CN107540417A - A kind of alga fertilizer preparation method using undaria pinnitafida as raw material - Google Patents
A kind of alga fertilizer preparation method using undaria pinnitafida as raw material Download PDFInfo
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- CN107540417A CN107540417A CN201710927170.8A CN201710927170A CN107540417A CN 107540417 A CN107540417 A CN 107540417A CN 201710927170 A CN201710927170 A CN 201710927170A CN 107540417 A CN107540417 A CN 107540417A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
A kind of alga fertilizer preparation method using undaria pinnitafida as raw material, undaria pinnitafida combines the pueraria root residue process for fixation of preprocess method and enzyme by colloid mill and high pressure homogenizer, obtained marine alga is homogenized through immobilised enzymes two step method enzymolysis process Biocatalytic Conversion, realize that algal polysaccharides and protein are degraded into the target of alga oligosaccharides and polypeptide respectively, the final continuous production processes established the undaria pinnitafida enzymolysis based on continuous flowing stirred tank and prepare alga fertilizer, the centrifuged supernatant of enzymolysis liquid and undaria pinnitafida residue, supernatant is through falling film evaporation into concentrate, ultrafiltrate is liquid seaweed extract；Undaria pinnitafida digests the common fermentation such as residue and cake of camellia oleifera seeds into solid organic fertilizer, reaches the purpose that the full cell of undaria pinnitafida utilizes.The present invention improves the undaria pinnitafida level of resources utilization, improves undaria pinnitafida process technology and the alga fertilizer level of production, realizes that enzyme is repeated and is used continuously, reduces the consumption of enzyme, reduce alga fertilizer production cost, product quality is good, pure natural, pollution-free degradable.
Description
Technical field
The invention belongs to chemical technology field, it is related to the preparation method of marine alga fertilizer.
Background technology
Marine alga fertilizer is the bio-feritlizer be processed into through science using sea-plant marine alga as primary raw material, mainly into
Divide is that the natural bioactivity substance for being advantageous to growth and development of plants and marine alga extracted from marine alga absorbs simultaneously richness from ocean
The mineral nutrient element of collection in vivo, including algal polysaccharides, phenols poly-compounds, mannitol, glycine betaine, plant growth are adjusted
Save material (basic element of cell division, gibberellin, auxin and abscisic acid etc.) and the micro member such as nitrogen, phosphorus, potassium and iron, boron, molybdenum, iodine
Element.In addition, being increase fertilizer efficiency and the chelation of fertilizer, appropriate humic acid and appropriate trace element have also been dissolved in.
Compared with conventional fertilizers, alga fertilizer has following distinguishing feature.
First, alga fertilizer is more comprehensive than conventional fertilizers nutrition, balanced, obvious effect of increasing production is grown after crop applying, and seldom go out
Existing nutritional deficiency disease.Alga fertilizer is advantageous to growth and development of plants using natural seaweed as primary raw material, containing what is largely extracted from marine alga
Natural bioactivity substance and marine alga absorbed from ocean and be enriched with mineral nutrient element in vivo, including marine alga is more
Sugar, phenols poly-compounds, mannitol, glycine betaine, plant growth regulating substance（The basic element of cell division, gibberellin, auxin and de-
Fall acid etc.）With the trace element such as nitrogen, phosphorus, potassium and iron, boron, molybdenum, iodine.In addition, be increase fertilizer efficiency and fertilizer chelation, part
Product has also dissolved in appropriate humic acid and appropriate trace element, can meet the needs of plant growth each stage is to nutrition.2nd,
Contain a large amount of disease-resistant factors and special composition, fertile medicine economic benefits and social benefits in alga fertilizer.Degeneration-resistant disease resistance significantly increases after crop applying, blade face
Pesticidal effects can be improved by spraying.Algal polysaccharides and oligosaccharide, mannitol, phenols poly-compounds, the beet contained in alga fertilizer
The materials such as alkali, alginic acid and natural antibiotics, there is significant inhibitory anti-virus, anthelminthic effect, really play fertile medicine effect, greatly
The width enhancing cold-resistant of crop, drought resisting, disease-resistant, resistant to lodging, salinity and alkalinity resistance, to epidemic disease, virosis, anthrax, downy mildew, grey mold, white
The stronger resistance of the generation such as powder disease, droop.In addition, the alginic acid in alga fertilizer can reduce the surface tension of water, in plant
Surface forms thin film, increases contact area, makes water-soluble substances be easier to enter plant through cauline leaf superficial cell film
Cell, plant is set most effectively to absorb the nutritional ingredient in seaweed extracted liquor.If alga fertilizer and insecticide, bactericide are mixed
Use, there is synergistic effect, sprinkling expense can be reduced.3rd, substantial amounts of high active ingredient is contained in alga fertilizer, plant easily absorbs.
