CN110541012B - Preparation method of amino acid liquid, amino acid water-soluble fertilizer and application thereof - Google Patents
Preparation method of amino acid liquid, amino acid water-soluble fertilizer and application thereof Download PDFInfo
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- CN110541012B CN110541012B CN201910789669.6A CN201910789669A CN110541012B CN 110541012 B CN110541012 B CN 110541012B CN 201910789669 A CN201910789669 A CN 201910789669A CN 110541012 B CN110541012 B CN 110541012B
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05C—NITROGENOUS FERTILISERS
- C05C11/00—Other nitrogenous fertilisers
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
Abstract
The invention provides a preparation method of amino acid liquid, the amino acid liquid, an amino acid water-soluble fertilizer and application thereof, and relates to the technical field of agricultural fertilizer manufacturing, wherein the preparation method comprises the following steps: (1) extracting the defatted soybean meal with water to obtain a soybean protein extracting solution; the mass ratio of the defatted soybean meal to water is 1 (25-35); (2) mixing the soybean protein extracting solution obtained in the step (1) with bacillus subtilis fermentation liquor for fermentation to obtain amino acid liquor; the using amount of the bacillus subtilis fermentation liquor accounts for 5-10% of the total volume of the soybean protein extracting solution; the number of effective viable bacteria in the bacillus subtilis fermentation liquor is more than or equal to 2 hundred million/mL. The preparation method of the invention can ensure that the conversion rate of the amino acid reaches more than 45 percent and the content of the free amino acid reaches more than 100 g/L.
Description
Technical Field
The invention relates to the technical field of agricultural fertilizer manufacturing, in particular to a preparation method of amino acid liquid, the amino acid liquid, an amino acid water-soluble fertilizer and application thereof.
Background
Due to continuous cropping planting of people in successive years, unreasonable use of chemical fertilizers for a long time and multiple flushing application of macroelement water-soluble fertilizers, severe soil hardening, soil acidification and soil salinization are caused to occur, increasingly severe soil fertility reduction and occurrence of soil-borne diseases are caused, and finally crop quality is reduced.
The resources such as bean pulp and the like are rich in amino acid, have wide sources and low cost, but do not achieve high-value utilization to a certain extent.
Disclosure of Invention
In view of the above, the invention aims to provide an amino acid solution, a preparation method of the amino acid solution, an amino acid water-soluble fertilizer and an application of the amino acid water-soluble fertilizer. The amino acid conversion rate in the amino acid solution prepared by the preparation method provided by the invention reaches more than 45%, and the content of free amino acid reaches more than 100 g/L.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of amino acid liquid, which comprises the following steps:
(1) extracting the defatted soybean meal with water to obtain a soybean protein extracting solution; the mass ratio of the defatted soybean meal to water is 1 (25-35);
(2) mixing the soybean protein extracting solution obtained in the step (1) with bacillus subtilis fermentation liquor for fermentation to obtain amino acid liquor; the using amount of the bacillus subtilis fermentation liquor accounts for 5-10% of the total volume of the soybean protein extraction liquid; the number of effective viable bacteria in the bacillus subtilis fermentation liquor is more than or equal to 2 hundred million/mL.
Preferably, the pH value extracted in the step (1) is 7-12.
Preferably, the extraction temperature in the step (1) is 40-80 ℃, and the extraction time is 50-100 min.
Preferably, the extraction in step (1) is microwave extraction.
Preferably, the power of microwave extraction is 300-1000W.
Preferably, the fermentation temperature in the step (2) is 25-30 ℃, and the fermentation time is 24-72 h.
The invention also provides the amino acid solution prepared by the preparation method.
The invention also provides an amino acid water-soluble fertilizer which is prepared from the amino acid liquid and trace elements.
Preferably, the trace elements comprise one or more of ferrous sulfate heptahydrate, zinc sulfate heptahydrate, boric acid, ethylenediamine tetraacetic acid iron sodium and ethylenediamine tetraacetic acid zinc sodium.
The invention also provides application of the amino acid water-soluble fertilizer in the technical scheme in improving crop yield.
The invention provides a preparation method of amino acid liquid, which takes defatted soybean meal as a raw material and utilizes bacillus subtilis rich in protease to carry out fermentation treatment, so that the conversion rate of amino acid reaches more than 45 percent, and the content of free amino acid reaches more than 100 g/L.
The amino acid water-soluble fertilizer provided by the invention can not only provide a nitrogen source for crops, but also promote photosynthesis of crops, regulate crop growth, relieve fertilizer damage and phytotoxicity and relieve toxic effects of heavy metal ions, so that the growth condition of the crops can be effectively improved, and the effect of improving the crop yield is achieved.
