Slow-release organic-inorganic compound fertilizer based on synergistic interaction of carrier immobilization enzyme and cofactors and preparation method of slow-release organic-inorganic compound fertilizer

Abstract

The invention provides a slow-release organic-inorganic compound fertilizer based on synergistic interaction of a carrier immobilized enzyme and a cofactor and a preparation method of the slow-release organic-inorganic compound fertilizer, and belongs to the technical field of organic fertilizers. According to the application, heat-insulation enzyme-loaded particles are added during pile building, a semi-coke-perlite low-heat-conduction framework forms a cold island in a reactor core, and a starch-polypeptide-sodium tripolyphosphate cross-linked membrane is used for firmly immobilizing enzyme, so that heat inactivation is prevented, fibrin is rapidly cracked, free ammonia is adsorbed, and the pile temperature is promoted to rapidly rise; then, dynamic humidity regulating particles are put, reversible water release-water absorption of montmorillonite is combined with glycerin, and a sorbitol hydrophilic film automatically and stably controls water content and supplements easily-degradable carbon, so that over-wet anaerobic or over-dry enzyme loss is avoided; the enzyme cofactor powder added in the later period slowly releases manganese, calcium and copper to serve as enzyme complement, alanyl glutamine serves as a molecular chaperone to repair damaged conformation, and copper and zinc are chelated to passivate heavy metal; the three agents sequentially realize'keep-alive, stable-alive ', prolong the high activity period of enzyme, accelerate lignin degradation, and reduce nitrogen volatilization, odor and heavy metal risks at the same time.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of organic fertilizer, and relates to a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor and a preparation method thereof. BACKGROUND

[0002] The current livestock and poultry manure composting process usually sprays liquid compound bacteria or compound enzyme at one time when building the pile, and then relies on the raw materials to enter the high temperature stage by self-heating. The temperature of the pile rapidly rises in a short time, and the added enzyme often encounters thermal shock as soon as it enters the constant temperature platform. The macromolecular substrate cannot be cracked in time, the soluble carbon and nitrogen are released slowly, the aerobic bacteria are weak after expansion, and the fermentation process is thus lengthened repeatedly.

[0003] The sharp fluctuation of temperature also affects the imbalance of water. The water content is high at the beginning of building the pile, the capillary water is driven to infiltrate and accumulate at the bottom by the heat of the pile core, and the local environment soon turns into anaerobic environment, producing pungent volatile acid gas. Then the continuous high temperature of the pile core causes the surface to rapidly lose water and crack, forming a hard shell to block oxygen. Anaerobic and dry cracking zones appear alternately in the pile, and the internal microorganism community is frequently imbalanced. The added enzyme cannot rely on the stable three-phase interface to play a role, and the activity further declines.

[0004] On the other hand, the metal cofactor required by the key enzyme system is easily complexed with organic acid and phosphate in the high-temperature and alkaline environment, or precipitated as carbonate and phosphate, so that the cofactor is "dislocated". At this time, even if the enzyme protein has not been completely denatured, it is also in the "enzyme invalid" state due to the loss of metal cooperation; the metal cycle dependent on lignin degradation is also inhibited, leading to further slow carbon chain breaking process.

[0005] With the increase of temperature and pH, the deamination reaction of protein is intensified. The escape of a large amount of ammonia not only causes direct loss of nitrogen, but also inhibits nitrifying bacteria, creating a new bottleneck for nitrogen cycle in the pile. The interweaving of ammonia, sulfide and organic acid promotes the transfer of heavy metals such as copper and zinc from relatively inert complex state to more active exchange state or dissolved state, bringing potential environmental migration risk. SUMMARY

[0006] To solve the above problems, the application aims to provide a slow-release type organic-inorganic compound fertilizer based on synergistic effect of carrier immobilized enzyme and cofactor and a preparation method thereof. The application takes heat-insulating enzyme-loaded particles, dynamic moisture-regulating particles and enzyme cofactor powder as cores to form a closed loop of "keep alive-stabilize alive-revive alive". The heat-insulating enzyme-loaded particles build a cold island in the core by a low thermal conductivity skeleton of semi-coke and perlite and firmly immobilize enzymes by a cross-linked membrane of starch, polypeptide and sodium tripolyphosphate, thereby preventing high-temperature inactivation and rapidly cracking fibers and proteins while adsorbing and releasing free ammonia to promote rapid heating of the pile; the dynamic moisture-regulating particles automatically and stably control the moisture content by reversibly releasing and absorbing water of montmorillonite in combination with a hydrophilic membrane of glycerol and sorbitol and provide easily degradable carbon to avoid anaerobic over-wetting or enzyme loss due to over-drying and maintain enzyme activity in the high-temperature section; and the enzyme cofactor powder slowly releases metal cofactors such as manganese, calcium and copper, alanyl glutamine acts as a molecular chaperone to revive damaged enzymes, and chelates copper, zinc and other heavy metals to passivate them. The three agents synergistically realize long-lasting enzyme activity, deep degradation of lignin, reduction of nitrogen loss and odor emission, improve fermentation efficiency and reduce the dependence on frequent turning or long-period natural cooling.

[0007] To achieve the above object, the application adopts the following technical solutions:

[0008] In a first aspect, the application provides a preparation method of a slow-release type organic-inorganic compound fertilizer based on synergistic effect of carrier immobilized enzyme and cofactor, which comprises the following steps:

[0009] The livestock and poultry manure is mixed with short-cut straw and rice husk to build a first pile, and the heat-insulating enzyme-loaded particles are added during the building of the pile; the pile is turned over on the 2nd day of building and dynamic moisture-regulating particles are added to obtain a second pile; the third pile is obtained after fermentation and turning over; the fourth pile is obtained after turning over and adding enzyme cofactor powder after the third pile is fermented at a constant temperature; and the slow-release type organic-inorganic compound fertilizer based on synergistic effect of carrier immobilized enzyme and cofactor is obtained after continuous fermentation and cooling.

[0010] The preparation method of the heat-insulating enzyme-loaded particles is as follows:

[0011] The shaving is subjected to anaerobic carbonization, pulverization and sieving to obtain semi-coke powder; corn starch dispersion is gelatinized and mixed with soybean polypeptide dispersion and sodium tripolyphosphate solution to obtain a cross-linking and bonding liquid; the semi-coke powder and expanded perlite are uniformly mixed, placed in a rotary granulator and sprayed with the cross-linking and bonding liquid and a complex enzyme liquid to obtain granules, which are dried to obtain the heat-insulating enzyme-loaded particles.

[0012] The preparation method of the dynamic moisture-regulating particles is as follows:

[0013] Montmorillonite, microcrystalline cellulose, glycerol and sorbitol are mixed, extruded by a screw extruder and granulated to obtain an initial product, which is dried to obtain the dynamic moisture-regulating particles.

[0014] The preparation method of the enzyme cofactor powder is as follows:

[0015] The mixed metal salt solution is prepared, the calcium lignosulfonate is mixed with the mixed metal salt solution to obtain a reaction solution, and the reaction solution is dried to obtain a dried product, and the dried product is mixed with alanyl glutamine to obtain an enzyme cofactor powder.

