Bacillus siamensis and application thereof in fermented rapeseed meal
Fermentation of rapeseed meal with Bacillus sicca BSJ-2 solved the problem of low protein digestibility, improved its utilization and nutritional value in animal feed, and achieved a significant enhancement of protein and amino acids.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- INST OF MICROBIOLOGY CHINESE ACAD OF SCI
- Filing Date
- 2024-12-17
- Publication Date
- 2026-06-23
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of microbial technology and relates to a strain of Bacillus sicca and its application in fermented rapeseed meal. Background Technology
[0002] Protein is a crucial component of animal feed. With the rapid development of my country's livestock and aquaculture industries and the continuous expansion of large-scale farming, the shortage of protein feed has become a prominent issue. Soybean meal, the most widely used protein feed ingredient, is heavily reliant on imports. Affected by market supply and demand imbalances, soybean meal prices have risen continuously, hindering the healthy development of my country's livestock industry. Rapeseed meal, a byproduct of rapeseed oil extraction, is rich in protein, amino acids, vitamins, and other trace elements, with a crude protein content of approximately 35%-45%. Under the "Soybean Meal Reduction and Substitution Action," rapeseed meal demonstrates significant application potential due to its wide availability, high protein content, balanced amino acid composition, and low price. However, currently, rapeseed meal has a lower protein digestibility compared to soybean meal, affecting protein absorption and utilization by animals. There is an urgent need to improve the protein digestibility of rapeseed meal and enhance its nutritional value for better application in animal feed. Microbial solid-state fermentation can improve the chemical composition and physicochemical properties of feed, and is economical and environmentally friendly, making it an ideal means to improve the nutritional value and digestibility of feed for animals.
[0003] Studies have shown that *Bacillus sicca* has potential applications in biocontrol, detoxification of mycotoxin-contaminated feed, and screening for high-protease-producing strains, warranting further research and exploration. While few conventional Bacillus strains can improve the protein utilization rate of rapeseed meal, identifying Bacillus strains capable of degrading recalcitrant proteins in rapeseed meal for fermentation pretreatment can effectively improve the nutritional quality and protein digestibility of rapeseed meal, providing a theoretical basis for its application, promotion, and efficiency improvement. Summary of the Invention
[0004] The technical problem solved by this invention is to provide a substance that improves the protein utilization rate of rapeseed meal.
[0005] To solve the above-mentioned technical problems, the first aspect of the present invention provides Bacillus sicca ( Bacillus siamensis The BSJ-2 strain has the accession number CGMCC No. 32638.
[0006] Secondly, the present invention provides the use of the strain or its culture medium or its bacterial suspension or its fermentation broth described in the first aspect in any of the following or in the preparation of products having any of the following: 1) Degradation of stored protein in rapeseed meal; 2) Improve the nutritional quality of rapeseed meal; 3) Improve the protein utilization rate of rapeseed meal; 4) Improve the composition and / or content of essential amino acids that are key to animal growth in rapeseed meal.
[0007] In the applications described above, the bacterial solution can also be the supernatant of the bacterial solution, specifically, the OD... 600 The value is approximately 1.0. Centrifuge the bacterial solution at 10,000 rpm for 10 minutes and collect the supernatant.
[0008] In the applications described above, the product can be a microbial agent.
[0009] Thirdly, the present invention provides a product in which the active ingredient is the strain described in the first aspect or its culture medium or its bacterial liquid or its fermentation broth.
[0010] The product described above has any of the following functions: 1) Degradation of stored protein in rapeseed meal; 2) Improve the nutritional quality of rapeseed meal; 3) Improve the protein utilization rate of rapeseed meal; 4) Improve the composition and / or content of essential amino acids that are key to animal growth in rapeseed meal.
[0011] Fourthly, the present invention provides the application of the strain or its culture medium or its bacterial liquid or its fermentation broth described in the first aspect or the product described in the third aspect in the preparation of rapeseed meal fermentation products; Alternatively, this invention provides the application of the strain or its culture medium or its bacterial liquid or its fermentation broth described in the first aspect, or the product described in the third aspect, in the preparation of animal feed using rapeseed meal as raw material. The above-mentioned preparation of rapeseed meal fermentation product involves adding BSJ-2 bacterial agent at an addition amount of 1*10^6 live bacteria. 8 CFU / g FM (wet weight) is evenly sprayed onto the rapeseed meal to be used, and stirred evenly to obtain a mixture. Take 30 g of the mixture and put it into a polyethylene bag. Cover the bag opening with sealing film and tie it tightly with rubber bands. Open the bag at 9:00 and 21:00 every day to stir and ventilate. Ferment for 3 days with an average indoor temperature between 20 and 25℃. Collect the rapeseed meal fermentation product.
