Stable fertilizer containing antibacterial peptide and preparation method thereof
By adding the antimicrobial peptide extracted from the Jerusalem artichoke residue to the fertilizer after enzymatically lying the protease, the problem of low nitrogen fertilizer utilization and failure to effectively utilize Jerusalem artichoke residue in the prior art is solved, and stable fertilizers and high-efficiency nitrogen fertilizer utilization are achieved without additional inhibitors.
Patent Information
- Application Number
- CN202510388124.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-31
- Publication Date
- 2025-06-13
AI Technical Summary
The prior art is difficult to improve the utilization rate of nitrogen fertilizer by not adding nitration inhibitors or urease inhibitors, and it is not effective to utilize the synergistic effect of erotic artichoke residue in fertilizers.
By adding the antimicrobial peptide extracted after protease to the fertilizer, a stable fertilizer containing antimicrobial peptide was prepared, meeting the standard of nitration inhibition rate ≥6%.
This method does not require additional inhibitors to meet the standards of stable fertilizers, improves the utilization rate of nitrogen fertilizers, and applies extracts to fertilizers through multi-enzyme lysis processes, significantly improving fertilizer efficiency.
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Abstract
Description
Technical Field
[0001] The present invention relates to a stable fertilizer containing antibacterial peptides and a preparation method thereof. Background Art
[0002] Nitrogen fertilizers have high fluidity and are easily lost after being applied to the soil. Therefore, locking up nitrogen fertilizers is the key to improving the utilization rate of fertilizers. At present, there are two types of fertilizers for improving the nitrogen utilization rate in China. One is a slow-release fertilizer using film coating or adding formaldehyde; the other is a stable fertilizer adding one or both of urease inhibitors or nitrification inhibitors. The above two types of fertilizers can improve the nitrogen utilization rate and have a good market response.
[0003] Jerusalem artichoke, also known as Helianthus tuberosus, is a plant of the genus Helianthus in the Compositae family. It has the characteristics of drought tolerance and salt tolerance, and is therefore often used for wind prevention and sand fixation. Jerusalem artichoke can be used to prepare inulin, and its by-product is Jerusalem artichoke residue. The yield of Jerusalem artichoke residue is huge, with a water content of about 83%. At the same time, it contains a large amount of cellulose, hemicellulose, protein and starch. At present, Jerusalem artichoke residue is mainly used for animal feed and rarely applied to agriculture, especially for fertilizers. Even when applied to fertilizers, it only plays the role of organic matter and cannot enhance the fertilizer efficiency well.
[0004] At present, there is no product or method that applies Jerusalem artichoke residue to fertilizers, can meet the requirements of stable fertilizers without adding additional nitrification inhibitors or urease inhibitors, and can improve the nitrogen utilization rate. Summary of the Invention
[0005] The present invention provides a stable fertilizer containing antibacterial peptides and a preparation method thereof. The technical problem to be solved is to apply Jerusalem artichoke residue to fertilizers, so that the prepared fertilizer can meet the standards of stable fertilizers without adding additional nitrification inhibitors or urease inhibitors, and can improve the nitrogen utilization rate.
[0006] To solve the above technical problems, the present invention adopts the following technical solutions: A stable fertilizer containing antibacterial peptides contains Jerusalem artichoke antibacterial peptides; the Jerusalem artichoke antibacterial peptides are products obtained by enzymatically hydrolyzing Jerusalem artichoke residue; the enzymatic hydrolysis is protease hydrolysis; The protease is an acidic protease or a basic protease; The requirement for the stable fertilizer is that the nitrification inhibition rate is ≥ 6%.
[0007] The enzymatic hydrolysis also includes one or several of cellulase hydrolysis, amylase hydrolysis and xylanase hydrolysis.
[0008] The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residue, water and a single enzyme is 1:8 - 25:0.00002 - 0.00005; The single enzyme is one of protease, cellulase, amylase and xylanase.
[0009] The enzymatic hydrolysis includes protease hydrolysis, cellulase hydrolysis, amylase hydrolysis and xylanase hydrolysis; The order of the enzymatic hydrolysis is to first perform cellulase hydrolysis and finally perform protease hydrolysis; The conditions for cellulase hydrolysis are a temperature of 30 - 60°C, a pH of 4 - 6, and a time of 3 - 6 h; The conditions for amylase hydrolysis are a temperature of 30 - 60°C, a pH of 4 - 6, and a time of 3 - 6 h; The conditions for xylanase hydrolysis are a temperature of 30 - 60°C, a pH of 4 - 6, and a time of 3 - 6 h; The conditions for protease hydrolysis are a temperature of 30 - 60°C, a pH of 3.5 - 5.5 or 12 - 14, and a time of 3 - 6 h; It also includes enzyme inactivation, and the conditions for enzyme inactivation are a temperature of 95 - 100°C and a time of 1 - 3 min.
[0010] It also includes filtration before protease hydrolysis. The obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for protease hydrolysis, and after protease hydrolysis, filtration is performed, and the obtained filtrate is the antibacterial peptide solution; The antibacterial peptide solution and the enzymatic hydrolysate are combined to obtain the Jerusalem artichoke antibacterial peptide solution; The antibacterial peptide solution is dried to obtain powdered antibacterial peptide; The Jerusalem artichoke antibacterial peptide solution is dried to obtain powdered Jerusalem artichoke antibacterial peptide; The Jerusalem artichoke antibacterial peptide includes one or two of the antibacterial peptide solution, powdered antibacterial peptide, Jerusalem artichoke antibacterial peptide solution and powdered Jerusalem artichoke antibacterial peptide; In protease hydrolysis, the mass ratio of the filter residue, water and protease is 1:8 - 20:0.00002 - 0.00005.