Detailed Description
The invention provides a preparation method of amino acid liquid, which comprises the following steps:
(1) extracting the defatted soybean meal with water to obtain a soybean protein extracting solution; the mass ratio of the defatted soybean meal to water is 1 (25-35);
(2) mixing the soybean protein extracting solution obtained in the step (1) with bacillus subtilis fermentation liquor for fermentation to obtain amino acid liquor; the using amount of the bacillus subtilis fermentation liquor accounts for 5-10% of the total volume of the soybean protein extraction liquid; the number of effective viable bacteria in the bacillus subtilis fermentation liquor is more than or equal to 2 hundred million/mL.
The method comprises the steps of extracting defatted soybean meal with water to obtain a soybean protein extracting solution; the mass ratio of the defatted soybean meal to water is 1 (25-35), and preferably 1: 30. The source of the defatted soybean meal is not particularly limited, and the defatted soybean meal can be obtained by adopting a conventional commercial product.
In the invention, the pH value of the extraction is preferably 7-12, and more preferably 8-9. In the invention, the extraction temperature is preferably 40-80 ℃, more preferably 50-70 ℃, and most preferably 60 ℃; the extraction time is preferably 50-100 min, more preferably 60-80 min, and most preferably 70 min.
In the invention, the extraction method is preferably microwave extraction, and the power of the microwave extraction is preferably 300-1000W, more preferably 600-900W, and most preferably 800W.
In the invention, the soybean protein extracting solution is preferably supernatant obtained by centrifugal impurity removal, and the centrifugal rotating speed is preferably 4000 r/min-6000 r/min, and more preferably 5000 r/min; the centrifugation time is preferably 10-30 min, more preferably 15-25 min, and most preferably 20 min.
Mixing the obtained soybean protein extracting solution with bacillus subtilis fermentation liquor for fermentation to obtain amino acid liquor; the using amount of the bacillus subtilis fermentation liquor accounts for 5-10%, preferably 6-8% of the total volume of the soybean protein extracting solution; the number of effective viable bacteria in the bacillus subtilis fermentation liquor is more than or equal to 2 hundred million/mL. The method for mixed fermentation is not particularly limited, and a conventional mixed fermentation method can be adopted.
In the invention, the Bacillus subtilis is preferably a Sanju-07 strain which is preserved in China general microbiological culture Collection center in 29 days 12 and 29 days 2014, the preservation address is No. 3 of No.1 Xilu Beichen of Chaoyang district in Beijing, and the preservation number is CGMCC No. 10248.
In the invention, the fermentation medium of the bacillus subtilis uses water as a solvent, and each liter of the fermentation medium comprises: 25-35 g of starch, 2-7 g of cane sugar, 1-5 g of monopotassium phosphate, 0.1-0.5 g of manganese sulfate and 0.2-2 g of yeast extract; the pH value of the fermentation medium is preferably 7.0-7.2. In the present invention, the fermentation medium of bacillus subtilis uses water as a solvent, and preferably comprises per liter: 30g of starch, 5g of cane sugar, 3g of monopotassium phosphate, 0.3g of manganese sulfate and 1g of yeast extract.
In the invention, the fermentation temperature is preferably 25-30 ℃, and more preferably 28 ℃; the fermentation time is preferably 24-72 h, more preferably 36-60 h, and most preferably 48 h.
The invention also provides an amino acid water-soluble fertilizer which is prepared from the amino acid liquid and trace elements. In the invention, the trace elements preferably comprise one or more of ferrous sulfate heptahydrate, zinc sulfate heptahydrate, boric acid, sodium iron ethylenediaminetetraacetate and sodium zinc ethylenediaminetetraacetate. In the invention, 20-40 g, more preferably 25-35 g, 70-100 g, more preferably 80-90 g of zinc sulfate heptahydrate, and 10-20 g, more preferably 12-18 g of boric acid are preferably added into 1L of the amino acid solution. The preparation method of the amino acid water-soluble fertilizer is not particularly limited, and the amino acid water-soluble fertilizer is prepared by stirring for 1-4 hours at the temperature of 15-40 ℃ and at the speed of 100-800 r/min; more preferably, the material is prepared by stirring for 2-3 hours at the temperature of 20-30 ℃ and the speed of 400-600 r/min.
The application method of the amino acid water-soluble fertilizer is not particularly limited, and the amino acid water-soluble fertilizer can not be preferably mixed with alkaline pesticide, alkaline fertilizer and phosphate fertilizer for use.
