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CN-121405530-B - Preparation method of integrated pesticide fertilizer rich in antibacterial peptide

CN121405530BCN 121405530 BCN121405530 BCN 121405530BCN-121405530-B

Abstract

The invention discloses a preparation method of an integrated medical fertilizer rich in antibacterial peptide, and discloses a preparation method of an integrated medical fertilizer with a sodium alginate-chitosan biphase embedding nucleation and a chemical fertilizer substrate sprayed at a low temperature to form a shell, by using the method, the encapsulation rate of the sodium alginate-chitosan microcapsule on thalli reaches 85-95%, the survival rate of the thalli stored for 6 months under the normal temperature of a core-shell structure is 80-90%, the activity of the antibacterial peptide is reserved for 60-80%, and the compressive strength of particles is improved Compared with the sodium alginate single-layer embedding technology, the method has remarkable advantages in the aspects of cell survival rate and antibacterial peptide activity maintenance.

Inventors

  • ZHAO GENHAI
  • WANG YI
  • LI WEIDONG
  • Yu Bentao
  • YANG YOUSONG
  • CHANG LU
  • CAI JUN

Assignees

  • 湖北丰乐生态肥业有限公司
  • 中国科学院合肥物质科学研究院

Dates

Publication Date
20260512
Application Date
20251226

Claims (6)

  1. 1. The preparation method of the integrated pesticide fertilizer rich in the antibacterial peptide comprises the following steps: step 1, preparing bacillus subtilis calcium alginate microspheres rich in antibacterial peptide; in the step 1, bacillus subtilis capable of secreting antimicrobial peptide is fermented and cultured, wet thalli are collected, trehalose and Tween-80 are resuspended and then fully mixed with sodium alginate to form a mixed solution, and the mixed solution is dripped into In the solution, controlling the particle size of calcium alginate microspheres, and collecting the calcium alginate microspheres, wherein the fermentation time of the bacillus subtilis is 36-60h, and the wet bacillus subtilis thalli are resuspended by 5.0-10.0% of trehalose and 0.05-0.1% of Tween-80; Step 2, preparing microcapsules coated with chitosan outside calcium alginate microspheres; in the step 2, chitosan and glycerol form a mixed solution, and the mixed solution is sprayed on the surfaces of calcium alginate microspheres by using a fluidized bed to prepare microcapsules; step 3, compounding the microcapsule and the core-shell structure of the fertilizer to obtain the integrated medicine fertilizer with the core-shell structure, In the step3, the fertilizer base material is urea and/or ammonium phosphate and/or potassium chloride, and is crushed into powder to prepare fertilizer slurry, and the fertilizer slurry and humic acid are alternately sprayed on the surfaces of the microcapsules by adopting a fluidized bed bottom spraying lamination process to obtain the integrated medical fertilizer with the core-shell structure.
  2. 2. The method for preparing an antibacterial peptide-enriched integrated pesticide fertilizer according to claim 1, wherein in the step 1, the concentration of sodium alginate is 2.0-3.0% W/V, and the concentration of bacillus subtilis in the mixed solution is maintained Sodium alginate.
  3. 3. The method for preparing the integrated medical fertilizer rich in the antibacterial peptide, which is characterized in that in the step 1, The concentration is 0.05-0.1M, the crosslinking time is 20-60 min, and the particle size of the calcium alginate microsphere is 200-500 mu m.
  4. 4. The method for preparing the integrated pesticide fertilizer rich in the antibacterial peptide, which is disclosed in claim 1, is characterized in that in the step 2, 0.5-1.5% w/v chitosan and 0.1-0.5% glycerol are used for forming a mixed solution, the coating time is 5-15 min, the thickness of the coating is controlled to be 50-80 mu m, the drying inlet temperature of a fluidized bed is 35-40 ℃, and the wind speed is 1.5m/s.
  5. 5. The method for preparing the integrated pesticide fertilizer rich in the antibacterial peptide, which is disclosed in claim 1, is characterized in that in the step 3, fertilizer base material urea and/or ammonium phosphate and/or potassium chloride are crushed to 80-100 meshes to prepare fertilizer slurry, and the solid content is 65-75%.
  6. 6. The method for preparing the integrated pesticide fertilizer rich in the antibacterial peptide, which is disclosed in claim 1, is characterized in that in the step 3, the concentration of humic acid is 1-3%, the air inlet temperature of a fluidized bed is 35-45 ℃, and the thickness of a shell layer is controlled to be 200-300 mu m.