[0011] Preparation method of a stability fertilizer containing antibacterial peptide. The Jerusalem artichoke antibacterial peptide is added to the fertilizer to obtain the stability fertilizer containing antibacterial peptide; The mass ratio of the Jerusalem artichoke antibacterial peptide in the fertilizer is 0.1% - 90%; The fertilizer is one or several of organic-inorganic fertilizer, bio-organic fertilizer, compound fertilizer and compound mixed fertilizer.
[0012] The addition of the Jerusalem artichoke antibacterial peptide to the fertilizer means mixing the Jerusalem artichoke antibacterial peptide and the fertilizer raw materials and granulating to obtain the stability fertilizer containing antibacterial peptide; The fertilizer raw materials are one or several of urea, ammonium chloride, ammonium sulfate, potassium chloride, potassium sulfate and monoammonium phosphate.
[0013] It also includes drying after granulation, and the drying temperature is 30 - 70°C.
[0014] The invention has the following beneficial technical effects: 1) In this application, by adding the product obtained from the enzymatic hydrolysis of Jerusalem artichoke residue with protease, where the protease is acidic protease or alkaline protease, the finally prepared fertilizer can meet the requirements of the stability fertilizer standard without the need to additionally add nitrification inhibitors or urease inhibitors, and the nitrogen utilization rate is mentioned.
[0015] 2) In this application, the Jerusalem artichoke residue is enzymatically hydrolyzed with protease, and also includes enzymatic hydrolysis with cellulase, amylase, and xylanase. Thus, the prepared extract has good fertilizer efficiency when applied to fertilizers and meets the requirements of the stability fertilizer standard.
[0016] 3) This application defines the order of enzymatic hydrolysis. First, enzymatic hydrolysis with cellulase is carried out, and finally, enzymatic hydrolysis with protease is carried out. Further, it is also defined that filtration is carried out before enzymatic hydrolysis with protease, and the obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for enzymatic hydrolysis with protease, and after enzymatic hydrolysis with protease, filtration is carried out, and the obtained filtrate is the antibacterial peptide solution. Adding this antibacterial solution to the fertilizer can meet the requirements of the stability fertilizer standard and can improve the nitrogen utilization rate. Detailed implementation mode
[0017] To better understand the above technical solution, the technical solution of the present invention will be described in detail below in combination with preferred examples. Example 1
[0018] A stability fertilizer containing antibacterial peptides, containing Jerusalem artichoke antibacterial peptides; the Jerusalem artichoke antibacterial peptides are the products obtained after enzymatic hydrolysis of Jerusalem artichoke residue; the enzymatic hydrolysis is enzymatic hydrolysis with acidic protease; specifically: Add Jerusalem artichoke residue and acidic protease to water for enzymatic hydrolysis, and the obtained product is Jerusalem artichoke antibacterial peptides; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residue, water, and acidic protease is 1:12:0.00002; The conditions for enzymatic hydrolysis with acidic protease are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after enzymatic hydrolysis with protease, and the conditions for inactivating the enzyme are a temperature of 95 °C and a time of 2 min.
[0019] Adding Jerusalem artichoke antibacterial peptides to the fertilizer means mixing Jerusalem artichoke antibacterial peptides and fertilizer raw materials, and granulating by extrusion to obtain a stability fertilizer containing antibacterial peptides; The mass ratio of Jerusalem artichoke antibacterial peptides and fertilizer raw materials is 4:96; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate, and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 2
[0020] A stability fertilizer containing antibacterial peptides, containing Jerusalem artichoke antibacterial peptides; the Jerusalem artichoke antibacterial peptides are the products obtained after enzymatic hydrolysis of Jerusalem artichoke residue; the enzymatic hydrolysis is sequentially enzymatic hydrolysis with cellulase and enzymatic hydrolysis with acidic protease; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residue, water, cellulase, and acid protease is 1:12:0.00002:0.00002; The conditions for cellulase enzymatic hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for acid protease enzymatic hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis. The conditions for enzyme inactivation are a temperature of 95 °C and a time of 2 min.
[0021] Adding Jerusalem artichoke antibacterial peptide to the fertilizer means mixing the Jerusalem artichoke antibacterial peptide and the fertilizer raw materials, and then extruding and granulating to obtain the antibacterial peptide-containing stable fertilizer; The mass ratio of Jerusalem artichoke antibacterial peptide to the fertilizer raw materials is 4:96; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate, and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 3
[0022] A stable fertilizer containing antibacterial peptide contains Jerusalem artichoke antibacterial peptide; the Jerusalem artichoke antibacterial peptide is a product obtained by enzymatic hydrolysis of Jerusalem artichoke residue; the enzymatic hydrolysis is sequentially cellulase enzymatic hydrolysis and acid protease enzymatic hydrolysis. It also includes filtering after cellulase enzymatic hydrolysis to obtain a filtrate and a filter residue. The obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for protease enzymatic hydrolysis, and after protease enzymatic hydrolysis, it is filtered. The obtained filtrate is the antibacterial peptide solution; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residue, water, and cellulase is 1:12:0.00002; The mass ratio of the filter residue, water, and acid protease is 1:12:0.00002; The conditions for cellulase enzymatic hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for acid protease enzymatic hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis. The conditions for enzyme inactivation are a temperature of 95 °C and a time of 2 min.
