Preparation method of agricultural jerusalem artichoke antibacterial peptide-containing extract and application of agricultural jerusalem artichoke antibacterial peptide-containing extract in agriculture
The preparation of antibacterial peptide extracts by enzymatic treatment of Jerusalem artichoke residues has been solved, which may cause bacterial colonization problems when Jerusalem artichoke residues are directly applied to agriculture, and has achieved high-yield agricultural applications that do not affect crop growth, and has improved the antibacterial and free radical scavenging effects.
Patent Information
- Application Number
- CN202510444631.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-10
- Publication Date
- 2025-07-01
AI Technical Summary
In the prior art, when Jerusalem artichoke residue is directly applied to agriculture, it is easy to cause colonization of harmful bacteria, affecting the growth of crop roots and reducing yield.
The extract of antimicrobial peptides containing Jerusalem artichoke for agriculture was prepared by enzymatic treatment of the Jerusalem artichoke residue, including cellulose, amylase and proteolytic, and applied to agriculture or added to fertilizers.
This method makes the application of Jerusalem artichoke residue not affecting crop growth and can improve crop yield. At the same time, the prepared antibacterial peptide extract has good antibacterial effect and free radical scavenging ability.
Smart Images
Figure SMS_1 
Figure SMS_2 
Figure SMS_3
Abstract
Description
Technical Field
[0001] The invention relates to a preparation method of an agricultural Jerusalem artichoke antimicrobial peptide extract and application thereof in agriculture. Background Art
[0002] Jerusalem artichoke, also known as Jerusalem artichoke and devil ginger, is a plant of the genus Helianthus in the Asteraceae family. It is drought-resistant and salt-alkali-resistant, and is therefore often used for windbreak and sand fixation. Jerusalem artichoke can be used to prepare inulin, and its byproduct is Jerusalem artichoke residue, which has a huge output and a water content of about 83%. It also contains a large amount of cellulose, hemicellulose, protein and starch. Currently, Jerusalem artichoke residue is mainly used as animal feed and is rarely used in agriculture. Since Jerusalem artichoke residue contains a large amount of water and starch, it is directly used in agriculture. The presence of harmful pathogens can easily cause colonization, affect the root growth of plants, and thus affect crop yields.
[0003] Currently, there are no products and methods for applying Jerusalem artichoke residue to agriculture without affecting crop growth and further increasing crop yield. Summary of the invention
[0004] The invention provides a preparation method of an agricultural Jerusalem artichoke antimicrobial peptide extract and application thereof in agriculture, and solves the technical problem of applying Jerusalem artichoke residues to agriculture to influence crop growth and further influence crop yield.
[0005] In order to solve the above technical problems, the present invention adopts the following technical solutions: A method for preparing an agricultural Jerusalem artichoke antimicrobial peptide extract is carried out according to the following steps: The Jerusalem artichoke residue is subjected to enzymatic hydrolysis to obtain an agricultural Jerusalem artichoke antimicrobial peptide extract; The enzymolysis includes protein hydrolysis, cellulose hydrolysis and starch hydrolysis; The enzymatic hydrolysis is carried out in the following order: cellulose enzymatic hydrolysis, starch enzymatic hydrolysis, and protein enzymatic hydrolysis; The agricultural Jerusalem artichoke antimicrobial peptide extract is used in agriculture.
[0006] Also included is enzymatic hydrolysis of xylan.
[0007] 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; The enzymatic hydrolysis sequence is cellulose hydrolysis, starch hydrolysis, xylan hydrolysis, and protein hydrolysis.
[0008] The protease used in the proteolysis is an acidic protease or an alkaline protease; The conditions for cellulose enzymatic 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; It also includes enzyme inactivation, which is carried out at a temperature of 95 to 100°C and a time of 1 to 3 minutes. The conditions for proteolysis are a temperature of 30 to 60°C, a pH of 3.5 to 5.5 or 12 to 14, and a time of 3 to 6 hours.