[0012] The above-mentioned BSJ-2 bacterial agent has a concentration of 3*10. 9 The BSJ-2 bacterial suspension (CFU / mL, water as solvent) was prepared as follows: The strain described in the first aspect was cultured in LB medium at 37°C and 220 rpm for 12 h with shaking to obtain the fermentation broth. The fermentation broth was plated on LB plates for counting, centrifuged, and resuspended in sterile water to obtain the BSJ-2 bacterial agent with a concentration of 3*10⁻⁶. 9 CFU / mL.
[0013] The rapeseed meal to be used above is obtained by adjusting the dry matter content of rapeseed meal to 40% with sterile distilled water.
[0014] Fifthly, the present invention provides a rapeseed meal fermentation product, which is prepared by a method comprising the following steps: adding the strain or its culture broth or its bacterial liquid or its fermentation broth or the product described in the first aspect to rapeseed meal, and fermenting to obtain the rapeseed meal fermentation product.
[0015] The fermentation time is 3 days; the fermentation temperature is 20~25℃.
[0016] The above-mentioned strains were added to rapeseed meal at a ratio of 1*10^9 live bacteria. 8 CFU / g FM (wet weight).
[0017] In a sixth aspect, the present invention provides an animal feed in which the active ingredient is the rapeseed meal fermentation product described in the fifth aspect.
[0018] In the above text, the storage proteins include cruciferin protein and / or napin protein.
[0019] The improvement in the nutritional quality of rapeseed meal described above is reflected in the increased content of crude protein (CP) and soluble protein (SP), the increased content of non-protein nitrogen (NPN) and fat (FAT), the significant decrease in the content of neutral detergent fiber (NDF), the slight increase in the content of acid detergent fiber (ADF), the decrease in neutral detergent insoluble crude protein (NDICP), and / or the increase in acid detergent insoluble crude protein (ADICP).
[0020] In the above text, the improvement in the protein utilization rate of rapeseed meal is reflected in the increase of nonprotein nitrogen (PA) and rapidly degradable rumen protein (PB1) components that are more easily digested and absorbed by the rumen; and / or the decrease of moderately degradable rumen protein (PB2) and slowly degradable rumen protein (PB3) components that are less easily digested and absorbed.
[0021] The essential amino acids that are crucial for animal growth mentioned above include serine, threonine, and / or isoleucine.
[0022] The products mentioned above may also be drugs, compositions, health products, functional foods, foods for special medical purposes, or other biological products.
[0023] The products described above may also include pharmaceutically acceptable carriers, which may be excipients, stabilizers, suspending agents or diluents, as is well known to those skilled in the art.
[0024] Furthermore, the carrier materials include, but are not limited to, water-soluble carrier materials (such as polyethylene glycol, polyvinylpyrrolidone, organic acids, etc.), poorly soluble carrier materials (such as ethyl cellulose, cholesterol stearate, etc.), and enteric-coated carrier materials (such as cellulose acetate phthalate and carboxymethyl ethyl cellulose, etc.). Using these materials, various dosage forms can be formulated, including but not limited to tablets, capsules, pellets, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal preparations, lozenges, suppositories, lyophilized powder injections, etc. These can be conventional formulations, sustained-release formulations, controlled-release formulations, and various microparticle delivery systems. To formulate unit-dose dosage forms into tablets, various carriers known in the art can be widely used. Examples of carriers include diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, and aluminum silicate; humectants and binders such as water, glycerin, polyethylene glycol, ethanol, propanol, starch paste, dextrin, syrup, honey, glucose solution, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, shellac, methyl cellulose, potassium phosphate, and polyvinylpyrrolidone; and disintegrants. Examples of carriers include dried starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene, sorbitol fatty acid esters, sodium dodecyl sulfate, methylcellulose, and ethylcellulose; disintegration inhibitors include sucrose, tristearate, cocoa butter, and hydrogenated oil; absorption enhancers include quaternary ammonium