[0023] Adding the antibacterial peptide solution to the fertilizer means mixing the Jerusalem artichoke antibacterial peptide and the fertilizer raw materials, and then extruding and granulating to obtain the antibacterial peptide-containing stable fertilizer; The mass ratio of the antibacterial peptide solution to the fertilizer raw materials is 4:96; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate, and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 4
[0024] A stable fertilizer containing an antibacterial peptide, which contains a Jerusalem artichoke antibacterial peptide; the Jerusalem artichoke antibacterial peptide is a product obtained by enzymatically hydrolyzing Jerusalem artichoke residues; the enzymatic hydrolysis is successively carried out by cellulase enzymatic hydrolysis and acid protease enzymatic hydrolysis, and also includes filtering after cellulase enzymatic hydrolysis to obtain a filtrate and a filter residue, and the obtained filtrate is an enzymatic hydrolysate; the obtained filter residue is used for protease enzymatic hydrolysis, and after protease enzymatic hydrolysis and filtration, the obtained filtrate is an antibacterial peptide solution; Mix the antibacterial peptide solution and the enzymatic hydrolysate to obtain a Jerusalem artichoke antibacterial peptide solution; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residues, water and cellulase is 1:12:0.00002; The mass ratio of the filter residue, water and acid protease is 1:12:0.00002; The conditions for cellulase enzymatic hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for acid protease enzymatic hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis, and the conditions for inactivating the enzyme are a temperature of 95 °C and a time of 2 min.
[0025] Adding the Jerusalem artichoke antibacterial peptide solution to the fertilizer means mixing the Jerusalem artichoke antibacterial peptide and the fertilizer raw materials and granulating by extrusion to obtain a stable fertilizer containing an antibacterial peptide; The mass ratio of the Jerusalem artichoke antibacterial peptide solution and the fertilizer raw materials is 8:92; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 5
[0026] A stable fertilizer containing an antibacterial peptide, which contains a Jerusalem artichoke antibacterial peptide; the Jerusalem artichoke antibacterial peptide is a product obtained by enzymatically hydrolyzing Jerusalem artichoke residues; the enzymatic hydrolysis is successively carried out by cellulase enzymatic hydrolysis, amylase enzymatic hydrolysis and acid protease enzymatic hydrolysis; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residues, water, cellulase, amylase and acid protease is 1:12:0.00002:0.00002:0.00002; The conditions for cellulase enzymatic hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for acid protease enzymatic hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for amylase enzymatic hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis, and the conditions for inactivating the enzyme are a temperature of 95 °C and a time of 2 min.
[0027] Adding the Jerusalem artichoke antibacterial peptide to the fertilizer means mixing the Jerusalem artichoke antibacterial peptide and the fertilizer raw materials and granulating by extrusion to obtain a stable fertilizer containing an antibacterial peptide; The mass ratio of the Jerusalem artichoke antibacterial peptide and the fertilizer raw materials is 4:96; The fertilizer raw material is a composition of urea, ammonium sulfate, potassium sulfate, and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 6
[0028] A stable fertilizer containing an antibacterial peptide, which contains the Jerusalem artichoke antibacterial peptide; the Jerusalem artichoke antibacterial peptide is a product obtained by enzymatically hydrolyzing Jerusalem artichoke residue; the enzymatic hydrolysis is sequentially carried out by cellulase hydrolysis, amylase hydrolysis, and acid protease hydrolysis, and also includes filtering after amylase hydrolysis to obtain a filtrate and a filter residue, and the obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for protease hydrolysis, and after protease hydrolysis and filtration, the obtained filtrate is the antibacterial peptide solution; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residue, water, cellulase, and amylase is 1:12:0.00002:0.00002; The mass ratio of the filter residue, water, and acid protease is 1:12:0.00002; The conditions for cellulase hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for acid protease hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for amylase hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis, and the conditions for inactivating the enzyme are a temperature of 95 °C and a time of 2 min.
[0029] Adding the antibacterial peptide solution to the fertilizer is to mix the Jerusalem artichoke antibacterial peptide and the fertilizer raw material, and then extrude and granulate to obtain the stable fertilizer containing antibacterial peptide; The mass ratio of the antibacterial peptide solution and the fertilizer raw material is 4:96; The fertilizer raw material is a composition of urea, ammonium sulfate, potassium sulfate, and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 7
[0030] A stable fertilizer containing an antibacterial peptide, which contains the Jerusalem artichoke antibacterial peptide; the Jerusalem artichoke antibacterial peptide is a product obtained by enzymatically hydrolyzing Jerusalem artichoke residue; the enzymatic hydrolysis is sequentially carried out by cellulase hydrolysis, amylase hydrolysis, and acid protease hydrolysis, and also includes filtering after amylase hydrolysis to obtain a filtrate and a filter residue, and the obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for protease hydrolysis, and after protease hydrolysis and filtration, the obtained filtrate is the antibacterial peptide solution; Merge the antibacterial peptide solution and the enzymatic hydrolysate to obtain the Jerusalem artichoke antibacterial peptide solution; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residue, water, cellulase, and amylase is 1:12:0.00002:0.00002; The mass ratio of the filter residue, water, and acid protease is 1:12:0.00002; The conditions for cellulase hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The enzymatic hydrolysis conditions for acid protease are a temperature of 40 °C, a pH of 4, and a time of 4 h; The enzymatic hydrolysis conditions for amylase are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis. The enzyme inactivation conditions are a temperature of 95 °C and a time of 2 min.