[0016] As a preferred technical solution of the application, in the preparation method of the slow-release organic-inorganic compound fertilizer based on carrier solidification enzyme and cofactor synergistic effect, the mass ratio of the livestock and poultry manure, the short-cut straw and the rice husk is (7-8):(1-1.5):(0.5-1), for example, it can be (7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9 or 8):(1.0, 1.05, 1.10, 1.15, 1.20, 1.25, 1.30, 1.35, 1.40, 1.45 or 1.5):(0.5, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95 or 1.0), but not limited to the listed values, other values not listed in the range are also applicable.

[0017] In some optional embodiments, the water content of the first pile body is 50-60%, for example, it can be 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59% or 60%, but not limited to the listed values, other values not listed in the range are also applicable.

[0018] In some optional embodiments, the feeding amount of the heat-insulating enzyme-loaded particles is 1-2% of the dry basis mass of the first pile body, for example, it can be 1.0%, 1-1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9% or 2.0%, but not limited to the listed values, other values not listed in the range are also applicable.

[0019] In some optional embodiments, the addition amount of the dynamic humidity control particles in the second pile body is 0.8-1.2% of the dry basis mass of the first pile body, for example, it can be 0.80%, 0.84%, 0.88%, 0.92%, 0.96%, 1.00%, 1.04%, 1.08%, 1.12%, 1.16% or 1.20%, but not limited to the listed values, other values not listed in the range are also applicable.

[0020] The frequency of turning is 1 time per day for 0-3 days after pile building, and 1 time every 2 days for 4-10 days.

[0021] In some optional embodiments, the third pile has a temperature of 55-65℃ for 42-52h, for example, it can be 55℃, 56℃, 57℃, 58℃, 59℃, 60℃, 61℃, 62℃, 63℃, 64℃ or 65℃ for 42h, 43h, 44h, 45h, 46h, 47h, 48h, 49h, 50h, 51h or 52h, but not only limited to the listed values, other values not listed in the range are also applicable.

[0022] In some optional embodiments, the fourth pile has an enzyme cofactor powder added in an amount of 0.6-1.0% of the dry basis mass of the first pile, for example, it can be 0.60%, 0.64%, 0.68%, 0.72%, 0.76%, 0.80%, 0.84%, 0.88%, 0.92%, 0.96% or 1.00%, but not only limited to the listed values, other values not listed in the range are also applicable.

[0023] In some optional embodiments, the fourth pile has a temperature of 55-65℃ for 3-5d, for example, it can be 55℃, 56℃, 57℃, 58℃, 59℃, 60℃, 61℃, 62℃, 63℃, 64℃ or 65℃ for 3.0d, 3.2d, 3.4d, 3.6d, 3.8d, 4.0d, 4.2d, 4.4d, 4.6d, 4.8d or 5.0d, but not only limited to the listed values, other values not listed in the range are also applicable.

[0024] As a preferred technical solution of the present application, in the preparation method of the heat-insulating enzyme-loaded particles, the temperature of the shaving anaerobic carbonization is 380-420℃, for example, it can be 380℃, 384℃, 388℃, 392℃, 396℃, 400℃, 404℃, 408℃, 412℃, 416℃ or 420℃, but not only limited to the listed values, other values not listed in the range are also applicable.

[0025] In some optional embodiments, the shaving anaerobic carbonization time is 60-90min, for example, it can be 60min, 63min, 66min, 69min, 72min, 75min, 78min, 81min, 84min, 87min or 90min, but not only limited to the listed values, other values not listed in the range are also applicable.

[0026] In some optional embodiments, the nitrogen flow rate during the shaving oxygen-deficient carbonization is 3-6 L / (min·kg), for example, it can be 3.0 L / (min·kg), 3.3 L / (min·kg), 3.6 L / (min·kg), 3.9 L / (min·kg), 4.2 L / (min·kg), 4.5 L / (min·kg), 4.8 L / (min·kg), 5.1 L / (min·kg), 5.4 L / (min·kg), 5.7 L / (min·kg) or 6.0 L / (min·kg), but not only limited to the listed values, other values not listed in the range are also applicable.

[0027] In some optional embodiments, the particle size of the semi-coke powder is 400-600 μm, for example, it can be 400 μtm, 420 μm, 440 μm, 460 μm, 480 μm, 500 μm, 520 μm, 540 μm, 560 μm, 580 μm or 600 μm, but not only limited to the listed values, other values not listed in the range are also applicable.

[0028] In some optional embodiments, the mass fraction of the corn starch dispersion is 6-8 wt.%, for example, it can be 6.0 wt.%, 6.2 wt.%, 6.4 wt.%, 6.6 wt.%, 6.8 wt.%, 7.0 wt.%, 7.2 wt.%, 7.4 wt.%, 7.6 wt.%, 7.8 wt.% or 8.0 wt.%, but not only limited to the listed values, other values not listed in the range are also applicable.

[0029] In some optional embodiments, the temperature for gelatinization of the corn starch dispersion is 85-95℃, for example, it can be 85℃, 86℃, 87℃, 88℃, 89℃, 90℃, 91℃, 92℃, 93℃, 94℃ or 95℃, but not only limited to the listed values, other values not listed in the range are also applicable.

[0030] In some optional embodiments, the time for gelatinization of the corn starch dispersion is 20-30 min, for example, it can be 20 min, 21 min, 22 min, 23 min, 24 min, 25 min, 26 min, 27 min, 28 min, 29 min or 30 min, but not only limited to the listed values, other values not listed in the range are also applicable.

[0031] In some alternative embodiments, the mass fraction of the soy polypeptide dispersion is 0.8-1.2 wt.%, for example, it can be 0.80 wt.%, 0.84 wt.%, 0.88 wt.%, 0.92 wt.%, 0.96 wt.%, 1.00 wt.%, 1.04 wt.%, 1.08 wt.%, 1.12 wt.%, 1.16 wt.%, or 1.20 wt.%, but is not limited to the listed values, and other values not listed in the range are also applicable.

[0032] In some alternative embodiments, the mass fraction of the sodium tripolyphosphate solution is 0.08-0.12 wt.%, for example, it can be 0.080 wt.%, 0.084 wt.%, 0.088 wt.%, 0.092 wt.%, 0.096 wt.%, 0.100 wt.%, 0.104 wt.%, 0.108 wt.%, 0.112 wt.%, 0.116 wt.%, or 0.120 wt.%, but is not limited to the listed values, and other values not listed in the range are also applicable.

[0033] In some alternative embodiments, the mass ratio of the corn starch, soy polypeptide, and sodium tripolyphosphate is 1:(0.1-0.18):(0.01-0.018), for example, it can be 1:(0.10, 0.108, 0.116, 0.124, 0.132, 0.140, 0.148, 0.156, 0.164, 0.172, or 0.180):(0.010, 0.0108, 0.0116, 0.0124, 0.0132, 0.0140, 0.0148, 0.0156, 0.0164, 0.0172, or 0.0180), but is not limited to the listed values, and other values not listed in the range are also applicable.