Growing way is vigorous after crop applying, can significantly improve yield and the quality of crop.Specific algal polysaccharides in marine alga, alginic acid,
The materials such as highly unsaturated fatty acid, there is very high bioactivity, the production of the nonspecific activity factor in plant can be stimulated
It is raw.Meanwhile also contain natural plants growth regulator in alga fertilizer, such as：Auxin, cytokinin-like substance and gibberellin
Deng each hormone ratio is close in its ratio and terrestrial plant, has the function that to adjust Endogenous hormone balance very well.Crop applying
Afterwards, yield and quality significantly improve, and the industrial crops quality such as tobacco, cotton, flowers can be significantly improved, especially to booth vegetable
Crop, the effect that increases both production and value are very notable.4th, alga fertilizer contains abundant organic matter and the sustained release factor, fertilizer efficiency length, can improve soil
Earth Tiny ecosystem, native promoting root growth living and anti-continuous cropping.Alga fertilizer can directly make soil or soil is increased organic matter, activation soil by plant
Various beneficial microbes in earth.These microorganisms can play a part of catalyst in plant, microbial metabolism circulation, make
The biopotency increase of soil.The metabolin of plant and edaphon can provide more nutrients for plant.Meanwhile marine alga is more
The sequestering system that sugar is formed can be such that nutrition slowly discharges, and extend fertilizer efficiency.In addition, the native compound such as sodium alginate that it contains
It is natural soil conditioner, the formation of soil granular structure can be promoted, improves soil internal void space, consolidate in coordination soil,
Liquid, gas proportions, recover due to soil over-burden and chemical contamination and lose lac balance, increase geobiont
Vigor, promote the release of available nutrient, be advantageous to root growth, improve the resistance of crop and anti-continuous cropping ability.5th, alga fertilizer
Natural safety nuisance free.Traditional chemical fertilizer fertilizer efficiency is single, seriously polluted, and long-term use can cause soil texture to be destroyed.
Alga fertilizer category natural seaweed extract, there is good compatibility with terrestrial plant, it is nontoxic to people, animal, it is environmentally safe,
With other any chemical fertilizer it is all incomparable the advantages of, be put into organic food special fertilizer material abroad.Alga fertilizer polymerize
Various active material in ocean, possess nutrition comprehensively, disease-resistant yield-increasing, it is living native, nuisanceless many advantages, such as, be really meet it is existing
For the good fertilizer in agricultural development direction.
China's alga fertilizer industry is started late, and phase late 1990s begins one's study, into 21 century after obtained it is fast
Speed development.In terms of the mode of production, mainly there are chemical method, physical method, 3 kinds of biological method.Currently, domestic most manufacturing enterprises
Main chemical leaching test production alga fertilizer simple using cheap, is that highly basic high temperature can destroy using the maximum inferior position of chemical method
The activity of marine alga endogenous substance；Physics extraction method is better than chemical leaching test, and it is thin that the technique cooled down using high pressure low temperature reaches marine alga
The broken purpose of cell wall, obtained macromolecular substances are difficult to directly be absorbed by crop, and fertilizer efficiency is not high；Most preferably biological method,
Extracted using enzyme hydrolysis, the activity of endogenous substance can be made to preserve to greatest extent.Due to the costliness of biology enzyme price
And the complexity of enzymolysis process, the country only have several families to take bioanalysis i.e. enzyme extraction.
Develop alga fertilizer, China has raw material advantage, and about 1,500,000 tons of the annual marine alga total output in China, 80% above is artificial
Cultivation, the marine alga quantity currently used for system fertilizer is seldom, no more than the 10% of marine alga total amount；And China is large agricultural country, fertilizer demand
Amount is very huge, therefore the development space of alga fertilizer industry is also very big.Undaria pinnitafida is the quality raw materials for producing alga fertilizer.On the one hand
For extension undaria pinnitafida industrial chain, improve undaria pinnitafida utilization ratio；On the other hand alga fertilizer production cost is reduced；Develop undaria pinnitafida life
Thing catalyzed conversion alga fertilizer technical meaning is great.
