Soil native flora propagation preparation based on compounding of protein and yeast metabolite

Abstract

The invention belongs to the technical field of soil improvement, and particularly relates to a soil native flora propagation preparation based on compounding of protein and yeast metabolite, which is prepared from the following raw materials in percentage by weight: 38-42% of protein substances, 35-40% of yeast metabolite and 18-22% of auxiliary components. The zein is applied to flora propagation for the first time, the spherical structure of the zein can wrap a slow-release nitrogen source, the nitrogen supply period is prolonged to 90 days (only 45 days in conventional urea), the compounding proportion of the protein and the metabolite accurately meets the metabolism requirement of soil microorganisms, flora inhibition caused by carbon and nitrogen imbalance is avoided, and the nitrogen supply efficiency is improved. The microbial community propagation efficiency is effectively improved, the diversity recovery of the soil is guaranteed, the original beneficial bacteria of the soil can be directly propagated, and a preparation with adaptive nutrition can be provided for the soil microbial community, so that the diversity and quantity of the original microbial community and soil organisms of the soil are recovered, and the sustainable restoration of the soil is realized; and the method can be suitable for saline-alkali soil, continuous cropping obstacles (strawberries and oranges) and mine restoration scenes, and is high in applicability.

Description

Technical Field

[0001] This invention belongs to the field of soil improvement technology, specifically relating to a soil native microbial community propagation preparation based on a compound of protein and yeast metabolites. Background Technology

[0002] In agricultural planting, the long-term and excessive use of chemical fertilizers and pesticides has led to a decrease in organic matter content, poor soil structure, and soil compaction, severely restricting sustainable agricultural development. Globally, 30% of arable land has degraded. The core manifestations of soil degradation are:

[0003] 1. Loss of microbial diversity: Long-term application of chemical fertilizers leads to a 50%-80% decrease in the abundance of functional microbial communities (such as nitrogen-fixing bacteria and phosphate-solubilizing bacteria); 2. Destruction of aggregate structure: More than 60% of soils have an organic matter content of less than 2% and a porosity of less than 30%, resulting in a loss of water and fertilizer retention capacity.

[0004] 3. Nutrient cycling imbalance: Traditional fertilization leads to an imbalance in the C / N ratio (generally >30:1), which inhibits microbial activity.

[0005] Existing technologies typically use soil conditioners to improve and restore contaminated soil. Traditional soil conditioners often rely on single microbial agents or organic fertilizers, which have problems such as low colonization rate of microbial communities, insufficient nutrient supply, and unstable effects. Summary of the Invention

[0006] The purpose of this invention is to provide a soil native microbial community propagation preparation based on a combination of protein and yeast metabolites. This preparation can directly propagate beneficial native soil bacteria and provide suitable nutrients for the soil microbial community, thereby restoring the diversity and quantity of native soil microbial community and soil organisms, and thus achieving sustainable soil remediation.

[0007] The specific technical solution adopted by this invention is as follows:

[0008] A soil native microbial community propagation preparation based on a protein and yeast metabolite compound is prepared from the following raw materials in percentage fractions:

[0009] Proteins 38-42%, yeast metabolites 35-40%, auxiliary components 18-22%, of which:

[0010] The protein components include 32-36% hydrolyzed soybean protein, 6-8% zein, and 2-4% animal protein;

[0011] Yeast metabolites include 30-35% of Saccharomyces cerevisiae metabolites and 5-10% of metabolites from other microorganisms;

[0012] The auxiliary ingredients include 12-15% humic acid and 6-7% glucose residue.

[0013] Furthermore, the animal protein source is fish meal or bone meal.

[0014] Furthermore, the other microbial metabolites contain Bacillus subtilis and Trichoderma.

[0015] Furthermore, the humic acid is derived from lignite.

[0016] A method for preparing a soil native microbial community propagation agent based on a protein and yeast metabolite compound includes the following steps:

[0017] S1. Raw material pretreatment:

[0018] Enzyme inactivation treatment of protein substances: Spread the mixed protein powder of soybean hydrolyzed protein, zein and animal protein in an evaporating dish of the corresponding percentage fractions, and place the evaporating dish in a vacuum oven. Control the temperature of the vacuum oven at 110±2℃, the vacuum degree at ≤10KPa, and the dry distillation time at 15±2min. After dry distillation, cool the mixed protein powder to room temperature and grind it through a 200-mesh sieve.