[0034] In some alternative embodiments, the mass ratio of the semi-coke powder and expanded perlite is 1:(0.16-0.25), for example, it can be 1:0.160, 1:0.169, 1:0.178, 1:0.187, 1:0.196, 1:0.205, 1:0.214, 1:0.223, 1:0.232, 1:0.241, or 1:0.250, but is not limited to the listed values, and other values not listed in the range are also applicable.

[0035] In some optional embodiments, the activity ratio of cellulase, hemicellulase and protease in the composite enzyme solution is 1:(0.3-0.5):(0.2-0.4), for example, it can be 1:(0.30, 0.32, 0.34, 0.36, 0.38, 0.40, 0.42, 0.44, 0.46, 0.48 or 0.50):(0.20, 0.22, 0.24, 0.26, 0.28, 0.30, 0.32, 0.34, 0.36, 0.38 or 0.40), but is not limited to the listed values, other unlisted values ​​within this range are also applicable.

[0036] In some optional embodiments, the total enzyme activity of the complex enzyme solution is 6 × 10⁻⁶. 4 ~1×10 5 U / kg, for example, could be 6.0 × 10 4 U / kg, 6.4×10 4 U / kg, 6.8×10 4 U / kg, 7.2×10 4 U / kg, 7.6×10 4 U / kg, 8.0×10 4 U / kg, 8.4×10 4 U / kg, 8.8×10 4 U / kg, 9.2×10 4 U / kg, 9.6×10 4 U / kg or 1.0×10 5 U / kg, but not limited to the listed values; other unlisted values ​​within this range also apply.

[0037] In some optional embodiments, the amount of crosslinking binder added is 6-10% of the total mass of semi-coke powder and expanded perlite, for example, it can be 6%, 6.4%, 6.8%, 7.2%, 7.6%, 8.0%, 8.4%, 8.8%, 9.2%, 9.6% or 10%, but is not limited to the listed values, and other unlisted values ​​within this range are also applicable.

[0038] In some optional embodiments, the amount of the compound enzyme solution fed is 2-4% of the total mass of the semi-coke powder and expanded perlite, for example, it can be 2%, 2.2%, 2.4%, 2.6%, 2.8%, 3.0%, 3.2%, 3.4%, 3.6%, 3.8% or 4%, but is not limited to the listed values, and other unlisted values ​​within this range are also applicable.

[0039] In some optional embodiments, the rotating speed of the rotary granulator is 40-60 rpm, for example, it can be 40 rpm, 42 rpm, 44 rpm, 46 rpm, 48 rpm, 50 rpm, 52 rpm, 54 rpm, 56 rpm, 58 rpm or 60 rpm, but not only limited to the listed values, other values not listed in the range are also applicable.

[0040] In some optional embodiments, the drying temperature of the granules is 40-50℃, for example, it can be 40℃, 41℃, 42℃, 43℃, 44℃, 45℃, 46℃, 47℃, 48℃, 49℃ or 50℃, but not only limited to the listed values, other values not listed in the range are also applicable.

[0041] In some optional embodiments, the drying time of the granules is 4-6h, for example, it can be 4.0h, 4.2h, 4.4h, 4.6h, 4.8h, 5.0h, 5.2h, 5.4h, 5.6h, 5.8h or 6.0h, but not only limited to the listed values, other values not listed in the range are also applicable.

[0042] As a preferred technical solution of the application, in the preparation method of the dynamic humidity regulating granules, the mass ratio of the montmorillonite, microcrystalline cellulose, glycerol and sorbitol is 1:(0.28-0.40):(0.14-0.22):(0.14-0.22), for example, it can be 1:(0.28, 0.292, 0.304, 0.316, 0.328, 0.340, 0.352, 0.364, 0.376, 0.388 or 0.40):(0.14, 0.148, 0.156, 0.164, 0.172, 0.180, 0.188, 0.196, 0.204, 0.212 or 0.222):(0.14, 0.148, 0.156, 0.164, 0.172, 0.180, 0.188, 0.196, 0.204, 0.212 or 0.22), but not only limited to the listed values, other values not listed in the range are also applicable.

[0043] In some optional embodiments, the air-drying temperature of the primary product is 25-35℃, for example, it can be 25℃, 26℃, 27℃, 28℃, 29℃, 30℃, 31℃, 32℃, 33℃, 34℃ or 35℃, but not only limited to the listed values, other values not listed in the range are also applicable.

[0044] In some optional embodiments, the initial product has a sun drying time of 24-48h, for example, it can be 24h, 26.4h, 28.8h, 31.2h, 33.6h, 36h, 38.4h, 40.8h, 43.2h, 45.6h or 48h, but not limited to the listed values, and other values not listed in the range are also applicable.

[0045] In some optional embodiments, the dynamic humidity regulating granules have a particle length of 2-4mm, for example, it can be 2mm, 2.2mm, 2.4mm, 2.6mm, 2.8mm, 3.0mm, 3.2mm, 3.4mm, 3.6mm, 3.8mm or 4mm, but not limited to the listed values, and other values not listed in the range are also applicable.

[0046] As a preferred technical solution of the present application, in the preparation method of the enzyme cofactor powder, the total concentration of the mixed metal salt solution is 0.2-0.4M, for example, it can be 0.20M, 0.22M, 0.24M, 0.26M, 0.28M, 0.30M, 0.32M, 0.34M, 0.36M, 0.38M or 0.40M, but not limited to the listed values, and other values not listed in the range are also applicable.

[0047] In some optional embodiments, the mixed metal salt includes magnesium chloride, calcium chloride and copper sulfate in a molar ratio of 1:(2.8-32):(0.25-0.35), for example, it can be 1:(2.80, 2.84, 2.88, 2.92, 2.96, 3.00, 3.04, 3.08, 3.12, 3.16 or 3.20):(0.25, 0.26, 0.27, 0.28, 0.29, 0.30, 0.31, 0.32, 0.33, 0.34 or 0.35), but not limited to the listed values, and other values not listed in the range are also applicable.

[0048] In some optional embodiments, the mass / volume ratio of the calcium lignosulfonate and the mixed metal salt solution is 1:(0.6-0.8)g / mL, for example, it can be 1:0.60g / mL, 1:0.62g / mL, 1:0.64g / mL, 1:0.66g / mL, 1:0.68g / mL, 1:0.70g / mL, 1:0.72g / mL, 1:0.74g / mL, 1:0.76g / mL, 1:0.78g / mL or 1:0.80g / mL, but not limited to the listed values, and other values not listed in the range are also applicable.

[0049] In some optional embodiments, the time for wet mixing the calcium lignosulfonate with the mixed metal salt solution is 20-30 min, for example, can be 20 min, 21 min, 22 min, 23 min, 24 min, 25 min, 26 min, 27 min, 28 min, 29 min or 30 min, but not only limited to the listed values, other values not listed in the range of values are also applicable.

