, bacillus amyloliquefaciens, pediococcus acidilactici and lactobacillus rhamnosus; the composite bacteria are prepared into composite bacteria liquid before fermentation; the composite bacteria liquid is composed of bacillus velezensis liquid, bacillus amyloliquefaciens liquid, pediococcus acidilactici liquid and lactobacillus rhamnosus liquid. By adopting the co-fermentation system of bacillus and lactic acid bacteria, the functions of enzymolysis, detoxification, pH adjustment and bacteriostasis are synergized, the retention rate of crude protein in the fermentation product is high, the contents of acid-soluble protein and small peptides are greatly increased, the removal effect of aflatoxin is remarkable, the pH is reduced to the acidic range, and the biological safety is improved.

Description

Technical Field

[0001] This invention belongs to the field of peanut meal fermentation technology, specifically the process of fermenting peanut meal with compound bacteria and its fermentation products. Background Technology

[0002] Peanut meal is a byproduct of peanut oil extraction. It is widely available, inexpensive, high in crude protein, and has a relatively balanced amino acid composition, making it a valuable plant protein raw material in the feed industry. However, untreated peanut meal presents several problems: firstly, its protein is mostly large-molecule protein, resulting in low digestibility and absorption, and it contains high levels of anti-nutritional factors; secondly, peanut meal is highly susceptible to contamination by Aspergillus flavus during storage and transportation, producing aflatoxin, which severely limits its safe and efficient use in livestock, poultry, and aquatic feed.

[0003] To improve the feed quality of peanut meal and reduce the risk of toxins, existing technologies commonly employ physical, chemical, and microbial fermentation methods. Among these, microbial fermentation has become the mainstream technology for the resource utilization of peanut meal due to its mild conditions, environmental safety, and ability to simultaneously achieve protein degradation, toxin removal, and quality improvement. Currently, most existing processes use Bacillus, lactic acid bacteria, or a simple combination of two bacteria for fermentation. However, the function of a single strain is limited, with insufficient ability to adjust the system's pH, completely degrade toxins, or simultaneously achieve the multiple objectives of high protein degradation rate, high small peptide content, high total acid content, and efficient detoxification. Summary of the Invention

[0004] To overcome the aforementioned technical problems, this invention provides a process for fermenting peanut meal using compound bacteria and its fermentation products. This invention achieves synergistic effects of enzymatic hydrolysis, detoxification, pH adjustment, and antibacterial activity by employing a co-fermentation system of Bacillus and lactic acid bacteria. The fermentation products exhibit high crude protein retention, significantly increased content of acid-soluble proteins and small peptides, remarkable aflatoxin removal, and improved pH and biocompatibility by reducing the pH to the acidic range.

[0005] The present invention solves the above-mentioned technical problems through the following technical solutions.

[0006] The process of fermenting peanut meal with compound bacteria includes the following steps: fermenting peanut meal mixture with compound bacteria; The compound bacteria consist of Bacillus belyssus, Bacillus amyloliquefaciens, Pediococcus lactis, and Lactobacillus rhamnosus.

[0007] The compound bacteria are prepared into a compound bacterial solution and then fermented; the compound bacterial solution is composed of Bacillus vesiculosus solution, Bacillus amyloliquefaciens solution, Pediococcus lactis solution and Lactobacillus rhamnosus solution.

[0008] According to some embodiments of the present invention, the activity of each bacterial strain in the Bacillus bellis solution, Bacillus amyloliquefaciens solution, Pediococcus lactis solution, and Lactobacillus rhamnosus solution is 0.5 × 10⁻⁶. 9 ~2.0×10 9 CFU / mL.

[0009] According to some embodiments of the present invention, the volume ratio of Bacillus vesiculosus solution, Bacillus amyloliquefaciens solution, Pediococcus lactis solution and Lactobacillus rhamnosus solution is 1~5:1~5:1:1, preferably 1~3:2~4:1:1.

