Dilute titanium alcohol yield increasing and growth promoting agent and preparation method thereof
By preparing dilute titanium alcohol production-enhancing agents, the problem of poor production-enhancing agents in the prior art is solved, and effective production-enhancing agents and increase yields of crops is achieved.
Patent Information
- Application Number
- CN202510025523.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-08
- Publication Date
- 2025-03-28
AI Technical Summary
In the prior art, growth promoters have poor effect on crop yield increase, affecting crop growth and yield.
By preparing a diluted titanium alcohol production-enhancing agent, including hemoglobin extraction, crude peptide preparation, chelate preparation and other steps, a diluted titanium alcohol solution with polypeptide, rare earth and sterol components is formed, and a production-enhancing agent is obtained by spray drying.
Effectively promote crop yield increase, improve crop photosynthesis capabilities, resist external adverse factors, accelerate nutritional growth rate, and increase yield.
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Abstract
Description
Technical Field
[0001] The invention relates to the technical field of crop growth and fertilizer, and in particular to a dilute titanium alcohol yield-increasing and growth-promoting agent and a preparation method thereof. Background Art
[0002] Crops refer to various plants cultivated in agriculture. They include food crops and cash crops (oil crops, vegetable crops, flowers, grasses, and trees). Fruits include pears, greengages, apples, peaches, apricots, walnuts, plums, cherries, strawberries, crab apples, and red dates. In the prior art, growth promoters have poor effects on increasing crop yields, are detrimental to crop growth, and affect crop yields. Based on this, the present invention further improves and processes them. Summary of the invention
[0003] In view of the defects of the prior art, the purpose of the present invention is to provide a dilute titanium alcohol production-increasing and growth-promoting agent and a preparation method to solve the problems raised in the above-mentioned background technology.
[0004] The present invention solves the technical problem by adopting the following technical solution: The present invention provides a method for preparing a dilute titanium alcohol yield-increasing and growth-promoting agent, comprising the following steps: (1) Hemoglobin extraction: add anticoagulant to fresh animal blood, collect the precipitate by centrifugation, and collect the supernatant hemoglobin after homogenization to obtain a hemoglobin solution with a mass concentration of 10-20%; (2) Preparation of crude peptide: Add enzyme to the hemoglobin solution and heat to 45-55°C for enzymatic hydrolysis for 4-6 hours. After the enzymatic hydrolysis is completed, the enzyme is inactivated; (3) Isolation and purification of crude peptides: The enzymatic hydrolysate in step (2) was centrifuged at 4°C to collect the supernatant, and filtered through a 20 nm nanofiltration membrane to obtain a peptide solution; (4) Preparation of chelate: The polypeptide liquid is ultrasonically pretreated before chelation, and a chelating ligand is added to the polypeptide liquid in step (3), wherein the chelating ligand accounts for 5-15% of the mass ratio of the polypeptide liquid. After the chelation reaction is completed, the mixture is concentrated and spray-dried to obtain an animal blood chelated peptide; (5) The pH of the animal blood chelating peptide is adjusted to 4.5, and then 8% EDTA rare earth of the animal blood chelating peptide is added, and the mixture is stirred at 40 rpm for 40 minutes at 37°C to obtain a chelating solution. After all the solution is dissolved, 5% stigmasterol of the animal blood chelating peptide is added and chelated and stirred for 50 minutes to obtain a dilute titanium alcohol solution. The dilute titanium alcohol solution is concentrated and spray-dried to obtain a dilute titanium alcohol yield-increasing and growth-promoting agent.
[0005] Preferably, the anticoagulant is 5-10% of the total amount of fresh animal blood, and the anticoagulant is 10-15 parts of trisodium citrate, 2-5 parts of sodium hexametaphosphate, 1-3 parts of sodium chloride, 1-2 parts of citric acid and 10-15 parts of water.
[0006] Preferably, the enzyme is composed of a mixture of Bacillus licheniformis alkaline protease, Bacillus subtilis neutral protease and Aspergillus oryzae composite flavor protease in a mass ratio of 3-5:4-3:4-3, and the added amount of the enzyme is 3-10% of the mass of the hemoglobin solution.
[0007] Preferably, the enzyme inactivation conditions are: enzyme inactivation temperature 90-95°C, enzyme inactivation time 10-15 min.
[0008] Preferably, the ultrasonic treatment conditions of the ultrasonic pretreatment are: ultrasonic frequency 20 kHz, ultrasonic power 200 W, ultrasonic time 5-20 min, ultrasonic temperature 45° C.-55° C., ultrasonic working time 10-20 s, and rest time 5-10 s.