The content of the invention
Remained high for alga fertilizer raw materials for production cost, undaria pinnitafida is not implemented full cell and utilized in alga fertilizer production process
The problems such as.It is an object of the invention to provide a kind of alga fertilizer preparation method using undaria pinnitafida as raw material, undaria pinnitafida is through colloid mill
It is anti-in continuously flowing agitator tank formula by establishing immobilised enzymes two step method enzymolysis process after high pressure homogenizer combination pretreatment
Answer and alga fertilizer continuous production and enzyme recycling are realized in device, improve production efficiency, reduce the cost of enzyme；Produced in enzymolysis process
Undaria pinnitafida residue and the fermenting and producing solid-state seaweed organic fertilizer such as cake of camellia oleifera seeds, this method realizes that the full cell of undaria pinnitafida utilizes, be big
The big consumption for reducing enzyme, alga fertilizer production cost is greatly reduced, produces that alga fertilizer is pure natural, degradable, quality is good, protection ring
Border, reduce the wasting of resources.
The purpose of the present invention is achieved through the following technical solutions.
A kind of alga fertilizer preparation method using undaria pinnitafida as raw material of the present invention, according to the following steps.
(1) undaria pinnitafida pre-processes：Undaria pinnitafida is cleaned with running water；Soaked 54 hours with running water again, during which every 6 is small
Shi Huanshui 1 time；Undaria pinnitafida after immersion is shredded with knife, and broken undaria pinnitafida crushes through colloid mill；Crush rear skirt band puree and distilled water
With volume ratio 1:2 ~ 4 are mixed, and suspension is handled through high pressure homogenizer；Homogenizer condition of work：60 ~ 80 MPas of pressure, circulation time
Number 3 ~ 5 times, 1000 ls/h of flow, room temperature condition.
(2) first step enzyme digestion reaction：Undaria pinnitafida homogenate and distilled water are with mass ratio 1 after pretreatment:2 ~ 5 mix, the company of being placed in
Afterflow is moved in stirred tank, and cellulase is placed in together with agitating shaft through pueraria root residue immobilization, immobilized cellulase
The wire netting inframe of rotation, according to undaria pinnitafida and cellulase mass ratio 5 ~ 20:1 addition immobilized cellulase, regulation pH to
5.0 ~ 6.0,160 ~ 320 revs/min of speed of agitator, after stirring 3 ~ 9 minutes, speed of agitator is adjusted to 40 ~ 120 revs/min, temperature
Control is at 40 ~ 60 DEG C；After immobilized cellulase Biocatalytic Conversion 2 ~ 4 hours, start continuous feed and discharging, reaction terminate
Stop charging and discharging within first 1 ~ 2 hour.
(3) second step enzyme digestion reaction：Discharging enters in another continuous flowing stirred tank in step (2), alkalescence
Protease is through pueraria root residue immobilization, and immobilization alkali protease is placed in the wire netting inframe rotated together with agitating shaft, by alkalescence
The mg/litre of protease concentration 12000 ~ 60000 add immobilization alkali protease, adjust pH to 6.0 ~ 7.5, by 170 ~ 340 turns/
After minute stirring 3 ~ 8 minutes, speed of agitator is adjusted to 60 ~ 120 revs/min, and temperature control is at 45 ~ 65 DEG C；Reaction starting 2 ~ 4 is small
Shi Hou, starts continuous feed and discharging, and reaction terminates to stop charging and discharging in first 1 ~ 2 hour.
(4) under the enzymolysis liquid room temperature condition for obtaining step (3), 6000 ~ 12000 revs/min, centrifuge 3 ~ 6 minutes, obtain
To supernatant and undaria pinnitafida residue, supernatant falling film evaporation technology is concentrated, 10 ~ 20 ls/h of evaporation capacity, temperature 45 ~ 65
DEG C, 0.02 ~ 0.08 MPa of pressure, cocnentration factor 2.1 ~ 3.5, concentrate is through ultrafiltration (0.02 micron of membrane aperture, temperature room temperature, pressure
0.4 MPa, the meter per second of cross-flow rate 3.6,800 liters/square m/h) it is to obtain liquid seaweed extract afterwards.
(5) by the undaria pinnitafida residue in step (4) and cake of camellia oleifera seeds, pericarp, waste dish, brewage waste residue, chicken manure with mass ratio
1:3~5:4~8:5~7:2~4:4 ~ 6 heap fermentations, 12 ~ 18 cubic metres of volume, 40 ~ 70 centimetres of height, 45 ~ 65 DEG C of temperature, pass through
Fermentation materials are translated into control temperature, fermentation time 30 ~ 50 days；Fermentation ends obtain high-quality solid organic fertilizer.
Continuous feed and discharge method are taken in step (2) of the present invention, speed is consistent, and is 4 ~ 6 kgs/hr.