[0010] Bacillus amyloliquefaciens has a rich enzyme production system and high enzyme yield, and can efficiently synthesize proteases, cellulases, and aflatoxin B1 degrading enzymes. The higher proportion of these enzymes can increase the concentration of degrading enzymes in the fermentation system, accelerating the decomposition of macromolecular proteins and crude fibers, as well as the degradation of aflatoxin. At the same time, this strain can rapidly consume oxygen in the fermentation system, creating an anaerobic environment for the growth and reproduction of anaerobic lactic acid bacteria. Therefore, its ratio is slightly higher than that of Bacillus belyss. As a Bacillus strain, Bacillus belyss can assist Bacillus amyloliquefaciens in producing a variety of active enzymes, enhancing the degradation effect of macromolecules. It can also produce secondary metabolites such as antimicrobial peptides, inhibiting the growth of mold in the fermentation material, and forming a synergistic system with Bacillus amyloliquefaciens. Pediococcus lactis can produce pediococcin to inhibit Gram-positive pathogenic bacteria, and Lactobacillus rhamnosus can efficiently synthesize lactic acid. The combination of the two can not only rapidly reduce the pH value of the fermentation system and extend the shelf life of the product, but also enhance the biosafety of the fermentation material. The above bacterial culture ratio ensures both the dominant role of Bacillus and the full metabolic effect of Lactic acid bacteria; ultimately, Bacillus initiates the environmental setup first, followed by Lactic acid bacteria to stabilize the system and improve quality.

[0011] According to some embodiments of the present invention, the fermentation is carried out at 30-40°C for 3-7 days; preferably, the fermentation is carried out at 35-39°C for 4-6 days.

[0012] According to some embodiments of the present invention, the pre-fermentation vacuum is applied to allow fermentation to proceed under anaerobic conditions.

[0013] According to some embodiments of the present invention, the fermentation system contains 130-150g of peanut meal mixture, 250-350μL of compound bacterial solution, and 50-80g of fermentation culture medium; preferably, the fermentation system contains 130-150g of peanut meal mixture, 250-300μL of compound bacterial solution, and 55-70g of fermentation culture medium.

[0014] According to some embodiments of the present invention, the peanut meal mixture is 80-95% peanut meal, with the remainder being peanut shells, wheat bran, or palm meal; preferably, the peanut meal mixture is 88-95% peanut meal, with the remainder being peanut shells, wheat bran, or palm meal.

[0015] According to some embodiments of the present invention, the fermentation medium contains 3-7% sucrose, 0.2-2.5% protease, and the balance water.

[0016] According to some embodiments of the present invention, the peanut meal mixture contains 47.0-50.5% crude protein, 1.8-2.3% acid-soluble protein, 3.9-4.3% small peptides and 0.3-0.55% total acid; The molecular weight of the small peptide is <5000 Da, preferably ≤1000 Da. Total acidity refers to the total amount of all acidic components in food, including free acidity and bound acidity.

[0017] According to some embodiments of the present invention, the aflatoxin content in the peanut meal mixture is ≥30 ppb.

[0018] According to some embodiments of the present invention, the preparation process of the composite bacterial solution includes the following steps: Bacillus bellis, Bacillus amyloliquefaciens, Pediococcus lactis and Lactobacillus rhamnosus are activated and inoculated respectively to obtain Bacillus bellis solution, Bacillus amyloliquefaciens solution, Pediococcus lactis solution and Lactobacillus rhamnosus solution respectively, and the composite bacterial solution is obtained by mixing the various bacterial solutions.

[0019] The inoculation process involves first culturing the culture at 28-32°C for 14-18 hours to obtain a primary bacterial culture, and then incubating it at 0.8-1.2% in shake flask culture medium at 28-32°C for 30-40 hours.

[0020] The *Bacillus berreatus* and the *Bacillus amyloliquefaciens* were both cultured on *Bacillus* agar plates during the inoculation process; the *Lactobacillus rhamnosus* was cultured on *Lactobacillus* agar plates during the inoculation process; and the *Pediococcus lactis* was cultured on *Pediococcus* agar plates during the inoculation process.