[0009] Preferably, the centrifugal separation conditions in step (3) are: centrifugal temperature 4-5°C, centrifugal speed 3500-4500g, and centrifugal time 10-15min.
[0010] Preferably, the preparation method of the chelating ligand is: Take 100 kg of fresh anticoagulated chicken blood, centrifuge at 3000g for 15 min, add 5 times the volume of water to the precipitate and stir for 40 min, centrifuge at 3000g for 10 min to obtain a hemoglobin solution, add 4‰ alkaline protease, 3‰ neutral protease and 3‰ composite flavor protease for enzymatic hydrolysis, the enzymatic hydrolysis time is 6 h, after the enzymatic hydrolysis, raise the temperature to 95℃ and maintain for 15 min to inactivate the enzyme to obtain an enzymatic solution, centrifuge the enzymatic solution in a 4000g tubular centrifuge for 10 min, filter the supernatant with a 20nm nanofiltration membrane for purification, and then ultrasonically treat for 15 min. The ultrasonic working conditions are working time of 10-20s, rest time of 5-10s, ultrasonic temperature of 55℃, frequency of 20kHz, and power of 200W.
[0011] Preferably, the rare titanium alcohol contains ≥60% polypeptide, ≥0.05% rare earth and ≥0.1% sterol.
[0012] The invention also provides a production-increasing and growth-promoting agent prepared by a method for preparing a dilute titanium alcohol production-increasing and growth-promoting agent.
[0013] Compared with the prior art, the present invention has the following beneficial effects: The present invention can be effectively used to promote the increase of crop yield, enhance crop photosynthesis, effectively resist and overcome external adverse factors on crop growth, and further promote the acceleration of crop nutritional growth and increase crop yield. DETAILED DESCRIPTION
[0014] The following is a clear and complete description of the technical solutions in the embodiments of the present invention in conjunction with specific embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0015] The preparation method of a dilute titanium alcohol production-increasing and growth-promoting agent of this embodiment comprises the following steps: (1) Hemoglobin extraction: add anticoagulant to fresh animal blood, collect the precipitate by centrifugation, and collect the supernatant hemoglobin after homogenization to obtain a hemoglobin solution with a mass concentration of 10-20%; (2) Preparation of crude peptides: Add enzymes to the hemoglobin solution and heat to 45-55°C for enzymatic hydrolysis for 4-6 hours. After the enzymatic hydrolysis is completed, the enzyme is inactivated. (3) Isolation and purification of crude peptides: The enzymatic hydrolysate in step (2) was centrifuged at 4°C to collect the supernatant, and filtered through a 20 nm nanofiltration membrane to obtain a peptide solution; (4) Preparation of chelate: The polypeptide liquid is ultrasonically pretreated before chelation, and a chelating ligand is added to the polypeptide liquid in step (3), wherein the chelating ligand accounts for 5-15% of the mass ratio of the polypeptide liquid. After the chelation reaction is completed, the mixture is concentrated and spray-dried to obtain an animal blood chelated peptide; (5) The pH of the animal blood chelating peptide is adjusted to 4.5, and then 8% EDTA rare earth of the animal blood chelating peptide is added, and the mixture is stirred at 40 rpm for 40 minutes at 37°C to obtain a chelating solution. After all the solution is dissolved, 5% stigmasterol of the animal blood chelating peptide is added and chelated and stirred for 50 minutes to obtain a dilute titanium alcohol solution. The dilute titanium alcohol solution is concentrated and spray-dried to obtain a dilute titanium alcohol yield-increasing and growth-promoting agent.
[0016] The anticoagulant of this embodiment is 5-10% of the total amount of fresh animal blood, and the anticoagulant is 10-15 parts of trisodium citrate, 2-5 parts of sodium hexametaphosphate, 1-3 parts of sodium chloride, 1-2 parts of citric acid and 10-15 parts of water.
[0017] The enzyme of this embodiment is composed of Bacillus licheniformis alkaline protease, Bacillus subtilis neutral protease and Aspergillus oryzae composite flavor protease mixed in a mass ratio of 3-5:4-3:4-3, and the added amount of the enzyme is 3-10% of the mass of the hemoglobin solution.
[0018] The enzyme inactivation conditions of this embodiment are: enzyme inactivation temperature 90-95° C., enzyme inactivation time 10-15 min.