In the invention, the application method of the amino acid water-soluble fertilizer comprises the following steps: and (3) seedling stage: spraying the liquid with the optimal concentration of 0.3-1.0L/mu and 500-1000 times; drip irrigation preferably by 0.5-1.0L/mu and 500-1000 times of liquid; growth period and fruiting period: spraying the solution with the optimal concentration of 0.3-2.0L/mu and 1000-2000 times; the drip irrigation is preferably 0.5-2.0L/mu, 500-1000 times of liquid.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without any inventive step, are within the scope of the present invention.
Example 1
Drying and crushing 1000g of defatted soybean meal, and mixing the materials according to a material-liquid ratio of 1:30, adding distilled water, adding an alkali liquor to adjust the pH value to 9, extracting the soybean protein by using microwaves of 750W at the temperature of 60 ℃ for 60min, centrifuging to remove impurities, collecting 25L of supernatant, and adding 5% of bacillus subtilis fermentation liquor at the temperature of 30 ℃ to perform fermentation treatment for 36h to obtain 450mL of amino acid liquor;
adding 30g of ferrous sulfate heptahydrate, 80g of zinc sulfate heptahydrate and 12g of boric acid into 1L of amino acid solution, and stirring at 25 ℃ and a stirring speed of 500r/min for 1h to obtain the amino acid-containing water-soluble fertilizer.
Example 2
Drying and crushing 1000g of defatted soybean meal, and mixing the raw materials according to a material-liquid ratio of 1:30, adding distilled water, adding an alkali liquor to adjust the pH value to 9, extracting the soybean protein by using microwaves of 750W at 60 ℃ for 60min, centrifuging to remove impurities, collecting 25L of supernatant, and adding 5% bacillus subtilis fermentation liquor at 30 ℃ for fermentation treatment for 48h to obtain 580mL of amino acid liquor;
adding 45g of EDTA-Fe, 65g of zinc sulfate heptahydrate and 30g of boric acid into 1L of amino acid solution, and stirring at 25 ℃ and a stirring speed of 500r/min for 1h to obtain the amino acid-containing water-soluble fertilizer.
Example 3
Drying and crushing 1000g of defatted soybean meal, and mixing the raw materials according to a material-liquid ratio of 1:30 adding distilled water, adding alkali liquor to adjust the pH value to 9, extracting soybean protein by microwave of 750W at 60 ℃ for 60min, centrifuging to remove impurities, collecting supernatant of 25L, and adding 5% bacillus subtilis at 30 ℃ for fermentation treatment for 60h to obtain 560g of amino acid liquor;
adding 55g of EDTA-Fe, 55g of EDTA-Zn and 55g of boric acid into 1L of amino acid solution, and stirring at 25 ℃ and a stirring speed of 500r/min for 1h to obtain the amino acid-containing water-soluble fertilizer.
Comparative example 1
A mixed amino acid liquid is prepared by using waste beer yeast, and amino acid and a plurality of trace elements are organically combined to form a novel amino acid fertilizer, which comprises the following specific steps:
mechanically crushing yeast: waste beer yeast paste: mixing water with the mass ratio of 2:1, and then performing high-pressure homogenization, wherein the temperature is controlled to be 20-30 ℃, and the homogenization time is 5-10 min;
heating enzymolysis: adding 0.8% of papain into the yeast crushing liquid obtained in the last step, maintaining the temperature of the yeast crushing liquid at 40-55 ℃, and continuously stirring for 6-8 hours;
③ inactivating enzyme: and (3) rapidly heating the enzymolysis liquid obtained in the last step to 95 ℃, continuously stirring, maintaining for 2 hours, and inactivating the papain.
Fourthly, centrifugal separation: continuously separating and separating the amino acid extracting solution after the enzyme deactivation in the last step to remove impurities such as cell debris and the like in the extracting solution, wherein the rotating speed of a centrifugal machine is 6000 rpm;
pressure reduction and concentration: concentrating the supernatant obtained in the last step under reduced pressure, maintaining the temperature at 95 deg.C and vacuum degree at 0.1 or less until the amino acid content in the concentrated solution is more than 100g/L
Cooling: and (4) cooling the amino acid concentrated solution obtained in the last step to room temperature to obtain the composite amino acid solution, and storing for later use.
Seventhly, 40-60g of zinc sulfate heptahydrate, 40-50g of boric acid, 10-15g of ammonium molybdate tetrahydrate and 20-30g of copper sulfate pentahydrate are added into 1L of the compound amino acid solution.