[0009] It also includes filtration after enzymatic hydrolysis of cellulose and starch, the obtained filtrate is the enzymatic hydrolysis liquid, the obtained filter residue is used for protein hydrolysis, and filtration after protein hydrolysis, and the obtained filtrate is the antimicrobial peptide liquid; or After enzymatic hydrolysis of cellulose, starch and xylan, the filtrate is filtered, and the obtained filtrate is the enzymatic hydrolysis liquid. The obtained filter residue is used for protein hydrolysis, and after protein hydrolysis, the filtrate is filtered, and the obtained filtrate is the antimicrobial peptide liquid. The antimicrobial peptide liquid and the enzymatic hydrolyzate are combined to obtain a liquid agricultural extract containing Jerusalem artichoke antimicrobial peptides; Drying the liquid Jerusalem artichoke antimicrobial peptide extract for agriculture to obtain a powdered Jerusalem artichoke antimicrobial peptide extract for agriculture; The Jerusalem artichoke antimicrobial peptide extract for agricultural use includes a liquid Jerusalem artichoke antimicrobial peptide extract for agricultural use and a powdered Jerusalem artichoke antimicrobial peptide extract for agricultural use; The mass ratio of filter residue, water and protease in proteinase hydrolysis is 1:8-20:0.00002-0.00005.
[0010] The use of Jerusalem artichoke antimicrobial peptide extracts in agriculture, and the direct use of Jerusalem artichoke antimicrobial peptide extracts in agriculture as nutrient solution; or Adding Jerusalem artichoke antimicrobial peptide extracts to fertilizers for agricultural use; The mass ratio of Jerusalem artichoke antimicrobial peptide extract in agricultural 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.
[0011] The method of adding the Jerusalem artichoke antimicrobial peptide extract to the fertilizer comprises adding the Jerusalem artichoke antimicrobial peptide extract to the fertilizer raw material, granulating and drying the extract to obtain the antimicrobial peptide-containing stable fertilizer; The drying temperature is 50-80°C.
[0012] The invention has the following beneficial technical effects: 1) In the present application, Jerusalem artichoke residue is subjected to enzymatic hydrolysis, and the enzymatic hydrolysis includes protein hydrolysis, cellulose hydrolysis and starch hydrolysis, and the agricultural Jerusalem artichoke antimicrobial peptide extract prepared by the invention will not affect the growth of crops and can further increase the crop yield when applied in agriculture.
[0013] 2) The enzymatic hydrolysis sequence of the present application is to first perform cellulose enzymatic hydrolysis, starch enzymatic hydrolysis, and then perform proteinase hydrolysis, and the protease used is acidic protease or alkaline protease. The Jerusalem artichoke antimicrobial peptide extract prepared in this way has a good antibacterial effect and can better scavenge free radicals.
[0014] 3) The present application also includes filtration after enzymatic hydrolysis of cellulose, enzymatic hydrolysis of starch and enzymatic hydrolysis of xylan, the obtained filtrate is the enzymatic hydrolysis liquid, the obtained filter residue is used for protein hydrolysis, and the obtained filtrate is filtered after protein hydrolysis, and the obtained filtrate is the antimicrobial peptide liquid; the antimicrobial peptide liquid and the enzymatic hydrolysis liquid are combined to obtain a liquid Jerusalem artichoke antimicrobial peptide extract for agricultural use; by filtering and separating the enzymatic hydrolysis liquid, a better Jerusalem artichoke antimicrobial peptide extract can be obtained. DETAILED DESCRIPTION
[0015] In order to better understand the above technical solution, the technical solution of the present invention is described in detail below in conjunction with preferred examples. Example 1
[0016] A method for preparing an agricultural Jerusalem artichoke antimicrobial peptide extract is carried out according to the following steps: Mix Jerusalem artichoke residue, water and cellulase, degrade for 4 hours at 45°C and pH 4, add amylase, degrade for 4 hours at 45°C and pH 4, add acid protease, degrade for 3 hours at 45°C and pH 4, filter, and the filtrate is a liquid Jerusalem artichoke antimicrobial peptide extract for agricultural use; The mass ratio of the enzymatic hydrolysis Jerusalem artichoke residue, water, cellulase, amylase and protease is 1:24:00002: 00002:0.00002.
[0017] The application of Jerusalem artichoke antimicrobial peptide extract in agriculture, the mass ratio of Jerusalem artichoke antimicrobial peptide extract in fertilizer is 1% Example 2
[0018] A method for preparing an agricultural Jerusalem artichoke antimicrobial peptide extract is carried out according to the following steps: Mix Jerusalem artichoke residue, water and cellulase, degrade for 4 hours at 45°C and pH 4, add amylase, degrade for 4 hours at 45°C and pH 4, add alkaline protease, degrade for 3 hours at 45°C and pH 4, filter, and the filtrate is a liquid Jerusalem artichoke antimicrobial peptide extract for agricultural use; The mass ratio of the enzymatic hydrolysis Jerusalem artichoke residue, water, cellulase, amylase and protease is 1:24:00002: 00002:0.00002.