[0019] The ultrasonic treatment conditions of the ultrasonic pretreatment in this embodiment are: ultrasonic frequency 20kHz, ultrasonic power 200W, ultrasonic time 5-20 min, ultrasonic temperature 45°C-55°C, ultrasonic working time 10-20 s, and rest time 5-10 s.
[0020] The centrifugal separation conditions in step (3) of this embodiment are: centrifugal temperature 4-5°C, centrifugal speed 3500-4500g, and centrifugal time 10-15min.
[0021] The preparation method of the chelate ligand of this embodiment is: Take 100 kg of fresh anticoagulated chicken blood, centrifuge at 3000g for 15 min, add 5 times the volume of water to the precipitate and stir for 40 min, centrifuge at 3000g for 10 min to obtain a hemoglobin solution, add 4‰ alkaline protease, 3‰ neutral protease and 3‰ composite flavor protease for enzymatic hydrolysis, the enzymatic hydrolysis time is 6 h, after the enzymatic hydrolysis, raise the temperature to 95℃ and maintain for 15 min to inactivate the enzyme to obtain an enzymatic solution, centrifuge the enzymatic solution in a 4000g tubular centrifuge for 10 min, filter the supernatant with a 20nm nanofiltration membrane for purification, and then ultrasonically treat for 15 min. The ultrasonic working conditions are working time of 10-20s, rest time of 5-10s, ultrasonic temperature of 55℃, frequency of 20kHz, and power of 200W.
[0022] In the rare titanium alcohol of this embodiment, the polypeptide is ≥60%, the rare earth is ≥0.05%, and the sterol is ≥0.1%.
[0023] The production-increasing and growth-promoting agent is prepared by a preparation method of a dilute titanium alcohol production-increasing and growth-promoting agent in this embodiment.
[0024] Embodiment 1: The preparation method of a dilute titanium alcohol production-increasing and growth-promoting agent of this embodiment comprises the following steps: (1) Hemoglobin extraction: add anticoagulant to fresh animal blood and collect the precipitate by centrifugation. After homogenization, collect the supernatant hemoglobin to obtain a hemoglobin solution with a mass concentration of 10%; (2) Preparation of crude peptides: Add enzymes to the hemoglobin solution and heat to 45°C for enzymatic hydrolysis for 4-6 hours. After the enzymatic hydrolysis is completed, the enzyme is inactivated. (3) Isolation and purification of crude peptides: The enzymatic hydrolysate in step (2) was centrifuged at 4°C to collect the supernatant, and filtered through a 20 nm nanofiltration membrane to obtain a peptide solution; (4) Preparation of chelate: The polypeptide liquid is ultrasonically pretreated before chelation, and a chelating ligand is added to the polypeptide liquid in step (3), wherein the chelating ligand accounts for 5% of the mass ratio of the polypeptide liquid. After the chelation reaction is completed, the mixture is concentrated and spray-dried to obtain an animal blood chelated peptide; (5) The pH of the animal blood chelating peptide is adjusted to 4.5, and then 8% EDTA rare earth of the animal blood chelating peptide is added, and the mixture is stirred at 40 rpm for 40 minutes at 37°C to obtain a chelating solution. After all the solution is dissolved, 5% stigmasterol of the animal blood chelating peptide is added and chelated and stirred for 50 minutes to obtain a dilute titanium alcohol solution. The dilute titanium alcohol solution is concentrated and spray-dried to obtain a dilute titanium alcohol yield-increasing and growth-promoting agent.
[0025] The anticoagulant in this embodiment is 5% of the total amount of fresh animal blood, and the anticoagulant is composed of 10 parts of trisodium citrate, 2 parts of sodium hexametaphosphate, 1 part of sodium chloride, 1 part of citric acid and 10 parts of water.
[0026] The enzyme of this embodiment is composed of Bacillus licheniformis alkaline protease, Bacillus subtilis neutral protease and Aspergillus oryzae composite flavor protease mixed in a mass ratio of 3:4:4, and the added amount of the enzyme is 3-10% of the mass of the hemoglobin solution.
[0027] The enzyme inactivation conditions of this example are: enzyme inactivation temperature 90° C., enzyme inactivation time 10 min.
[0028] The ultrasonic treatment conditions of the ultrasonic pretreatment in this embodiment are: ultrasonic frequency 20 kHz, ultrasonic power 200 W, ultrasonic time 5 min, ultrasonic temperature 45° C., ultrasonic working time 10 s, and rest time 5-10 s.