[0021] Further, the Bacillus agar plate culture medium contains 4-6 g / L yeast extract, 8-13 g / L tryptone, 8-13 g / L sodium chloride, and 16-25 g / L agar powder; preferably, the Bacillus agar culture medium contains 4.5-5.5 g / L yeast extract, 9-11 g / L tryptone, 9-11 g / L sodium chloride, and 18-22 g / L agar powder. Furthermore, both the Lactobacillus agar plate and the Pediococcus agar plate contain 5.5–4.5 g / L yeast extract, 8–13 g / L peptone, 15–25 g / L glucose, 0.1–0.3 g / L magnesium sulfate, 4.5–5.5 g / L beef meal, 0.03–0.08 g / L manganese sulfate, 4–6 g / L sodium acetate, 1.5–2.5 g / L triammonium citrate, 0.5–1.5 g / L Tween-80, 1.5–2.5 g / L dipotassium hydrogen phosphate, and 17–25 g / L... Agar powder; preferably, the lactobacillus culture medium contains 3.8~4.2 g / L yeast powder, 9~11 g / L peptone, 18~22 g / L glucose, 0.18~0.22 g / L magnesium sulfate, 4.8~5.2 g / L beef meal, 0.04~0.06 g / L manganese sulfate, 4.8~5.2 g / L sodium acetate, 1.8~2.2 g / L triammonium citrate, 0.9~1.1 g / L Tween-80, 1.8~2.2 g / L dipotassium hydrogen phosphate and 18~22 g / L agar powder.

[0022] The activation process involves culturing at 28-32°C for 20-30 hours.

[0023] The *Bacillus belye* and *Bacillus amyloliquefaciens* were cultured in shake flasks using *Bacillus* culture medium during the inoculation process; *Lactobacillus rhamnosus* was cultured in shake flasks using *Lactobacillus* culture medium during the inoculation process; and *Pediococcus lactis* was cultured in shake flasks using *Pediococcus* culture medium during the inoculation process.

[0024] Further, the Bacillus shake flask culture medium contains 4.5-5.5 g / L yeast extract, 8-13 g / L tryptone, and 8-13 g / L sodium chloride; preferably, the Bacillus shake flask culture medium contains 4.8-5.2 g / L yeast extract, 9-11 g / L tryptone, and 9-11 g / L sodium chloride.

[0025] Furthermore, both the Lactobacillus shake flask culture medium and the Pediococcus shake flask culture medium contain 3.5–4.5 g / L yeast extract, 7–13 g / L peptone, 15–25 g / L glucose, 0.15–0.25 g / L magnesium sulfate, 4.5–5.5 g / L beef meal, 0.03–0.08 g / L manganese sulfate, 4.5–5.5 g / L sodium acetate, 1.5–2.5 g / L triammonium citrate, 0.7–1.5 g / L Tween-80, and 1.5–2.5 g / L... Dipotassium hydrogen phosphate; preferably, the Lactobacillus shake flask culture medium contains 3.8-4.2 g / L yeast extract, 9-11 g / L peptone, 18-22 g / L glucose, 0.18-0.22 g / L magnesium sulfate, 4.8-5.2 g / L beef meal, 0.04-0.06 g / L manganese sulfate, 4.8-5.2 g / L sodium acetate, 1.8-2.2 g / L triammonium citrate, 0.9-1.1 g / L Tween-80 and 1.8-2.2 g / L dipotassium hydrogen phosphate.

[0026] The fermentation product disclosed in this invention is obtained by the aforementioned process.

[0027] According to some embodiments of the present invention, the fermentation product contains 47-52% crude protein, 6.0-8.0% acid-soluble protein, 12-15% small peptides and 2.0-2.8% total acid; preferably, the fermentation product contains 48-51% crude protein, 6.5-7.5% acid-soluble protein, 13-14.6% small peptides and 2.1-2.7% total acid.

[0028] According to some embodiments of the present invention, the aflatoxin content in the fermentation product is ≤50 ppb, preferably ≤30 ppb, for example 10~30 ppb.

[0029] This invention also discloses the application of compound bacterial solution in feed preparation or water quality improvement.

[0030] The feed includes daily basic feed, moldy feed, or aquatic feed.