[0019] Yeast metabolite filtration: After the mixed solution of brewing metabolites and other microbial metabolites of the corresponding percentage fraction is pre-filtered through 0.45μm, it enters the ceramic membrane system and is filtered according to the parameters of pressure 0.1-0.3MPa and flow rate 5-10L / h. The ceramic membrane system is backwashed every 30 minutes to prevent membrane clogging.

[0020] S2, Gradient Mixing:

[0021] First stage: Pre-crush the glucose residue to a particle size ≤1mm, then put the crushed glucose residue, enzyme-inactivated protein substances, and filtered yeast metabolites into a magnetic stirrer and stir at 50-70rpm for 10-15min to form mixed solution one.

[0022] Second stage: Dissolve humic acid in a small amount of deionized water and slowly add it dropwise to the above mixed solution to form mixed solution two. Then, use a high pressure homogenizer to shear mixed solution two at a shear rate of 1500-2000 rpm for 5-8 minutes. After shearing, let it stand for 30 minutes to form a uniform microcapsule suspension.

[0023] S3, Dynamic Fermentation:

[0024] Aerobic stage: Add the microcapsule suspension to a constant temperature fermenter and ferment for 15 days according to the following parameters: temperature 35-38℃, aeration rate 0.5vvm (volume ratio), stirring rate 200rpm, and turning frequency 3 times / week (mechanical turner).

[0025] Anaerobic stage: Nitrogen gas is introduced into the fermenter for 30 minutes to replace the oxygen in the fermenter and create an anaerobic environment. Fermentation is then carried out for 21 days according to the parameters of temperature gradient control (decreasing by 2℃ per week) and nitrogen flow rate of 0.1L / min.

[0026] S4. Post-processing:

[0027] Ultrafine grinding: airflow grinding to D90≤50μm;

[0028] Coating treatment: Sodium alginate-chitosan composite membrane (coating rate ≥95%);

[0029] S5. Quality Inspection:

[0030] Laser particle size analysis (D50 = 18-25 μm);

[0031] FTIR spectroscopy verifies functional group integrity;

[0032] High-throughput sequencing monitors bacterial community activity.

[0033] The technical effects achieved by this invention are as follows:

[0034] This invention is the first to apply zein to microbial community propagation. Its spherical structure can encapsulate a slow-release nitrogen source, extending the nitrogen supply cycle to 90 days (compared to only 45 days for conventional urea). Furthermore, the ratio of protein to metabolites precisely matches the metabolic needs of soil microorganisms, avoiding microbial community inhibition caused by carbon-nitrogen imbalance. This effectively improves microbial community propagation efficiency and ensures the restoration of soil diversity. It is a formulation that can directly propagate native beneficial bacteria in the soil and provide suitable nutrients for soil microorganisms, thereby restoring the diversity and quantity of native soil microorganisms and soil organisms, thus achieving sustainable soil remediation. Moreover, yeast metabolites and other microbial metabolites are not equivalent to extracts, ensuring that this formulation does not introduce foreign microorganisms into the soil. With the technical goal of restoring the diversity and quantity of native soil microorganisms and soil organisms, it achieves sustainable restoration of the soil micro-ecosystem. It is also applicable to saline-alkali land, continuous cropping obstacles (strawberry, citrus), and mine remediation scenarios, demonstrating strong applicability. Detailed Implementation

[0035] To make the objectives and advantages of this invention clearer, the invention will be specifically described below with reference to embodiments. It should be understood that the following text is merely used to describe one or more specific embodiments of the invention and does not strictly limit the scope of protection specifically claimed by the invention.

[0036] A soil native microbial community propagation preparation based on a protein and yeast metabolite compound is prepared from the following raw materials in percentage fractions:

[0037] Proteins 38-42%, yeast metabolites 35-40%, auxiliary components 18-22%, of which:

[0038] The protein components include 32-36% hydrolyzed soybean protein, 6-8% zein, and 2-4% animal protein. Among them, hydrolyzed soybean protein contains 18 kinds of amino acids (including rare amino acids such as ornithine) to provide a slow-release nitrogen source, while zein is rich in hydrophobic amino acids (leucine and isoleucine) to form a nanoscale slow-release carrier. When zein is applied to the propagation of bacterial communities, its spherical structure can encapsulate the slow-release nitrogen source and extend the nitrogen supply cycle to 90 days (compared to only 45 days for conventional urea).