[0019] The application of Jerusalem artichoke antimicrobial peptide extract in agriculture, the mass ratio of Jerusalem artichoke antimicrobial peptide extract in fertilizer is 1% Example 3
[0020] A method for preparing an agricultural Jerusalem artichoke antimicrobial peptide extract is carried out according to the following steps: Mix Jerusalem artichoke residue, water and cellulase, degrade for 4 hours at 45°C and pH 4, add amylase, degrade for 4 hours at 45°C and pH 4, filter, and the obtained filtrate is the enzymatic solution, add water and acid protease to the filter residue, degrade for 3 hours at 45°C and pH 4, filter, and the obtained filtrate is the antimicrobial peptide solution; The antimicrobial peptide liquid and the enzymatic hydrolyzate are combined to obtain a liquid agricultural extract containing Jerusalem artichoke antimicrobial peptides. The mass ratio of the enzymatic hydrolysis Jerusalem artichoke residue, water, cellulase and amylase is 1:12:00002:00002; The mass ratio of the filter residue, water and acid protease is 1:12:00002.
[0021] The application of Jerusalem artichoke antimicrobial peptide extract in agriculture, the mass ratio of Jerusalem artichoke antimicrobial peptide extract in fertilizer is 1%. Example 4
[0022] A method for preparing an agricultural Jerusalem artichoke antimicrobial peptide extract is carried out according to the following steps: Mix Jerusalem artichoke residue, water and cellulase, degrade for 4 hours at 45°C and pH 4, add amylase, degrade for 4 hours at 45°C and pH 4, filter, and the obtained filtrate is the enzymatic solution, add water and alkaline protease to the residue, degrade for 3 hours at 45°C and pH 4, filter, and the obtained filtrate is the antimicrobial peptide solution; The antimicrobial peptide liquid and the enzymatic hydrolyzate are combined to obtain a liquid agricultural extract containing Jerusalem artichoke antimicrobial peptides. The mass ratio of the enzymatic hydrolysis Jerusalem artichoke residue, water, cellulase and amylase is 1:12:00002:00002; The mass ratio of the filter residue, water and alkaline protease is 1:12:00002.
[0023] The application of Jerusalem artichoke antimicrobial peptide extract in agriculture, the mass ratio of Jerusalem artichoke antimicrobial peptide extract in fertilizer is 10%. Example 5
[0024] A method for preparing an agricultural Jerusalem artichoke antimicrobial peptide extract is carried out according to the following steps: Mix Jerusalem artichoke residue, water and cellulase, degrade for 4 hours at 45°C and pH 4, add amylase, degrade for 4 hours at 45°C and pH 4, add xylanase, degrade for 3 hours at 50°C and pH 4.5, filter, and the obtained filtrate is the enzymatic solution, add water and acid protease to the filter residue, degrade for 3 hours at 45°C and pH 4, and filter, and the obtained filtrate is the antimicrobial peptide solution; The antimicrobial peptide liquid and the enzymatic hydrolyzate are combined to obtain a liquid agricultural extract containing Jerusalem artichoke antimicrobial peptides; The mass ratio of the enzymatic hydrolysis Jerusalem artichoke residue, water, cellulase, amylase and xylanase is 1:12:00002:00002:00002; The mass ratio of the filter residue, water and acid protease is 1:12:00002.
[0025] The application of Jerusalem artichoke antimicrobial peptide extract in agriculture, the mass ratio of Jerusalem artichoke antimicrobial peptide extract in fertilizer is 20%. Example 6
[0026] A method for preparing an agricultural Jerusalem artichoke antimicrobial peptide extract is carried out according to the following steps: Mix Jerusalem artichoke residue, water and cellulase, degrade for 3 hours at 55°C and pH 4, add amylase, degrade for 4 hours at 45°C and pH 4.5, filter, and the obtained filtrate is the enzymatic solution, add water and acid protease to the filter residue, degrade for 3 hours at 50°C and pH 4, filter, and the obtained filtrate is the antimicrobial peptide solution; The antimicrobial peptide liquid and the enzymatic hydrolyzate are combined to obtain a liquid agricultural extract containing Jerusalem artichoke antimicrobial peptides. The mass ratio of the enzymatic hydrolysis Jerusalem artichoke residue, water, cellulase and amylase is 1:14:00003:00003; The mass ratio of the filter residue, water and acid protease is 1:14:00003.