[0031] Adding the Jerusalem artichoke antimicrobial peptide solution to the fertilizer means mixing the Jerusalem artichoke antimicrobial peptide and the fertilizer raw materials, and then obtaining the antimicrobial peptide-containing stable fertilizer through extrusion granulation; The mass ratio of the Jerusalem artichoke antimicrobial peptide solution to the fertilizer raw materials is 8:92; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate, and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 8
[0032] A stable fertilizer containing antimicrobial peptides contains Jerusalem artichoke antimicrobial peptides; the Jerusalem artichoke antimicrobial peptides are the products obtained after enzymatic hydrolysis of Jerusalem artichoke residues; the enzymatic hydrolysis is successively carried out by cellulase enzymatic hydrolysis, amylase enzymatic hydrolysis, xylanase enzymatic hydrolysis, and acid protease enzymatic hydrolysis. It also includes filtering after xylanase enzymatic hydrolysis to obtain a filtrate and a filter residue. The obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for protease enzymatic hydrolysis, and after filtering the protease enzymatic hydrolysis, the obtained filtrate is the antimicrobial peptide solution; Merging the antimicrobial peptide solution and the enzymatic hydrolysate gives the Jerusalem artichoke antimicrobial peptide solution; The mass ratio of the Jerusalem artichoke residue, water, cellulase, amylase, and xylanase for enzymatic hydrolysis is 1:12:0.00002:0.00002:0.00002; The mass ratio of the filter residue, water, and acid protease is 1:12:0.00002; The cellulase enzymatic hydrolysis conditions are a temperature of 40 °C, a pH of 4, and a time of 4 h; The acid protease enzymatic hydrolysis conditions are a temperature of 40 °C, a pH of 4, and a time of 4 h; The amylase enzymatic hydrolysis conditions are a temperature of 40 °C, a pH of 4, and a time of 4 h; The xylanase enzymatic hydrolysis conditions are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis. The enzyme inactivation conditions are a temperature of 95 °C and a time of 2 min.
[0033] Adding the Jerusalem artichoke antimicrobial peptide solution to the fertilizer means mixing the Jerusalem artichoke antimicrobial peptide and the fertilizer raw materials, and then obtaining the antimicrobial peptide-containing stable fertilizer through extrusion granulation; The mass ratio of the Jerusalem artichoke antimicrobial peptide solution to the fertilizer raw materials is 8:92; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate, and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 9
[0034] A stable fertilizer containing antibacterial peptides, which contains Jerusalem artichoke antibacterial peptides; the Jerusalem artichoke antibacterial peptides are products obtained by enzymatically hydrolyzing Jerusalem artichoke residues; the enzymatic hydrolysis is alkaline protease hydrolysis; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residues, water and alkaline protease is 1:12:0.00002; The conditions for alkaline protease hydrolysis are a temperature of 40 °C, a pH of 13, and a time of 4 h; It also includes inactivating the enzyme after protease hydrolysis, and the conditions for inactivating the enzyme are a temperature of 95 °C and a time of 2 min.
[0035] Adding the Jerusalem artichoke antibacterial peptides to the fertilizer is to mix the Jerusalem artichoke antibacterial peptides and fertilizer raw materials, and then obtain the stable fertilizer containing antibacterial peptides through extrusion granulation; The mass ratio of the Jerusalem artichoke antibacterial peptides and fertilizer raw materials is 4:96; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 10
[0036] A stable fertilizer containing antibacterial peptides, which contains Jerusalem artichoke antibacterial peptides; the Jerusalem artichoke antibacterial peptides are products obtained by enzymatically hydrolyzing Jerusalem artichoke residues; the enzymatic hydrolysis is successively cellulase hydrolysis and alkaline protease hydrolysis; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residues, water, cellulase, and alkaline protease is 1:12:0.00002:0.00002; The conditions for cellulase hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for alkaline protease hydrolysis are a temperature of 40 °C, a pH of 13, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis, and the conditions for inactivating the enzyme are a temperature of 95 °C and a time of 2 min.
[0037] Adding the Jerusalem artichoke antibacterial peptides to the fertilizer is to mix the Jerusalem artichoke antibacterial peptides and fertilizer raw materials, and then obtain the stable fertilizer containing antibacterial peptides through extrusion granulation; The mass ratio of the Jerusalem artichoke antibacterial peptides and fertilizer raw materials is 4:96; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 11
[0038] A stable fertilizer containing antibacterial peptides, which contains Jerusalem artichoke antibacterial peptides; the Jerusalem artichoke antibacterial peptides are products obtained by enzymatically hydrolyzing Jerusalem artichoke residues; the enzymatic hydrolysis is successively cellulase hydrolysis and alkaline protease hydrolysis, and it also includes filtering after cellulase hydrolysis to obtain a filtrate and a filter residue, the obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for protease hydrolysis, and after protease hydrolysis, filtration is carried out, and the obtained filtrate is the antibacterial peptide solution; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residue, water and cellulase is 1:12:0.00002; The mass ratio of the filter residue, water and alkaline protease is 1:12:0.00002; The conditions for cellulase hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for alkaline protease hydrolysis are a temperature of 40 °C, a pH of 13, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis, and the conditions for enzyme inactivation are a temperature of 95 °C and a time of 2 min.