[0050] In some optional embodiments, the drying temperature of the reaction solution is 45-55℃, for example, can be 45℃, 46℃, 47℃, 48℃, 49℃, 50℃, 51℃, 52℃, 53℃, 54℃ or 55℃, but not only limited to the listed values, other values not listed in the range of values are also applicable.

[0051] In some optional embodiments, the drying time of the reaction solution is 4-6h, for example, can be 4.0h, 4.2h, 4.4h, 4.6h, 4.8h, 5.0h, 5.2h, 5.4h, 5.6h, 5.8h or 6.0h, but not only limited to the listed values, other values not listed in the range of values are also applicable.

[0052] In some optional embodiments, the mass ratio of the dried product to alanyl glutamine is 1:(0.4-0.6), for example, can be 1:0.40, 1:0.42, 1:0.44, 1:0.46, 1:0.48, 1:0.50, 1:0.52, 1:0.54, 1:0.56, 1:0.58 or 1:0.60, but not only limited to the listed values, other values not listed in the range of values are also applicable.

[0053] In a second aspect, the present application provides a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and coenzyme.

[0054] The present application adds heat-insulating enzyme-loaded particles at the beginning of the pile building. The heat-insulating enzyme-loaded particles use the low thermal conductivity of the semi-coke powder and the double-pore structure of the expanded perlite to form a cold island micro-zone throughout the core, so that the cellulase, hemicellulase and protease fixed on the pore wall are protected from the impact of direct heat flow above 50°C, thereby delaying inactivation. At the same time, the outer layer is coated with a starch-polypeptide-sodium tripolyphosphate ternary cross-linked glue film. The starch-polypeptide-sodium tripolyphosphate cross-linked network anchors the enzyme molecules in the inner wall of the skeleton through multiple actions of hydrogen bonds, ionic association and phosphate ester bonds, preventing desorption and conformational loosening caused by high temperature, high humidity and shear. The enzyme protein is protected while being in a permeable pore environment, which can continuously contact fiber, semi-fiber and protein substrates, break down macromolecular substrates into oligosaccharides and free amino acids, increase the concentration of soluble carbon and nitrogen, provide bait and receptors for the exponential expansion of aerobic microorganisms, and ensure that the pile can enter the hot state in a short time. Due to the strong adsorption of the semi-coke skeleton itself, part of the free amino acids released in the early stage can be captured and re-released, avoiding the phenomenon of ammonia shock inhibiting the bacterial population.

[0055] When the pile temperature enters the high temperature platform, the dynamic humidity adjusting particles added in the present application begin to play the function of "water constant device". The unique interlayer hydration-dehydration reversible process of montmorillonite can continuously release water without overflow between 50-65°C, and quickly reabsorb water when the temperature temporarily drops at night or during turning. Glycerol and sorbitol form a hydrophilic plasticized film on the surface of microcrystalline cellulose, which can stabilize the water film in the particle micropore and reduce the evaporation speed of free water, while itself is gradually metabolized by microorganisms as a carbon source, making the humidity adjusting function coupled with carbon supply. With this dynamic water release-absorption mechanism, the overall moisture content of the pile is locked in the appropriate range, avoiding the common problem of "anaerobic in the early stage due to excessive moisture, and enzyme loss in the later stage due to excessive dryness" in aerobic composting, and ensuring the continuous activity of enzymes in the high temperature stage.

[0056] After a certain period of high temperature, even with the double insurance of heat insulation and humidity adjustment, part of the enzyme will inevitably produce metal cofactor dislocation or local conformation collapse, at which time the enzyme cofactor powder added in the present application acts as a "reviver". The calcium lignosulfonate-metal complex slowly dissociates under the driving of the pH and temperature gradient of the pile body, releasing manganese, calcium, copper and other ions to the surrounding. These metals are essential cofactors for key enzymes such as cellulase family and polyphenol oxidase; the release rate is controlled by the sulfonic acid ligand, and there will be no ion shock. The synergistic alanyl glutamine is a stable dipeptide molecular chaperone, whose carbonyl and amide groups can insert into the hydrophobic domain of the heat-denatured enzyme molecule, guiding the refolding of the protein through hydrogen bond rearrangement and hydrophobic shielding, thereby achieving secondary "activation" in the enzyme semi-inactivation stage. The dipeptide and metal-sulfonic acid network can also weakly chelate free copper and zinc, so that heavy metals are first converted to organic-residue state in the pile body, reducing the risk of subsequent migration in the soil.

[0057] The accurate connection of the three particles on the time axis forms a closed-loop mechanism of "keeping alive-stabilizing alive-reviving": the heat-insulating enzyme-loaded particles first avoid heat flow impact, protect enzyme activity, and quickly open the substrate; the dynamic moisture control particles then stabilize moisture and temperature, so that the enzyme stays in the thermal stable zone for the longest time; and the enzyme cofactor powder finally re-energizes part of the heat-damaged enzyme and synchronously passivates heavy metals. Because the semi-coke powder and montmorillonite have physical-chemical dual capture effect on ammonia, and the enzyme cofactor powder can provide acidic sites while mismatching metal-sulfonic acid, ammonia gas and sulfides are synchronously reduced. The three materials are accurately put into the pile according to different stages of biochemical needs, reducing the dependence of traditional composting on natural cooling and repeated turning, and improving enzyme activity retention rate, lignin degradation rate and nitrogen retention rate in the high-temperature stage, while reducing the bioavailability of heavy metals and odor emission.

[0058] Compared with the prior art, the application has the following beneficial effects:

[0059] In the application, the heat-insulating enzyme-loaded particles form a cold island in the core of the pile by using the semi-coke-perlite double-hole low-thermal-conductivity framework to avoid the inactivation of fixed enzymes due to high temperature; and the starch-polypeptide-sodium tripolyphosphate cross-linked membrane firmly anchors the enzymes to prevent desorption. The protected enzyme proteins efficiently crack fibers and proteins in the permeable pores, rapidly release soluble carbon and nitrogen, promote explosive proliferation of microorganisms, and rapidly heat the pile; and the semi-coke powder simultaneously adsorbs and releases free ammonia to inhibit ammonia gas impact.

[0060] In the application, the dynamic moisture control particles use the reversible water release and water absorption of montmorillonite and the hydrophilic film of glycerol and sorbitol to automatically release water at high temperature and absorb water at low temperature, stabilize the moisture content of the pile, avoid over-wet anaerobic and enzyme loss in the later stage, and incidentally provide available carbon sources for microorganisms, thereby ensuring long-term stability of enzyme activity in the high-temperature stage.

[0061] In the application, the enzyme cofactor powder releases essential metals such as manganese, calcium and copper in the late high-temperature stage to repair enzyme cofactors that have been displaced by heat, and alanyl glutamine as a molecular chaperone promotes the refolding and recovery of damaged enzymes; the metal-sulfonic acid-dipeptide system also chelates free copper and zinc to reduce the activity of heavy metals, and the whole system plays a dual role of "reviving enzyme activity and passivating heavy metals".