Pueraria root residue described in step (2) of the present invention and (3), preprocess method are as follows：Caused by kudzu-vine root powder production process
Pueraria root residue distilled water flushing 3 ~ 5 times, is placed in beaker, and pueraria root residue is with 40% ~ 50% Eusol with mass ratio 1:1 ~ 3 is mixed
It is even, vibrated in shaking table, after being vibrated 2 ~ 6 minutes by 180 ~ 360 revs/min, adjustment of rotational speed is 40 ~ 120 revs/min, temperature control
System is at 20 ~ 40 DEG C, duration of oscillation 36 ~ 54 hours, and vibration terminates rear pueraria root residue distilled water flushing 3 ~ 5 times.
Immobilized cellulase described in step (2) of the present invention, process for fixation are as follows：Cellulase is dissolved in 0.01
~ 0.04 mol/L phosphate buffer, pH of cushioning fluid are 5.0 ~ 6.0, and enzyme concentration is 60 ~ 90 g/l；Pueraria root residue after pretreatment
With cellulase solution with mass ratio 1:2 ~ 5 mix, and are vibrated in shaking table, after being vibrated 2 ~ 6 minutes by 180 ~ 360 revs/min, turn
Speed is adjusted to 40 ~ 120 revs/min, and for temperature control at 20 ~ 40 DEG C, duration of oscillation 36 ~ 54 hours, vibration secures fibre after terminating
Tie up the pueraria root residue of plain enzyme distilled water flushing 3 ~ 5 times.
Continuous feed and discharge method are taken in step (3) of the present invention, speed is consistent, and is 5 ~ 7 kgs/hr.
Immobilization alkali protease described in step (3) of the present invention, enzyme immobilizatio method are as follows：Alkali protease is molten
For solution in 0.01 ~ 0.04 mol/L phosphate buffer, pH of cushioning fluid is 6.5 ~ 7.5, and enzyme concentration is 70 ~ 100 g/l；Pretreatment
Pueraria root residue and alkaline protease solution are with mass ratio 1 afterwards:2 ~ 4 mix, vibrated in shaking table, by 190 ~ 380 revs/min vibration 3 ~
After 6 minutes, adjustment of rotational speed is 60 ~ 120 revs/min, and temperature control is at 30 ~ 40 DEG C, and duration of oscillation 54 ~ 72 hours, vibration terminates
The pueraria root residue of alkali protease distilled water flushing 3 ~ 5 times is secured afterwards.
First step enzyme digestion reaction described in step (2) of the present invention is mainly more by the marine alga in the structures such as undaria pinnitafida cell membrane
Sugar, which changes into the products such as the high oligosaccharides of the utilization ratios such as crop, alginic acid, can induce the expression of the differential proteins such as crop, preventing and treating
The related disease such as crop.
Second step enzyme digestion reaction described in step (3) of the present invention, primarily to undaria pinnitafida protein is catalytically converted into
The products such as specific polypeptide, amino acid.
Pericarp described in step (5) of the present invention is banana skin, watermelon peel or apple skin, waste dish be Chinese cabbage, romaine lettuce or
Lettuce waste dish, it is brewed spirit waste residue to brewage waste residue.
The present invention establishes a kind of alga fertilizer preparation method using undaria pinnitafida as raw material, is catalyzed based on immobilised enzymes two step method
Undaria pinnitafida is converted, finally establishes the continuous production processes that undaria pinnitafida enzymolysis prepares alga fertilizer, enzymolysis liquid kills centrifuged after enzyme
Supernatant and undaria pinnitafida residue, supernatant through falling film evaporation into concentrate be liquid seaweed extract；Undaria pinnitafida digests residue and oil tea
The common fermentations such as grouts reach the purpose that the full cell of undaria pinnitafida utilizes into solid organic fertilizer.The undaria pinnitafida level of resources utilization is improved, is protected
Environment has been protected, has realized that enzyme is repeated and is used continuously, greatly reduces the consumption of enzyme, greatly reduce alga fertilizer production cost, it is practical
Property is very strong, is easy to scale, and it is good and be all-natural product to produce alga fertilizer quality, pollution-free degradable, is that one kind meets work
Industry demand, environmentally friendly new method.
Embodiment
The present invention will be described further by following examples.
Undaria pinnitafida and cake of camellia oleifera seeds used in following examples are commercially available.
Pericarp and waste dish used in following examples are respectively collected from fruit wholesale market and the market of farm produce.
Chicken manure used in following examples is collected from chicken farm.
Cellulase and alkali protease used in following examples are purchased from Sigma-Aldrich companies.
Embodiment 1.
