CN117209312A - Fertilizer synergist containing fish protein oligopeptide and preparation method thereof - Google Patents
Fertilizer synergist containing fish protein oligopeptide and preparation method thereof Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title claims abstract description 26
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 58
- 239000000243 solution Substances 0.000 claims description 49
- 238000003756 stirring Methods 0.000 claims description 41
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 40
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- 238000000034 method Methods 0.000 claims description 18
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- 239000004365 Protease Substances 0.000 claims description 17
- 150000001875 compounds Chemical class 0.000 claims description 16
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- 238000004140 cleaning Methods 0.000 claims description 15
- 238000010438 heat treatment Methods 0.000 claims description 15
- 239000013049 sediment Substances 0.000 claims description 15
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- 210000001835 viscera Anatomy 0.000 claims description 15
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- 239000011575 calcium Substances 0.000 claims description 12
- 229910052791 calcium Inorganic materials 0.000 claims description 12
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 11
- 239000001110 calcium chloride Substances 0.000 claims description 11
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 11
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- 239000002131 composite material Substances 0.000 claims description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 10
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Abstract
The invention relates to the technical field of fertilizer synergists, in particular to a fertilizer synergist containing fish protein oligopeptide and a preparation method thereof.
Description
Technical Field
The invention relates to the technical field of fertilizer synergists, in particular to a fertilizer synergist containing fish protein oligopeptide and a preparation method thereof.
Background
The fertilizer synergist is an active substance for increasing the nutrient availability, can activate nitrogen, phosphorus and potassium in soil, meets the requirements of crops on the nutrients, and simultaneously adjusts the physiological functions of the crops, improves the fertilizer utilization rate, reduces environmental pollution, improves the yield increasing efficiency of unit fertilizer on the crops, promotes the optimization and upgrading of the fertilizer industry structure, and drives the environmental improvement and the recovery and the promotion of the soil fertility, so that the research of the influence of the fertilizer synergist on the growth and the development of the crops is of great significance.
Natural substances such as alginic acid, amino acid, humic acid and the like can improve the utilization rate of the fertilizer, are environment-friendly, and have obvious advantages when added into the fertilizer. Fish protein contains abundant amino acid and oligopeptide micromolecules, the minced fillet is decomposed through an acid-base hydrolysis method and an enzymolysis method, the amino acid and oligopeptide micromolecules contained in the minced fillet can be extracted, and the minced fillet is prepared into a fertilizer synergist which can play a role in improving soil and assisting crop growth, but the production process of the fertilizer synergist containing fish protein extracts on the market is not mature enough, and how to improve the production process becomes a technical problem to be solved urgently by a person skilled in the art.
Disclosure of Invention
Technical problem to be solved
Aiming at the defects existing in the prior art, the invention provides a fertilizer synergist containing fish protein oligopeptide and a preparation method thereof, aiming at improving soil and promoting crop growth by the prepared fertilizer synergist containing fish protein oligopeptide.