[0031] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0032] Compared with the prior art, the beneficial effects of the present invention are: This invention employs a fermentation process using a combination of *Bacillus belye*, *Bacillus amyloliquefaciens*, *Pediococcus lactis*, and *Lactobacillus rhamnosus* to process peanut meal. *Bacillus belye* and *Bacillus amyloliquefaciens* form the core of enzymatic hydrolysis and environmental construction. They not only synergistically produce various active enzymes such as proteases, cellulases, and aflatoxin B1 degrading enzymes, significantly improving the degradation efficiency of macromolecules and the removal of aflatoxin, but also create an anaerobic and sterile environment suitable for lactic acid bacteria growth through rapid oxygen consumption and antimicrobial peptide secretion. *Pediococcus lactis* and *Lactobacillus rhamnosus* achieve acid reduction and antibacterial effects; the large-scale synthesis of lactic acid rapidly lowers the system pH, inhibiting the growth of contaminating bacteria and extending the product's shelf life, while the secretion of pediococcal peptides further enhances the biosafety of the fermentation product.

[0033] The fermentation process of this invention achieves the technical objectives of high protein degradation rate, high small molecule nutrient content, efficient detoxification, and high biocompatibility. The crude protein content in the fermentation product remains at a high level, while the content of acid-soluble proteins and small peptides is significantly increased. These small molecule nutrients are more easily digested and absorbed by livestock, poultry, and aquatic animals, significantly improving the feed value of peanut meal. The aflatoxin removal effect is significant, solving the problem of toxic contamination during peanut meal storage. Simultaneously, the pH value of the fermentation product is reduced to the acidic range, and combined with the antibacterial substances produced by the microorganisms, the shelf life and biocompatibility of the product are greatly improved. It can be directly used as a high-quality plant protein raw material in the feed industry, realizing the resource utilization and high-value utilization of peanut meal. Attached Figure Description

[0034] To facilitate understanding by those skilled in the art, the present invention will be further described below with reference to the accompanying drawings.

[0035] Figure 1 This is a comparison diagram of the fermentation system before and after fermentation in the example. Detailed Implementation

[0036] To facilitate understanding of the present invention, the present invention will be described more fully and in detail below with reference to preferred embodiments, but the scope of protection of the present invention is not limited to the following specific embodiments.

[0037] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those skilled in the art. The technical terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the scope of the invention.

[0038] The "range" disclosed in this invention is defined by a lower limit and an upper limit. A given range is defined by selecting a lower limit and an upper limit, which define the boundaries of a particular range. Ranges defined in this way can include or exclude endpoints and can be combined arbitrarily; that is, any lower limit can be combined with any upper limit to form a range. For example, if ranges of 60-120 and 80-110 are listed for specific parameters, it is understood that ranges of 60-110 and 80-120 are also expected. Furthermore, if minimum range values ​​1 and 2 are listed, and if maximum range values ​​3, 4, and 5 are listed, then the following ranges are all expected: 1-3, 1-4, 1-5, 2-3, 2-4, and 2-5. In this invention, unless otherwise stated, the numerical range "ab" represents a shortened representation of any combination of real numbers between a and b, where a and b are real numbers. For example, the numerical range "0-5" means that all real numbers between "0-5" have been listed herein; "0-5" is merely a shortened representation of these numerical combinations. Furthermore, when a parameter is described as an integer greater than or equal to 2, it is equivalent to disclosing that the parameter is, for example, an integer such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, etc.

[0039] Unless otherwise specified, all embodiments and optional embodiments of the present invention can be combined with each other to form new technical solutions.

[0040] Unless otherwise specified, all technical features and optional technical features of this invention can be combined to form new technical solutions.

[0041] Unless otherwise specified, all steps of the present invention may be performed sequentially or randomly, preferably sequentially. For example, the method includes steps (a) and (b), indicating that the method may include steps (a) and (b) performed sequentially, or it may include steps (b) and (a) performed sequentially. For example, the mention that the method may also include step (c) indicates that step (c) may be added to the method in any order; for example, the method may include steps (a), (b), and (c), or it may include steps (a), (c), and (b), or it may include steps (c), (a), and (b), etc.