Undaria pinnitafida is cleaned with running water, then soaked 54 hours with running water, changes water 1 time within during which every 6 hours；After immersion
Undaria pinnitafida is shredded (wide with knife：0.5 cm, it is long：5 cms), broken undaria pinnitafida crushes through colloid mill.Undaria pinnitafida after crushing
Mud is with distilled water with volume ratio 1:2 are mixed, and suspension is handled through high pressure homogenizer；Homogenizer condition of work：60 MPas of pressure, is followed
Ring number 3 times, 1000 ls/h of flow, room temperature condition.Pueraria root residue preprocess method caused by kudzu-vine root powder production process：With steaming
Distilled water is rinsed 3 times, is placed in beaker, and pueraria root residue is with 40% Eusol with mass ratio 1:1 mixes, and vibrates, presses in shaking table
After 180 revs/min vibrate 2 minutes, adjustment of rotational speed is 40 revs/min, and temperature control duration of oscillation 36 hours, vibrates at 20 DEG C
Pueraria root residue distilled water flushing 3 times after end, for immobilised enzymes.Cellulase immobilization method：Cellulase is dissolved in
0.01 mol/L phosphate buffer, pH of cushioning fluid 5.0, enzyme concentration are 60 g/l；Pueraria root residue and cellulose after pretreatment
Enzyme solutions are with mass ratio 1:2 mix, and are vibrated in shaking table, and after being vibrated 2 minutes by 180 revs/min, adjustment of rotational speed is 40 revs/min
Clock, for temperature control at 20 DEG C, duration of oscillation 36 hours, vibration secures the pueraria root residue distilled water flushing of cellulase after terminating
3 times.The process for fixation of alkali protease is as follows：Alkali protease is dissolved in 0.01 mol/L phosphate buffer, pH of buffer
It is worth for 6.5, enzyme concentration is 70 g/l；Pretreated pueraria root residue is with alkaline protease solution with mass ratio 1:2 mix, and are shaking
Vibrated in bed, after 190 revs/min of vibrations 3 minutes, adjustment of rotational speed is 60 revs/min, and temperature control is at 30 DEG C, duration of oscillation
54 hours, vibration secured the pueraria root residue of alkali protease distilled water flushing 3 times after terminating.
Undaria pinnitafida homogenate and distilled water are with mass ratio 1 after pretreatment:2 mix, and it is anti-to be placed in 50 liters of continuous flowing agitator tank formulas
Answer in device, immobilized cellulase is placed in the wire netting inframe rotated together with agitating shaft, according to undaria pinnitafida and enzyme mass ratio 5:1
Immobilised enzymes is added, adjusts pH to 5.0,160 revs/min of speed of agitator, after stirring 3 minutes, speed of agitator is adjusted to 40 revs/min
Clock, temperature control is at 40 DEG C；After immobilised enzymes Biocatalytic Conversion 2 hours, start continuous feed and discharging, input and output material speed are equal
For 4 kgs/hr, reaction terminates to stop charging and discharging in first 1 hour.It is anti-that discharging enters another 50 liters continuous flowing agitator tank formulas
Answer in device, immobilization alkali protease is placed in the wire netting inframe rotated together with agitating shaft, by the mg/litre of enzyme concentration 12000
Immobilised enzymes is added, adjusts pH to 6.0, after being stirred 3 minutes by 170 revs/min, speed of agitator is adjusted to 60 revs/min, temperature
Control is at 45 DEG C；After reaction starting 2 hours, it is 5 kgs/hr to start continuous feed and discharging, input and output material speed, reaction knot
Stop charging and discharging within 1 hour before beam.The enzymolysis liquid of discharging is through centrifugation（6000 revs/min, 3 minutes, room temperature condition）Obtain
Clear liquid and undaria pinnitafida residue, supernatant falling film evaporation technology are concentrated, 10 ls/h of evaporation capacity, temperature 45 C, pressure 0.02
MPa, cocnentration factor 2.1, concentrate is through ultrafiltration (0.02 micron of membrane aperture, temperature room temperature, 0.4 MPa of pressure, cross-flow rate 3.6
Meter per second, 800 liters/square m/h) it is to obtain liquid seaweed extract afterwards.Undaria pinnitafida residue and cake of camellia oleifera seeds, banana skin, Chinese cabbage
Waste dish, brewed spirit waste residue, chicken manure are with mass ratio 1:3:4:5:2:4 heap fermentations, 12 cubic metres of volume, 40 centimetres of height, temperature
45 DEG C of degree, temperature, fermentation time 30 days are controlled by translating into fermentation materials.Fermentation ends obtain high-quality solid organic fertilizer.
Embodiment 2.