[0039] Adding the antibacterial peptide solution to the fertilizer means mixing the Jerusalem artichoke antibacterial peptide and the fertilizer raw materials, and then performing extrusion granulation to obtain the antibacterial peptide-containing stable fertilizer; The mass ratio of the antibacterial peptide solution to the fertilizer raw materials is 4:96; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 12
[0040] A stable fertilizer containing antibacterial peptides contains Jerusalem artichoke antibacterial peptides; the Jerusalem artichoke antibacterial peptides are products obtained by enzymatically hydrolyzing Jerusalem artichoke residues; the enzymatic hydrolysis is sequentially cellulase hydrolysis and alkaline protease hydrolysis, and it also includes filtering after cellulase hydrolysis to obtain a filtrate and a filter residue, and the obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for protease hydrolysis, and after protease hydrolysis, filtration is performed, and the obtained filtrate is the antibacterial peptide solution; Merging the antibacterial peptide solution and the enzymatic hydrolysate to obtain the Jerusalem artichoke antibacterial peptide solution; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residue, water and cellulase is 1:12:0.00002; The mass ratio of the filter residue, water and alkaline protease is 1:12:0.00002; The conditions for cellulase hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for alkaline protease hydrolysis are a temperature of 40 °C, a pH of 13, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis, and the conditions for enzyme inactivation are a temperature of 95 °C and a time of 2 min.
[0041] Adding the Jerusalem artichoke antibacterial peptide solution to the fertilizer means mixing the Jerusalem artichoke antibacterial peptide and the fertilizer raw materials, and then performing extrusion granulation to obtain the antibacterial peptide-containing stable fertilizer; The mass ratio of the Jerusalem artichoke antibacterial peptide solution to the fertilizer raw materials is 8:92; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 13
[0042] A stable fertilizer containing an antibacterial peptide, which contains a Jerusalem artichoke antibacterial peptide; the Jerusalem artichoke antibacterial peptide is a product obtained by enzymatically hydrolyzing Jerusalem artichoke residues; the enzymatic hydrolysis is successively carried out by cellulase hydrolysis, amylase hydrolysis and alkaline protease hydrolysis; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residues, water, cellulase, amylase and alkaline protease is 1:12:0.00002:0.00002:0.00002; The conditions for cellulase hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for alkaline protease hydrolysis are a temperature of 40 °C, a pH of 13, and a time of 4 h; The conditions for amylase hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis, and the conditions for inactivating the enzyme are a temperature of 95 °C and a time of 2 min.
[0043] Adding the Jerusalem artichoke antibacterial peptide to the fertilizer is to mix the Jerusalem artichoke antibacterial peptide and the fertilizer raw materials, and then extrude and granulate to obtain the stable fertilizer containing the antibacterial peptide; The mass ratio of the Jerusalem artichoke antibacterial peptide and the fertilizer raw materials is 4:96; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 14
[0044] A stable fertilizer containing an antibacterial peptide, which contains a Jerusalem artichoke antibacterial peptide; the Jerusalem artichoke antibacterial peptide is a product obtained by enzymatically hydrolyzing Jerusalem artichoke residues; the enzymatic hydrolysis is successively carried out by cellulase hydrolysis, amylase hydrolysis and alkaline protease hydrolysis, and it also includes filtering after amylase hydrolysis to obtain a filtrate and a filter residue, and the obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for protease hydrolysis, and after protease hydrolysis, filtration is carried out, and the obtained filtrate is the antibacterial peptide solution; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residues, water, cellulase and amylase is 1:12:0.00002:0.00002; The mass ratio of the filter residue, water and alkaline protease is 1:12:0.00002; The conditions for cellulase hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; The conditions for alkaline protease hydrolysis are a temperature of 40 °C, a pH of 13, and a time of 4 h; The conditions for amylase hydrolysis are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis, and the conditions for inactivating the enzyme are a temperature of 95 °C and a time of 2 min.
[0045] Adding the antibacterial peptide solution to the fertilizer is to mix the Jerusalem artichoke antibacterial peptide and the fertilizer raw materials, and then extrude and granulate to obtain the stable fertilizer containing the antibacterial peptide; The mass ratio of the antibacterial peptide solution and the fertilizer raw materials is 4:96; The fertilizer raw material is a composition of urea, ammonium sulfate, potassium sulfate, and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 15
[0046] A stable fertilizer containing an antibacterial peptide, containing Jerusalem artichoke antibacterial peptide; the Jerusalem artichoke antibacterial peptide is a product obtained by enzymatically hydrolyzing Jerusalem artichoke residue; the enzymatic hydrolysis is successively carried out by cellulase hydrolysis, amylase hydrolysis, and alkaline protease hydrolysis, and also includes filtering after amylase hydrolysis to obtain a filtrate and a filter residue, and the obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for protease hydrolysis, and after protease hydrolysis and filtration, the obtained filtrate is the antibacterial peptide solution; Merge the antibacterial peptide solution and the enzymatic hydrolysate to obtain the Jerusalem artichoke antibacterial peptide solution; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residue, water, cellulase, and amylase is 1:12:0.00002:0.00002; The mass ratio of the filter residue, water, and alkaline protease is 1:12:0.00002; The cellulase hydrolysis conditions are a temperature of 40 °C, a pH of 4, and a time of 4 h; The alkaline protease hydrolysis conditions are a temperature of 40 °C, a pH of 13, and a time of 4 h; The amylase hydrolysis conditions are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis, and the enzyme inactivation conditions are a temperature of 95 °C and a time of 2 min.