[0029] The centrifugal separation conditions in step (3) of this embodiment are: centrifugal temperature 4°C, centrifugal speed 3500g, and centrifugal time 10min.
[0030] The preparation method of the chelate ligand of this embodiment is: Take 100 kg of fresh anticoagulated chicken blood, centrifuge it at 3000g for 15 min, add 5 times the volume of water to the precipitate and stir for 40 min, centrifuge it at 3000g for 10 min to obtain a hemoglobin solution, add 4‰ alkaline protease, 3‰ neutral protease and 3‰ composite flavor protease for enzymatic hydrolysis for 6 h, after the enzymatic hydrolysis, raise the temperature to 95℃ and maintain it for 15 min to inactivate the enzyme to obtain an enzymatic solution, centrifuge the enzymatic solution in a 4000g tubular centrifuge for 10 min, filter the supernatant with a 20nm nanofiltration membrane for purification, and then ultrasonically treat it for 15 min. The ultrasonic working conditions are working time of 10 s, rest time of 5-s, ultrasonic temperature of 55℃, frequency of 20 kHz, and power of 200W.
[0031] In the rare titanium alcohol of this embodiment, the polypeptide is ≥60%, the rare earth is ≥0.05%, and the sterol is ≥0.1%.
[0032] The production-increasing and growth-promoting agent is prepared by a preparation method of a dilute titanium alcohol production-increasing and growth-promoting agent in this embodiment.
[0033] Embodiment 2: The preparation method of a dilute titanium alcohol production-increasing and growth-promoting agent of this embodiment comprises the following steps: (1) Hemoglobin extraction: add anticoagulant to fresh animal blood and collect the precipitate by centrifugation. After homogenization, collect the supernatant hemoglobin to obtain a hemoglobin solution with a mass concentration of 20%; (2) Preparation of crude peptide: Add enzyme to the hemoglobin solution and heat to 55°C for enzymatic hydrolysis for 6 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated. (3) Isolation and purification of crude peptides: The enzymatic hydrolysate in step (2) was centrifuged at 4°C to collect the supernatant, and filtered through a 20 nm nanofiltration membrane to obtain a peptide solution; (4) Preparation of chelate: The polypeptide liquid is ultrasonically pretreated before chelation, and a chelating ligand is added to the polypeptide liquid in step (3), wherein the chelating ligand accounts for 15% of the mass ratio of the polypeptide liquid. After the chelation reaction is completed, the mixture is concentrated and spray-dried to obtain an animal blood chelated peptide; (5) The pH of the animal blood chelating peptide is adjusted to 4.5, and then 8% EDTA rare earth of the animal blood chelating peptide is added, and the mixture is stirred at 40 rpm for 40 minutes at 37°C to obtain a chelating solution. After all the solution is dissolved, 5% stigmasterol of the animal blood chelating peptide is added and chelated and stirred for 50 minutes to obtain a dilute titanium alcohol solution. The dilute titanium alcohol solution is concentrated and spray-dried to obtain a dilute titanium alcohol yield-increasing and growth-promoting agent.
[0034] The anticoagulant in this embodiment is 10% of the total amount of fresh animal blood, and the anticoagulant is 15 parts of trisodium citrate, 5 parts of sodium hexametaphosphate, 3 parts of sodium chloride, 2 parts of citric acid and 15 parts of water.
[0035] The enzyme of this example is composed of a mixture of Bacillus licheniformis alkaline protease, Bacillus subtilis neutral protease and Aspergillus oryzae composite flavor protease in a mass ratio of 5:3:3, and the added amount of the enzyme is 10% of the mass of the hemoglobin solution.
[0036] The enzyme inactivation conditions of this example are: enzyme inactivation temperature 95° C., enzyme inactivation time 15 min.
[0037] The ultrasonic treatment conditions of the ultrasonic pretreatment in this embodiment are: ultrasonic frequency 20 kHz, ultrasonic power 200 W, ultrasonic time 20 min, ultrasonic temperature 55° C., ultrasonic working time 20 s, and rest time 10 s.
[0038] The centrifugal separation conditions in step (3) of this embodiment are: centrifugal temperature 5°C, centrifugal speed 4500g, and centrifugal time 15min.