Undaria pinnitafida is cleaned with running water, then soaked 54 hours with running water, changes water 1 time within during which every 6 hours；After immersion
Undaria pinnitafida is shredded (wide with knife：0.5 cm, it is long：5 cms), broken undaria pinnitafida crushes through colloid mill.Undaria pinnitafida after crushing
Mud is with distilled water with volume ratio 1:4 are mixed, and suspension is handled through high pressure homogenizer；Homogenizer condition of work：80 MPas of pressure, is followed
Ring number 5 times, 1000 ls/h of flow, room temperature condition.Pueraria root residue preprocess method caused by kudzu-vine root powder production process：With steaming
Distilled water is rinsed 5 times, is placed in beaker, and pueraria root residue is with 50% Eusol with mass ratio 1:3 mix, and vibrate, press in shaking table
After 360 revs/min vibrate 6 minutes, adjustment of rotational speed is 120 revs/min, and temperature control duration of oscillation 54 hours, vibrates at 40 DEG C
Pueraria root residue distilled water flushing 5 times after end, for immobilised enzymes.Cellulase immobilization method：Cellulase is dissolved in
0.04 mol/L phosphate buffer, pH of cushioning fluid 6.0, enzyme concentration are 90 g/l；Pueraria root residue and cellulose after pretreatment
Enzyme solutions are with mass ratio 1:5 mix, and are vibrated in shaking table, and after being vibrated 6 minutes by 360 revs/min, adjustment of rotational speed is 120 revs/min
Clock, for temperature control at 40 DEG C, duration of oscillation 54 hours, vibration secures the pueraria root residue distilled water flushing of cellulase after terminating
5 times.The process for fixation of alkali protease is as follows：Alkali protease is dissolved in 0.04 mol/L phosphate buffer, pH of buffer
It is worth for 7.5, enzyme concentration is 100 g/l；Pretreated pueraria root residue is with alkaline protease solution with mass ratio 1:4 mix, and are shaking
Vibrated in bed, after 380 revs/min of vibrations 6 minutes, adjustment of rotational speed is 120 revs/min, and temperature control is at 40 DEG C, duration of oscillation
72 hours, vibration secured the pueraria root residue of alkali protease distilled water flushing 5 times after terminating.
Undaria pinnitafida homogenate and distilled water are with mass ratio 1 after pretreatment:5 mix, and it is anti-to be placed in 50 liters of continuous flowing agitator tank formulas
Answer in device, immobilized cellulase is placed in the wire netting inframe rotated together with agitating shaft, according to undaria pinnitafida and enzyme mass ratio 20:
1 addition immobilised enzymes, adjust pH to 6.0,320 revs/min of speed of agitator, stirring 9 minutes after, speed of agitator be adjusted to 120 turns/
Minute, temperature control is at 60 DEG C；After immobilised enzymes Biocatalytic Conversion 4 hours, start continuous feed and discharging, input and output material speed
It it is 6 kgs/hr, reaction terminates to stop charging and discharging in first 2 hours.Discharging enters another 50 liters continuous flowing agitator tank formulas
In reactor, immobilization alkali protease is placed in the wire netting inframe rotated together with agitating shaft, by 60000 milligrams of enzyme concentration/
Addition immobilised enzymes is risen, adjusts pH to 7.5, after being stirred 8 minutes by 340 revs/min, speed of agitator is adjusted to 120 revs/min,
Temperature control is at 65 DEG C；After reaction starting 4 hours, it is 6 kgs/hr to start continuous feed and discharging, input and output material speed, instead
It should terminate to stop charging and discharging in first 2 hours.The enzymolysis liquid of discharging is through centrifugation（12000 revs/min, 6 minutes, room temperature condition）
To supernatant and undaria pinnitafida residue, supernatant falling film evaporation technology is concentrated, 20 ls/h of evaporation capacity, 65 DEG C of temperature, pressure
0.08 MPa of power, cocnentration factor 3.5, concentrate is through ultrafiltration (0.02 micron of membrane aperture, temperature room temperature, 0.4 MPa of pressure, cross-flow speed
The meter per second of rate 3.6,800 liters/square m/h) it is to obtain liquid seaweed extract afterwards.Undaria pinnitafida residue and cake of camellia oleifera seeds, apple skin,
Lettuce waste dish, brewed spirit waste residue, chicken manure are with mass ratio 1:5:8:7:4:6 heap fermentations, 18 cubic metres of volume, 70 lis of height
Rice, 65 DEG C of temperature, temperature, fermentation time 50 days is controlled by translating into fermentation materials.It is organic that fermentation ends obtain high-quality solid-state
Fertilizer.
Embodiment 3.