Technical proposal
In order to achieve the above purpose, the invention is realized by the following technical scheme:
a fertilizer synergist containing fish protein oligopeptide is prepared from the following raw materials: crucian protein extraction component, snakehead protein extraction component, compound protease, calcium chloride and fish protein amino acid fermentation liquor;
the preparation method of the crucian protein extraction component comprises the following steps:
step 1, removing heads, scales and viscera of fresh crucian carp, cleaning with deionized water, mincing, and performing grinding according to the following steps of 1: adding deionized water into the solution with the feed liquid ratio of 5w/v, homogenizing for 10min at the rotating speed of 1000r/min by using a homogenizer, and obtaining the minced fillet;
step 2, adjusting the pH value of the minced fillet in the step 1 to 3 by using a hydrochloric acid solution, standing for 10min at normal temperature, centrifuging for 20min at a rotating speed of 10000r/min, removing the uppermost layer of grease and the lowermost layer of sediment after centrifuging, and taking a protein solution in the middle layer;
step 3, adjusting the pH value of the protein solution in the step 2 to 5-6 by using a sodium hydroxide solution, and centrifuging for 20min at a rotating speed of 10000r/min to obtain a precipitate, namely a crucian protein extraction component;
the preparation method of the snakehead protein extraction component comprises the following steps:
step a, removing heads, scales and viscera of fresh snakeheads, cleaning the fresh snakeheads by deionized water, and mincing the fresh snakeheads according to the following formula 1: dispersing the feed liquid ratio of 2w/v in deionized water, heating to 50 ℃ in water bath, and placing in an oscillator for oscillating for 8-10min to obtain the black minced fillet;
and b, regulating the pH value of the snakehead paste in the step a to 7 by using a phosphate buffer, adding alkaline protease with the mass of 1% of that of the snakehead paste, carrying out enzymolysis for 6-8h at the temperature of 55 ℃, heating in a boiling water bath for 20min after the enzymolysis is finished, and centrifuging for 10min at the rotating speed of 10000r/min to obtain a supernatant which is the snakehead protein extraction component.
Further, the concentration of the hydrochloric acid solution in the step 2 is 2mol/L, and the concentration of the sodium hydroxide solution in the step 3 is 2mol/L.
Further, the composite protease is prepared by mixing pepsin, neutral protease and trypsin according to the weight ratio.
Further, the preparation method of the fish protein amino acid fermentation broth comprises the following steps: removing head, scale and viscera of the green-occupied fish, cleaning with deionized water, crushing and homogenizing to obtain green-occupied surimi, pouring the green-occupied surimi into a fermentation tank, adding 5% by weight of composite zymophyte, stirring at a rotating speed of 300-400r/min for 5min, stirring uniformly, placing at 15-25 ℃ for sealing fermentation for 10d, filtering after fermentation, and removing impurities to obtain the fish protein amino acid fermentation liquor.
Still further, the composite zymophyte is bacillus subtilis, lactobacillus acidophilus and bifidobacterium bifidum according to 4:2:3 weight ratio, and mixing.
A preparation method of a fertilizer synergist containing fish protein oligopeptide comprises the following steps:
s1, extracting a crucian protein extract component, a snakehead protein extract component and deionized water according to a ratio of 1:1:2, then regulating the pH value to 6-7, adding compound protease with the weight of 2 percent of the compound protease, heating in water bath to 40 ℃, preserving at constant temperature for 2 hours for enzymolysis, and obtaining the oligopeptide mixed component after enzymolysis;
s2, extracting a crucian protein extract component, a snakehead protein extract component and calcium chloride according to a ratio of 1:1:4, adding deionized water with the weight being 15 times of that of the supernatant, mixing and stirring, regulating the pH value to 9, placing the mixture in a constant-temperature water bath shaking table at 50 ℃ for chelation for 20min, centrifuging at the rotating speed of 4000r/min for 10min after chelation is finished to remove sediment, adding absolute ethyl alcohol with the volume being 90% of that of the supernatant for alcohol precipitation after rotary evaporation and concentration, and finally centrifuging at the rotating speed of 4000r/min for 20min to obtain sediment which is recorded as a fish protein peptide chelated calcium component;
and S3, weighing 50 parts by weight of fish protein amino acid fermentation liquor, adding 10-15 parts by weight of oligopeptide mixed component in S1 and 8-10 parts by weight of fish protein peptide chelated calcium component in S2, and mixing and stirring to obtain the fertilizer synergist containing the fish protein oligopeptide.
Further, the mixing and stirring speed of the uniform mixing in the step S1 is 400-500r/min, and the stirring time is 8-10min.
Further, the pH value is adjusted by using a hydrochloric acid solution or a sodium hydroxide solution with the concentration of 1mol/L in the step S1.
Further, the pH value is adjusted by using a hydrochloric acid solution or a sodium hydroxide solution with the concentration of 1mol/L in the step S2.
