CN115745674A - Microbial oligosaccharide chelated trace element water-soluble fertilizer and preparation method thereof - Google Patents
Microbial oligosaccharide chelated trace element water-soluble fertilizer and preparation method thereof Download PDFInfo
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- 235000013619 trace mineral Nutrition 0.000 title claims abstract description 58
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- 238000002360 preparation method Methods 0.000 title claims abstract description 21
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- 238000000034 method Methods 0.000 claims abstract description 10
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Abstract
The invention discloses a microbial oligosaccharide chelating trace element water-soluble fertilizer and a preparation method thereof, and relates to the technical field of water-soluble fertilizer preparation. Various trace elements are added in the form of culture medium components before fermentation, and are chelated with polysaccharide under the biological action of microorganisms to form a stable polysaccharide-trace element complex, and then the oligosaccharide-trace element liquid fertilizer is finally obtained through enzymolysis; the product of the invention has more stable performance and simpler and more convenient preparation process than other methods.
Description
Technical Field
The invention relates to the technical field of water-soluble fertilizer preparation, in particular to a microbial oligosaccharide chelated trace element water-soluble fertilizer and a preparation method thereof.
Background
Oligosaccharides (also called oligosaccharides), generally refer to saccharides with low degree of polymerization composed of 2 to 10 monosaccharides, and generally have a molecular weight of less than 3000Da, and are important functional substances in vivo. Compared with high molecular weight substances such as polysaccharide and the like, the oligosaccharide has higher solubility and utilization degree, can be better absorbed and utilized, and is widely applied to the fields of food, health care products, medicines, additives and the like at present. In recent years, the application of oligosaccharide substances in the agricultural field is attracting more and more attention in recent years, and as important signal molecules in plants, the oligosaccharide substances can promote the growth of plants, improve the resistance of the plants to diseases, enhance the adaptability of the plants to the environment, improve the yield and quality of crops, and have wide application prospects in the aspects of improving the stress resistance of the plants and promoting the growth of the plants. According to a new edition of pesticide management regulations newly implemented in 2017, 8, 1 and 1, saccharides used in agricultural production are mainly classified into natural plant resistance inducers and natural plant growth regulators.
The deep processing of oligosaccharide substances to obtain derivative products is another research focus in the field in recent years. Among them, studies on chelation of trace elements by oligosaccharides to improve their biological activities and utilization efficiency have been increasing. The trace elements are important nutrients for growth and development of animals and plants, are also important components of various enzymes in the bodies of animals and plants, and play an important role in metabolism and healthy development of organisms. At present, trace element additives of elements such as oligosaccharide chelating zinc and the like are proved to have better utilization effect in the breeding industry, but research for applying the technology in the agricultural field basically stays in the laboratory stage. Because metal elements in the inorganic salt are easily fixed in soil and become insoluble substances to lose the effect, the fertilizer efficiency is seriously influenced; and due to the antagonism among elements, the fertilizer is easy to be adsorbed and fixed by soil colloid and has poor absorption effect. People find that better application effect can be obtained after inorganic salt of trace elements is converted into an organic chelate state, for example, the effectiveness of the amino acid chelated trace element fertilizer in soil is better than that of an inorganic trace element fertilizer, the amino acid chelated trace element fertilizer is easier to be absorbed by wheat, and the growth of the wheat is promoted, so the development of chelated trace elements is gradually increased in recent years.
In the prior art, raw materials such as chitin, seaweed and the like rich in polysaccharide are degraded into oligosaccharide through enzymolysis or chemical treatment, and then chelated with trace elements or compounded, or the raw materials rich in polysaccharide are fermented by microorganisms and then chelated with the trace elements.
Such as patent: a alginic acid chitosan oligosaccharide chelating medium trace element biotechnological water soluble fertilizer and a preparation method thereof; application No.: CN201710895784.2; application publication No.: CN107805154A; the patent mainly uses chemical chelation, and the chitosan oligosaccharide and trace elements are simply mixed and heated, so that the chelation efficiency is low.
Such as the patent: a method for preparing a liquid fertilizer containing seaweed oligosaccharide trace elements by combining chemical reaction with multi-stage fermentation; application No.: CN201610977803.1; publication No.: CN106588251A; the patent mainly adopts chemical chelation, seaweed oligosaccharide is prepared by a fermentation method, but the chelation with trace elements at the later stage still adopts a simple mixing and heating mode, and the chelation efficiency is lower.