Undaria pinnitafida is cleaned with running water, then soaked 54 hours with running water, changes water 1 time within during which every 6 hours；After immersion
Undaria pinnitafida is shredded (wide with knife：0.5 cm, it is long：5 cms), broken undaria pinnitafida crushes through colloid mill.Undaria pinnitafida after crushing
Mud is with distilled water with volume ratio 1:3 are mixed, and suspension is handled through high pressure homogenizer；Homogenizer condition of work：70 MPas of pressure, is followed
Ring number 4 times, 1000 ls/h of flow, room temperature condition.Pueraria root residue preprocess method caused by kudzu-vine root powder production process：With steaming
Distilled water is rinsed 4 times, is placed in beaker, and pueraria root residue is with 45% Eusol with mass ratio 1:2 mix, and vibrate, press in shaking table
After 270 revs/min vibrate 4 minutes, adjustment of rotational speed is 80 revs/min, and temperature control duration of oscillation 45 hours, vibrates at 30 DEG C
Pueraria root residue distilled water flushing 4 times after end, for immobilised enzymes.Cellulase immobilization method：Cellulase is dissolved in
0.03 mol/L phosphate buffer, pH of cushioning fluid 5.5, enzyme concentration are 75 g/l；Pueraria root residue and cellulose after pretreatment
Enzyme solutions are with mass ratio 1:4 mix, and are vibrated in shaking table, and after being vibrated 4 minutes by 270 revs/min, adjustment of rotational speed is 80 revs/min
Clock, for temperature control at 30 DEG C, duration of oscillation 45 hours, vibration secures the pueraria root residue distilled water flushing of cellulase after terminating
4 times.The process for fixation of alkali protease is as follows：Alkali protease is dissolved in 0.03 mol/L phosphate buffer, pH of buffer
It is worth for 7.0, enzyme concentration is 85 g/l；Pretreated pueraria root residue is with alkaline protease solution with mass ratio 1:3 mix, and are shaking
Vibrated in bed, after 275 revs/min of vibrations 5 minutes, adjustment of rotational speed is 90 revs/min, and temperature control is at 35 DEG C, duration of oscillation
63 hours, vibration secured the pueraria root residue of alkali protease distilled water flushing 4 times after terminating.
Undaria pinnitafida homogenate and distilled water are with mass ratio 1 after pretreatment:4 mix, and it is anti-to be placed in 50 liters of continuous flowing agitator tank formulas
Answer in device, immobilized cellulase is placed in the wire netting inframe rotated together with agitating shaft, according to undaria pinnitafida and enzyme mass ratio 12:
1 addition immobilised enzymes, adjust pH to 5.5,240 revs/min of speed of agitator, stirring 6 minutes after, speed of agitator be adjusted to 80 turns/
Minute, temperature control is at 50 DEG C；After immobilised enzymes Biocatalytic Conversion 3 hours, start continuous feed and discharging, input and output material speed
It it is 5 kgs/hr, reaction terminates to stop charging and discharging in first 1.5 hours.Discharging enters another 50 liters continuous flowing agitator tanks
In formula reactor, immobilization alkali protease is placed in the wire netting inframe rotated together with agitating shaft, by the milli of enzyme concentration 35000
G/l addition immobilised enzymes, adjust pH to 6.5, by 250 revs/min stirring 5 minutes after, speed of agitator is adjusted to 90 revs/min
Clock, temperature control is at 50 DEG C；After reaction starting 3 hours, starting continuous feed and discharging, input and output material speed is 5 kilograms/and it is small
When, reaction terminates to stop charging and discharging in first 1.5 hours.The enzymolysis liquid of discharging is through centrifugation（9000 revs/min, 4 minutes, room temperature
Condition）Supernatant and undaria pinnitafida residue are obtained, supernatant falling film evaporation technology is concentrated, 15 ls/h of evaporation capacity, temperature
55 DEG C, 0.05 MPa of pressure, cocnentration factor 2.8, concentrate through ultrafiltration (0.02 micron of membrane aperture, temperature room temperature, 0.4 MPa of pressure,
The meter per second of cross-flow rate 3.6,800 liters/square m/h) it is to obtain liquid seaweed extract afterwards.Undaria pinnitafida residue and cake of camellia oleifera seeds, west
Melon skin, romaine lettuce waste dish, brewed spirit waste residue, chicken manure are with mass ratio 1:4:6:6:3:5 heap fermentations, 15 cubic metres of volume, height
55 centimetres, 55 DEG C of temperature, temperature, fermentation time 40 days are controlled by translating into fermentation materials.Fermentation ends obtain high-quality solid-state
Organic fertilizer.
Comparative example 1.