[0039] Yeast metabolites contain 30-35% of Saccharomyces cerevisiae metabolites and 5-10% of metabolites from other microbial species. Among them, yeast metabolites contain glutathione (≥100mg / g) and nucleotides, which enhance the stress resistance of the microbial community.

[0040] The auxiliary ingredients include 12-15% humic acid and 6-7% glucose residue. The glucose residue serves as a short-term carbon source supplement, promoting the rapid reproduction of microorganisms.

[0041] The animal protein sources are fish meal and bone meal. Animal protein provides essential amino acids and minerals, enhancing the metabolic diversity of the gut microbiota.

[0042] The other microbial metabolites contain Bacillus subtilis and Trichoderma, which synergistically activate the indigenous microbial community and enhance stress resistance.

[0043] The humic acid is extracted from lignite and is used to improve soil aggregate structure and provide carboxyl and phenolic hydroxyl active sites.

[0044] A method for preparing a soil native microbial community propagation agent based on a protein and yeast metabolite compound includes the following steps:

[0045] S1. Raw material pretreatment:

[0046] Enzyme inactivation treatment of protein substances: Spread the mixed protein powder of soybean hydrolyzed protein, zein and animal protein in an evaporating dish of the corresponding percentage fractions, and place the evaporating dish in a vacuum oven. Control the temperature of the vacuum oven at 110±2℃, the vacuum degree at ≤10KPa, and the dry distillation time at 15±2min. After dry distillation, cool the mixed protein powder to room temperature and grind it through a 200-mesh sieve.

[0047] Yeast metabolite filtration: After the mixed solution of brewing metabolites and other microbial metabolites of the corresponding percentage fraction is pre-filtered through 0.45μm, it enters the ceramic membrane system and is filtered according to the parameters of pressure 0.1-0.3MPa and flow rate 5-10L / h. The ceramic membrane system is backwashed every 30 minutes to prevent membrane clogging.

[0048] S2, Gradient Mixing:

[0049] First stage: Pre-crush the glucose residue to a particle size ≤1mm, then put the crushed glucose residue, enzyme-inactivated protein substances, and filtered yeast metabolites into a magnetic stirrer and stir at 50-70rpm for 10-15min to form mixed solution one.

[0050] Second stage: Dissolve humic acid in a small amount of deionized water and slowly add it dropwise to the above mixed solution to form mixed solution two. Then, use a high pressure homogenizer to shear mixed solution two at a shear rate of 1500-2000 rpm for 5-8 minutes. After shearing, let it stand for 30 minutes to form a uniform microcapsule suspension.

[0051] S3, Dynamic Fermentation:

[0052] Aerobic stage: Add the microcapsule suspension to a constant temperature fermenter and ferment for 15 days according to the following parameters: temperature 35-38℃, aeration rate 0.5vvm (volume ratio), stirring rate 200rpm, and turning frequency 3 times / week (mechanical turner).

[0053] Anaerobic stage: Nitrogen gas is introduced into the fermenter for 30 minutes to replace the oxygen in the fermenter and create an anaerobic environment. Fermentation is then carried out for 21 days according to the parameters of temperature gradient control (decreasing by 2℃ per week) and nitrogen flow rate of 0.1L / min.

[0054] S4. Post-processing:

[0055] Ultrafine grinding: airflow grinding to D90≤50μm;

[0056] Coating treatment: Sodium alginate-chitosan composite membrane (coating rate ≥95%);

[0057] S5. Quality Inspection:

[0058] Laser particle size analysis (D50 = 18-25 μm);

[0059] FTIR spectroscopy verifies functional group integrity;

[0060] High-throughput sequencing monitors bacterial community activity.

[0061] This formulation utilizes the above-described preparation process to construct a three-tiered nutritional system: rapid-acting, sustained-release, and controlled-release.

[0062] Rapid-acting layer (0-7 days): Monosaccharides (glucose ≤2%) in yeast metabolites rapidly activate indigenous bacteria;

[0063] Slow-release layer (15-60 days): Soy protein hydrolysates provide a continuous nitrogen source (C / N = 12:1);

[0064] Controlled-release layer (60-180 days): Corn gliadin nanoparticles slowly release amino acids.

[0065] Example 1:

[0066] A method for preparing a soil native microbial community propagation agent based on a protein and yeast metabolite compound includes the following steps:

[0067] S1. Raw material pretreatment:

[0068] Enzyme inactivation treatment of protein substances: Spread the mixed protein powder of soybean hydrolyzed protein, zein and animal protein in an evaporating dish with the corresponding percentage fractions, and place the evaporating dish in a vacuum oven. Control the temperature of the vacuum oven to 112℃, the vacuum degree to 10KPa, and the dry distillation time to 17min. After dry distillation, cool the mixed protein powder to room temperature and grind it through a 200-mesh sieve.