[0039] The preparation method of the chelate ligand of this embodiment is: Take 100 kg of fresh anticoagulated chicken blood, centrifuge at 3000g for 15 min, add 5 times the volume of water to the precipitate and stir for 40 min, centrifuge at 3000g for 10 min to obtain a hemoglobin solution, add 4‰ alkaline protease, 3‰ neutral protease and 3‰ composite flavor protease for enzymatic hydrolysis, the enzymatic hydrolysis time is 6 h, after the enzymatic hydrolysis, raise the temperature to 95℃ and maintain for 15 min to inactivate the enzyme to obtain an enzymatic solution, centrifuge the enzymatic solution in a 4000g tubular centrifuge for 10 min, filter the supernatant with a 20nm nanofiltration membrane for purification, and then ultrasonically treat for 15 min. The ultrasonic working conditions are working time of 20 s, rest time of 10 s, ultrasonic temperature of 55℃, frequency of 20 kHz, and power of 200 W.
[0040] In the rare titanium alcohol of this embodiment, the polypeptide is ≥60%, the rare earth is ≥0.05%, and the sterol is ≥0.1%.
[0041] The production-increasing and growth-promoting agent is prepared by a preparation method of a dilute titanium alcohol production-increasing and growth-promoting agent in this embodiment.
[0042] Embodiment 3: The preparation method of a dilute titanium alcohol production-increasing and growth-promoting agent of this embodiment comprises the following steps: (1) Hemoglobin extraction: add anticoagulant to fresh animal blood and collect the precipitate by centrifugation. After homogenization, collect the supernatant hemoglobin to obtain a hemoglobin solution with a mass concentration of 15%; (2) Preparation of crude peptides: Add enzymes to the hemoglobin solution and heat to 50°C for enzymatic hydrolysis for 5 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated. (3) Isolation and purification of crude peptides: The enzymatic hydrolysate in step (2) was centrifuged at 4°C to collect the supernatant, and filtered through a 20 nm nanofiltration membrane to obtain a peptide solution; (4) Preparation of chelate: The polypeptide liquid is ultrasonically pretreated before chelation, and a chelating ligand is added to the polypeptide liquid in step (3), wherein the chelating ligand accounts for 5-15% of the mass ratio of the polypeptide liquid. After the chelation reaction is completed, the mixture is concentrated and spray-dried to obtain an animal blood chelated peptide; (5) The pH of the animal blood chelating peptide is adjusted to 4.5, and then 8% EDTA rare earth of the animal blood chelating peptide is added, and the mixture is stirred at 40 rpm for 40 minutes at 37°C to obtain a chelating solution. After all the solution is dissolved, 5% stigmasterol of the animal blood chelating peptide is added and chelated and stirred for 50 minutes to obtain a dilute titanium alcohol solution. The dilute titanium alcohol solution is concentrated and spray-dried to obtain a dilute titanium alcohol yield-increasing and growth-promoting agent.
[0043] The anticoagulant in this embodiment is 7.5% of the total amount of fresh animal blood, and the anticoagulant is 12.5 parts of trisodium citrate, 3.5 parts of sodium hexametaphosphate, 2 parts of sodium chloride, 1-2 parts of citric acid and 12.5 parts of water.
[0044] The enzyme of this example is composed of a mixture of Bacillus licheniformis alkaline protease, Bacillus subtilis neutral protease and Aspergillus oryzae composite flavor protease in a mass ratio of 4:3:3, and the added amount of the enzyme is 6.5% of the mass of the hemoglobin solution.
[0045] The enzyme inactivation conditions of this example are: enzyme inactivation temperature 92.5° C., enzyme inactivation time 12.5 min.
[0046] The ultrasonic treatment conditions of the ultrasonic pretreatment in this embodiment are: ultrasonic frequency 20 kHz, ultrasonic power 200 W, ultrasonic time 10 min, ultrasonic temperature 50° C., ultrasonic working time 15 s, and rest time 7.5 s.
[0047] The centrifugal separation conditions in step (3) of this embodiment are: centrifugal temperature 5°C, centrifugal speed 3800g, and centrifugal time 12.5min.
[0048] The preparation method of the chelate ligand of this embodiment is: Take 100 kg of fresh anticoagulated chicken blood, centrifuge at 3000g for 15 min, add 5 times the volume of water to the precipitate and stir for 40 min, centrifuge at 3000g for 10 min to obtain a hemoglobin solution, add 4‰ alkaline protease, 3‰ neutral protease and 3‰ composite flavor protease for enzymatic hydrolysis, the enzymatic hydrolysis time is 6 h, after the enzymatic hydrolysis, raise the temperature to 95℃ and maintain for 15 min to inactivate the enzyme to obtain an enzymatic solution, centrifuge the enzymatic solution in a 4000g tubular centrifuge for 10 min, filter the supernatant with a 20nm nanofiltration membrane for purification, and then ultrasonically treat for 15 min. The ultrasonic working conditions are working time of 15 s, rest time of 7.5 s, ultrasonic temperature of 55℃, frequency of 20 kHz, and power of 200 W.