The amino acid solutions prepared in examples 1 to 3 and comparative examples were subjected to performance tests, and specific results are shown in table 1.
Table 1 results of performance test of amino acid solutions prepared in examples 1 to 3 and comparative example
| Example 1 | Example 2 | Example 3 | Comparative example | Detection standard | |
| Conversion ratio of amino acid (%) | 45.6 | 58.2 | 56.7 | 35.2 | Formaldehyde titration method |
| Content of free amino acids (g/L) | 102.4 | 112.1 | 108.3 | 45.5 | NY/T1975 |
As can be seen from Table 1, the amino acid conversion rate of the amino acid solution provided by the invention is higher than the comparative ratio by more than 10%, and the content of free amino acid is higher than 50g/L, which indicates that the amino acid-containing water-soluble fertilizer provided by the invention has higher content of free amino acid, can provide nutrition for crops, promotes photosynthesis of the crops, and thus improves the yield of the crops.
Example 4
The amino acid-containing water-soluble fertilizer prepared in example 1 was applied to pakchoi, and the control example was set as follows:
treatment 1: spraying the amino acid-containing water-soluble fertilizer of example 1 + conventional fertilization;
and (3) treatment 2: spraying equivalent clear water and applying fertilizer conventionally;
the test is carried out by arranging random block groups, setting 2 treatments and repeating for 3 times, wherein the area of each cell is 30m2。
The method for spraying the amino acid-containing water-soluble fertilizer on the pakchoi comprises the following steps: spraying Chinese cabbage at seedling stage (3 months and 27 days), using 600 times of diluent, and spraying at a speed of 666.7m2Uniformly spraying 500mL of product mixed with 300kg of water, and converting into uniformly spraying 22.5mL of product mixed with 13.5kg of water in each cell (30 square meters); spraying at interval of 15 days (4 months and 11 days) for 1 time, and spraying for 2 times, each time of 666.7m2The total amount is 1.0L.
Sowing pakchoi in 2019, 3 months and 17 days; treatment 1, 2 conventional fertilization: applying 40% to each cell at 27 days of 3 monthsFertilizer special for vegetable (N: P)2O5∶K2O is 20:7:13) and 2.25kg (amounting to 666.7m each)250kg is applied); applying fertilizer for the second time in 11 months and 5 days in 2018, and applying 40% of special fertilizer for vegetables (N: P) to each cell2O5∶K2O is 20:7:13)2.7kg (amounting to 666.7m each260kg was applied). Other management methods are consistent during growth. The Chinese cabbage is collected in 2019, 4 months and 25 days, and the yield is measured actually.
1. Influence of different treatments on growth of pakchoi
Field investigations showed (table 2): after the treatment 1 is sprayed with the amino acid-containing water-soluble fertilizer, the root system of the pakchoi grows longer and the leaves grow vigorously; the leaf of the pakchoi treated in the treatment 1 is thicker than that of the pakchoi treated in the treatment 2, and the leaf color is dark green; the average plant height of the treated plants 1 is 22.1cm, and the plant height is increased by 0.9cm compared with that of the treated plants 2; the weight of the individual plant in treatment 1 was 41.2g on average, which was 3.4g higher than that in treatment 2.
TABLE 2 survey table of field growth conditions of Chinese cabbage treated differently
2 Effect of different treatments on the yield of Brassica rapa
The yield of Chinese cabbage treated differently (see Table 3), 1 m per 666.7m2The average yield was 2535.7kg, which was specific for treatment 2 at 666.7m2The average yield of 2324.6kg is increased by 211.1kg and 9.08 percent.
TABLE 3 Chinese cabbage yield for different treatments
Analysis of variance of yield for the experimental treatment cells (see Table 4), F-15.017 > F0.0110.92, indicating that the yield difference between treatments reached a very significant level.
TABLE 4 ANOVA TABLE
| Source of variation | Sum of squares | Degree of freedom | Mean square | F value | F0.05 | F0.01 |
| Between blocks | 25.660 | 2 | 12.830 | 3.831 | 5.14 | 10.92 |
| Treatment room | 100.584 | 2 | 50.292 | 15.017** | 5.14 | 10.92 |
| Error of the measurement | 13.396 | 4 | 3.349 | |||
| Total variation | 139.640 | 8 |
Note: indicates that the difference reaches a significant level, and indicates that the difference reaches a very significant level.
TABLE 5 multiple comparisons
Note: LSD 0.05-5.71 kg/cell, LSD 0.01-8.83 kg/cell, different lower case letters indicate significant difference by LSD test, and different upper case letters indicate very significant difference by LSD test.