Such as the patent: a chitosan oligosaccharide compound preparation for improving the yield of wheat and application thereof; application No.: CN201310689965.1; publication No.: CN104710231A; the patent directly mixes oligosaccharide and trace elements, and the chelating efficiency is not high enough.
In conclusion, the preparation process of the oligosaccharide chelating trace elements mainly has the following defects:
(1) Mainly uses simple chemical chelation, namely, the oligosaccharide and the trace elements are directly compounded and mixed according to the proportion, so that the chelation efficiency is low;
(2) Most of the oligosaccharide sources are crustaceans or seaweeds, and complex pretreatment is required.
Disclosure of Invention
The invention aims to at least solve one of the technical problems in the prior art and provides a microbial oligosaccharide chelated trace element water-soluble fertilizer and a preparation method thereof.
The technical solution of the invention is as follows:
a preparation method of a microbial oligosaccharide chelated trace element water-soluble fertilizer comprises the steps of fermenting enterococcus faecium, adding excessive trace elements while fermenting to form a polysaccharide-trace element compound, adding an enzyme solution after the fermentation is finished, degrading, and performing centrifugal filtration to obtain the oligosaccharide-containing trace element liquid fertilizer.
Specifically, the method comprises the following steps:
s1: culturing enterococcus faecium by adopting an MRS culture medium, and standing for culture to obtain an enterococcus faecium bacterial liquid;
s2: adding a fermentation culture medium into a fermentation tank, adding trace elements, adjusting the pH value in the tank to 7.5-7.8, heating to 115-121 ℃, sterilizing and cooling;
s3: inoculating enterococcus faecium liquid into a fermentation tank, keeping the fermentation temperature at 28-33 ℃, standing for culture, and intermittently stirring the fermentation liquid for 36-40 hours;
s4: after fermentation, adding the compound enzyme solution into the fermentation liquor in the S3, keeping the temperature at 30-35 ℃, and reacting for 8-10 hours;
s5: and (4) obtaining the liquid fertilizer containing the microbial oligosaccharide and the trace elements through centrifugal filtration.
In a preferred embodiment of the present invention, in step S1, the culture time is 8 to 12 hours and the culture temperature is 30 to 34 ℃.
In a preferable embodiment of the invention, in step S2, the trace element is at least one of zinc sulfate, manganese sulfate and magnesium sulfate, and the addition amount is 0.1-0.5wt% of the fermentation medium.
As a preferable embodiment of the present invention, in step S2, the fermentation medium specifically includes: 2 to 3.0 weight percent of cane sugar, 0.5 to 1.0 weight percent of peptone, 0.02 to 0.05 weight percent of monopotassium phosphate, 0.02 to 0.05 weight percent of dipotassium phosphate, 0.1 to 0.5 weight percent of magnesium sulfate, 0.001 to 0.002 weight percent of manganese sulfate and the balance of water.
As a preferable scheme of the invention, in the step S3, the inoculation amount of the enterococcus faecium bacterial liquid is 3-5% of the volume of the material.
In a preferred embodiment of the present invention, the complex enzyme solution includes at least one of amylase and glycosidase.
As a preferable scheme of the invention, the addition amount of the compound enzyme liquid is 0.005-0.01% of the volume of the fermentation liquid.
A microbial oligosaccharide chelated trace element water-soluble fertilizer is prepared by adopting the preparation method.
The beneficial effects of the invention are:
various trace elements are added in the form of culture medium components before fermentation, and are chelated with polysaccharide under the biological action of microorganisms to form a stable polysaccharide-trace element complex, and then the oligosaccharide-trace element liquid fertilizer is finally obtained through enzymolysis; the product of the invention has more stable performance and simpler and more convenient preparation process compared with other methods.
Detailed Description
The technical solution of the present invention is further illustrated by the following specific examples.