[0062] In the application, the heat-insulating enzyme-loaded particles first insulate and fix enzymes and rapidly crack the substrate; the dynamic moisture control particles then automatically release and absorb water to stabilize temperature and humidity, prolonging the high-activity period of the enzyme; and the enzyme cofactor powder finally releases metals and molecular chaperones to revive damaged enzymes and passivate heavy metals. Through the closed loop of "keeping alive-stabilizing alive-reviving", ammonia is adsorbed and sulfides are inhibited, realizing long-lasting enzyme activity, deep lignin degradation, low nitrogen loss and pollution reduction, and reducing the dependence of traditional composting on natural cooling and repeated turning. DETAILED DESCRIPTION

[0063] The technical solutions of the present application will be described in detail below with reference to specific embodiments. The embodiments described herein are specific embodiments of the present application, which are used to illustrate the concept of the present application; these descriptions are all explanatory and exemplary, and should not be understood as limiting the embodiments of the present application and the protection scope of the present application. In addition to the embodiments described herein, those skilled in the art can also employ other technical solutions that are obvious based on the content disclosed in the claims and the specification of the present application, which include technical solutions that employ any obvious substitutions and modifications to the embodiments described herein.

[0064] The chemical reagents used in the examples and comparative examples of the present application are all commercially available products, which are not further purified or treated.

[0065] Example 1

[0066] The present embodiment provides a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor and a preparation method thereof. The preparation method of the slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor specifically comprises the following steps:

[0067] The livestock and poultry manure is mixed with short-cut straw and rice husk at a mass ratio of 7.3:1.5:0.9 to obtain a first heap, wherein the moisture content of the first heap is 54%, and heat-insulating enzyme-loaded particles with a feeding amount of 1.7% of the dry basis mass of the first heap are added during the heap building; the second heap is obtained by turning over the heap on the 2nd day of the heap building and adding dynamic humidity adjusting particles with a mass of 1.2% of the dry basis mass of the first heap; the third heap is obtained after fermentation and turning over; wherein the turning over frequency is 1 time per day for 0-3 days and 1 time every 2 days for 4-10 days; the fourth heap is obtained by turning over and adding enzyme cofactor powder with a mass of 0.9% of the dry basis mass of the first heap when the temperature of the third heap is maintained at 65℃ for 48h; the heap is continuously maintained at 58℃ for 5 days, and the slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor is obtained after cooling;

[0068] The preparation method of the heat-insulating enzyme-loaded particles is as follows:

[0069] The shavings are carbonized for 60 min under anoxic conditions at 380℃, with a nitrogen atmosphere flow rate of 3 L / (min·kg), crushed, and sieved to obtain semi-coke powder with a particle size of 400 μm; a 6wt.% corn starch dispersion is gelatinized for 30 min at 95℃, then mixed with a 0.8wt.% soybean polypeptide dispersion and a 0.08wt.% sodium tripolyphosphate solution to obtain a cross-linking and binding liquid, wherein the mass ratio of corn starch, soybean polypeptide, and sodium tripolyphosphate is 1:0.1:0.01; the semi-coke powder is mixed with expanded perlite at a mass ratio of 1:0.16, placed in a rotary granulator, and sprayed with the cross-linking and binding liquid and a composite enzyme liquid, wherein the enzyme activity ratio of cellulase Celluclast 1.5L, hemicellulase Pentopan Mono BG, and protease Alcalase 2.4L FG in the composite enzyme liquid is 1:0.5:0.2, and the total enzyme activity of the composite enzyme liquid is 6×10 4 U / kg, the cross-linking and binding liquid is fed at a rate of 6% of the total mass of the semi-coke powder and expanded perlite, and the composite enzyme liquid is fed at a rate of 2% of the total mass of the semi-coke powder and expanded perlite; the granules are obtained at a rotation speed of 55 rpm, and dried at 48℃ for 6 h to obtain heat-insulating enzyme-loaded granules;

[0070] The preparation method of the dynamic humidity-controlling granules is as follows:

[0071] Montmorillonite, microcrystalline cellulose, glycerol, and sorbitol are mixed at a mass ratio of 1:0.35:0.17:0.19, extruded and granulated by a screw extruder to obtain a primary product, and then dried at 32℃ for 36 h to obtain dynamic humidity-controlling granules with a particle length of 4 mm;

[0072] The preparation method of the enzyme cofactor powder is as follows:

[0073] A mixed metal salt solution with a total concentration of 0.35 M is prepared, wherein the mixed metal salt includes magnesium chloride, calcium chloride, and copper sulfate at a molar ratio of 1:3.0:0.3; calcium lignosulfonate is mixed with the mixed metal salt solution at a mass / volume ratio of 1:0.7 g / mL for 28 min to obtain a reaction liquid, which is dried at 52℃ for 4 h to obtain a dried product; the dried product is mixed with alanyl glutamine at a mass ratio of 1:0.6 to obtain the enzyme cofactor powder.

[0074] Example 2

[0075] The present embodiment provides a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier immobilized enzymes and cofactors and a preparation method thereof. The preparation method of the slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier immobilized enzymes and cofactors specifically includes the following steps:

[0076] Mix livestock and poultry manure with short-cut straw and rice husk at a mass ratio of 8:1:0.5 and build a heap to obtain a first heap, wherein the moisture content of the first heap is 60%, and a heat-insulating enzyme carrier particle with a dosage of 2% of the dry mass of the first heap is added during the building of the heap; the heap is turned over on the 2nd day of building, and a dynamic humidity adjusting particle with a dosage of 0.8% of the dry mass of the first heap is added to obtain a second heap, and a third heap is obtained after fermentation and turning over, wherein the turning over frequency is 1 time per day for 0-3 days and 1 time every 2 days for 4-10 days; when the temperature of the third heap is maintained at 55°C for 52 hours, the heap is turned over and an enzyme cofactor powder with a dosage of 0.6% of the dry mass of the first heap is added to obtain a fourth heap, and the heap is continuously maintained at 65°C for 3 days to obtain a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier immobilized enzyme and cofactor;

[0077] The preparation method of the heat-insulating enzyme carrier particle is as follows:

[0078] The shavings are carbonized under anoxic conditions at 420°C for 90 minutes, wherein the nitrogen atmosphere flow rate is 6 L / (min·kg), and the semi-coke powder with a particle size of 600 μm is obtained by crushing and sieving; a corn starch dispersion solution with a mass fraction of 7.2 wt.% is gelatinized at 85°C for 20 minutes, and then mixed with a soybean polypeptide dispersion solution with a mass fraction of 1.2 wt.% and a sodium tripolyphosphate solution with a mass fraction of 0.12 wt.% to obtain a cross-linking and bonding liquid, wherein the mass ratio of corn starch, soybean polypeptide and sodium tripolyphosphate is 1:0.18:0.018; the semi-coke powder and expanded perlite are mixed at a mass ratio of 1:0.25, placed in a rotary granulator, and the cross-linking and bonding liquid and a composite enzyme liquid are sprayed, wherein the enzyme activity ratio of cellulase Celluclast 1.5L, hemicellulase Pentopan Mono BG and protease Alcalase 2.4L FG in the composite enzyme liquid is 1:0.3:0.4, and the total enzyme activity of the composite enzyme liquid is 1×10 5 U / kg, the dosage of the cross-linking and bonding liquid is 10% of the total mass of the semi-coke powder and expanded perlite, and the dosage of the composite enzyme liquid is 4% of the total mass of the semi-coke powder and expanded perlite; the granules are obtained at a rotation speed of 40 rpm, and the heat-insulating enzyme carrier particles are obtained by drying at 40°C for 4 hours;