Further, the stirring speed of the mixing and stirring in the step S3 is 300-500r/min, and the stirring time is 15-20min.
Advantageous effects
The invention provides a fertilizer synergist containing fish protein oligopeptide and a preparation method thereof, and compared with the prior art, the fertilizer synergist has the following beneficial effects:
according to the invention, proteins in crucian are extracted by an acidolysis method, proteins and polypeptides in snakehead are extracted by an enzymolysis method, the crucian protein extraction component and the snakehead protein extraction component are decomposed into oligopeptide micromolecules by an enzymolysis method after being mixed, the crucian protein extraction component, the snakehead protein extraction component and calcium chloride are chelated by a chelation method to prepare fish protein peptide chelated calcium, the oligopeptide micromolecules and the fish protein peptide chelated calcium are mixed into fish protein amino acid fermentation liquor, and can be absorbed by crops to synthesize nucleic acid or endogenous hormone, so that the metabolic function and stress resistance of the crops are enhanced, and the physical and chemical properties of soil are improved by the action of amino acid in the fish protein amino acid fermentation liquor, the water and fertilizer retention capacity and the air permeability of the soil are improved, and the effect of improving the growth of the crops is achieved to a certain extent.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. It will be apparent that the described embodiments are some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The invention is further described below with reference to examples.
Example 1
The fertilizer synergist containing the fish protein oligopeptide of the embodiment comprises the following raw materials: crucian protein extraction component, snakehead protein extraction component, compound protease, calcium chloride and fish protein amino acid fermentation liquor;
the preparation method of the crucian protein extraction component comprises the following steps:
step 1, removing heads, scales and viscera of fresh crucian carp, cleaning with deionized water, mincing, and performing grinding according to the following steps of 1: adding deionized water into the solution with the feed liquid ratio of 5w/v, homogenizing for 10min at the rotating speed of 1000r/min by using a homogenizer, and obtaining the minced fillet;
step 2, adjusting the pH value of the minced fillet in the step 1 to 3 by using a hydrochloric acid solution, standing for 10min at normal temperature, centrifuging for 20min at a rotating speed of 10000r/min, removing the uppermost layer of grease and the lowermost layer of sediment after centrifuging, and taking a protein solution in the middle layer;
step 3, adjusting the pH value of the protein solution in the step 2 to 5 by using a sodium hydroxide solution, and centrifuging for 20min at a rotating speed of 10000r/min to obtain a precipitate, namely a crucian protein extraction component;
the preparation method of the snakehead protein extraction component comprises the following steps:
step a, removing heads, scales and viscera of fresh snakeheads, cleaning the fresh snakeheads by deionized water, and mincing the fresh snakeheads according to the following formula 1: dispersing the feed liquid ratio of 2w/v in deionized water, heating to 50 ℃ in a water bath, and then placing in an oscillator for oscillating for 8min to obtain the minced fillet;
and b, regulating the pH value of the snakehead paste in the step a to 7 by using a phosphate buffer, adding alkaline protease with the mass of 1% of that of the snakehead paste, carrying out enzymolysis for 6 hours at the temperature of 55 ℃, heating in a boiling water bath for 20 minutes after the enzymolysis is finished, and centrifuging for 10 minutes at the rotating speed of 10000r/min to obtain a supernatant which is the snakehead protein extraction component.
The concentration of the hydrochloric acid solution in the step 2 is 2mol/L, and the concentration of the sodium hydroxide solution in the step 3 is 2mol/L.
The compound protease is prepared by mixing pepsin, neutral protease and trypsin according to the weight ratio.
The preparation method of the fish protein amino acid fermentation broth comprises the following steps: removing heads, scales and viscera of the green-occupied fish, cleaning with deionized water, crushing and homogenizing to obtain green-occupied minced fillet, pouring the green-occupied minced fillet into a fermentation tank, adding 5% by weight of composite zymophyte, stirring at a rotating speed of 300r/min for 5min, stirring uniformly, placing at 15 ℃ for sealing fermentation for 10d, and filtering to remove impurities after fermentation is finished to obtain the fish protein amino acid fermentation liquor.