[0042] Unless otherwise specified, the terms "comprising" and "including" as used in this invention can be open-ended or closed-ended. For example, "comprising" and "including" can mean that other components not listed may also be included, or that only the listed components may be included.

[0043] Unless otherwise specified, the term "or" is inclusive in this invention. For example, the phrase "A or B" means "A, B, or both A and B". More specifically, the condition "A or B" is satisfied by any of the following conditions: A is true (or exists) and B is false (or does not exist); A is false (or does not exist) and B is true (or exists); or both A and B are true (or exist).

[0044] The raw material information used in the following examples is as follows: Bacillus belesiensis ( Bacillus velezensis LPF421, deposited at the Guangdong Provincial Center for Microbial Culture Collection on March 5, 2025, with accession number GDMCC No: 65977; this strain is disclosed in patent application number CN202510515716.3; Pediococcus acidilactici ( Pediococcus acidilactici The strain originated from the Institute of Microbiology, Chinese Academy of Sciences, with strain number CGMCC1.2696 and an activity of 10 billion CFU / g. Lactobacillus rhamnosus ( Lactobacillus rhamnosus Originating from Xi'an Xihai Biotechnology Co., Ltd., with an activity of 10 billion CUF / g; Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens (Source: Shandong Yixiou Biotechnology Co., Ltd., activity 10 billion CFU / g;) The peanut meal was purchased from Guangdong Yuehai Feed Group, and the production areas were Guangdong and Guangxi respectively. Wheat bran powder comes from Xiangyuan Feed Comprehensive Service Center in Yuanshi County; Palm meal comes from Lingshou County Senzhuo Agricultural Products Co., Ltd.; The protease was purchased from Xiasheng Biotechnology Co., Ltd. SFG-2441, which is a (alkaline) protease preparation refined from Bacillus subtilis fermentation. Bacillus agar plate medium: yeast extract (Angel yeast extract powder FM902) 5g / L, tryptone 10g / L, sodium chloride 10g / L, agar powder 20g / L; Lactobacillus agar plate and Pediococcus agar plate: yeast extract 4 g / L, peptone 10 g / L, glucose 20 g / L, magnesium sulfate 0.2 g / L, beef extract 5 g / L, manganese sulfate 0.05 g / L, sodium acetate 5 g / L, triammonium citrate 2 g / L, Tween-80 1 g / L, dipotassium hydrogen phosphate 2 g / L, agar powder 20 g / L; Bacillus shake flask culture medium and Pediococcus shake flask culture medium: yeast extract 5g / L, tryptone 10g / L, sodium chloride 10g / L; Lactobacillus shake flask culture medium: yeast extract 4g / L, peptone 10g / L, glucose 20g / L, magnesium sulfate 0.2g / L, beef meal 5g / L, manganese sulfate 0.05g / L, sodium acetate 5g / L, triammonium citrate 2g / L, Tween-80 1g / L, dipotassium hydrogen phosphate 2g / L; This includes, but is not limited to, the models from the above manufacturers.