[0079] The preparation method of the dynamic humidity adjusting particle is as follows:

[0080] Montmorillonite, microcrystalline cellulose, glycerol and sorbitol are mixed at a mass ratio of 1:0.28:0.22:0.14, extruded by a screw extruder and granulated to obtain a primary product, and the dynamic humidity adjusting particles with a particle length of 2.5 mm are obtained by drying at 25°C for 48 hours;

[0081] The preparation method of the enzyme cofactor powder is as follows:

[0082] A mixed metal salt solution with a total concentration of 0.2M was prepared, wherein the mixed metal salt included magnesium chloride, calcium chloride and copper sulfate in a molar ratio of 1:2.8:0.25; the calcium lignosulfonate and the mixed metal salt solution were mixed at a mass / volume ratio of 1:0.6 g / mL for 30 min to obtain a reaction solution, and the reaction solution was dried at 45℃ for 5h to obtain a dried product; the dried product was mixed with alanyl glutamine at a mass ratio of 1:0.4 to obtain an enzyme cofactor powder.

[0083] Example 3

[0084] The present embodiment provides a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor and a preparation method thereof, and the preparation method of the slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor specifically includes the following steps:

[0085] The livestock and poultry manure was mixed with the short-cut straw and rice husk at a mass ratio of 7:1.35:1 to obtain a first heap, wherein the moisture content of the first heap was 50%, and the heat-insulating enzyme-loaded particles with a feeding amount of 1% of the dry basis mass of the first heap were added during the heap building; the heap was turned over on the 2nd day of the heap building, and the dynamic humidity adjusting particles with a feeding amount of 1.1% of the dry basis mass of the first heap were added to obtain a second heap, and a third heap was obtained after fermentation and turning over, wherein the turning over frequency was 1 time per day for 0-3 days and 1 time every 2 days for 4-10 days; when the temperature of the third heap was maintained at 62℃ for 42h, the heap was turned over and the enzyme cofactor powder with a feeding amount of 1% of the dry basis mass of the first heap was added to obtain a fourth heap, and the heap temperature was maintained at 55℃ for 4d, and a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor was obtained after cooling;

[0086] The preparation method of the heat-insulating enzyme-loaded particles is as follows:

[0087] The shavings were carbonized under anoxic conditions at 400℃ for 72 min, wherein the nitrogen atmosphere flow rate was 4 L / (min·kg), and the semi-coke powder with a particle size of 580 μm was obtained by crushing and sieving; a corn starch dispersion solution with a mass fraction of 8 wt.% was gelatinized at 92℃ for 25 min, and then mixed with a soybean polypeptide dispersion solution with a mass fraction of 1.1 wt.% and a sodium tripolyphosphate solution with a mass fraction of 0.1 wt.% to obtain a cross-linking and bonding liquid, wherein the mass ratio of corn starch, soybean polypeptide and sodium tripolyphosphate was 1:0.14:0.012; the semi-coke powder and expanded perlite were mixed at a mass ratio of 1:0.22, and then placed in a rotary granulator, and the cross-linking and bonding liquid and a composite enzyme liquid were sprayed, wherein the enzyme activity ratio of cellulase Celluclast 1.5L, hemicellulase Pentopan Mono BG and protease Alcalase 2.4L FG in the composite enzyme liquid was 1:0.45:0.25, and the total enzyme activity of the composite enzyme liquid was 8×10 4U / kg, the feeding amount of the crosslinking binding liquid is 9% of the total mass of the semi-coke powder and the expanded perlite, and the feeding amount of the compound enzyme liquid is 3.5% of the total mass of the semi-coke powder and the expanded perlite; the granules are obtained at a rotating speed of 60 rpm, and the heat-insulating enzyme-loaded granules are obtained by drying at 50℃ for 5h;

[0088] The preparation method of the dynamic humidity adjusting granules is as follows:

[0089] Montmorillonite, microcrystalline cellulose, glycerol and sorbitol are mixed in a mass ratio of 1:0.4:0.14:0.22, extruded and granulated by a screw extruder to obtain a primary product, and the primary product is dried at 35℃ for 24h to obtain the dynamic humidity adjusting granules with a particle length of 3.2mm;

[0090] The preparation method of the enzyme cofactor powder is as follows:

[0091] A mixed metal salt solution with a total concentration of 0.4M is prepared, wherein the mixed metal salt includes magnesium chloride, calcium chloride and copper sulfate in a molar ratio of 1:3.2:0.28; the calcium lignosulfonate and the mixed metal salt solution are mixed wet at a mass / volume ratio of 1:0.8g / mL for 20min to obtain a reaction liquid, and the reaction liquid is dried at 55℃ for 6h to obtain a dried product; the dried product and alanyl glutamine are mixed in a mass ratio of 1:0.5 to obtain the enzyme cofactor powder.

[0092] Example 4

[0093] The present embodiment provides a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor and a preparation method thereof. The preparation method of the slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor specifically includes the following steps:

[0094] Livestock and poultry manure, short-cut straw and rice husk are mixed in a mass ratio of 7.6:1.15:0.7 to obtain a first heap, wherein the moisture content of the first heap is 57%, and the heat-insulating enzyme-loaded granules with a feeding amount of 1.3% of the dry basis mass of the first heap are added during the building of the heap; the second heap is obtained by turning over the heap on the second day and adding the dynamic humidity adjusting granules with an amount of 0.9% of the dry basis mass of the first heap; the third heap is obtained after fermentation and turning over; wherein the turning over frequency is 1 time per day for 0-3 days and 1 time every 2 days for 4-10 days; the fourth heap is obtained by turning over the heap after the temperature of the third heap is maintained at 58℃ for 45h and adding the enzyme cofactor powder with an amount of 0.75% of the dry basis mass of the first heap; the slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor is obtained by continuously maintaining the heap at 62℃ for 3.5d and cooling;

[0095] The preparation method of the heat-insulating enzyme-loaded granules is as follows:

