CN109984149A - A method of biostimulant is prepared using amino acid fermentation tail washings - Google Patents
A method of biostimulant is prepared using amino acid fermentation tail washings Download PDFInfo
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- CN109984149A CN109984149A CN201711491412.XA CN201711491412A CN109984149A CN 109984149 A CN109984149 A CN 109984149A CN 201711491412 A CN201711491412 A CN 201711491412A CN 109984149 A CN109984149 A CN 109984149A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N61/00—Biocides, pest repellants or attractants, or plant growth regulators containing substances of unknown or undetermined composition, e.g. substances characterised only by the mode of action
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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- Agronomy & Crop Science (AREA)
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Abstract
The invention belongs to amino acid fermentation tail washings process fields, disclose a kind of method for preparing biostimulant using amino acid fermentation tail washings, it includes the following steps: step 1) separation, drying and crushes, step 2 concentration, step 3) microorganism enzymolysis processing, step 4) prepares marine algae extract, and step 5) prepares biostimulant.The present invention handles fermentation tailing liquor, by carrying out reasonable compatibility to concentrate, enzymolysis liquid, humic acid and marine algae extract, prepares biostimulant, can regulate and control plant physiology process, promote plant strain growth, improve product quality.
Description
Technical field
The invention belongs to amino acid fermentation tail washings process fields, and in particular to a kind of to utilize the preparation life of amino acid fermentation tail washings
The method of object stimulin.
Background technique
Biostimulant is usually natural, plays promotion work well containing bioactive substance, for plant
A kind of product, neither pesticide, is less conventional fertilizers；The target of biostimulant is crops itself, it can change
The Physiology and biochemistry state of kind plant plays an extraordinary facilitation to the physiological system of inside plants, improves pesticide effect
The utilization rate of fruit and fertilizer improves the level that crops resist adverse circumstance, also improves the ultimate output and quality of agricultural product of crops.
2013, biostimulant has been divided into 8 classifications in a survey report of European Union, including humic acid, meet it is organic
Substance, non-organic mineral (including phosphite ester), marine algae extract, chitin and chitosan derivative, resists beneficial chemical element
Rising agent, free amino acid.
Amino acid fermentation tail washings be in glutamic acid commercial process extract product after liquid, have CODCr high,
BOD5 high, mycoprotein content are high, sulfate radical (sulfuric acid tune pH) content is high, ammonia-nitrogen content is high and pH value (1.5-3.2) low " five is high
One is low " the characteristics of, processing difficulty is larger, higher cost, it is difficult to obtain the relatively high byproduct of sexual valence.Studies have shown that amino
Acid has apparent facilitation to the growth of plant.Before applicant patented technology " 2014105587445, a kind of utilization
The method of glutamic acid wastewater production liquid fertilizer " discloses a kind of method for preparing liquid fertilizer using Amino Acid Fermentation Wastewater,
This process employs waste, the fertilizer efficiency of preparation is good, wherein using the concentrated hydrochloric acid of 6-8mol/L, in 60 DEG C of temperature
Lower stirring hydrolyzes 12-16 hours, prepares amino acid to hydrolyze mycoprotein, and this method can get 50% or so percent hydrolysis, but
It is that the hydrochloric acid of high concentration causes heavy corrosion to equipment, equipment updating decision has aggravated business burden.How above-mentioned technique is improved,
Preparing biostimulant using fermentation tailing liquor is our technical issues that need to address.
Summary of the invention
Have the research of patented technology based on applicant, on this basis, is sent out the present invention provides a kind of using amino acid
The method that ferment tail washings prepares biostimulant.
The present invention is using technical solution as described below:
A method of biostimulant being prepared using amino acid fermentation tail washings comprising following steps: step 1) separation, drying
And crush, step 2 concentration, step 3) microorganism enzymolysis processing, step 4) prepares marine algae extract, step 5) preparation biology
Stimulin.