The LSD multiple comparisons of the test treatment cell yields (see table 5) showed that: the yield difference between the treatment 1 and the treatment 2 reaches an extremely obvious level, which shows that the Chinese cabbage sprayed with the amino acid-containing water-soluble fertilizer has extremely obvious yield increasing effect in the soil with zinc and boron deficiency (the effective zinc is less than 1.0mg/kg, and the effective boron is less than 0.5 mg/kg). The fertilizer is prepared by adding the trace elements such as amino acid, zinc, boron and the like into the fertilizer, and the nutrient elements required by growth of the pakchoi are absorbed by the leaves to improve the nutrient condition of the pakchoi, so that the growth of the leaves and the root system of the pakchoi can be better promoted, and the yield of the pakchoi is improved.
3 yield and ratio of pakchoi in different treatment rooms
The different treatment output values are calculated according to the selling price of the pakchoi of 5.0 yuan/kg and the amino acid-containing water-soluble fertilizer of 40 yuan/L. Treatment 1 at 666.7m intervals2The average yield of pakchoi is increased by 211.1kg compared with treatment 2, the yield value is increased by 1055.5 yuan, and the fertilizer cost is deducted by 40 yuan (treatment 1 is 666.7m per treatment)2Co-applying amino acid-containing water soluble fertilizer 1.0L, 40 yuan per 666.7m2Increase yield to 1015.5 yuan.
Treatment 1 vs. treatment 2 at 666.7m intervals2The cost of the amino acid-containing water-soluble fertilizer is 40 yuan, the yield is increased by 1055.5 yuan, and the yield-to-yield ratio is 26.4:1 (see table 6).
TABLE 6 economic benefit to production to input ratio of Chinese cabbage treated differently
After the pakchoi is sprayed with the amino acid-containing water-soluble fertilizer, the root system of the pakchoi grows longer and the leaves grow vigorously; the leaf of the pakchoi treated in the treatment 1 is thicker than that of the pakchoi treated in the treatment 2, and the leaf color is dark green; the average plant height of the treated plants 1 is 22.1cm, and the plant height is increased by 0.9cm compared with that of the treated plants 2; the weight of the individual plant in treatment 1 was 41.2g on average, which was 3.4g higher than that in treatment 2.
The effect of increasing the yield of the pakchoi by spraying the amino acid-containing water-soluble fertilizer and conventional fertilization is very obvious compared with the effect of spraying equivalent clear water and conventional fertilization, and the yield is increased by 9.08%.
The economic benefit of the Chinese cabbage sprayed with the amino acid-containing water-soluble fertilizer and the conventional fertilizer application is higher, and compared with the method of spraying the same amount of clear water and the conventional fertilizer application, the economic benefit is higher, and the fertilizer is applied every 666.7m2The yield is increased by 1015.5 yuan, and the production-input ratio is 26.4: 1.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (5)
1. The preparation method of the amino acid solution is characterized by comprising the following steps:
(1) extracting the defatted soybean meal with water to obtain a soybean protein extracting solution; the mass ratio of the defatted soybean meal to water is 1 (25-35);
(2) mixing the soybean protein extracting solution obtained in the step (1) with bacillus subtilis fermentation liquor for fermentation to obtain amino acid liquor; the using amount of the bacillus subtilis fermentation liquor accounts for 5-10% of the total volume of the soybean protein extracting solution; the number of effective viable bacteria in the bacillus subtilis fermentation liquor is more than or equal to 2 hundred million/mL;
the fermentation temperature in the step (2) is 25-30 ℃, and the fermentation time is 24-72 hours.
2. The method according to claim 1, wherein the pH value extracted in the step (1) is 7 to 12.
3. The method according to claim 1, wherein the temperature of the extraction in the step (1) is 40 to 80 ℃ and the extraction time is 50 to 100 min.
4. The method according to claim 3, wherein the extraction in step (1) is performed by microwave extraction.
5. The method according to claim 4, wherein the microwave extraction power is 300-1000W.
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| CN113862324A (en) * | 2021-09-30 | 2021-12-31 | 广东拉多美化肥有限公司 | Preparation method of mixed liquid of polypeptide and amino acid, amino acid foliar fertilizer and polypeptide water-soluble fertilizer |
| CN117362084A (en) * | 2023-10-27 | 2024-01-09 | 中山市承铭农业技术开发有限公司 | Amino acid water-soluble fertilizer prepared from soybeans and production process |
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