[0027] The application of Jerusalem artichoke antimicrobial peptide extract in agriculture, the mass ratio of Jerusalem artichoke antimicrobial peptide extract in fertilizer is 60%. Example 7
[0028] A method for preparing an agricultural Jerusalem artichoke antimicrobial peptide extract is carried out according to the following steps: Mix Jerusalem artichoke residue, water and cellulase, degrade for 4 hours at 45°C and pH 4, add amylase, degrade for 3 hours at 50°C and pH 4.5, add xylanase, degrade for 3 hours at 50°C and pH 4.5, Filter, the filtrate is the enzymatic solution, add water and alkaline protease to the residue, degrade for 3 hours at 55°C and pH 4, and filter the filtrate to obtain the antimicrobial peptide solution. The antimicrobial peptide liquid and the enzymatic hydrolyzate are combined to obtain a liquid agricultural extract containing Jerusalem artichoke antimicrobial peptides; The mass ratio of the enzymatic hydrolysis Jerusalem artichoke residue, water, cellulase, amylase and xylanase is 1:15:00003:00003:00002; The mass ratio of the filter residue, water and alkaline protease is 1:15:00003.
[0029] The application of Jerusalem artichoke antimicrobial peptide extract in agriculture, the mass ratio of Jerusalem artichoke antimicrobial peptide extract in fertilizer is 90%.
[0030] The beneficial effects of the present invention are further illustrated below in conjunction with test data: Experiment 1 1.1 Experimental location: Gansu Smart Agriculture Engineering Technology Research Center.
[0031] 1.2 Experimental testing: hydroxyl radical scavenging rate (%) and antibacterial diameter (mm).
[0032] 1.3 Test materials: Comparison 1 (except that cellulase, amylase and acid protease were added together, the rest were the same as in Example 1), Comparison 2 (except that the enzymatic hydrolysis order is cellulase hydrolysis, protease hydrolysis, and amylase hydrolysis, the others are consistent with Example 1), Comparison 3 (except that the enzymatic hydrolysis order is cellulase hydrolysis, amylase hydrolysis, protease hydrolysis, and xylanase hydrolysis, the others are consistent with Example 5), Comparison 4 (except that neutral protease is used and the pH during enzymatic hydrolysis is 7, the others are consistent with Example 1), the liquid agricultural Jerusalem artichoke antimicrobial peptide extract prepared in Examples 1 to 7, the antimicrobial peptide liquid prepared in Comparison 5 (the antimicrobial peptide liquid prepared in Example 3), and Comparison 6 (the enzymatic hydrolyzate prepared in Example 3).
[0033] 1.4 Detection method: Hydroxyl free radical scavenging rate (%): Take 2 ml of 6 mmol / L ferrous sulfate, liquid agricultural Jerusalem artichoke antimicrobial peptide extract (or antimicrobial peptide liquid), and 6 mmol / L hydrogen peroxide, let it stand for 10 minutes, then add 6 mmol / L salicylic acid, let it stand for 30 minutes, and measure the absorbance at 510 nm. Scavenging rate (%) = (A0-(A1-A2)) / A0*100%; where A0: absorbance without adding liquid agricultural Jerusalem artichoke antimicrobial peptide extract (or antimicrobial peptide liquid), A1: absorbance with adding liquid agricultural Jerusalem artichoke antimicrobial peptide extract (or antimicrobial peptide liquid), A2: absorbance without adding salicylic acid; Inhibition diameter (mm): Take the bacteria that have been matured and cultured for 24 hours with an inoculation loop 2-3 times, then spread it evenly on the surface of the culture medium with a coating rod, spot it on a 6mm wide filter paper, place it at 37℃ for one day, and measure the diameter (mm) of the inhibition zone with a vernier caliper.
[0034] The bacteria used for the inhibition diameter were nitrite bacteria, nitrobacter, and Escherichia coli; Note: Since cellulose is the main component of plant cell walls and provides structural support and protection for plants, this application first performs cellulase degradation according to convention to dissolve the cell contents, and then performs other enzymatic hydrolysis.