[0045] Example 1 The process for fermenting peanut meal using compound bacteria in this embodiment is as follows: S1. Inoculation: Bacillus belye, Bacillus amyloliquefaciens, Pediococcus lactis and Lactobacillus rhamnosus were activated on their respective agar plates at 30°C for 24 h to obtain individual colonies; Activation: The four individual colonies were inoculated into shake flask culture medium and cultured for 16 hours at 30°C and 220 rpm. Then, they were inoculated again into shake flask culture medium at a 1% inoculum and cultured for 36 hours at 30°C and 220 rpm. The resulting cultures were Bacillus belyssus, Bacillus amyloliquefaciens, Pediococcus lactis, and Lactobacillus rhamnosus. S2. Preparation of the composite bacterial solution: 56 μL of *Bacillus belyssioides* solution, 112 μL of *Bacillus amyloliquefaciens* solution, 56 μL of *Pediococcus lactis* solution, and 56 μL of *Lactobacillus rhamnosus* solution were separately taken and mixed to obtain a composite bacterial solution (total 280 μL); the activity of all four bacteria in the composite bacterial solution was 1.5 × 10⁻⁶. 9 CFU / mL; S3. Fermentation: Weigh 130g of peanut meal (from Guangdong, passed through a 20-mesh sieve) and 10g of peanut shell powder (passed through a 10-mesh sieve) and mix them to obtain peanut meal mixture. The peanut meal mixture contains 48% crude protein, 1.94% acid-soluble protein, 4.04% small peptides and 0.42% total acid, with a pH of 6.41 and an aflatoxin content of 35.09 ppb. Add fermentation medium (containing 0.24g sucrose, 0.06g protease and 59.3mL water) to the peanut meal mixture, then add the above-mentioned compound bacterial solution and mix well. Place the mixture in a double-sided threaded vacuum packaging bag, vacuum seal it, and store it at 37℃ for 5 days until the fermentation is complete. Figure 1 The images show a comparison of the fermentation system before and after fermentation in this embodiment. The left image shows the fermentation system before fermentation, and the right image shows the fermentation system after fermentation. Before fermentation, the liquid in the system was a cloudy, milky yellow color with low transparency; after fermentation, the liquid was significantly clearer, light transparent, with only a small amount of fine suspended matter remaining, and the amount of solid matter was reduced.

[0046] Example 2 The difference between this embodiment and Embodiment 1 is as follows: The peanut meal raw material in the peanut meal mixture was changed; in this embodiment, the peanut meal sourced from Guangxi was used. The peanut meal mixture contains 49.10% crude protein, 2.02% acid-soluble protein, 4.11% small peptides and 0.5% total acid, with a pH of 6.45 and an aflatoxin content of 119.56 ppb. The other raw materials, steps and parameters are the same as in Example 1.

[0047] Example 3 The difference between this embodiment and Embodiment 1 is as follows: S2. Preparation of compound bacterial solution: Take 80 μL of Bacillus vesiculosus solution, 120 μL of Bacillus amyloliquefaciens solution, 40 μL of Pediococcus lactis solution and 40 μL of Lactobacillus rhamnosus solution respectively, and mix the individual bacterial solutions to obtain a compound bacterial solution (total 280 μL). The other raw materials, steps and parameters are the same as in Example 1.

[0048] Example 4 The difference between this embodiment and Embodiment 1 is as follows: S2. Preparation of composite bacterial solution: Take 70 μL of Bacillus vesiculosus solution, 70 μL of Bacillus amyloliquefaciens solution, 70 μL of Pediococcus lactis solution and 70 μL of Lactobacillus rhamnosus solution respectively, and mix the individual bacterial solutions to obtain a composite bacterial solution (total 280 μL). The other raw materials, steps and parameters are the same as in Example 1.

[0049] Example 5 The difference between this embodiment and Embodiment 1 is as follows: S3 fermentation medium contains 0.3g sucrose and 59.3mL water; the fermentation medium does not contain trypsin. The other raw materials, steps and parameters are the same as in Example 1.

[0050] Example 6 The difference between this embodiment and Embodiment 1 is as follows: S3 fermentation is carried out at 39℃ for 4 days; The other raw materials, steps and parameters are the same as in Example 1.

[0051] Comparative Example 1 The difference between this comparative example and Example 1 is as follows: S2. Preparation of compound bacterial solution: Take 140 μL of Bacillus vesiculosus solution and 140 μL of Pediococcus lactis solution respectively, and mix the two solutions to obtain a compound bacterial solution (total 280 μL). The other raw materials, steps and parameters are the same as in Example 1.

[0052] Comparative Example 2 The difference between this comparative example and Example 1 is as follows: S2. Preparation of composite bacterial solution: Take 90 μL of Bacillus vesiculosus solution, 90 μL of Bacillus amyloliquefaciens solution, and 90 μL of Pediococcus lactis solution respectively, and mix the individual bacterial solutions to obtain a composite bacterial solution (total 270 μL). The other raw materials, steps and parameters are the same as in Example 1.

[0053] Comparative Example 3 The difference between this comparative example and Example 1 is as follows: S2. Preparation of composite bacterial solution: Take 90 μL of Bacillus vesiculosus solution, 90 μL of Pediococcus lactis solution and 90 μL of Lactobacillus rhamnosus solution respectively, and mix the individual bacterial solutions to obtain a composite bacterial solution (total 270 μL). The other raw materials, steps and parameters are the same as in Example 1.