The composite zymocyte is bacillus subtilis, lactobacillus acidophilus and bifidobacterium bifidum according to the following 4:2:3 weight ratio, and mixing.
A preparation method of a fertilizer synergist containing fish protein oligopeptide comprises the following steps:
s1, extracting a crucian protein extract component, a snakehead protein extract component and deionized water according to a ratio of 1:1:2, then regulating the pH value to 6, adding compound protease with the weight of 2 percent, heating in water bath to 40 ℃, preserving at constant temperature for 2 hours for enzymolysis, and obtaining the oligopeptide mixed component after enzymolysis;
s2, extracting a crucian protein extract component, a snakehead protein extract component and calcium chloride according to a ratio of 1:1:4, adding deionized water with the weight being 15 times of that of the supernatant, mixing and stirring, regulating the pH value to 9, placing the mixture in a constant-temperature water bath shaking table at 50 ℃ for chelation for 20min, centrifuging at the rotating speed of 4000r/min for 10min after chelation is finished to remove sediment, adding absolute ethyl alcohol with the volume being 90% of that of the supernatant for alcohol precipitation after rotary evaporation and concentration, and finally centrifuging at the rotating speed of 4000r/min for 20min to obtain sediment which is recorded as a fish protein peptide chelated calcium component;
and S3, weighing 50 parts by weight of fish protein amino acid fermentation liquor, adding 10 parts by weight of oligopeptide mixed components in the S1 and 8 parts by weight of fish protein peptide chelated calcium components in the S2, and mixing and stirring to obtain the fertilizer synergist containing the fish protein oligopeptide.
And S1, uniformly mixing and stirring at a speed of 400r/min for 8min.
In S1, hydrochloric acid solution or sodium hydroxide solution with the concentration of 1mol/L is used for regulating the pH value.
In S2, hydrochloric acid solution or sodium hydroxide solution with the concentration of 1mol/L is used for regulating the pH value.
And S3, mixing and stirring at a stirring speed of 300r/min for 15min.
Example 2
The fertilizer synergist containing the fish protein oligopeptide of the embodiment comprises the following raw materials: crucian protein extraction component, snakehead protein extraction component, compound protease, calcium chloride and fish protein amino acid fermentation liquor;
the preparation method of the crucian protein extraction component comprises the following steps:
step 1, removing heads, scales and viscera of fresh crucian carp, cleaning with deionized water, mincing, and performing grinding according to the following steps of 1: adding deionized water into the solution with the feed liquid ratio of 5w/v, homogenizing for 10min at the rotating speed of 1000r/min by using a homogenizer, and obtaining the minced fillet;
step 2, adjusting the pH value of the minced fillet in the step 1 to 3 by using a hydrochloric acid solution, standing for 10min at normal temperature, centrifuging for 20min at a rotating speed of 10000r/min, removing the uppermost layer of grease and the lowermost layer of sediment after centrifuging, and taking a protein solution in the middle layer;
step 3, adjusting the pH value of the protein solution in the step 2 to 6 by using a sodium hydroxide solution, and centrifuging for 20min at a rotating speed of 10000r/min to obtain a precipitate, namely a crucian protein extraction component;
the preparation method of the snakehead protein extraction component comprises the following steps:
step a, removing heads, scales and viscera of fresh snakeheads, cleaning the fresh snakeheads by deionized water, and mincing the fresh snakeheads according to the following formula 1: dispersing the feed liquid ratio of 2w/v in deionized water, heating to 50 ℃ in a water bath, and then placing the mixture in an oscillator to oscillate for 10min to obtain the minced fillet;
and b, regulating the pH value of the snakehead paste in the step a to 7 by using a phosphate buffer, adding alkaline protease with the mass of 1% of that of the snakehead paste, carrying out enzymolysis for 8 hours at the temperature of 55 ℃, heating in a boiling water bath for 20 minutes after the enzymolysis is finished, and centrifuging for 10 minutes at the rotating speed of 10000r/min to obtain a supernatant which is the snakehead protein extraction component.