[0035] Except for the experimental treatment, other operations were the same in this experiment.
[0036] 2 Results and analysis Hydroxyl radical scavenging rate (%) and antibacterial diameter (mm), where hydroxyl radical scavenging rate (%) is recorded as scavenging rate (%), see Table 1
[0037] From Comparison 1 in Table 1 (except that cellulase, amylase and protease are added together, everything else is the same as Example 1), Comparison of the data of Comparison 2 (except that the order of enzymatic hydrolysis is cellulase enzymatic hydrolysis, protease enzymatic hydrolysis, and amylase enzymatic hydrolysis, all other aspects are consistent with Example 1) and Example 1, as well as Comparison 3 (except that the order of enzymatic hydrolysis is cellulase enzymatic hydrolysis, amylase enzymatic hydrolysis, protease enzymatic hydrolysis, and xylanase enzymatic hydrolysis, all other aspects are consistent with Example 5) and Example 5 shows that the order of adding the enzymes affects the effect of the product.
[0038] From the comparison of the data of Comparative 4 (which is consistent with Example 1 except that neutral protease is used and the pH value during enzymatic hydrolysis is 7), Example 1 and Example 2, it can be seen that Example 1 and Example 2 which use acidic protease and alkaline protease for enzymatic hydrolysis have better antibacterial effects.
[0039] By comparing the data of Comparative 5 (the antimicrobial peptide solution prepared in Example 3) and Example 3, it can be seen that the antimicrobial solution of Comparative 5 alone has the best clearance rate and antimicrobial effect.
[0040] From Comparative Example 6 (the enzymatic hydrolysate prepared in Example 3), it can be seen that the enzymatic hydrolysate has a certain effect of scavenging free radicals, but has no antibacterial effect.
[0041] From the data comparison between Example 1 and Example 3, and Example 2 and Example 4, it can be seen that Example 3 and Example 4, in which the enzymatic hydrolyses of cellulase and amylase are separated by filtration and then subjected to protease hydrolysis, are better than Example 1 and Example 2 in terms of clearance rate and antibacterial effect.
[0042] From the comparison of the data of Example 3 and Example 5, it can be seen that the addition of xylanase to further degrade the water-soluble organic matter may affect the concentration of antimicrobial peptides, thereby reducing the clearance rate and antimicrobial effect.
[0043] The test data of Jerusalem artichoke residue used in this experiment (on a dry basis, %) are shown in Table 2
[0044] Experiment 2 1.1 Experimental location: Gansu Smart Agriculture Engineering Technology Research Center.
[0045] 1.2 Experimental test: root activity (mg / gh).
[0046] 1.3 Test materials: blank (untreated Jerusalem artichoke residue), control 1 (except for the addition of cellulase, amylase and protease The invention relates to comparative examples 1 to 2 (except that the enzymatic hydrolysis order is cellulase hydrolysis, protease hydrolysis and amylase hydrolysis, and the others are consistent with Example 1), comparative example 2 (except that the enzymatic hydrolysis order is cellulase hydrolysis, amylase hydrolysis, protease hydrolysis and xylanase hydrolysis, and the others are consistent with Example 5), comparative example 4 (except that neutral protease is used and the pH value during enzymatic hydrolysis is 7, and the others are consistent with Example 1), the liquid agricultural antimicrobial peptide extract containing Jerusalem artichoke prepared in Examples 1 to 7, the antimicrobial peptide liquid prepared in Comparison 5 (the antimicrobial peptide liquid prepared in Example 3), and the enzymatic hydrolyzate prepared in Comparison 6 (the enzymatic hydrolyzate prepared in Example 3).
[0047] 1.4 Experimental method: Take soil that has not been used with agricultural film for 4 years, remove stones, branches and roots, and place it in an oven at 80℃. Dry for 30 minutes, pass through a 1mm sieve, and put into pots, 1.5kg per pot. Transplant the lettuce seedlings into the pots, and control the soil moisture content at about 65% by drip irrigation with the diluted solution of each treatment. Plant three plants in each pot. The selected lettuces are of the same size to the naked eye. Harvest after 45 days of growth and measure the root activity after harvest. The planting experiment was carried out in a greenhouse, and the experiment was done in 4 parallels.
[0048] The dilution solution of each treatment is the dilution solution obtained by diluting each treatment with water at a mass ratio of 1:500; the amount of dilution solution used in each drip irrigation of each treatment in the experiment is the same.