[0096] The shavings are carbonized for 85 min under anoxic conditions at 410℃, with a nitrogen atmosphere flow rate of 5.5 L / (min·kg), and are crushed and sieved to obtain semi-coke powder with a particle size of 500 μm; a 6.8wt.% corn starch dispersion is gelatinized at 90℃ for 22 min, then mixed with a 0.95wt.% soybean polypeptide dispersion and a 0.095wt.% sodium tripolyphosphate solution to obtain a cross-linking binder solution, wherein the mass ratio of corn starch, soybean polypeptide and sodium tripolyphosphate is 1:0.16:0.015; the semi-coke powder is mixed with expanded perlite at a mass ratio of 1:0.18, placed in a rotary granulator, and sprayed with the cross-linking binder solution and a composite enzyme solution, wherein the enzyme activity ratio of cellulase Celluclast 1.5L, hemicellulase Pentopan MonoBG and protease Alcalase 2.4L FG in the composite enzyme solution is 1:0.32:0.33, and the total enzyme activity of the composite enzyme solution is 9×10 4 U / kg, the cross-linking binder solution is fed at 7% of the total mass of the semi-coke powder and expanded perlite, and the composite enzyme solution is fed at 2.8% of the total mass of the semi-coke powder and expanded perlite; the granules are obtained at a rotation speed of 47 rpm, and are dried at 45℃ for 4.5 h to obtain heat-insulating enzyme-loaded granules;

[0097] The preparation method of the dynamic humidity-controlling granules is as follows:

[0098] Montmorillonite, microcrystalline cellulose, glycerol and sorbitol are mixed at a mass ratio of 1:0.30:0.20:0.16, extruded and granulated by a screw extruder to obtain a primary product, which is then dried at 28℃ for 42 h to obtain dynamic humidity-controlling granules with a particle length of 2 mm;

[0099] The preparation method of the enzyme cofactor powder is as follows:

[0100] A mixed metal salt solution with a total concentration of 0.25 M is prepared, wherein the mixed metal salt comprises magnesium chloride, calcium chloride and copper sulfate at a molar ratio of 1:2.9:0.35; calcium lignosulfonate is mixed with the mixed metal salt solution at a mass / volume ratio of 1:0.65 g / mL for 25 min to obtain a reaction solution, which is then dried at 48℃ for 4.5 h to obtain a dried product; the dried product is mixed with alanyl glutamine at a mass ratio of 1:0.55 to obtain the enzyme cofactor powder.

[0101] Comparative Example 1

[0102] This comparative example provides a slow-release organic-inorganic compound fertilizer based on synergistic effects of carrier- immobilized enzymes and cofactors, which is different from Example 1 in that no heat-insulating enzyme-loaded granules, dynamic humidity-controlling granules and enzyme cofactor powder are added, and the other operation steps and process parameters are exactly the same as those of Example 1.

[0103] Comparative Example 2

[0104] The comparative example provides a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor, which is different from example 1 in that only the heat-insulating enzyme-loaded particles are added, and other operation steps and process parameters are completely same as example 1.

[0105] Comparative example 3

[0106] The comparative example provides a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor, which is different from example 1 in that no heat-insulating enzyme-loaded particles are added, and other operation steps and process parameters are completely same as example 1.

[0107] Comparative example 4

[0108] The comparative example provides a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor, which is different from example 1 in that only the dynamic humidity-controlling particles are added, and other operation steps and process parameters are completely same as example 1.

[0109] Comparative example 5

[0110] The comparative example provides a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor, which is different from example 1 in that no dynamic humidity-controlling particles are added, and other operation steps and process parameters are completely same as example 1.

[0111] Comparative example 6

[0112] The comparative example provides a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor, which is different from example 1 in that only the enzyme cofactor powder is added, and other operation steps and process parameters are completely same as example 1.

[0113] Comparative example 7

[0114] The comparative example provides a slow-release organic-inorganic compound fertilizer based on synergistic effect of carrier solidified enzyme and cofactor, which is different from example 1 in that no enzyme cofactor powder is added, and other operation steps and process parameters are completely same as example 1.

[0115] The slow-release organic-inorganic compound fertilizers based on synergistic effect of carrier solidified enzyme and cofactor of examples 1-4 and comparative examples 1-7 are tested for performance, and the specific process is as follows:

[0116] The "Hongshen No. 1" carrot planting base is selected, and the test area is randomly selected; 3 experimental areas are selected for each organic fertilizer as a repeated control, the same amount of organic fertilizer is applied to the selected experimental area, and the same and reasonable management is carried out, and the carrot growth and yield of each area are counted, wherein the average value of 3 experimental areas of each organic fertilizer is the final result.

[0117] The test results are shown in Table 1.

[0118] Table 1 Performance test results of Examples 1-4 and Comparative Examples 1-7

[0119]

[0120] From the test results of Example 1 and Comparative Example 1, it can be seen that, without the addition of the heat-insulating enzyme-loaded particles, the dynamic moisture-regulating particles and the enzyme cofactor powder, there is a lack of heat insulation protection during the fermentation process, the early-stage complex enzymes are rapidly inactivated at high temperature; without dynamic moisture regulation, the pile is prone to anaerobic in the early stage and prone to water loss in the later stage, and the microbial activity fluctuates greatly; at the same time, there is a lack of metal-molecular chaperone compensation, the heat-damaged enzymes cannot be revived, the lignin and protein are not deeply degraded, the soluble carbon and nitrogen supply is minimal, and the effective nutrient and humic acid content is the lowest, so that the crop nutrition is limited, and after being applied to the soil, the carrot has a small leaf area, a weak plant body, a slow dry matter accumulation, and a decreased yield.

[0121] From the test results of Example 1 and Comparative Example 2, it can be seen that, only the heat-insulating enzyme-loaded particles are added, the heat-insulating framework provides cold island protection, so that the enzyme activity is retained and the substrate is rapidly cracked in the early stage of high temperature; however, the dynamic moisture-regulating particles are lacking, the moisture in the pile fluctuates greatly in the middle and late stages, and the enzyme activity period is compressed; and the enzyme cofactor powder is also lacking, so that the displaced enzymes cannot be reconstructed in the late stage of high temperature, the lignin deep degradation and nitrogen remineralization efficiency are low, resulting in insufficient nutrient supply in the later stage, and the dry matter content and yield are decreased.

[0122] From the test results of Example 1 and Comparative Example 3, it can be seen that, without the addition of the heat-insulating enzyme-loaded particles, the moisture-regulating system maintains the appropriate moisture content, and the enzyme cofactor powder repairs part of the enzyme activity in the late stage of high temperature, so that the fertilizer maturity and effective nitrogen are improved in the middle and late stages; however, the heat-insulating protection is lacking in the early stage of high temperature, a large amount of enzyme activity is lost, the substrate cracking is slow, the early-stage nutrient release is lagged, the leaf area of the carrot is insufficient after application, the single-plant biomass growth is slow, and the dry matter accumulation and yield are decreased in the later stage.

[0123] From the test results of Example 1 and Comparative Example 4, it can be seen that, only the dynamic moisture-regulating particles are added, the dynamic moisture-regulating particles maintain the stability of the moisture content and ventilation of the pile, avoiding anaerobic in the early stage and over-drying in the later stage, so that the soluble nutrient release is relatively smooth; however, the heat-insulating enzyme-loaded particles are lacking, a large amount of enzyme activity is lost in the high-temperature stage, and the substrate cracking rate is low; and the enzyme cofactor powder is also lacking, so that the damaged enzymes cannot be revived in the late stage of high temperature, resulting in a decreased leaf area of the crop, a significantly decreased single-plant weight, dry matter and yield.