The concentration of the hydrochloric acid solution in the step 2 is 2mol/L, and the concentration of the sodium hydroxide solution in the step 3 is 2mol/L.
The compound protease is prepared by mixing pepsin, neutral protease and trypsin according to the weight ratio.
The preparation method of the fish protein amino acid fermentation broth comprises the following steps: removing heads, scales and viscera of the green-occupied fish, cleaning with deionized water, crushing and homogenizing to obtain green-occupied minced fillet, pouring the green-occupied minced fillet into a fermentation tank, adding 5% by weight of composite zymophyte, stirring at 400r/min for 5min, stirring uniformly, placing at 25 ℃ for sealing fermentation for 10d, and filtering to remove impurities after fermentation is finished to obtain the fish protein amino acid fermentation liquor.
The composite zymocyte is bacillus subtilis, lactobacillus acidophilus and bifidobacterium bifidum according to the following 4:2:3 weight ratio, and mixing.
A preparation method of a fertilizer synergist containing fish protein oligopeptide comprises the following steps:
s1, extracting a crucian protein extract component, a snakehead protein extract component and deionized water according to a ratio of 1:1:2, then regulating the pH value to 7, adding compound protease with the weight of 2 percent, heating in water bath to 40 ℃, preserving at constant temperature for 2 hours for enzymolysis, and obtaining the oligopeptide mixed component after enzymolysis;
s2, extracting a crucian protein extract component, a snakehead protein extract component and calcium chloride according to a ratio of 1:1:4, adding deionized water with the weight being 15 times of that of the supernatant, mixing and stirring, regulating the pH value to 9, placing the mixture in a constant-temperature water bath shaking table at 50 ℃ for chelation for 20min, centrifuging at the rotating speed of 4000r/min for 10min after chelation is finished to remove sediment, adding absolute ethyl alcohol with the volume being 90% of that of the supernatant for alcohol precipitation after rotary evaporation and concentration, and finally centrifuging at the rotating speed of 4000r/min for 20min to obtain sediment which is recorded as a fish protein peptide chelated calcium component;
and S3, weighing 50 parts by weight of fish protein amino acid fermentation liquor, adding 15 parts by weight of oligopeptide mixed components in the S1 and 10 parts by weight of fish protein peptide chelated calcium components in the S2, and mixing and stirring to obtain the fertilizer synergist containing the fish protein oligopeptide.
The mixing and stirring speed of the uniform mixing in the step S1 is 500r/min, and the stirring time is 10min.
In S1, hydrochloric acid solution or sodium hydroxide solution with the concentration of 1mol/L is used for regulating the pH value.
In S2, hydrochloric acid solution or sodium hydroxide solution with the concentration of 1mol/L is used for regulating the pH value.
And S3, mixing and stirring at a stirring speed of 500r/min for 20min.