salts and sodium dodecyl sulfate; and lubricants include talc, silica, corn starch, stearates, boric acid, liquid paraffin, and polyethylene glycol. Tablets can also be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets. Various carriers known in the art can be widely used to formulate unit-dose dosage forms into pills. Examples of carriers include diluents and absorbents such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, kaolin, and talc; binders such as gum arabic, tragacanth, gelatin, ethanol, honey, liquid sugar, rice paste, or flour paste; and disintegrants such as agar powder, dried starch, alginate, sodium dodecyl sulfonate, methylcellulose, and ethylcellulose. For preparing unit-dose dosage forms into suppositories, a wide variety of carriers known in the art can be used. Examples of carriers include polyethylene glycol, lecithin, cocoa butter, higher alcohols, esters of higher alcohols, gelatin, and semi-synthetic glycerides. For preparing unit-dose dosage forms into injectable formulations such as solutions, emulsions, lyophilized powders for injection, and suspensions, all diluents commonly used in the art can be used, such as water, ethanol, polyethylene glycol, 1,3-propanediol, ethoxylated isostearyl alcohol, polyoxyethylene isostearyl alcohol, and polyoxyethylene sorbitan fatty acid esters. In addition, to prepare isotonic injection solutions, appropriate amounts of sodium chloride, glucose, or glycerol can be added to the injectable formulation. Furthermore, conventional solubilizers, buffers, pH adjusters, etc., can also be added.In addition, colorants, preservatives, flavorings, tasters, sweeteners or other materials may be added to pharmaceutical preparations if necessary.
[0025] The experiments of this invention demonstrate that this invention has discovered Bacillus sicca ( Bacillus siamensis The BSJ-2 strain's fermentation products can degrade stored protein in rapeseed meal, improve the nutritional quality of rapeseed meal, increase the utilization rate of rapeseed meal protein, and can be used to prepare animal feed.
[0026] Preservation Instructions Strain name: BSJ-2 Latin name: Bacillus siamensis Preservation Institution: China General Microbiological Culture Collection Center, China Microbiological Culture Collection Committee Collection institution abbreviation: CGMCC Address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing Deposit date: November 14, 2024 CGMCC Registration Number: 32638 Classification and nomenclature: Bacillus sicca Bacillus siamensis Attached Figure Description Figure 1 To detect the activity of rapeseed meal protein degraded by BSJ-2 bacterial supernatant.
[0027] Figure 2 Comparison of the nutritional quality of fermented rapeseed meal (* represents BSJ-2 vs. FR, # represents BSJ-2 vs. CK).
[0028] Figure 3 Changes in CNCPS protein composition of fermented rapeseed meal.
[0029] Figure 4 Principal component analysis of amino acids in fermented rapeseed meal.
[0030] Figure 5 A heatmap of amino acid content in fermented rapeseed meal (FR vs. CK vs. BSJ-2).
[0031] Figure 6 A heatmap of amino acid content in fermented rapeseed meal (CK vs. BSJ-2).
[0032] Figure 7 The absolute amino acid content of fermented rapeseed meal (CK vs. BSJ-2). Detailed Implementation The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.
[0033] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.
[0034] Unless otherwise specified, the quantitative experiments in the following examples are all repeated three times, and the results are averaged.
[0035] Example 1: Isolation and Identification of Bacillus BSJ-2 (Siamese Bacillus) 1. Separation Weigh 10g of soil, add 90ml of sterile water, shake on a shaker for 30min, dilute the liquid and spread it on LB agar plates, incubate overnight upside down, and record the single colonies that grow as strain BSJ-2. Pick a single colony of strain BSJ-2 and transfer it to 3ml of LB liquid medium, and incubate at 37℃ with shaking at 220rpm for 24h.
[0036] 2. Molecular identification DNA was extracted from strain BSJ-2 and used as a template for amplification with primers 27F and 1492R to obtain PCR products of 16S rRNA.
[0037] 27F: 5'-AGA GTT TGA TCC TGG CTC AG-3' 1492R:5'-GGT TAC CTT GTT ACG ACT T-3 After sequencing, the full sequence of the 16S rRNA is shown in Sequence 1.
[0038] The strain BSJ-2 was deposited on November 14, 2024, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing; accession number CGMCC No. 32638; and classified as Bacillus sicca. Bacillus siamensis .