Specifically, described method includes following steps:
Step 1) separation, drying and crushing: amino acid fermentation tail washings is separated by disc separator, revolving speed is
5000rpm, centrifugation time 3min collect mycoprotein precipitating and waste mother liquor, then mycoprotein are precipitated and is dried, then powder
Broken machine is ground into mycoprotein powder；
Step 2 concentration: waste mother liquor is concentrated into the 1/5 of original volume, obtains concentrate；
Step 3) microorganism enzymolysis processing: aspergillus niger ATCC 16404 being inoculated into seed tank culture base and is cultivated, temperature
At 30 DEG C, culture to bacteria concentration is 1.0 × 10 for control8Cfu/ml obtains seed liquor；Mycoprotein powder obtained by step 1) is added
It being added in the seed liquor of 10 times of weight, adjustment pH is 3.5, and temperature is 30 DEG C, and 100rpm stirs 42h, tourmaline powder is then added,
It stirs evenly, while adjusting cultivation temperature is 41 DEG C, continues to cultivate 6h, obtains enzymolysis liquid；
Step 4) prepares marine algae extract: seaweed being dried, powder is then comminuted into, is then added to the 6wt%'s of double weight
In sodium bicarbonate solution, 30-50min is impregnated, then be warming up to 50 DEG C, under heat-retaining condition, is ultrasonically treated, is then down to room temperature, mistake
Filtrate is collected in filter, is condensed into the 1/3 of original volume to get marine algae extract；
Step 5) prepares biostimulant: by concentrate, enzymolysis liquid, humic acid and marine algae extract by being added to agitator tank
In, use potassium hydroxide adjust pH be 7, then 100rpm stir 30min, be finally placed in 4 DEG C preservations to get.
Preferably,
The ingredient of the seed tank culture base are as follows: 20g/L glucose, 6g/L corn pulp, 2g/L peptone, 0.2g/L biphosphate
Potassium, 0.2g/L dipotassium hydrogen phosphate, 0.1g/L magnesium sulfate, 0.1g/L manganese sulfate, solvent are water
Preferably,
The additive amount of the tourmaline powder accounts for the 2wt% of mycoprotein powder.
Preferably,
The ultrasonic time is 10-15min, supersonic frequency 20KHz.
Preferably,
The concentrate, enzymolysis liquid, humic acid and marine algae extract mass ratio be 30-50:20-30:10-15:7-9.
The beneficial effect that the present invention obtains mainly includes but is not limited to the following aspects:
The present invention handles fermentation tailing liquor, by closing to concentrate, enzymolysis liquid, humic acid and marine algae extract
Compatibility is managed, biostimulant is prepared, plant physiology process can be regulated and controled, promotes plant strain growth, improves product quality；
The present invention digests mycoprotein using the enzyme that aspergillus niger generates, and avoids using hydrochloric acid solution, will not generate to equipment rotten
Erosion, and aspergillus niger has cracking larger molecular organics and slightly solubility inorganic matter, is absorbed and utilized convenient for crop, improves soil knot
Structure；
The present invention improves the percent hydrolysis of mycoprotein powder by optimizing to aspergillus niger condition of enzyme production；Tourmaline
Anion can be discharged automatically, anion has stronger oxidisability, also persistently occurs DC static, released mineral matter and micro
Element plays a driving role to microbial reproduction and producing enzyme；The producing enzyme energy of bacterial strain can be improved in the heat treatment of appropriate time and temperature
Power and permeability of cell membrane, so that the output increased of acid protease, so that the ability of protein hydrolysate is accordingly promoted；
The present invention is by microbion zymolysis, by mycoprotein enzymatic hydrolysis at Free Amino Acids, can complexation of metal ions,
Microelement required for plant is carried in plant, additionally it is possible to directly provide required carbon source and nitrogen source for crop, promote
Into the development of plant strain growth and root system；Microorganism in soil can utilize Amino acid synthesis various plants during own metabolism
Growth regulator promotes plant strain growth to regulate and control plant physiology process；
Marine algae extract contains inorganic nutrients necessary to plant growth, and cromoci rich in, amine, indoles
The plant growth substances such as compound, the metabolism of tunable crop and physiological function promote crop root system growth and overground part
The increase of Biomass and yield solves phytotoxicity, pre- frost damage prevention cold damage, to arid, improvement agricultural product flavor and alleviation salt marsh harm
Promote crop root growth effect obvious.
Specific embodiment
In order to make those skilled in the art better understand the technical solutions in the application, specifically real below in conjunction with the application
Example is applied, the present invention is more clearly and completely described, it is clear that described embodiment is only that the application a part is real
Example is applied, instead of all the embodiments.Based on the embodiment in the application, those of ordinary skill in the art are not making creation
Property labour under the premise of every other embodiment obtained, should fall within the scope of the present invention.