Example 1
A preparation method of a microbial oligosaccharide chelating microelement water-soluble fertilizer comprises the following steps:
s1: adopts MRS culture medium (peptone 10.0g, beef extract 10.0g, yeast 5.0g, diammonium hydrogen citrate [ (NH) 4 ) 2 HC 6 H 5 O 7 ]2.0g, glucose (C) 6 H 12 O 6 ·H 2 O) 20.0g, tween 80.0 mL, sodium acetate (CH) 3 COONa·3H 2 O) 5.0g, dipotassium hydrogen phosphate (K) 2 HPO 4 ·3H 2 O) 2.0g, magnesium sulfate (MgSO) 4 ·7H 2 O) 0.58g, manganese sulfate (MnSO) 4 ·H 2 O) 0.25g, 1000mL of distilled water and pH 6.2-6.6), and culturing the enterococcus faecium for 12 hours at the culture temperature of 30 ℃;
s2: adding a fermentation culture medium (3.0% of sucrose, 1.0% of peptone, 0.05% of potassium dihydrogen phosphate, 0.05% of dipotassium hydrogen phosphate, 0.5% of magnesium sulfate, 0.002% of manganese sulfate and the balance of water in all proportions by mass) into a fermentation tank, adding trace elements (0.3% of zinc sulfate, 0.1% of manganese sulfate and 0.3% of magnesium sulfate by mass, based on 100% of the total mass of the fermentation culture medium), adjusting the pH value in the tank to be 7.5, heating to 120 ℃, sterilizing for 20min, and cooling to 30 ℃;
s3: inoculating a bacterial liquid of enterococcus faecium into a fermentation tank, wherein the inoculation amount is 3% of the volume of the material, the fermentation temperature is 30 ℃, standing for culture, intermittently stirring the fermentation liquid, and the fermentation time is 36 hours;
s4: after fermentation, adding a complex enzyme solution (containing alpha-amylase and glycosidase, the volume ratio of the alpha-amylase to the glycosidase is 1, and the volume ratio of the complex enzyme solution to the fermentation broth is 0.01%) into the fermentation broth, keeping the temperature at 35 ℃ and reacting for 10 hours; s5: and (3) performing centrifugal filtration to obtain the liquid fertilizer containing microbial oligosaccharide-trace element, adjusting the oligosaccharide content to be 0.1 percent, and taking the rest components as solvents.
Example 2
A preparation method of a microbial oligosaccharide chelated trace element water-soluble fertilizer comprises the following steps:
s1: adopts MRS culture medium (peptone 10.0g, beef extract 10.0g, yeast 5.0g, diammonium hydrogen citrate [ (NH) 4 ) 2 HC 6 H 5 O 7 ]2.0g, glucose (C) 6 H 12 O 6 ·H 2 O) 20.0g, tween 80.0 mL, sodium acetate (CH) 3 COONa·3H 2 O) 5.0g, dipotassium hydrogen phosphate (K) 2 HPO 4 ·3H 2 O) 2.0g, magnesium sulfate (MgSO) 4 ·7H 2 O) 0.58g, manganese sulfate (MnSO) 4 ·H 2 O) 0.25g, 1000mL of distilled water and pH 6.2-6.6), and culturing the enterococcus faecium for 10 hours at the culture temperature of 34 ℃;
s2: adding a fermentation medium (3.0% of sucrose, 1.0% of peptone, 0.05% of potassium dihydrogen phosphate, 0.05% of dipotassium hydrogen phosphate, 0.5% of magnesium sulfate, 0.002% of manganese sulfate and the balance of water in all proportions by mass) into a fermentation tank, adding trace elements (0.2 wt% of zinc sulfate, 0.1wt% of manganese sulfate and 0.3wt% of magnesium sulfate based on 100% of the total mass of the fermentation medium), adjusting the pH value in the tank to be 7.5, heating to 116 ℃, sterilizing for 15min, and cooling to 30 ℃;
s3: inoculating a bacterial liquid of enterococcus faecium into a fermentation tank, wherein the inoculation amount is 4% of the volume of the material, the fermentation temperature is 30 ℃, standing for culture, intermittently stirring the fermentation liquid, and the fermentation time is 40 hours;
s4: after fermentation, adding a complex enzyme solution (containing amylase and glycosidase, the volume ratio of the two is 1, and the volume ratio of the complex enzyme solution to the fermentation broth is 0.01%) into the fermentation broth, keeping the temperature at 35 ℃ and reacting for 10 hours; s5: and (3) performing centrifugal filtration to obtain the liquid fertilizer containing microbial oligosaccharide-trace element, adjusting the oligosaccharide content to be 0.1 percent, and taking the rest components as solvents.