[0049] Note: Blank (untreated Jerusalem artichoke residue) was used as basal fertilizer and mixed well with soil.
[0050] 1.5 Detection method: Root activity was determined based on the triphenyltetrazolium chloride (TTC) method.
[0051] Except for the experimental treatment, other operations were the same in this experiment.
[0052] 2 Results and analysis Root activity (mg / gh), recorded as activity (mg / gh) in the table, see Table 3 and Table 4 From the data in Table 1, Table 3 and Table 4, it can be found that although Comparison 5 has better free radical scavenging and antibacterial effects, Comparison 6 has average free radical scavenging and no antibacterial effect, but Comparison 5 has better effect in terms of root activity.
[0053] The other data in Table 1, Table 3 and Table 4 are basically consistent, that is, the preparation process affects the use effect.
[0054] Experiment 3 1.1 Test location: Dingxi City, Gansu Province. Potato test variety: Wotu No. 5.
[0055] 1.2 Experimental test: plant height (cm), number of single potatoes (pieces), average yield (kg / mu).
[0056] 1.3 Test materials: blank (untreated Jerusalem artichoke residue), control 1 (except for the addition of cellulase, amylase and protease The invention relates to comparative examples 1 to 2 (except that the enzymatic hydrolysis order is cellulase hydrolysis, protease hydrolysis and amylase hydrolysis, and the others are consistent with Example 1), comparative example 2 (except that the enzymatic hydrolysis order is cellulase hydrolysis, amylase hydrolysis, protease hydrolysis and xylanase hydrolysis, and the others are consistent with Example 5), comparative example 4 (except that neutral protease is used and the pH value during enzymatic hydrolysis is 7, and the others are consistent with Example 1), the liquid agricultural antimicrobial peptide extract containing Jerusalem artichoke prepared in Examples 1 to 7, the antimicrobial peptide liquid prepared in Comparison 5 (the antimicrobial peptide liquid prepared in Example 3), and the enzymatic hydrolyzate prepared in Comparison 6 (the enzymatic hydrolyzate prepared in Example 3).
[0057] 1.4 Experimental method: On April 14, 2024, the soil was turned over and the blank basal fertilizer was applied at a rate of 100 kg / mu. The other treatments were drip irrigation. Drip irrigation, the operation method is to dilute each treatment with water, the dilution multiple is 500 times, each drip irrigation time is 40 minutes, and drip irrigation is carried out every 20 days. Each treatment corresponds to 1 mu, and potatoes are sown on April 27, 2024, and the film is used for planting. The harvest time is October 5, 2024.
[0058] Except for the experimental treatment, other operations were the same in this experiment.
[0059] 2 Results and analysis Plant height (cm), number of single potatoes (pieces), and yield (kg / mu) are all average values, see Table 5
[0060] Potatoes were selected for the experiment to verify the effect on the underground part, wherein Table 3 is the effect on the root system, and Table 4 is the effect on the tuber. By comparing the blank, comparative example and embodiment in Table 1 in Table 3 and Table 4, it can be seen that directly applying Jerusalem artichoke residue to agriculture does not have a good effect.
[0061] From the comparison of the data of Comparison 1 (except that cellulase, amylase and protease are added together, everything else is consistent with Example 1), Comparison 2 (except that the enzymatic hydrolysis order is cellulase hydrolysis, protease hydrolysis, and amylase hydrolysis, everything else is consistent with Example 1) and Example 1 and Comparison 3 (except that the enzymatic hydrolysis order is cellulase hydrolysis, amylase hydrolysis, protease hydrolysis, and xylanase hydrolysis, everything else is consistent with Example 5) and Example 5, it can be seen that the order of enzyme addition affects the effect of the agricultural Jerusalem artichoke antimicrobial peptide extract.
[0062] From the comparison of the data of Comparative 4 (which is consistent with Example 1 except that neutral protease is used), Example 1 and Example 2, it can be seen that the selected protease directly affects the use effect.
[0063] From the comparison of the data of Comparative 5 (the antimicrobial peptide solution prepared in Example 3), Comparative 6 (the enzymatic hydrolysate prepared in Example 3) and Example 3, it can be seen that the agricultural antimicrobial peptide extract containing Jerusalem artichoke prepared by combining the antimicrobial peptide solution and the enzymatic hydrolysate in Example has the best effect.