Embodiment 1
A method of biostimulant is prepared using amino acid fermentation tail washings comprising following steps:
Step 1) separation, drying and crushing: amino acid fermentation tail washings is separated by disc separator, revolving speed is
5000rpm, centrifugation time 3min collect mycoprotein precipitating and waste mother liquor, then bacterial sediment are dried, then pulverizer
It is ground into mycoprotein powder；
Step 2 concentration: waste mother liquor is concentrated into 1/5th of original volume, obtains concentrate；
Step 3) microorganism enzymolysis processing: aspergillus niger ATCC 16404 being inoculated into seed tank culture base and is cultivated, temperature
At 30 DEG C, culture to bacteria concentration is 1.0 × 10 for control8Cfu/ml obtains seed liquor；The ingredient of the seed tank culture base are as follows:
20g/L glucose, 6g/L corn pulp, 2g/L peptone, 0.2g/L potassium dihydrogen phosphate, 0.2g/L dipotassium hydrogen phosphate, 0.1g/L sulphur
Sour magnesium, 0.1g/L manganese sulfate, solvent are water；Mycoprotein powder is added in the seed liquor of 10 times of weight, adjustment pH is
3.5, temperature is 30 DEG C, and 100rpm stirs 42h, and then addition accounts for the tourmaline powder of mycoprotein powder 2wt%, is stirred evenly, together
When adjustment cultivation temperature be 41 DEG C, continue cultivate 6h, obtain enzymolysis liquid；
Step 4) prepares marine algae extract: seaweed being dried, powder is then comminuted into, is then added to the 6wt%'s of double weight
In sodium bicarbonate solution, 30min is impregnated, then be warming up to 50 DEG C, under heat-retaining condition, be ultrasonically treated 10min, supersonic frequency is
20KHz is down to room temperature, and filtrate is collected in filtering, is condensed into the 1/3 of original volume to get marine algae extract；
Step 5) prepares biostimulant: by concentrate, enzymolysis liquid, humic acid and marine algae extract according to 30:20:10:7's
Mass ratio is added in agitator tank, use potassium hydroxide adjust pH be 7, then 100rpm stirring 30min, be finally placed in 4 DEG C
Save to get.
Embodiment 2
A method of biostimulant is prepared using amino acid fermentation tail washings comprising following steps:
Step 1) separation, drying and crushing: amino acid fermentation tail washings is separated by disc separator, revolving speed is
5000rpm, centrifugation time 3min collect mycoprotein precipitating and waste mother liquor, then bacterial sediment are dried, then pulverizer
It is ground into mycoprotein powder；
Step 2 concentration: waste mother liquor is concentrated into 1/5th of original volume, obtains concentrate；
Step 3) microorganism enzymolysis processing: aspergillus niger ATCC 16404 being inoculated into seed tank culture base and is cultivated, temperature
At 30 DEG C, culture to bacteria concentration is 1.0 × 10 for control8Cfu/ml obtains seed liquor；The ingredient of the seed tank culture base are as follows:
20g/L glucose, 6g/L corn pulp, 2g/L peptone, 0.2g/L potassium dihydrogen phosphate, 0.2g/L dipotassium hydrogen phosphate, 0.1g/L sulphur
Sour magnesium, 0.1g/L manganese sulfate, solvent are water；Mycoprotein powder is added in the seed liquor of 10 times of weight, adjustment pH is
3.5, temperature is 30 DEG C, and 100rpm stirs 42h, and then addition accounts for the tourmaline powder of mycoprotein powder 2wt%, is stirred evenly, together
When adjustment cultivation temperature be 41 DEG C, continue cultivate 6h, obtain enzymolysis liquid；
Step 4) prepares marine algae extract: seaweed being dried, powder is then comminuted into, is then added to the 6wt%'s of double weight
In sodium bicarbonate solution, 30-50min is impregnated, then be warming up to 50 DEG C, under heat-retaining condition, be ultrasonically treated 15min, supersonic frequency is
20KHz is down to room temperature, and filtrate is collected in filtering, is condensed into the 1/3 of original volume to get marine algae extract；
Step 5) prepares biostimulant: by concentrate, enzymolysis liquid, humic acid and marine algae extract according to 50:30:15:9's
Mass ratio is added in agitator tank, use potassium hydroxide adjust pH be 7, then 100rpm stirring 30min, be finally placed in 4 DEG C
Save to get.