Example 3
A preparation method of a microbial oligosaccharide chelated trace element water-soluble fertilizer comprises the following steps:
s1: adopts MRS culture medium (peptone 10.0g, beef extract 10.0g, yeast 5.0g, diammonium hydrogen citrate [ (NH) 4 ) 2 HC 6 H 5 O 7 ]2.0g, glucose (C) 6 H 12 O 6 ·H 2 O) 20.0g, tween 80.0 mL, sodium acetate (CH) 3 COONa·3H 2 O) 5.0g, dipotassium hydrogen phosphate (K) 2 HPO 4 ·3H 2 O) 2.0g, magnesium sulfate (MgSO) 4 ·7H 2 O) 0.58g, manganese sulfate (MnSO) 4 ·H 2 O) 0.25g, 1000mL of distilled water and pH 6.2-6.6), and culturing the enterococcus faecium for 8-12 hours at the culture temperature of 30-34 ℃;
s2: adding a fermentation culture medium (3.0% of sucrose, 1.0% of peptone, 0.05% of potassium dihydrogen phosphate, 0.05% of dipotassium hydrogen phosphate, 0.5% of magnesium sulfate, 0.002% of manganese sulfate and the balance of water in all proportions by mass percent) into a fermentation tank, adding trace elements (0.2% of zinc sulfate, 0.3% of manganese sulfate and 0.4% of magnesium sulfate by mass percent based on 100% of the total mass of the fermentation culture medium), adjusting the pH value in the tank to be 7.8, heating to 121 ℃, sterilizing for 30min, and cooling to 35 ℃;
s3: inoculating a bacterial liquid of enterococcus faecium into a fermentation tank, wherein the inoculation amount is 5% of the volume of the material, the fermentation temperature is 30 ℃, standing for culture, intermittently stirring the fermentation liquid, and the fermentation time is 36-40 hours;
s4: after fermentation, adding a complex enzyme solution (containing amylase and glycosidase, the volume ratio of the two is 1, and the volume ratio of the complex enzyme solution to the fermentation broth is 0.01%) into the fermentation broth, keeping the temperature at 35 ℃ and reacting for 10 hours; s5: and (3) performing centrifugal filtration to obtain the liquid fertilizer containing microbial oligosaccharide-trace element, adjusting the oligosaccharide content to 0.1-0.2%, and taking the rest components as a solvent.
Control group 1: in contrast to example 1, no trace elements were added to the fermentation medium.
Control group 2: unlike example 1, the same concentration of trace elements was dissolved in water without fermentation medium.
Test 1:
test groups: spraying the water-soluble fertilizer containing oligosaccharide-trace elements (containing zinc, manganese and magnesium) prepared in the example 1 on hybrid late rice;
control group 1: spraying a water-soluble fertilizer containing microbial polysaccharide in a control group 1 on the hybrid late rice;
control group 2: spraying trace element fertilizer (containing zinc, manganese and magnesium) of the control group 2 on the hybrid late rice;
test site: in a practical training base of biological science and engineering colleges of Jiangxi agriculture universities, spraying is started after 20 days of rice cultivation, spraying is carried out for 3 times at intervals of 10 days, and various indexes are measured in the mature period of rice. Spraying with a spraying kettle, wherein 20L of the spraying agent is sprayed per mu.
TABLE 1 influence of oligosaccharide-trace element water-soluble fertilizer on improvement of economic traits of hybrid late rice
From the test results, compared with a control group 2 (pure trace elements), the average plant height of the test group is increased by 1.9cm, the effective spike is increased by 0.6 ten thousand spikes/mu, the number of grains per spike is increased by 3.5 grains, the seed setting rate is improved by 2.0 percentage points, and the thousand-grain weight is increased by 0.5 g.
Test 2
Control group 3: in contrast to example 2, no trace elements were added to the fermentation medium.
Control group 4: unlike example 2, the same concentration of trace elements was dissolved in water without fermentation medium.
Test groups: spraying water-soluble fertilizer containing oligosaccharide-trace elements (containing zinc, manganese and magnesium) prepared in example 2 on the pakchoi;
control group 3: spraying a water-soluble fertilizer containing microbial polysaccharide in a control group 3 on the pakchoi;
control group 4: spraying microelement fertilizer (containing zinc, manganese and magnesium) of a control group 4 on the pakchoi;
test site: a practical training base of biological science and engineering colleges of the Jiangxi agriculture university is implemented in a pot culture mode (the diameter of a pot is 20cm, 40kg of soil is filled), 4 th true leaves of the pakchoi grow out, spraying is carried out for 4 times at intervals of 7 days, and various indexes are measured after the last spraying is carried out for 1 week. The specific dosage is 8mL per spraying.