[0064] From the data comparison between Example 1 and Example 3, and Example 2 and Example 4, it can be seen that the effects of Example 3 and Example 4 in which the enzymatic hydrolysis of cellulase and amylase was separated by filtration and then subjected to protease hydrolysis were better than those of Example 1 and Example 2 in which the enzymatic hydrolysis was not separated by filtration. This may be because the enzymatic hydrolysis products of cellulase and amylase reacted with the enzymatic hydrolysis products of protease, thereby affecting the use effect.
[0065] From the comparison of the data of Example 3 and Example 5, it can be seen that the product after enzymatic hydrolysis with xylanase has better effect.
Claims
1. A method for preparing an agricultural antimicrobial peptide extract containing Jerusalem artichoke, characterized in that: Follow the steps below, The Jerusalem artichoke residue is subjected to enzymatic hydrolysis to obtain an agricultural Jerusalem artichoke antimicrobial peptide extract; The enzymolysis includes protein hydrolysis, cellulose hydrolysis and starch hydrolysis; The enzymatic hydrolysis sequence is cellulose enzymatic hydrolysis, starch enzymatic hydrolysis, and protein enzymatic hydrolysis; The agricultural Jerusalem artichoke antimicrobial peptide extract is used in agriculture.
2. The method for preparing the Jerusalem artichoke antimicrobial peptide extract for agricultural use as claimed in claim 1, characterized in that: Also included is enzymatic hydrolysis of xylan.
3. The method for preparing an agricultural Jerusalem artichoke antimicrobial peptide extract 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; The enzymatic hydrolysis sequence is cellulose hydrolysis, starch hydrolysis, xylan hydrolysis, and protein hydrolysis.
4. The method for preparing the Jerusalem artichoke antimicrobial peptide extract for agricultural use according to claim 3, characterized in that: The protease used in the proteolysis is an acidic protease or an alkaline protease; The conditions for cellulose enzymatic 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 proteolysis are a temperature of 30 to 60°C, a pH of 3.5 to 5.5 or 12 to 14, and a time of 3 to 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 method for preparing the Jerusalem artichoke antimicrobial peptide extract for agricultural use according to claim 3, characterized in that: It also includes filtration after enzymatic hydrolysis of cellulose and starch, the obtained filtrate is the enzymatic hydrolysis liquid, the obtained filter residue is used for protein hydrolysis, and filtration after protein hydrolysis, and the obtained filtrate is the antimicrobial peptide liquid; or After enzymatic hydrolysis of cellulose, starch and xylan, the filtrate is filtered, and the obtained filtrate is the enzymatic hydrolysis liquid. The obtained filter residue is used for protein hydrolysis, and after protein hydrolysis, the filtrate is filtered, and the obtained filtrate is the antimicrobial peptide liquid. The antimicrobial peptide liquid and the enzymatic hydrolyzate are combined to obtain a liquid agricultural extract containing Jerusalem artichoke antimicrobial peptides; Drying the liquid Jerusalem artichoke antimicrobial peptide extract for agriculture to obtain a powdered Jerusalem artichoke antimicrobial peptide extract for agriculture; The Jerusalem artichoke antimicrobial peptide extract for agricultural use includes a liquid Jerusalem artichoke antimicrobial peptide extract for agricultural use and a powdered Jerusalem artichoke antimicrobial peptide extract for agricultural use; The mass ratio of filter residue, water and protease in proteinase hydrolysis is 1:8-20:0.00002-0.00005.
6. The use of the Jerusalem artichoke antimicrobial peptide extract in agriculture as claimed in claims 1 to 5, characterized in that: Agricultural use of Jerusalem artichoke antimicrobial peptide extracts as nutrient solution directly applied to agriculture; or Adding Jerusalem artichoke antimicrobial peptide extracts to fertilizers for agricultural use; The mass ratio of Jerusalem artichoke antimicrobial peptide extract in agricultural 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 use of the Jerusalem artichoke antimicrobial peptide extract in agriculture as claimed in claim 6, wherein the Jerusalem artichoke antimicrobial peptide extract is added to the fertilizer, and the Jerusalem artichoke antimicrobial peptide extract is added to the fertilizer raw material, and then granulated and dried to obtain the antimicrobial peptide-containing stable fertilizer; The drying temperature is 50-80°C.