[0039] Example 2: Application of Bacillus sicca BSJ-2 I. Bacillus sicca BSJ-2 has the activity of degrading stored protein in rapeseed meal. The rapeseed meal was purchased from Inner Mongolia Jinshengxiang Agricultural and Animal Husbandry Technology Co., Ltd. The majority of the protein in rapeseed meal is storage protein, with over 90% being cruciferin and napin, accounting for 60% and 30% respectively. Cruciferin is a complex composed of six monomeric proteins, with GenBank numbers 3KGL_A, 3KGL_B, 3KGL_C, 3KGL_D, 3KGL_E, and 3KGL_F, all submitted on November 1, 2023. Napin's GenBank number is P24565.1, submitted on March 27, 2024.
[0040] Weigh 1g of rapeseed meal sample, add 10mL of 10mM sodium chloride aqueous solution (pH=7), and extract by shaking at 37℃ and 180rpm for 1h. Centrifuge the resulting mixture at 10,000rpm for 10min. The supernatant obtained is obtained by filtering through a sterile 0.22μm filter membrane to obtain rapeseed meal protein extract.
[0041] BSJ-2 was streaked onto LB agar plates and incubated overnight at 37°C with the plates inverted. A single colony was then transferred to 3 mL of liquid LB medium and incubated at 37°C with shaking at 220 rpm for 12 hours. The OD of the bacterial culture was measured. 600 The value was approximately 1.0. The culture was centrifuged at 10,000 rpm for 10 min, and the supernatant was collected. After filtration through a sterile 0.22 μm filter membrane, it was mixed with rapeseed meal protein extract at a volume ratio of 1:1 and incubated at 37℃ for 5 h to obtain the test sample. A blank LB medium mixed with rapeseed meal protein extract at a volume ratio of 1:1 served as the control group.
[0042] Take 40 μL of the sample to be tested, add 10 μL of SDS-PAGE loading buffer (product number B1030-5), mix well by pipetting, boil in a water bath for 10 min, centrifuge at 12000 rpm for 5 min, and take 20 μL of the supernatant for SDS-PAGE detection (product number T202109).
[0043] The results are as follows Figure 1 As shown, the supernatant of BSJ-2 bacterial culture can effectively degrade the cruciferin subunit (approximately 50 KD) and napin protein (approximately 15 KD).
[0044] The above results indicate that Bacillus sicca BSJ-2 has the activity of degrading rapeseed meal storage proteins.
[0045] II. Fermentation with Bacillus sicca BSJ-2 improves the protein utilization rate and nutritional quality of rapeseed meal. 1. Rapeseed meal fermentation method 1) Adjust the dry matter content of rapeseed meal to 40% with sterile distilled water and set aside for later use. 2) Activated bacterial suspension: The BSJ-2 obtained in Example 1 was streaked onto LB agar plates and incubated overnight at 37°C with the plates inverted. Single colonies were then picked and transferred to 3 mL of liquid LB medium. After incubation at 37°C with shaking at 220 rpm for 12 h, 1% of the culture was transferred to 1.5 L of LB liquid medium and cultured at 37°C with shaking at 220 rpm for 12 h. The resulting BSJ-2 bacterial suspension was plated and counted on LB agar plates. After centrifugation, the suspension was resuspended in sterile water to obtain the BSJ-2 bacterial agent with a concentration of 3*10⁻⁶. 9 CFU / mL.
[0046] 3) Experimental group (BSJ-2): The BSJ-2 bacterial agent obtained in step 2) was added at a concentration of 1*10⁻⁶ viable bacteria. 8 CFU / gFM (wet weight) is evenly sprayed onto the rapeseed meal obtained in step 1), and stirred evenly to obtain a mixture. 30 g of the mixture is placed into a polyethylene bag, the bag opening is wrapped with sealing film and tied tightly with rubber bands. The bag is opened at 9:00 and 21:00 every day for stirring and ventilation. Fermentation is carried out for 3 days with an average indoor temperature between 20 and 25°C. The fermentation products are then collected.
[0047] Control group (CK): The only difference from the experimental group was that no BSJ-2 inoculant was added, and the fermentation products were collected.
[0048] Fresh sample group (FR): Unadjusted dry rapeseed meal without fermentation. Each group has 3 replicates.
[0049] 2. Detection of rapeseed meal fermented with Bacillus sicca BSJ-2 1) Nutritional quality related indicators Nutritional quality-related indicators were tested by the CVAS Feed Analysis China Service Center.