[0124] From the test results of Example 1 and Comparative Example 5, without the addition of dynamic humidity adjusting particles, the heat-insulating enzyme-loaded particles effectively protect enzyme activity in the initial stage of high temperature, and the enzyme cofactor powder repairs part of the enzyme system in the later stage, so that the overall degradation degree of lignin and protein is high; however, without the dynamic humidity adjusting particles, the water content of the pile fluctuates greatly: the early stage is prone to excessive moisture to inhibit aerobic bacteria, and the later stage is prone to water loss to shorten the enzyme activity window. Water imbalance weakens the continuous degradation and nitrogen remineralization process, leading to insufficient nutrient supply in the later stage, insufficient leaf area, slow plant growth, and reduced dry matter content and final yield.

[0125] From the test results of Example 1 and Comparative Example 6, only the addition of enzyme cofactor powder, the manganese, calcium, copper and molecular chaperone released by the enzyme cofactor powder can partially repair heat-damaged enzymes in the later stage of high temperature, but lack of heat protection, enzymes are largely inactivated in the initial stage of temperature rise, and lack of dynamic humidity adjusting particles leads to water imbalance, substrate cleavage rate and nitrogen fixation efficiency are always limited, nutrient release is low and lagging, and crops show weak growth and reduced yield.

[0126] From the test results of Example 1 and Comparative Example 7, without the addition of enzyme cofactor powder, the double insurance of heat insulation and humidity adjustment makes the enzyme activity time longer, and the water and temperature are in the ideal interval, so that the leaf area and single plant weight slightly decrease; however, without enzyme cofactor powder, the enzyme metal center deficiency and conformation recession in the later stage of high temperature cannot be repaired, the deep degradation of lignin and the nitrogen remineralization efficiency decrease, and the heavy metal passivation effect is weakened, the dry matter accumulation speed slows down in the later stage, and the final yield decreases.

[0127] The above merely describes specific embodiments of the present application, but the protection scope of the present application is not limited thereto. It should be understood by those skilled in the art that any changes or replacements within the technical scope disclosed by the present application can be easily conceived by those skilled in the art, and all such changes and replacements fall within the protection scope and disclosure scope of the present application.

Claims

1. A method for preparing a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier-immobilized enzyme and cofactor, characterized in that, The preparation method includes: Livestock and poultry manure is mixed with chopped straw and rice husks and piled up to obtain the first pile. Insulation enzyme granules are added during pile building. On the second day of pile building, the pile is turned over and dynamic humidity regulating granules are added to obtain the second pile. After fermentation and turning over, the third pile is obtained. After the first pile undergoes constant temperature fermentation, it is turned over and enzyme cofactor powder is added to obtain the fourth pile. Constant temperature fermentation continues, and after cooling, a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier-immobilized enzyme and cofactor is obtained.

2. The method for preparing a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier-immobilized enzyme and cofactor as described in claim 1, characterized in that: The mass ratio of the livestock and poultry manure, chopped straw and rice husk is (7-8):(1-1.5):(0.5-1); The amount of the heat-insulating enzyme-carrying particles added is 1-2% of the dry matrix of the first pile. The amount of dynamically conditioned particles added to the second pile is 0.8-1.2% of the dry basis mass of the first pile; The amount of enzyme cofactor powder added to the fourth pile is 0.6-1% of the dry basis mass of the first pile.

3. The method for preparing a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier-immobilized enzyme and cofactor as described in claim 1, characterized in that, The method for preparing the heat-insulating enzyme-carrying particles is as follows: The wood shavings are carbonized in an anaerobic environment, then crushed and sieved to obtain semi-coke powder. The corn starch dispersion is gelatinized and mixed with soybean polypeptide dispersion and sodium tripolyphosphate solution to obtain cross-linking binder. The semi-coke powder is mixed with expanded perlite, placed in a rotary granulator, and the cross-linking binder and composite enzyme solution are sprayed in. The mixture is stirred to obtain granules, and then dried to obtain heat-insulated enzyme-carrying granules.

4. The method for preparing a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier-immobilized enzyme and cofactor as described in claim 3, characterized in that, In the preparation method of the heat-insulated enzyme-carrying particles: The mass ratio of corn starch, soybean polypeptide, and sodium tripolyphosphate is 1:(0.1-0.18):(0.01-0.018); The mass ratio of the semi-coke powder to expanded perlite is 1:(0.16-0.25); The enzyme activity ratio of cellulase, hemicellulase and protease in the compound enzyme solution is 1:(0.3-0.5):(0.2-0.4).

5. The method for preparing a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier-immobilized enzyme and cofactor as described in claim 3, characterized in that, In the preparation method of the heat-insulated enzyme-carrying particles: The amount of cross-linking binder added is 6-10% of the total mass of semi-coke powder and expanded perlite; The amount of the compound enzyme solution added is 2-4% of the total mass of the semi-coke powder and expanded perlite.

6. The method for preparing a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier-immobilized enzyme and cofactor as described in claim 1, characterized in that, The preparation method of the dynamic humidity regulating particles is as follows: Montmorillonite, microcrystalline cellulose, glycerol, and sorbitol were mixed, extruded and granulated using a screw extruder to obtain the initial product, which was then air-dried to obtain dynamically conditioned granules.

7. The method for preparing a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier-immobilized enzyme and cofactor as described in claim 6, characterized in that, In the preparation method of the dynamic humidity-regulating particles: The mass ratio of montmorillonite, microcrystalline cellulose, glycerol and sorbitol is 1:(0.28-0.4):(0.14-0.22):(0.14-0.22).

8. The method for preparing a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier-immobilized enzyme and cofactor as described in claim 1, characterized in that, The preparation method of the enzyme cofactor powder is as follows: Prepare a mixed metal salt solution, wet mix calcium lignosulfonate with the mixed metal salt solution to obtain a reaction solution, dry to obtain a dried product, mix the dried product with alanine glutamine to obtain enzyme cofactor powder.

9. The method for preparing a slow-release organic-inorganic compound fertilizer based on the synergistic effect of carrier-immobilized enzyme and cofactor as described in claim 8, characterized in that, In the preparation method of the enzyme cofactor powder: The mixed metal salt solution comprises magnesium chloride, calcium chloride, and copper sulfate in a molar ratio of 1:(2.8-3.2):(0.25-0.35). The mass / volume ratio of the calcium lignosulfonate to the mixed metal salt solution is 1:(0.6-0.8) g / mL; The mass ratio of the dried product to alanine glutamine is 1:(0.4-0.6).

10. A slow-release organic-inorganic compound fertilizer prepared by the preparation method according to any one of claims 1-9, based on the synergistic effect of carrier-immobilized enzyme and cofactor.

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