[0047] Adding the Jerusalem artichoke antibacterial peptide solution to the fertilizer is to mix the Jerusalem artichoke antibacterial peptide and the fertilizer raw material, and after extrusion granulation, the stable fertilizer containing antibacterial peptide is obtained; The mass ratio of the Jerusalem artichoke antibacterial peptide solution to the fertilizer raw material is 8:92; The fertilizer raw material is a composition of urea, ammonium sulfate, potassium sulfate, and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 16
[0048] A stable fertilizer containing an antibacterial peptide, containing Jerusalem artichoke antibacterial peptide; the Jerusalem artichoke antibacterial peptide is a product obtained by enzymatically hydrolyzing Jerusalem artichoke residue; the enzymatic hydrolysis is successively carried out by cellulase hydrolysis, amylase hydrolysis, xylanase hydrolysis, and alkaline protease hydrolysis, and also includes filtering after xylanase hydrolysis to obtain a filtrate and a filter residue, and the obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for protease hydrolysis, and after protease hydrolysis and filtration, the obtained filtrate is the antibacterial peptide solution; Merge the antibacterial peptide solution and the enzymatic hydrolysate to obtain the Jerusalem artichoke antibacterial peptide solution; The mass ratio of the enzymatically hydrolyzed Jerusalem artichoke residue, water, cellulase, amylase, and xylanase is 1:12:0.00002:0.00002:0.00002; The mass ratio of the filter residue, water, and alkaline protease is 1:12:0.00002; The enzymatic hydrolysis conditions for cellulase are a temperature of 40 °C, a pH of 4, and a time of 4 h; The enzymatic hydrolysis conditions for alkaline protease are a temperature of 40 °C, a pH of 13, and a time of 4 h; The enzymatic hydrolysis conditions for amylase are a temperature of 40 °C, a pH of 4, and a time of 4 h; The enzymatic hydrolysis conditions for xylanase are a temperature of 40 °C, a pH of 4, and a time of 4 h; It also includes inactivating the enzyme after each enzymatic hydrolysis. The enzyme inactivation conditions are a temperature of 95 °C and a time of 2 min.
[0049] Adding the Jerusalem artichoke antimicrobial peptide solution to the fertilizer means mixing the Jerusalem artichoke antimicrobial peptide and the fertilizer raw materials, and then obtaining the antimicrobial peptide-containing stable fertilizer through extrusion granulation; The mass ratio of the Jerusalem artichoke antimicrobial peptide solution to the fertilizer raw materials is 8:92; The fertilizer raw materials are a composition of urea, ammonium sulfate, potassium sulfate, and monoammonium phosphate in a mass ratio of 55:10:20:15. Example 17
[0050] A stable fertilizer containing an antimicrobial peptide contains the Jerusalem artichoke antimicrobial peptide; the Jerusalem artichoke antimicrobial peptide is a product obtained by enzymatic hydrolysis of Jerusalem artichoke residue; the enzymatic hydrolysis is successively cellulase enzymatic hydrolysis, amylase enzymatic hydrolysis, xylanase enzymatic hydrolysis, and alkaline protease enzymatic hydrolysis; it also includes filtering after xylanase enzymatic hydrolysis to obtain a filtrate and a filter residue. The obtained filtrate is the enzymatic hydrolysate; the obtained filter residue is used for protease enzymatic hydrolysis, and after filtering the protease enzymatic hydrolysis, the obtained filtrate is the antimicrobial peptide solution; Merging the antimicrobial peptide solution and the enzymatic hydrolysate gives the Jerusalem artichoke antimicrobial peptide solution; The mass ratio of the Jerusalem artichoke residue for enzymatic hydrolysis, water, cellulase, amylase enzymatic hydrolysis, and xylanase enzymatic hydrolysis is 1:20:0.00004:0.00003:0.00002; The mass ratio of the filter residue, water, and alkaline protease is 1:15:0.00003; The enzymatic hydrolysis conditions for cellulase are a temperature of 50 °C, a pH of 4.5, and a time of 3 h; The enzymatic hydrolysis conditions for amylase are a temperature of 50 °C, a pH of 4.2, and a time of 4 h; The enzymatic hydrolysis conditions for xylanase are a temperature of 50 °C, a pH of 5, and a time of 3 h; The enzymatic hydrolysis conditions for alkaline protease are a temperature of 50 °C, a pH of 12, and a time of 5 h; It also includes inactivating the enzyme after each enzymatic hydrolysis. The enzyme inactivation conditions are a temperature of 100 °C and a time of 1 min.
[0051] Adding the Jerusalem artichoke antimicrobial peptide solution to the fertilizer means mixing the Jerusalem artichoke antimicrobial peptide and the fertilizer raw materials, and then obtaining the antimicrobial peptide-containing stable fertilizer through extrusion granulation; The mass ratio of the Jerusalem artichoke antimicrobial peptide solution to the fertilizer raw materials is 9:91; The fertilizer raw materials are a composition of urea, ammonium chloride, potassium chloride, and monoammonium phosphate in a mass ratio of 40:20:20:20.
[0052] The beneficial effects of the present invention are further illustrated below in conjunction with test data: Experiment 1 1.1 Test location: Gansu Smart Agriculture Engineering Technology Research Center.
[0053] 1.2 Experimental detection: Nitrification inhibition rate (%).