[0050] The fermentation products of the experimental group (BSJ-2) and the control group (CK) and the fresh sample group (FR) were tested for nutritional quality-related indicators.
[0051] The results are as follows Figure 2The results showed no significant differences in any indicators between the fresh sample group (FR) and the control group (CK) without fermentation. However, compared to the FR and CK groups, the BSJ-2 fermentation group significantly increased the content of crude protein (CP) and soluble protein (SP), as well as non-protein nitrogen (NPN) and fat (FAT). At the same time, it significantly reduced the content of neutral detergent fiber (NDF), while the content of acid detergent fiber (ADF) increased slightly. The BSJ-2 fermentation group had a lower level of neutral detergent-insoluble crude protein (NDICP) than the CK group, while the BSJ-2 fermentation group had a higher level of acid detergent-insoluble crude protein (ADICP) than both the CK and FR groups.
[0052] These indicators combined show that BSJ-2 fermentation can significantly improve the nutritional quality of rapeseed meal.
[0053] 2) Protein component analysis The fermentation products of the experimental and control groups in section 1 above, as well as the Cornell Net Carbohydrate and Net Protein System (CNCPS), ADF, NDF, and other indicators of the fresh sample group (FR) were detected.
[0054] Crude protein (CP), acid detergent fiber (ADF), and neutral detergent fiber (NDF) were determined according to the official analytical chemist association method described in the following literature: Horwitz, W. & AOAC International (Eds.). Official methods of analysis of AOAC International (18. ed., current through rev. 1, 2006).
[0055] Non-protein nitrogen (NPN) was determined using the TCA precipitation method, as described in the following literature: Latimer, GW (Ed.). (2023). AOAC Official Method 941.04 Urea and Ammoniacal Nitrogen in Animal Feed: Urease Method. In Official Methods of Analysis of AOACINTERNATIONAL (22nd ed.). Oxford University Press. The method for measuring soluble protein (SP) is described in the following literature: Krishnamoorthy, U., Muscato, TV, Sniffen, CJ, & Van Soest, PJ (1982). Nitrogen Fractions in Selected Feedstuffs. Journal of Dairy Science, 65(2), 217–225. The residues from the ADF and NDF procedures were used to determine acid detergent-insoluble protein (ADIP) and neutral detergent-insoluble protein (NDIP), with the methods described in the following literature: Horwitz, W. & AOAC International (Eds.). Official methods of analysis of AOAC International (18. ed., current throughrev. 1, 2006). The protein component content of CNCPS is calculated according to the following formula: The calculation method of each component of CNCPS (PA, PB1, PB2, PB3, PC) and the corresponding relationship with the index are as follows: PA=NPN; PB1=SP-PA; PB2=CP-SP-NDICP; PB3=NDIP-ADICP; PC=ADIP.
[0056] The results are as follows Figure 3As shown, the nonprotein nitrogen (PA) and rapidly degradable rumen protein (PB1) components in the fermentation products of the BSJ-2 fermentation group were significantly increased, reaching more than three times that of the fresh sample and the CK group, while the rumen medium-degradable protein (PB2) and slow-degradable rumen protein (PB3) components, which are not easily digested and absorbed, were significantly reduced.
[0057] This indicates that fermentation with Bacillus sicca BSJ-2 improves the protein utilization and nutritional quality of rapeseed meal.
[0058] 3) Fermentation with Bacillus sicca BSJ-2 can increase the content of important amino acids in rapeseed meal. The fermentation products of the experimental group and the control group in section 1 above, as well as the fresh sample group (FR), were used as the test samples.
[0059] Weigh 20 mg of each sample and add 400 μL of pre-cooled 80% methanol aqueous solution. Add two medium-sized steel balls and homogenize at low temperature in a tissue homogenizer. Add 600 μL of pre-cooled 80% methanol aqueous solution, vortex to mix, sonicate in an ice bath for 20 min, let stand at -20℃ for 1.5 h, centrifuge at 16000g at 4℃ for 20 min, collect the supernatant, and evaporate the supernatant to dryness using a high-speed vacuum centrifuge to obtain the sample. Redissolve the sample in 100 μL of pre-cooled 50% methanol aqueous solution, centrifuge at 20000g at 4℃ for 15 min, and collect the supernatant for UHPLC and mass spectrometry analysis.