Example 3
The fertilizer synergist containing the fish protein oligopeptide of the embodiment comprises the following raw materials: crucian protein extraction component, snakehead protein extraction component, compound protease, calcium chloride and fish protein amino acid fermentation liquor;
the preparation method of the crucian protein extraction component comprises the following steps:
step 1, removing heads, scales and viscera of fresh crucian carp, cleaning with deionized water, mincing, and performing grinding according to the following steps of 1: adding deionized water into the solution with the feed liquid ratio of 5w/v, homogenizing for 10min at the rotating speed of 1000r/min by using a homogenizer, and obtaining the minced fillet;
step 2, adjusting the pH value of the minced fillet in the step 1 to 3 by using a hydrochloric acid solution, standing for 10min at normal temperature, centrifuging for 20min at a rotating speed of 10000r/min, removing the uppermost layer of grease and the lowermost layer of sediment after centrifuging, and taking a protein solution in the middle layer;
step 3, adjusting the pH value of the protein solution in the step 2 to 5 by using a sodium hydroxide solution, and centrifuging for 20min at a rotating speed of 10000r/min to obtain a precipitate, namely a crucian protein extraction component;
the preparation method of the snakehead protein extraction component comprises the following steps:
step a, removing heads, scales and viscera of fresh snakeheads, cleaning the fresh snakeheads by deionized water, and mincing the fresh snakeheads according to the following formula 1: dispersing the feed liquid ratio of 2w/v in deionized water, heating to 50 ℃ in a water bath, and placing in an oscillator for oscillating for 9min to obtain the minced fillet;
and b, regulating the pH value of the snakehead paste in the step a to 7 by using a phosphate buffer, adding alkaline protease with the mass of 1% of that of the snakehead paste, carrying out enzymolysis for 7 hours at the temperature of 55 ℃, heating in a boiling water bath for 20 minutes after the enzymolysis is finished, and centrifuging for 10 minutes at the rotating speed of 10000r/min to obtain a supernatant which is the snakehead protein extraction component.
The concentration of the hydrochloric acid solution in the step 2 is 2mol/L, and the concentration of the sodium hydroxide solution in the step 3 is 2mol/L.
The compound protease is prepared by mixing pepsin, neutral protease and trypsin according to the weight ratio.
The preparation method of the fish protein amino acid fermentation broth comprises the following steps: removing heads, scales and viscera of the green-occupied fish, cleaning with deionized water, crushing and homogenizing to obtain green-occupied minced fillet, pouring the green-occupied minced fillet into a fermentation tank, adding 5% by weight of composite zymophyte, stirring at 400r/min for 5min, stirring uniformly, placing at 20 ℃ for sealing fermentation for 10d, and filtering to remove impurities after fermentation is finished to obtain the fish protein amino acid fermentation liquor.
The composite zymocyte is bacillus subtilis, lactobacillus acidophilus and bifidobacterium bifidum according to the following 4:2:3 weight ratio, and mixing.
A preparation method of a fertilizer synergist containing fish protein oligopeptide comprises the following steps:
s1, extracting a crucian protein extract component, a snakehead protein extract component and deionized water according to a ratio of 1:1:2, then regulating the pH value to 7, adding compound protease with the weight of 2 percent, heating in water bath to 40 ℃, preserving at constant temperature for 2 hours for enzymolysis, and obtaining the oligopeptide mixed component after enzymolysis;
s2, extracting a crucian protein extract component, a snakehead protein extract component and calcium chloride according to a ratio of 1:1:4, adding deionized water with the weight being 15 times of that of the supernatant, mixing and stirring, regulating the pH value to 9, placing the mixture in a constant-temperature water bath shaking table at 50 ℃ for chelation for 20min, centrifuging at the rotating speed of 4000r/min for 10min after chelation is finished to remove sediment, adding absolute ethyl alcohol with the volume being 90% of that of the supernatant for alcohol precipitation after rotary evaporation and concentration, and finally centrifuging at the rotating speed of 4000r/min for 20min to obtain sediment which is recorded as a fish protein peptide chelated calcium component;
and S3, weighing 50 parts by weight of fish protein amino acid fermentation liquor, adding 13 parts by weight of oligopeptide mixed components in the S1 and 9 parts by weight of fish protein peptide chelated calcium components in the S2, and mixing and stirring to obtain the fertilizer synergist containing the fish protein oligopeptide.
The mixing and stirring speed of the uniform mixing in the step S1 is 500r/min, and the stirring time is 9min.
In S1, hydrochloric acid solution or sodium hydroxide solution with the concentration of 1mol/L is used for regulating the pH value.
In S2, hydrochloric acid solution or sodium hydroxide solution with the concentration of 1mol/L is used for regulating the pH value.