[0054] 1.3 Test materials: Blank (except that the added is unenzymatically hydrolyzed Jerusalem artichoke residue, and the others are the same as in Example 1), Comparative 1 (except that the protease is neutral protease and the pH during enzymatic hydrolysis is 7, and the others are the same as in Example 1), and the fertilizers prepared in Examples 1 to 17.
[0055] 1.4 Detection method: Conducted in accordance with the stability fertilizer standard of GB / T 35113-2017.
[0056] Except for different experimental treatments, other operations in this experiment are the same.
[0057] 2 Results and analysis The nitrification inhibition rate (%), denoted as inhibition rate (%) in the table, is shown in Table 1 Table 1
[0058] From the comparison of the blank (except that the added is unenzymatically hydrolyzed Jerusalem artichoke residue, and the others are the same as in Example 1), Comparative 1 (except that the protease is neutral protease and the pH during enzymatic hydrolysis is 7, and the others are the same as in Example 1), Example 1, and Example 9 in Table 1, it can be seen that Comparative 1 prepared by blank and neutral protease enzymatic hydrolysis does not meet the requirements of stability fertilizers, while Example 1 and Example 9 with added acidic protease both meet the requirements of stability fertilizers, that is, the choice of protease affects the use effect.
[0059] From the data comparison of Example 1, Example 2, Example 3, Example 4 and Example 9, Example 10, Example 11, Example 12, it can be seen that the effects of Example 1 and Example 9 directly using protease hydrolysis are significantly inferior to those of Example 3 and Example 11 adding antibacterial peptide solution prepared by first hydrolyzing with cellulase and then with protease, that is, the hydrolysis sequence of protease affects the use effect; from the data comparison of Example 2, Example 4 and Example 10, Example 12, it can be seen that the effects of Example 4 and Example 12 prepared by filtering after cellulase hydrolysis and then performing protease hydrolysis are better than those of Example 2 and Example 10 directly performing protease hydrolysis without filtering after cellulase hydrolysis. From the result comparison of Example 3, Example 4 and Example 11, Example 12, it can be seen that the effects of Example 3 and Example 11 only adding antibacterial peptide solution are slightly better than those of Example 4 and Example 12 combining the enzyme hydrolysis solution and antibacterial peptide solution, that is, the preparation process affects the use effect.
[0060] Similarly, Example 5, Example 6, Example 7, Example 8 and Example 13, Example 14, Example 15, Example 16 also illustrate that the preparation process affects the use effect.
[0061] Experiment Two 1.1 Test site: Shangying Township, Linzhao County, Dingxi City, Gansu Province. Maize test variety: Dongfanghong No. 3 1.2 Experimental detection: Total nitrogen content of corn straw (% ), total nitrogen content of corn kernels (% ), and average yield (kg / mu).
[0062] 1.3 Test materials: Blank (except that the added is unhydrolyzed Jerusalem artichoke residue, and the others are the same as in Example 1), Comparison 1 (except that the protease is neutral protease and the pH during hydrolysis is 7, and the others are the same as in Example 1), and fertilizers prepared in Examples 1 to 17.
[0063] 1.4 Experimental method: Select 17 mu of corn fields with similar plots, divide them into 17 plots, set isolation belts between plots, and randomly correspond to the blank (except that the added is unhydrolyzed Jerusalem artichoke residue, and the others are the same as in Example 1), Comparison 1 (except that the protease is neutral protease and the pH during hydrolysis is 7, and the others are the same as in Example 1), and fertilizers prepared in Examples 1 to 17.
[0064] In this experiment, except for different experimental treatments, other operations are the same.
[0065] 2 Results and Analysis The total nitrogen content of corn straw (% ), total nitrogen content of corn kernels (% ), and average yield (kg / mu) are respectively recorded as total nitrogen in straw, total nitrogen in grains, and yield, and the average values are taken, as shown in Table 2 Table 2
[0066] By combining Table 1 and Table 2, it can be found that the nitrification inhibition rate in Table 1 is not completely consistent with the nitrogen content and yield in straw and grains in Table 2. The inventor believes that the organic substances produced by cellulase hydrolysis, amylase hydrolysis, and xylanase hydrolysis may have affected nitrogen absorption and plant growth, thereby affecting the nitrogen content in straw and grains and the final yield, resulting in the inconsistent final results between Table 1 and Table 2.
[0067] From the comparison of the blank in Table 2 (except that the added Jerusalem artichoke residue is not hydrolyzed, and the others are the same as in Example 1), Comparative Example 1 (except that the protease is neutral protease and the pH during hydrolysis is 7, and the others are the same as in Example 1), Example 1, and Example 9, it can be seen that the effects of Example 1 and Example 9 are better, that is, the selection of protease affects the use effect, which is consistent with Table 1.
[0068] From the comparison of the data of Example 1, Example 2, Example 3, Example 4, Example 9, Example 10, Example 11, and Example 12 in Table 2, it can be seen that the effects of Example 1 and Example 9 directly using protease hydrolysis are worse than those of Example 3 and Example 11 that are first hydrolyzed by cellulase and then by protease. It can be seen that the hydrolysis sequence of protease affects the use effect, which is consistent with Table 1.
[0069] From the comparison of the data of Example 2, Example 4, Example 10, and Example 12, it can be seen that the effects of Example 4 and Example 12 prepared by filtering after cellulase hydrolysis and then performing protease hydrolysis are better than those of Example 2 and Example 10 that directly perform protease hydrolysis without filtering after cellulase hydrolysis, which is consistent with Table 1.