Description

Preparation method of integrated pesticide fertilizer rich in antibacterial peptide Technical Field The invention relates to the technical field of agricultural fertilizers, in particular to a preparation method of an integral fertilizer rich in antibacterial peptide. Background The traditional chemical fertilizer is widely applied in agricultural production, but the soil problem and microorganism unbalance are easily caused by long-term use, the problems are prominently represented by soil hardening, acidification/salinization, organic matter reduction, the diversity of soil microorganisms is destroyed, the disease suppression capability and the nutrient conversion capability (such as nitrogen fixation, phosphorus dissolution and potassium dissolution bacteria reduction) of the soil are weakened, and the serious plant diseases and insect pests are further caused. The antibacterial peptide is taken as a special organic macromolecule, plays a role in killing various bacteria, fungi, partial viruses and parasites mainly by destroying microbial cell membranes or acting on intracellular targets, is not easy to generate drug resistance, is a novel, green and low-residue biopesticide and biostimulant, can be used for preventing and treating soil-borne diseases, leaf diseases and the like, and can also induce the disease resistance potential of plants and promote plant growth, so that the antibacterial peptide and the traditional chemical fertilizer are integrated and used in agricultural production and have wide application prospects. In the integrated preparation process of the antibacterial peptide and the chemical fertilizer, the antibacterial peptide structure can be damaged by early mixing granulation, mechanical stress, high temperature (the steam temperature is usually higher than 100 ℃), and the like, and the high-salt environment (the chemical fertilizer usually contains high-concentration inorganic salt ions such asEtc.) may also cause denaturation, aggregation or conformational change of the antimicrobial peptide, and secondly, after the fertilizer is applied to the soil, soil particles, especially clay minerals, organic matters, etc., may adsorb the antimicrobial peptide, reduce the bioavailability and migration and diffusion capacity thereof, reduce the effect, and protease, redox environment, etc. in the soil may also cause loss of the antimicrobial peptide activity. The antibacterial peptide is unstable and easy to inactivate in a complex fertilizer system and a severe environment (production, storage and soil). Disclosure of Invention The invention aims to provide a preparation method of an integrated medical fertilizer rich in antibacterial peptide, which adopts sodium alginate-chitosan biphasic embedding, the method combines a core-shell low-temperature granulation technical scheme, and has remarkable advantages in the aspects of cell survival rate and antibacterial peptide activity retention compared with a sodium alginate single-layer embedding technology. The technical solution adopted by the invention for solving the technical problems is as follows: the preparation method of the integrated pesticide fertilizer rich in the antibacterial peptide comprises the following steps: step 1, preparing bacillus subtilis calcium alginate microspheres rich in antibacterial peptide; Step 2, preparing microcapsules coated with chitosan outside calcium alginate microspheres; and 3, compounding the microcapsule with a core-shell structure of the fertilizer to obtain the core-shell structure integrated medical fertilizer. In step 1, bacillus subtilis capable of secreting antimicrobial peptide is fermented and cultured, wet thallus is collected, trehalose and Tween-80 are resuspended and then fully mixed with sodium alginate to form a mixed solution, and the mixed solution is dripped intoIn the solution, controlling the particle size of calcium alginate microspheres, and collecting the calcium alginate microspheres. Preferably, in the step 1, the fermentation time of the bacillus subtilis is 36-60 hours, and the wet bacillus subtilis thalli are resuspended by 5.0-10.0% of trehalose and 0.05-0.1% of Tween-80. Preferably, in the step 1, the concentration of the sodium alginate is 2.0-3.0% (w/v), and the concentration of the bacillus subtilis in the mixed solution is maintainedSodium alginate. Preferably, in step 1, the process comprises,The concentration is 0.05-0.1M, the crosslinking time is 20-60 min, and the particle size of the calcium alginate microsphere is 200-500 mu m. In the step 2, chitosan and glycerin form a mixed solution, and the mixed solution is sprayed on the surface of calcium alginate microspheres by using a fluidized bed to prepare the microcapsule. Preferably, in the step 2, 0.5-1.5% (w/v) chitosan and 0.1-0.5% glycerol are mixed to form a mixed solution, the coating time is 5-15 min, the coating thickness is controlled to be 50-80 mu m, the drying inlet temperature of a fluidized bed is 35