[0069] Yeast metabolite filtration: The mixed solution of brewing metabolites and other microbial metabolites of the corresponding percentage fraction is pre-filtered through 0.45μm and then enters the ceramic membrane system. The filtration is carried out according to the parameters of pressure 0.3MPa and flow rate 10L / h, and the ceramic membrane system is backwashed every 30 minutes to prevent membrane clogging.

[0070] S2, Gradient Mixing:

[0071] First stage: Pre-crush the glucose residue to a particle size ≤1mm, then put the crushed glucose residue, enzyme-inactivated protein substances, and filtered yeast metabolites into a magnetic stirrer and stir at 50rpm for 15min to form a mixed solution.

[0072] Second stage: Dissolve humic acid in a small amount of deionized water and slowly add it dropwise to the above mixed solution to form mixed solution two. Then, use a high pressure homogenizer to shear mixed solution two at a shear rate of 1500 rpm for 8 minutes. After shearing, let it stand for 30 minutes to form a uniform microcapsule suspension.

[0073] S3, Dynamic Fermentation:

[0074] Aerobic stage: The microcapsule suspension was added to a constant temperature fermenter and fermented for 15 days according to the following parameters: temperature 35℃, aeration rate 0.5vvm (volume ratio), stirring rate 200rpm, and turning frequency 3 times / week (mechanical turner).

[0075] Anaerobic stage: Nitrogen gas is introduced into the fermenter for 30 minutes to replace the oxygen in the fermenter and create an anaerobic environment. Fermentation is then carried out for 21 days according to the parameters of temperature gradient control (decreasing by 2℃ per week) and nitrogen flow rate of 0.1L / min.

[0076] S4. Post-processing:

[0077] Ultrafine grinding: airflow grinding to D90≤50μm;

[0078] Coating treatment: Sodium alginate-chitosan composite membrane (coating rate ≥95%);

[0079] S5. Quality Inspection:

[0080] Laser particle size analysis (D50 = 18-25 μm);

[0081] FTIR spectroscopy verifies functional group integrity;

[0082] High-throughput sequencing monitors bacterial community activity.

[0083] Example 2:

[0084] A method for preparing a soil native microbial community propagation agent based on a protein and yeast metabolite compound includes the following steps:

[0085] S1. Raw material pretreatment:

[0086] Enzyme inactivation treatment of protein substances: Spread the mixed protein powder of soybean hydrolyzed protein, zein and animal protein in an evaporating dish with the corresponding percentage fractions, and place the evaporating dish in a vacuum oven. Control the temperature of the vacuum oven to 108℃, the vacuum degree to 8KPa, and the dry distillation time to 13min. After dry distillation, cool the mixed protein powder to room temperature and grind it through a 200-mesh sieve.

[0087] Yeast metabolite filtration: The mixed solution of brewing metabolites and other microbial metabolites of the corresponding percentage fraction is pre-filtered through 0.45μm and then enters the ceramic membrane system. The filtration is carried out according to the parameters of pressure 0.1MPa and flow rate 5L / h, and the ceramic membrane system is backwashed every 30 minutes to prevent membrane clogging.

[0088] S2, Gradient Mixing:

[0089] First stage: Pre-crush the glucose residue to a particle size ≤1mm, then put the crushed glucose residue, enzyme-inactivated protein substances, and filtered yeast metabolites into a magnetic stirrer and stir at 70rpm for 10min to form mixed solution one.

[0090] Second stage: Dissolve humic acid in a small amount of deionized water and slowly add it dropwise to the above mixed solution to form mixed solution two. Then, use a high pressure homogenizer to shear mixed solution two at a shear rate of 2000 rpm for 5 minutes. After shearing, let it stand for 30 minutes to form a uniform microcapsule suspension.

[0091] S3, Dynamic Fermentation:

[0092] Aerobic stage: The microcapsule suspension was added to a constant temperature fermenter and fermented for 15 days according to the following parameters: temperature 38℃, aeration rate 0.5vvm (volume ratio), stirring rate 200rpm, and turning frequency 3 times / week (mechanical turner).