[0070] From the comparison of the results of Example 3, Example 4, Example 11, and Example 12 in terms of the antibacterial rate when combining Table 1 and Table 2, it can be seen that the effects of Example 3 and Example 11 with only antibacterial peptide solution added are slightly better than those of Example 4 and Example 12 with the combined enzyme hydrolysis solution and antibacterial peptide solution. However, in Table 2, it shows that Example 3 and Example 11 are worse than Example 4 and Example 12 in terms of the nitrogen content and yield of straw and grains, that is, the data in Table 2 are inconsistent with those in Table 1. The inventor believes that it may be the influence of the organic matter in the enzyme hydrolysis solution added in Example 4 and Example 12, that is, adding the enzyme hydrolysis solution to the antibacterial solution will have a positive effect.
[0071] Similarly, the data of Example 5, Example 6, Example 7, and Example 8 in Table 2 are basically consistent with the conclusions of Example 1 to 4; the conclusions of Example 13 to 16 are consistent with those of Example 9 to 12.
[0072] In summary, although the data in Table 1 and Table 2 are slightly inconsistent, it can be proved that the final effect is related to the choice of enzyme, the enzymatic hydrolysis sequence, and the enzymatic hydrolysis process. Moreover, the focus of this application is to prepare fertilizers that meet the requirements of the stability fertilizer standard of GB / T 35113-2017, and then select a better product plan based on field trials.
Claims
1. A stable fertilizer containing antimicrobial peptides, characterized in that: Contains Jerusalem artichoke antimicrobial peptides; the Jerusalem artichoke antimicrobial peptides are products obtained by enzymatic hydrolysis of Jerusalem artichoke residues; the enzymatic hydrolysis is protease hydrolysis; The protease is an acidic protease or an alkaline protease; The requirement for the stable fertilizer is that the nitrification inhibition rate is ≥ 6%.
2. The stable fertilizer containing antimicrobial peptides according to claim 1, characterized in that: The enzymatic hydrolysis also includes one or more of cellulase hydrolysis, starch hydrolysis and xylan hydrolysis.
3. The stable fertilizer containing antimicrobial peptide according to claim 1 or 2, characterized in that: The mass ratio of the enzymatic hydrolysis Jerusalem artichoke residue, water and single enzyme is 1:8-25:0.00002-0.00005; The single enzyme is one of protease, cellulase, amylase and xylanase.
4. The stable fertilizer containing antimicrobial peptide according to claim 3, characterized in that: The enzymolysis includes protease enzymolysis, cellulase enzymolysis, starch enzymolysis and xylan enzymolysis; The enzymatic hydrolysis sequence is to first perform cellulase hydrolysis and then perform protease hydrolysis; The conditions for cellulase hydrolysis are temperature 30-60°C, pH 4-6, and time 3-6h; The conditions for starch enzymatic hydrolysis are temperature 30-60°C, pH 4-6, and time 3-6h; The conditions for xylan hydrolysis are temperature 30-60°C, pH 4-6, and time 3-6h; The conditions for protease hydrolysis are temperature 30-60°C, pH 3.5-5.5 or 12-14, and time 3-6 hours; The process also includes inactivating the enzyme, and the conditions for inactivating the enzyme are a temperature of 95 to 100° C. and a time of 1 to 3 minutes.
5. The stable fertilizer containing antimicrobial peptide according to claim 4, characterized in that: The invention also includes filtering before protease hydrolysis, and the obtained filtrate is the hydrolysis liquid; the obtained filter residue is used for protease hydrolysis, and filtering after protease hydrolysis, and the obtained filtrate is the antimicrobial peptide liquid; The antimicrobial peptide solution and the enzymatic hydrolyzate are combined to obtain the Jerusalem artichoke antimicrobial peptide solution; Drying the antimicrobial peptide liquid to obtain powdered antimicrobial peptide; Drying the Jerusalem artichoke antimicrobial peptide liquid to obtain Jerusalem artichoke antimicrobial peptide powder; The Jerusalem artichoke antimicrobial peptide comprises one or two of antimicrobial peptide liquid, antimicrobial peptide powder, Jerusalem artichoke antimicrobial peptide liquid and Jerusalem artichoke antimicrobial peptide powder; The mass ratio of filter residue, water and protease in protease hydrolysis is 1:8-20:0.00002-0.00005.
6. The method for preparing a stable fertilizer containing antimicrobial peptides according to claims 1 to 5, characterized in that: Adding Jerusalem artichoke antimicrobial peptides into fertilizers to obtain stable fertilizers containing antimicrobial peptides; The mass ratio of Jerusalem artichoke antimicrobial peptides in fertilizer is 0.1% to 90%; The fertilizer is one or more of organic and inorganic fertilizers, bio-organic fertilizers, compound fertilizers and composite fertilizers.
7. The method for preparing a stable fertilizer containing antimicrobial peptides according to claim 6, characterized in that: The step of adding the Jerusalem artichoke antimicrobial peptide to the fertilizer is to mix the Jerusalem artichoke antimicrobial peptide and the fertilizer raw material, and then granulate the mixture to obtain a stable fertilizer containing the antimicrobial peptide. The fertilizer raw material is one or more of urea, ammonium chloride, ammonium sulfate, potassium chloride, potassium sulfate and monoammonium phosphate.
8. The method for preparing a stable fertilizer containing antimicrobial peptides according to claim 7, characterized in that: The method further comprises drying after granulation, wherein the drying temperature is 30 to 70°C.