[0054] Test case The fermentation products obtained from the above examples and comparative examples were subjected to the following tests, and the test results are shown in Table 1. The crude protein test method shall be performed in accordance with GB / T 6432-2018; The test method for acid-soluble proteins shall be performed in accordance with Appendix B, section B.4.1 of GB / T 22492-2008; The small peptide testing method shall be performed in accordance with GB / T 22492-2008; The total acid test method shall refer to GB / T 12456-2021; The aflatoxin test was performed using the Longke New Domain Tianjin Aflatoxin Detection Kit.

[0055]

[0056] Unless otherwise specified, all raw materials, reagents, instruments, and equipment used in this invention can be purchased commercially or prepared using existing methods. The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of this invention. It should be understood that the above descriptions are merely specific embodiments of this invention and are not intended to limit the invention. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of this invention should be included within the scope of protection of this invention.

Claims

1. A compound microbial fermentation process for peanut meal, characterized in that, Use a compound microbial fermentation process to ferment peanut meal mixture; The compound bacteria consist of Bacillus belyssus, Bacillus amyloliquefaciens, Pediococcus lactis, and Lactobacillus rhamnosus.

2. The compound microbial fermentation process for peanut meal as described in claim 1, characterized in that, The compound bacteria are prepared into a compound bacterial solution before fermentation; the compound bacterial solution consists of Bacillus vesiculosus solution, Bacillus amyloliquefaciens solution, Pediococcus lactis solution and Lactobacillus rhamnosus solution.

3. The compound microbial fermentation process for peanut meal as described in claim 2, characterized in that, The bacterial activities of *Bacillus belyssioides*, *Bacillus amyloliquefaciens*, *Pediococcus lactis*, and *Lactobacillus rhamnosus* in the composite bacterial solution were independently 0.5 × 10⁻⁶. 9 ~2.0×10 9 CFU / mL.

4. The compound microbial fermentation process for peanut meal as described in claim 3, characterized in that, The volume ratio of Bacillus vesiculosus, Bacillus amyloliquefaciens, Pediococcus lactis, and Lactobacillus rhamnosus in the compound bacterial solution is 1~5:1~5:1:1, preferably 1~3:2~4:1:

1.

5. The compound microbial fermentation process for peanut meal as described in claim 1, characterized in that, The fermentation is carried out at 30-40°C for 3-7 days; preferably, the fermentation is carried out at 35-39°C for 4-6 days. And / or, the pre-fermentation vacuum is applied.

6. The compound microbial fermentation process for peanut meal as described in claim 1, characterized in that, The fermentation system contains 130-150g of peanut meal mixture, 250-350μL of compound bacterial solution, and 50-80g of fermentation culture medium.

7. The compound microbial fermentation process for peanut meal as described in claim 6, characterized in that, The peanut meal mixture consists of 80-95% peanut meal, with the remainder being peanut shells, wheat bran, or palm meal. And / or, the fermentation medium contains 3-7% sucrose, 0.2-2.5% protease, and the remainder water.

8. The compound microbial fermentation process for peanut meal as described in claim 7, characterized in that, The peanut meal mixture contains 47.0-50.5% crude protein, 1.8-2.3% acid-soluble protein, 3.9-4.3% small peptides and 0.3-0.55% total acid; And / or, the aflatoxin content in the peanut meal mixture is ≥30 ppb.

9. A fermentation product, characterized in that, It is prepared by any one of the processes described in claims 1 to 8.

10. The fermentation product according to claim 9, characterized in that, The fermentation product contains 47-52% crude protein, 6.0-8.0% acid-soluble protein, 12-15% small peptides and 2.0-2.8% total acid; preferably, the fermentation product contains 48-51% crude protein, 6.5-7.5% acid-soluble protein, 13-14.6% small peptides and 2.1-2.7% total acid; The fermentation product contains ≤50 ppb of aflatoxin, preferably ≤30 ppb.

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