Embodiment 3
Influence of the different disposal method to mycoprotein degree of hydrolysis:
Each component is following (butt %) in amino acid fermentation tail washings of the present invention: mycoprotein 35.2, amino acid 1 4.1, polysaccharide object
Matter 5.3, ammonia nitrogen 9.9, sulphur 9.0, calcium 3.2, potassium 1.3, iron 0.9, surplus are other.
Group is set:
Using embodiment 1 as experimental group；
Control group 1: hydrolysis is stirred at a temperature of 60 DEG C using the hydrochloric acid of 6mol/L 16 hours to hydrolyze mycoprotein, remaining is operated
With embodiment 1；
Control group 2: hydrolysis is stirred at a temperature of 60 DEG C using the hydrochloric acid of 2mol/L 32 hours to hydrolyze mycoprotein, remaining is operated
With embodiment 1；
Control group 3: not adding tourmaline powder, remaining operation is the same as embodiment 1；
Control group 4: temperature is controlled at 30 DEG C, does not improve temperature, remaining operation is the same as embodiment 1.
Test method:
Using the content of Kjeldahl nitrogen determination protein；Ninhydrin colorimetry measures the content of amino acid in hydrolyzate；
The measurement of sample degree of hydrolysis:
Degree of hydrolysis (DH) 100%=(amino acid content/mycoprotein total amino acid content in hydrolyzate) × 100%；
The other degree of hydrolysis of each group and acid protease enzyme activity are shown in Table 1:
Table 1
| Group | Degree of hydrolysis (DH) % | Enzyme activity U/ml |
| Experimental group | 68.4 | 3504 |
| Control group 1 | 52.9 | ---- |
| Control group 2 | 45.7 | ---- |
| Control group 3 | 58.6 | 2891 |
| Control group 4 | 64.3 | 3166 |
Conclusion: as shown in table 1, by comparison, it was found that, experimental group makes the degree of hydrolysis of mycoprotein more using microbion zymolysis
It is high, hence it is evident that better than the Hydrochloric Acid Hydrolysis Method of control group 1-2 routine, and it is corrosion-free to equipment；Compared with control group 3-4, experimental group
Degree of hydrolysis and enzyme activity greatly improve, illustrate tourmaline powder and properly increase temperature yield of enzyme can be improved, to improve
Aspergillus niger increases the content of amino acid to the degree of hydrolysis of mycoprotein.
Embodiment 4
Influence of the biostimulant prepared by the present invention to vintage and quality.
Experimental group: the product of sprinkling 2 technique of embodiment of the present invention preparation；
The product of control group 1: preparation process is hydrolysis to be stirred at a temperature of 60 DEG C 16 hours using the hydrochloric acid of 6mol/L to hydrolyze
Mycoprotein, remaining operating procedure is the same as embodiment 2；Spray the product；
The product of control group 2: preparation process is hydrolysis to be stirred at a temperature of 60 DEG C 32 hours using the hydrochloric acid of 2mol/L to hydrolyze
Mycoprotein, remaining operating procedure is the same as embodiment 2；Spray the product；
Control group 3: sprinkling equivalent amount of water.
It is (the N P and K 1:0.5:1, according to per acre on the basis of routinely applying base manure for the huge peak of triennial for examination grape
50kg application), biostimulant (experimental group and control group 1-2) is applied when the grape spring, bud expanded, amount of application is 80kg per acre,
Control group 3 applies the water of isodose, other planting patterns are identical.The results are shown in Table 2.
Table 2
Conclusion: as shown in table 2, compared with control group 3, the product of experimental group and control group 1-2 can effectively promote grape
Strain yield and single fruit weight, but the promotion effect of experimental group is best；Experiment set product can also promote the quality of grape, including solvable
Property sugar and ascorbic specific gravity；As it can be seen that product of the present invention can greatly promote vintage and quality.