[0049] In the rare titanium alcohol of this embodiment, the polypeptide is ≥60%, the rare earth is ≥0.05%, and the sterol is ≥0.1%.
[0050] The production-increasing and growth-promoting agent is prepared by a preparation method of a dilute titanium alcohol production-increasing and growth-promoting agent in this embodiment.
[0051] Embodiment 4: The preparation method of a dilute titanium alcohol production-increasing and growth-promoting agent of this embodiment comprises the following steps: (1) Hemoglobin extraction: add anticoagulant to fresh animal blood, collect the precipitate by centrifugation, and collect the supernatant hemoglobin after homogenization to obtain a hemoglobin solution with a mass concentration of 12%; (2) Preparation of crude peptides: Add enzymes to the hemoglobin solution and heat to 47°C for enzymatic hydrolysis for 5 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated. (3) Isolation and purification of crude peptides: The enzymatic hydrolysate in step (2) was centrifuged at 4°C to collect the supernatant, and filtered through a 20 nm nanofiltration membrane to obtain a peptide solution; (4) Preparation of chelate: The polypeptide liquid is ultrasonically pretreated before chelation, and a chelating ligand is added to the polypeptide liquid in step (3), wherein the chelating ligand accounts for 7% of the mass ratio of the polypeptide liquid. After the chelation reaction is completed, the mixture is concentrated and spray-dried to obtain an animal blood chelated peptide; (5) The pH of the animal blood chelating peptide is adjusted to 4.5, and then 8% EDTA rare earth of the animal blood chelating peptide is added, and the mixture is stirred at 40 rpm for 40 minutes at 37°C to obtain a chelating solution. After all the solution is dissolved, 5% stigmasterol of the animal blood chelating peptide is added and chelated and stirred for 50 minutes to obtain a dilute titanium alcohol solution. The dilute titanium alcohol solution is concentrated and spray-dried to obtain a dilute titanium alcohol yield-increasing and growth-promoting agent.
[0052] The anticoagulant in this embodiment is 6% of the total amount of fresh animal blood, and the anticoagulant is 12 parts of trisodium citrate, 3 parts of sodium hexametaphosphate, 2 parts of sodium chloride, 1 part of citric acid and 12 parts of water.
[0053] The enzyme of this example is composed of a mixture of Bacillus licheniformis alkaline protease, Bacillus subtilis neutral protease and Aspergillus oryzae composite flavor protease in a mass ratio of 3:44, and the added amount of the enzyme is 5% of the mass of the hemoglobin solution.
[0054] The enzyme inactivation conditions of this example are: enzyme inactivation temperature 92° C., enzyme inactivation time 12 min.
[0055] The ultrasonic treatment conditions of the ultrasonic pretreatment in this embodiment are: ultrasonic frequency 20 kHz, ultrasonic power 200 W, ultrasonic time 8 min, ultrasonic temperature 48° C., ultrasonic working time 12 s, and rest time 7 s.
[0056] The centrifugal separation conditions in step (3) of this embodiment are: centrifugal temperature 5°C, centrifugal speed 3700g, and centrifugal time 12min.
[0057] The preparation method of the chelate ligand of this embodiment is: Take 100 kg of fresh anticoagulated chicken blood, centrifuge it at 3000g for 15 min, add 5 times the volume of water to the precipitate, stir for 40 min, and centrifuge it at 3000g for 10 min to obtain a hemoglobin solution, add 4‰ alkaline protease, 3‰ neutral protease and 3‰ composite flavor protease for enzymatic hydrolysis for 6 h. After the enzymatic hydrolysis, raise the temperature to 95℃ and maintain it for 15 min to inactivate the enzyme to obtain an enzymatic solution. Centrifuge the enzymatic solution at 4000g for 10 min in a tubular centrifuge, filter and purify the supernatant with a 20nm nanofiltration membrane, and then ultrasonically treat it for 15 min. The ultrasonic working conditions are working time of 13 s, rest time of 6 s, ultrasonic temperature of 55℃, frequency of 20 kHz, and power of 200 W.