[0060] The parameters and detection conditions are as follows: Ultra-high performance liquid chromatography (UHPLC) conditions A Shimadzu Nexera X2 A8-30AD high-performance liquid chromatography (HPLC) system was used. The column was a Waters UPA8 BEHAmide column (1.7µm, 2.1x100mm). The mobile phases were: Solution A was a 0.1% formic acid aqueous solution containing 10mM ammonium formate; Solution B was an acetonitrile solution containing 0.1% formic acid. The sample was placed in an autosampler at 4℃, the column temperature was 40℃, the flow rate was 300µL / min, and the injection volume was 2µL. The relevant HPLC gradients were as follows: 0–1 min, Solution B maintained at 90%; 1–9 min, Solution B linearly changed from 90% to 40%; 9–10.5 min, Solution B maintained at 40%; 10.5–10.6 min, Solution B linearly changed from 40% to 90%; 10.6–12 min, Solution B maintained at 90%. Mass spectrometry analysis Mass spectrometry analysis was performed using a QTRAP 5500 mass spectrometer (AB SCIEX) in positive ion mode. The QTRAP ESI source parameters are as follows: Positive ion mode: Source Temperature 550℃, Ion SourceGas1 (GAS1): 40, Ion Source Gas2 (GAS2): 50, Curtain Gas (CUR): 35, Ion SprayVoltage Floating (ISVF) 5500V. The analyte ion pairs are detected using MRM mode.
[0061] PCA principal component analysis results are as follows: Figure 4 As shown, the amino acid composition of the FR, CK and BSJ-2 groups differs significantly, and the repeatability among the repeats within each group is good.
[0062] The abundance heatmap of the three amino acid groups is as follows: Figure 5 The results showed that, compared with the FR group, the amino acid content of both the CK group and the BSJ-2 group was significantly increased.
[0063] Further analysis was conducted on the CK group (fermentation without BSJ-2) and the BSJ-2 group. Data from the CK and BSJ groups were compared separately, and the results are as follows: Figure 6 , 7 As shown, the results indicate that, compared with the CK group, the BSJ-2 group had significantly higher levels of essential amino acids crucial for animal growth, including serine, threonine, and isoleucine.
[0064] The above Figure 5 and Figure 6 Use the ComplexHeatmap package from Bioconductor version 2.22.0. Figure 7 The graphpad prism version 8.01 was used, and the significance analysis was performed using two-way anova (P < 0.05).
[0065] Therefore, Bacillus sicca BSJ-2 fermentation improves the composition and content of amino acids that are important for animal growth in rapeseed meal. The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims.
Claims
1. Bacillus sicca ( Bacillus siamensis The BSJ-2 strain has the accession number CGMCC No. 32638.
2. The use of the strain of claim 1 or its culture medium or its bacterial suspension or its fermentation broth in any of the following or in the preparation of a product having any of the following characteristics: 1) Degradation of stored protein in rapeseed meal; 2) Improve the nutritional quality of rapeseed meal; 3) Improve the protein utilization rate of rapeseed meal; 4) Improve the composition and / or content of essential amino acids that are key to animal growth in rapeseed meal.
3. The application according to claim 2, characterized in that: The product is a microbial agent.
4. A product, wherein the active ingredient is the strain of claim 1 or its culture medium or its bacterial culture or its fermentation broth.
5. The product according to claim 4, characterized in that: The product has any of the following functions: 1) Degradation of stored protein in rapeseed meal; 2) Improve the nutritional quality of rapeseed meal; 3) Improve the protein utilization rate of rapeseed meal; 4) Improve the composition and / or content of essential amino acids that are key to animal growth in rapeseed meal.
6. The use of the strain of claim 1 or its culture medium or its bacterial liquid or its fermentation broth, or the product of claim 4 or 5, in the preparation of rapeseed meal fermentation products; The application of the strain or its culture medium or its bacterial liquid or its fermentation broth as described in claim 1, or the product as described in claim 4 or 5, in the preparation of animal feed made from rapeseed meal.
7. A rapeseed meal fermentation product, which is prepared by a method comprising the following steps: adding the strain of claim 1 or its culture medium or its bacterial liquid or its fermentation liquid or the product of claim 4 or 5 to rapeseed meal, and fermenting to obtain the rapeseed meal fermentation product.
8. An animal feed, wherein the active ingredient is the rapeseed meal fermentation product as described in claim 7.