And S3, mixing and stirring at a stirring speed of 400r/min for 18min.
Performance detection
The fertilizer potentiators containing fish protein oligopeptides prepared in examples 1 to 3 were designated as examples 1 to 3, and fertilizer potentiators randomly purchased from the market were designated as comparative examples, and the potentiating ability of examples 1 to 3 and comparative examples were examined;
the specific detection method comprises the following steps:
1) Examples 1-3 and comparative examples were first prepared according to 1:10000 mass ratio, then
Placing 100 wheat seeds in 4 trays with the same size respectively, then pouring diluted examples 1-3 and comparative examples into the 4 trays respectively, and performing water culture until the three-leaf period is finished;
2) After the water planting is finished, 30 wheat seedlings are randomly selected from each tray, the wheat seedling height, the whole fresh weight, the fresh weight of wheat seedling stems and leaves, the root length and the root weight are measured after seed coats are removed, and the average value is obtained after the measurement.
The data obtained are recorded in the following table:
as shown by the data in the table above, the growth vigor of wheat seedlings added with the fertilizer synergist containing the fish protein oligopeptide in the examples 1-3 is obviously better than that of wheat seedlings added with the fertilizer synergist containing the fish protein oligopeptide in the comparative examples, and the plant height, the whole plant fresh weight, the fresh weight of wheat seedlings stem leaves, the root length and the root weight of the wheat seedlings added with the fertilizer synergist containing the fish protein oligopeptide in the examples 1-3 are obviously better than those of the comparative examples, so that the fertilizer synergist containing the fish protein oligopeptide prepared by the invention has better market application prospect.
It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one … …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element.
The above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (10)
1. The fertilizer synergist containing the fish protein oligopeptide is characterized by comprising the following raw materials: crucian protein extraction component, snakehead protein extraction component, compound protease, calcium chloride and fish protein amino acid fermentation liquor;
the preparation method of the crucian protein extraction component comprises the following steps:
step 1, removing heads, scales and viscera of fresh crucian carp, cleaning with deionized water, mincing, and performing grinding according to the following steps of 1: adding deionized water into the solution with the feed liquid ratio of 5w/v, homogenizing for 10min at the rotating speed of 1000r/min by using a homogenizer, and obtaining the minced fillet;
step 2, adjusting the pH value of the minced fillet in the step 1 to 3 by using a hydrochloric acid solution, standing for 10min at normal temperature, centrifuging for 20min at a rotating speed of 10000r/min, removing the uppermost layer of grease and the lowermost layer of sediment after centrifuging, and taking a protein solution in the middle layer;
step 3, adjusting the pH value of the protein solution in the step 2 to 5-6 by using a sodium hydroxide solution, and centrifuging for 20min at a rotating speed of 10000r/min to obtain a precipitate, namely a crucian protein extraction component;
the preparation method of the snakehead protein extraction component comprises the following steps:
step a, removing heads, scales and viscera of fresh snakeheads, cleaning the fresh snakeheads by deionized water, and mincing the fresh snakeheads according to the following formula 1: dispersing the feed liquid ratio of 2w/v in deionized water, heating to 50 ℃ in water bath, and placing in an oscillator for oscillating for 8-10min to obtain the black minced fillet;
and b, regulating the pH value of the snakehead paste in the step a to 7 by using a phosphate buffer, adding alkaline protease with the mass of 1% of that of the snakehead paste, carrying out enzymolysis for 6-8h at the temperature of 55 ℃, heating in a boiling water bath for 20min after the enzymolysis is finished, and centrifuging for 10min at the rotating speed of 10000r/min to obtain a supernatant which is the snakehead protein extraction component.
2. The fertilizer synergist containing fish protein oligopeptide according to claim 1, wherein the concentration of the hydrochloric acid solution in the step 2 is 2mol/L, and the concentration of the sodium hydroxide solution in the step 3 is 2mol/L.