[0058] In the rare titanium alcohol of this embodiment, the polypeptide is ≥60%, the rare earth is ≥0.05%, and the sterol is ≥0.1%.
[0059] The production-increasing and growth-promoting agent is prepared by a preparation method of a dilute titanium alcohol production-increasing and growth-promoting agent in this embodiment.
[0060] Purpose of the test Weihai Soil and Fertilizer Station conducted experiments on apples using the diluted titanium alcohol, dosage form (powder), and technical indicators (peptide ≥ 60%, rare earth ≥ 0.05%, and sterol ≥ 0.1%) of Example 3 to verify its yield-increasing effect, providing a scientific basis for the promotion of the fertilizer.
[0061] Time and Place Experimental period: June to October 2022 Test location: Mengjiazhuang, Qiaotou Town, Weihai City Materials and Methods Test soil: Soil nutrient conditions are shown in Table 1.
[0062] Table 1 Soil nutrient status years Organic matter (g / kg) Alkaline nitrogen (mg / kg) Available phosphorus (mg / kg) Available potassium (mg / kg) pH 2022 11.2 94.1 46.1 91.6 5.6 Test materials: dilute titanium alcohol, dosage form (powder), technical indicators (peptide ≥ 60%, rare earth ≥ 0.05%, sterol ≥ 0.1%).
[0063] Test varieties and cultivation methods: The test product is Yanfu No. 3, with a tree age of 12 years. The planting density is about 45 plants / 667㎡. On the third and sixth days after spraying, 10 samples were randomly selected from each treatment and the fruit circumference was accurately measured with a vernier caliper. According to the investigation, the average increase of a single fruit in the test area sprayed with dilute titanium alcohol was 3.151mm after 3 days, and the average increase of a single fruit after 6 days reached 5.714mm; the average increase of a single fruit in the control test area was 2.834mm after 3 days, and the average increase of a single fruit after 6 days was 4.46mm.
[0064] Test methods Experimental design: The experiment has two treatments and three replications, with a total of 6 experimental plots, 10 trees in each plot, and each plot is randomly arranged with protection rows.
[0065] Treatment 1: conventional fertilization + spraying of diluted titanium alcohol Treatment 2: Conventional fertilization + spraying with equal amount of water Fertilization method: Conventional fertilization was to apply 2.5 kg of compound fertilizer and 50 kg of high-quality decomposed farmyard manure per plant on March 24; 80 kg of high-quality farmyard manure was applied per plant on June 24; Treatment 1 was based on conventional fertilization, and on June 10, 250 g of diluted titanium alcohol was sprayed per mu, diluted 1000 times, for a total of 1 spraying; Treatment 2 was sprayed with an equal amount of clean water at the same time as conventional fertilizers.
[0066] Results and Analysis Effects of different treatments on the biological shape of apples.
[0067] The test shows that the leaves of fruit trees in the test area sprayed with dilute titanium alcohol are dark green, the branches are strong, the fruits swell quickly, the color is good, and the ability to resist diseases and pests is strong.
[0068] The effects of different treatments on apple quality are shown in Table 2.
[0069] Table 2 Comparison of different treatment quality conditions Output and input-output ratio are shown in Table 3.
[0070] Table 3 Comparison of output and input-output ratio deal with Yield kg / mu Output value yuan / mu Yield increase kg / mu compared with the control Yield increase rate compared with control (%) Compared with the control, the value-added is RMB / mu Investment per mu (yuan) Net added value per mu (yuan) Input-output ratio Process 1 3561 21712 261 7.9 1846 70 1776 1:25.4 Process 2 3300 19866 in conclusion Dilute titanium alcohol has a significant yield-increasing effect on apples. The plots sprayed with the test fertilizer increased their yield by 261 kg / mu compared with the control, with a yield increase rate of 7.9%, a first-grade fruit rate increased by 15%, and an input-output ratio of 1:25.4.
[0071] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be considered exemplary and non-restrictive in all respects, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention.
[0072] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.