[0093] Anaerobic stage: Nitrogen gas is introduced into the fermenter for 30 minutes to replace the oxygen in the fermenter and create an anaerobic environment. Fermentation is then carried out for 21 days according to the parameters of temperature gradient control (decreasing by 2℃ per week) and nitrogen flow rate of 0.1L / min.

[0094] S4. Post-processing:

[0095] Ultrafine grinding: airflow grinding to D90≤50μm;

[0096] Coating treatment: Sodium alginate-chitosan composite membrane (coating rate ≥95%);

[0097] S5. Quality Inspection:

[0098] Laser particle size analysis (D50 = 18-25 μm);

[0099] FTIR spectroscopy verifies functional group integrity;

[0100] High-throughput sequencing monitors bacterial community activity.

[0101] The above description is merely a preferred embodiment of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention. Structures, devices, and operating methods not specifically described or explained in this invention are implemented according to conventional methods in the art unless otherwise specified or limited.

Claims

1. A soil native microbial community propagation preparation based on a compound of protein and yeast metabolites, characterized in that: It is prepared from the following percentage fractions of raw materials: Proteins 38-42%, yeast metabolites 35-40%, auxiliary components 18-22%, of which: The protein components include 32-36% hydrolyzed soybean protein, 6-8% zein, and 2-4% animal protein; Yeast metabolites include 30-35% of Saccharomyces cerevisiae metabolites and 5-10% of metabolites from other microorganisms; The auxiliary ingredients include 12-15% humic acid and 6-7% glucose residue.

2. The soil native microbial community propagation preparation based on a protein and yeast metabolite compound according to claim 1, characterized in that: The animal protein sources are fish meal and bone meal.

3. The soil native microbial community propagation preparation based on a protein and yeast metabolite compound according to claim 1, characterized in that: The other microbial metabolites contain Bacillus subtilis and Trichoderma.

4. The soil native microbial community propagation preparation based on a protein and yeast metabolite compound according to claim 1, characterized in that: The humic acid is derived from lignite.

5. A method for preparing a soil native microbial community propagation agent based on a compound of protein and yeast metabolites according to any one of claims 1-4, characterized in that: Includes the following steps: S1. Raw material pretreatment: Enzyme inactivation treatment of protein substances: Spread the mixed protein powder of soybean hydrolyzed protein, zein and animal protein in an evaporating dish of the corresponding percentage fractions, and place the evaporating dish in a vacuum oven. Control the temperature of the vacuum oven at 110±2℃, the vacuum degree at ≤10KPa, and the dry distillation time at 15±2min. After dry distillation, cool the mixed protein powder to room temperature and grind it through a 200-mesh sieve. Yeast metabolite filtration: After the mixed solution of brewing metabolites and other microbial metabolites of the corresponding percentage fraction is pre-filtered through 0.45μm, it enters the ceramic membrane system and is filtered according to the parameters of pressure 0.1-0.3MPa and flow rate 5-10L / h. The ceramic membrane system is backwashed every 30 minutes to prevent membrane clogging. S2, Gradient Mixing: First stage: Pre-crush the glucose residue to a particle size ≤1mm, then put the crushed glucose residue, enzyme-inactivated protein substances, and filtered yeast metabolites into a magnetic stirrer and stir at 50-70rpm for 10-15min to form mixed solution one. Second stage: Dissolve humic acid in a small amount of deionized water and slowly add it dropwise to the above mixed solution to form mixed solution two. Then, use a high pressure homogenizer to shear mixed solution two at a shear rate of 1500-2000 rpm for 5-8 minutes. After shearing, let it stand for 30 minutes to form a uniform microcapsule suspension. S3, Dynamic Fermentation: Aerobic stage: Add the microcapsule suspension to a constant temperature fermenter and ferment for 15 days according to the following parameters: temperature 35-38℃, aeration rate 0.5vvm (volume ratio), stirring rate 200rpm, and turning frequency 3 times / week (mechanical turner). Anaerobic stage: Nitrogen gas is introduced into the fermenter for 30 minutes to replace the oxygen in the fermenter and create an anaerobic environment. Fermentation is then carried out for 21 days according to the parameters of temperature gradient control (decreasing by 2℃ per week) and nitrogen flow rate of 0.1L / min. S4. Post-processing: Ultrafine grinding: airflow grinding to D90≤50μm; Coating treatment: Sodium alginate-chitosan composite membrane (coating rate ≥95%); S5. Quality Inspection: Laser particle size analysis (D50 = 18-25 μm); FTIR spectroscopy verifies functional group integrity; High-throughput sequencing monitors bacterial community activity.