The present invention is also tested using peanut and corn as trial crops, wherein the peanut florescence initial stage according to
60g/ mus of amount sprays product of the present invention, and 10-15% can be improved in peanut yield；It is sprayed at corn heading stage according to 60g/ mus of amount
Product of the present invention, corn yield can be improved 20% or so.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or spy described in conjunction with this embodiment or example
Point is included at least one embodiment or example of the invention.In the present specification, schematic expression of the above terms are not
It must be directed to identical embodiment or example.Although the embodiments of the present invention has been shown and described above, Ke Yili
Solution, above-described embodiment are exemplary, are not considered as limiting the invention, and those skilled in the art are at this
It can make changes, modifications, alterations, and variations to the above described embodiments in the range of invention.
Claims (7)
1. a kind of method for preparing biostimulant using amino acid fermentation tail washings comprising following steps: step 1) separation is dried
Dry and crushing, step 2 concentration, step 3) microorganism enzymolysis processing, step 4) prepare marine algae extract, step 5) preparation life
Object stimulin.
2. the method according to claim 1, wherein described method includes following steps:
Step 1) separation, drying and crushing: amino acid fermentation tail washings is separated by disc separator, collects thallus egg
White precipitating and waste mother liquor, then mycoprotein is precipitated and is dried, it is then comminuted into mycoprotein powder；
Step 2 concentration: waste mother liquor obtained by step 1) is concentrated into the 1/5 of original volume, obtains concentrate；
Step 3) microorganism enzymolysis processing: aspergillus niger ATCC 16404 being inoculated into seed tank culture base and is cultivated, temperature
At 30 DEG C, culture to bacteria concentration is 1.0 × 10 for control8Cfu/ml obtains seed liquor；Mycoprotein powder obtained by step 1) is added
It being added in the seed liquor of 10 times of weight, adjustment pH is 3.5, and temperature is 30 DEG C, and 100rpm stirs 42h, tourmaline powder is then added,
Adjusting temperature simultaneously is 41 DEG C, continues to obtain enzymolysis liquid with 100rpm stir culture 6h；
Step 4) prepares marine algae extract: seaweed being dried, powder is ground into, is then added to the carbonic acid of the 6wt% of double weight
In hydrogen sodium solution, 30-50min is impregnated, then be warming up to 50 DEG C, under heat-retaining condition, be ultrasonically treated, be then down to room temperature, filtered, received
Collect filtrate, is condensed into the 1/3 of original volume to get marine algae extract；
Step 5) prepares biostimulant: by enzymolysis liquid obtained by concentrate obtained by step 2, step 3), humic acid and step 4)
Gained marine algae extract by being added in agitator tank, use potassium hydroxide adjust pH be 7, then 100rpm stirring 30min, finally
Be placed in 4 DEG C save to get.
3. according to the method described in claim 2, it is characterized in that, the ingredient of the seed tank culture base are as follows: 20g/L grape
Sugar, 6g/L corn pulp, 2g/L peptone, 0.2g/L potassium dihydrogen phosphate, 0.2g/L dipotassium hydrogen phosphate, 0.1g/L magnesium sulfate, 0.1g/
L manganese sulfate, solvent are water.
4. according to the method described in claim 2, it is characterized in that, the additive amount of the tourmaline powder accounts for mycoprotein powder
2wt%。
5. according to the method described in claim 2, supersonic frequency is it is characterized in that, the ultrasonic time is 10-15min
20KHz。
6. according to the method described in claim 2, it is characterized in that, concentrate obtained by step 2, enzymolysis liquid, corruption obtained by step 3)
The mass ratio of marine algae extract obtained by phytic acid and step 4) is 30-50:20-30:10-15:7-9.
7. according to the method described in claim 2, it is characterized in that, the revolving speed of the disc separator is 5000rpm, when centrifugation
Between be 3min.
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| CN117776816A (en) * | 2023-12-22 | 2024-03-29 | 中化农业（临沂）研发中心有限公司 | Biostimulant composition for targeted promotion of nitrogen assimilation of plants, preparation method thereof and fertilizer |
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| CN112409057A (en) * | 2020-12-30 | 2021-02-26 | 赵兰坤 | Method for preparing organic fertilizer by using amino acid fermentation waste |
| CN117776816A (en) * | 2023-12-22 | 2024-03-29 | 中化农业（临沂）研发中心有限公司 | Biostimulant composition for targeted promotion of nitrogen assimilation of plants, preparation method thereof and fertilizer |
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