3. The fertilizer synergist containing fish protein oligopeptide according to claim 1, wherein the composite protease is prepared by mixing pepsin, neutral protease and trypsin according to an equal weight ratio.
4. The fertilizer synergist containing fish protein oligopeptide according to claim 1, wherein the preparation method of the fish protein amino acid fermentation broth is as follows: removing head, scale and viscera of the green-occupied fish, cleaning with deionized water, crushing and homogenizing to obtain green-occupied surimi, pouring the green-occupied surimi into a fermentation tank, adding 5% by weight of composite zymophyte, stirring at a rotating speed of 300-400r/min for 5min, stirring uniformly, placing at 15-25 ℃ for sealing fermentation for 10d, filtering after fermentation, and removing impurities to obtain the fish protein amino acid fermentation liquor.
5. The fertilizer synergist containing fish protein oligopeptide according to claim 4, wherein the compound zymocyte is bacillus subtilis, lactobacillus acidophilus and bifidobacterium bifidum according to 4:2:3 weight ratio, and mixing.
6. The method for preparing a fertilizer synergist containing fish protein oligopeptide according to any one of claims 1-5, wherein the preparation method comprises the following steps:
s1, extracting a crucian protein extract component, a snakehead protein extract component and deionized water according to a ratio of 1:1:2, then regulating the pH value to 6-7, adding compound protease with the weight of 2 percent of the compound protease, heating in water bath to 40 ℃, preserving at constant temperature for 2 hours for enzymolysis, and obtaining the oligopeptide mixed component after enzymolysis;
s2, extracting a crucian protein extract component, a snakehead protein extract component and calcium chloride according to a ratio of 1:1:4, adding deionized water with the weight being 15 times of that of the supernatant, mixing and stirring, regulating the pH value to 9, placing the mixture in a constant-temperature water bath shaking table at 50 ℃ for chelation for 20min, centrifuging at the rotating speed of 4000r/min for 10min after chelation is finished to remove sediment, adding absolute ethyl alcohol with the volume being 90% of that of the supernatant for alcohol precipitation after rotary evaporation and concentration, and finally centrifuging at the rotating speed of 4000r/min for 20min to obtain sediment which is recorded as a fish protein peptide chelated calcium component;
and S3, weighing 50 parts by weight of fish protein amino acid fermentation liquor, adding 10-15 parts by weight of oligopeptide mixed component in S1 and 8-10 parts by weight of fish protein peptide chelated calcium component in S2, and mixing and stirring to obtain the fertilizer synergist containing the fish protein oligopeptide.
7. The method for preparing a fertilizer synergist containing fish protein oligopeptide according to claim 6, wherein the mixing and stirring speed of the uniformly mixed in the step S1 is 400-500r/min, and the stirring time is 8-10min.
8. The method for preparing a fertilizer synergist containing fish protein oligopeptide according to claim 6, wherein the pH value is adjusted by using 1mol/L hydrochloric acid solution or sodium hydroxide solution in S1.
9. The method for preparing a fertilizer synergist containing fish protein oligopeptide according to claim 6, wherein the pH value is adjusted by using 1mol/L hydrochloric acid solution or sodium hydroxide solution in S2.
10. The method for preparing a fertilizer synergist containing fish protein oligopeptide according to claim 6, wherein the stirring speed of mixing and stirring in the step S3 is 300-500r/min, and the stirring time is 15-20min.
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| CN115925473A (en) * | 2022-08-30 | 2023-04-07 | 仲恺农业工程学院 | Fish protein polypeptide water-soluble fertilizer and preparation method thereof |
| CN116283434A (en) * | 2022-12-02 | 2023-06-23 | 河北福赛农业科技有限公司 | Double-decomposition process of fish protein water-soluble fertilizer |
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| CN106866782A (en) * | 2017-02-07 | 2017-06-20 | 浙江省海洋水产研究所 | A kind of method for extracting Fish protein |
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