TABLE 2 influence of oligosaccharide-microelement water-soluble fertilizer on the improvement of pakchoi character
As can be seen from the test results, compared with the control group 2 (pure trace elements), the average plant height of the test group is increased by 1.8cm, the fresh weight is increased by 3.85 g/plant, and the vitamin C content is increased by 22.0mg/kg.
In conclusion, the water soluble fertilizer can improve the growth performance of crops such as rice and the like, and meanwhile, the disease and insect resistance is remarkably improved.
The above additional technical features can be freely combined and used in superposition by those skilled in the art without conflict.
The above description is only a preferred embodiment of the present invention, and the technical solutions that achieve the objects of the present invention by substantially the same means are within the protection scope of the present invention.
Claims (9)
1. A preparation method of a microbial oligosaccharide chelated trace element water-soluble fertilizer is characterized by comprising the steps of fermenting enterococcus faecium, adding excessive trace elements while fermenting to form a polysaccharide-trace element compound, adding an enzyme solution after the fermentation is finished, degrading, and performing centrifugal filtration to obtain the oligosaccharide-containing trace element liquid fertilizer.
2. A preparation method of a microbial oligosaccharide chelated trace element water-soluble fertilizer is characterized by comprising the following steps:
s1: adopting MRS culture medium to culture enterococcus faecium, and standing for culture to obtain enterococcus faecium bacterial liquid;
s2: adding a fermentation culture medium into a fermentation tank, adding trace elements, adjusting the pH value in the tank to 7.5-7.8, heating to 115-121 ℃, sterilizing and cooling;
s3: inoculating enterococcus faecium liquid into a fermentation tank, keeping the fermentation temperature at 28-33 ℃, standing for culture, and intermittently stirring the fermentation liquid for 36-40 hours;
s4: after fermentation is finished, adding the compound enzyme solution into the fermentation liquor in the S3, keeping the temperature at 30-35 ℃, and reacting for 8-10 hours;
s5: and (4) performing centrifugal filtration to obtain the liquid fertilizer containing the microbial oligosaccharide and the trace elements.
3. The method for preparing a water-soluble fertilizer containing microbial oligosaccharide and trace elements as claimed in claim 2, wherein the culture time in step S1 is 8-12 hours, and the culture temperature is 30-34 ℃.
4. The method for preparing a water-soluble fertilizer containing microbial oligosaccharide chelating trace elements as claimed in claim 2, wherein in step S2, the trace elements are at least one of zinc sulfate, manganese sulfate and magnesium sulfate, and the addition amount of the trace elements is 0.1-0.5wt% of the fermentation medium.
5. The microbial oligosaccharide chelating trace element water-soluble fertilizer and the preparation method thereof as claimed in claim 2, wherein in the step S2, the fermentation medium specifically comprises: 2 to 3.0 weight percent of cane sugar, 0.5 to 1.0 weight percent of peptone, 0.02 to 0.05 weight percent of monopotassium phosphate, 0.02 to 0.05 weight percent of dipotassium phosphate, 0.1 to 0.5 weight percent of magnesium sulfate, 0.001 to 0.002 weight percent of manganese sulfate and the balance of water.
6. The method for preparing a water-soluble fertilizer containing microbial oligosaccharide and chelated trace elements as claimed in claim 2, wherein in step S3, the inoculation amount of enterococcus faecium liquid is 3-5% of the volume of the material.
7. The method for preparing a water-soluble fertilizer with microbial oligosaccharide and chelated trace elements as claimed in claim 2, wherein the complex enzyme solution comprises at least one of amylase and glycosidase.
8. The method for preparing a water-soluble fertilizer with microbial oligosaccharide and chelated trace elements as claimed in claim 2, wherein the amount of the complex enzyme solution added is 0.005-0.01% of the volume of the fermentation broth.
9. A water-soluble fertilizer with microbial oligosaccharide chelated trace elements, which is characterized by being prepared by the preparation method of any one of claims 1 to 8.
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