Claims
1. A method for preparing a dilute titanium alcohol production-increasing and growth-promoting agent, characterized in that: The following steps are involved: (1) Hemoglobin extraction: add anticoagulant to fresh animal blood, collect the precipitate by centrifugation, and collect the supernatant hemoglobin after homogenization to obtain a hemoglobin solution with a mass concentration of 10-20%; (2) Preparation of crude peptides: Add enzymes to the hemoglobin solution and heat to 45-55°C for enzymatic hydrolysis for 4-6 hours. After the enzymatic hydrolysis is completed, the enzyme is inactivated. (3) Isolation and purification of crude peptides: The enzymatic hydrolysate in step (2) was centrifuged at 4°C to collect the supernatant, and filtered through a 20 nm nanofiltration membrane to obtain a peptide solution; (4) Preparation of chelate: The polypeptide liquid is ultrasonically pretreated before chelation, and a chelating ligand is added to the polypeptide liquid in step (3), wherein the chelating ligand accounts for 5-15% of the mass ratio of the polypeptide liquid. After the chelation reaction is completed, the mixture is concentrated and spray-dried to obtain an animal blood chelated peptide; (5) The pH of the animal blood chelating peptide is adjusted to 4.5, and then 8% EDTA rare earth of the animal blood chelating peptide is added, and the mixture is stirred at 40 rpm for 40 minutes at 37°C to obtain a chelating solution. After all the solution is dissolved, 5% stigmasterol of the animal blood chelating peptide is added and chelated and stirred for 50 minutes to obtain a dilute titanium alcohol solution. The dilute titanium alcohol solution is concentrated and spray-dried to obtain a dilute titanium alcohol yield-increasing and growth-promoting agent.
2. The method for preparing a dilute titanium alcohol production-increasing and growth-promoting agent according to claim 1, characterized in that: The anticoagulant is 5-10% of the total amount of fresh animal blood, and the anticoagulant is composed of 10-15 parts of trisodium citrate, 2-5 parts of sodium hexametaphosphate, 1-3 parts of sodium chloride, 1-2 parts of citric acid and 10-15 parts of water.
3. The method for preparing a dilute titanium alcohol production-increasing and growth-promoting agent according to claim 1, characterized in that: The enzyme is composed of alkaline protease of Bacillus licheniformis, neutral protease of Bacillus subtilis and composite flavor protease of Aspergillus oryzae in a mass ratio of 3-5:4-3:4-3, and the added amount of the enzyme is 3-10% of the mass of the hemoglobin solution.
4. The method for preparing a dilute titanium alcohol production-increasing and growth-promoting agent according to claim 1, characterized in that: The enzyme inactivation conditions are: enzyme inactivation temperature 90-95° C., enzyme inactivation time 10-15 min.
5. The method for preparing a dilute titanium alcohol production-increasing and growth-promoting agent according to claim 1, characterized in that: The ultrasonic treatment conditions of the ultrasonic pretreatment are: ultrasonic frequency 20kHz, ultrasonic power 200W, ultrasonic time 5-20 min, ultrasonic temperature 45°C-55°C, ultrasonic working time 10-20 s, and rest time 5-10 s.
6. The method for preparing a dilute titanium alcohol production-increasing and growth-promoting agent according to claim 1, characterized in that: The centrifugal separation conditions in step (3) are: centrifugal temperature 4-5°C, centrifugal speed 3500-4500g, and centrifugal time 10-15min.
7. The method for preparing a dilute titanium alcohol production-increasing and growth-promoting agent according to claim 1, characterized in that: The preparation method of the chelate ligand is as follows: Take 100 kg of fresh anticoagulated chicken blood, centrifuge at 3000g for 15 min, add 5 times the volume of water to the precipitate and stir for 40 min, centrifuge at 3000g for 10 min to obtain a hemoglobin solution, add 4‰ alkaline protease, 3‰ neutral protease and 3‰ composite flavor protease for enzymatic hydrolysis, the enzymatic hydrolysis time is 6 h, after the enzymatic hydrolysis, raise the temperature to 95℃ and maintain for 15 min to inactivate the enzyme to obtain an enzymatic solution, centrifuge the enzymatic solution in a 4000g tubular centrifuge for 10 min, filter the supernatant with a 20nm nanofiltration membrane for purification, and then ultrasonically treat for 15 min. The ultrasonic working conditions are working time of 10-20s, rest time of 5-10s, ultrasonic temperature of 55℃, frequency of 20kHz, and power of 200W.
8. The method for preparing a dilute titanium alcohol production-increasing and growth-promoting agent according to claim 1, characterized in that: The rare titanium alcohol contains ≥60% polypeptide, ≥0.05% rare earth and ≥0.1% sterol.
9. A production-increasing and growth-promoting agent prepared by the method for preparing a dilute titanium alcohol production-increasing and growth-promoting agent as described in any one of claims 1 to 8.