Undaria pinnitafida is cleaned with running water, then soaked 54 hours with running water, changes water 1 time within during which every 4 hours；After immersion
Undaria pinnitafida is shredded (wide with knife：0.5 cm, it is long：5 cms).Broken undaria pinnitafida is with distilled water with mass ratio 1:2 mix,
It is placed in 50 liters of continuous flowing stirred tanks, does not add immobilised enzymes, adjust pH to 5.0,160 revs/min of speed of agitator
Clock, after stirring 3 minutes, speed of agitator is adjusted to 40 revs/min, and temperature control is at 40 DEG C；After reaction starting 2 hours, start to connect
Continuous charging and discharging, input and output material speed is 4 kgs/hr, and reaction terminates to stop charging and discharging in first 1 hour.Discharging enters
In another 50 liters continuous flowing stirred tanks, enzyme is not added, adjusts pH to 6.0, is stirred 3 minutes by 170 revs/min
Afterwards, speed of agitator is adjusted to 60 revs/min, and temperature control is at 45 DEG C；After reaction starting 2 hours, start continuous feed and discharging,
Input and output material speed is 5 kgs/hr, and reaction terminates to stop charging and discharging in first 1 hour.Out-feed liquid is through centrifugation（6000 revs/min
Clock, 3 minutes, room temperature condition）Supernatant and undaria pinnitafida residue are obtained, supernatant falling film evaporation technology is concentrated, evaporation capacity 10
L/h, temperature 45 C, 0.02 MPa of pressure, cocnentration factor 2.1, concentrate through ultrafiltration (0.02 micron of membrane aperture, temperature room temperature,
0.4 MPa of pressure, the meter per second of cross-flow rate 3.6,800 liters/square m/h) it is to obtain liquid seaweed extract, undaria pinnitafida residue afterwards
It is directly discarded.
Comparative example 2.
Undaria pinnitafida is cleaned with running water, then soaked 54 hours with running water, changes water 1 time within during which every 6 hours；After immersion
Undaria pinnitafida is shredded (wide with knife：0.5 cm, it is long：5 cms), broken undaria pinnitafida is with distilled water with mass ratio 1:4 mix,
It is placed in 50 liters of continuous flowing stirred tanks, does not add enzyme, adjust pH to 6.0,320 revs/min of speed of agitator, stirring
After 9 minutes, speed of agitator is adjusted to 120 revs/min, and temperature control is at 60 DEG C；After reaction starting 4 hours, start continuous feed
And discharging, input and output material speed are 6 kgs/hr, reaction terminates to stop charging and discharging in first 2 hours.Discharging enters another company
Afterflow is moved in stirred tank, does not add enzyme, adjusts pH to 7.5, after being stirred 8 minutes by 340 revs/min, speed of agitator
90 revs/min are adjusted to, temperature control is at 55 DEG C；After reaction starting 3 hours, start continuous feed and discharging, input and output material speed
It it is 6 kgs/hr, reaction terminates to stop charging and discharging in first 1.5 hours.Out-feed liquid is through centrifugation（9000 revs/min, 4 points
Clock, room temperature condition）Supernatant and undaria pinnitafida residue are obtained, supernatant falling film evaporation technology is concentrated, 15 liters of evaporation capacity/small
When, 55 DEG C of temperature, 0.05 MPa of pressure, cocnentration factor 2.8, concentrate is through ultrafiltration (0.02 micron of membrane aperture, temperature room temperature, pressure
0.4 MPa, the meter per second of cross-flow rate 3.6,800 liters/square m/h) it is to obtain liquid seaweed extract afterwards.Undaria pinnitafida residue is direct
It is discarded.
Comparative example 3.
Commercially available common foliar fertilizer.
Above example 1-3 and comparative example 1-3 application effect are as follows.
Crop field grape manure trial.
Test site：The horizontal Jiang Zhen in Jiangxi Province Ji'an City Jian County.Belong to middle subtropical zone hills mountain city monsoon moist climate, weather
Gently, sunshine is enriched, and makes a clear distinction between the four seasons, and summer in winter length, spring and autumn is short, and abundant rainfall, frost-free period is longer, average temperature of the whole year 17.5-
18.6℃。
For trying soil：Red soil, nutrient content are：The g kg of organic matter 9.80, the full mg/kg of nitrogen 186.25 are quick-acting
The mg/kg of phosphorus 5.89, the mg/kg of available potassium 45.66, soil pH 5.8.
Experimental cultivar：Kyoto grape.
Experimental design：This experiment sets 6 processing altogether, and specific processing is shown in Table 1, and often processing is repeated 3 times, random district's groups arrangement,
Experimental plot area is 667 square metres.
Test and carried out in 2016, cell sets section ridging, vine 200 plants/acre of specification of plantation, 750 grams of Seedling Stage
Foliar fertilizer, 150 kilograms are watered, is stirred, foliage-spray is once；The Berry Growth phase, 1500 grams of foliar fertilizers, 150 kilograms are watered,
Foliage-spray stir once.Base manure and each processing field management method are identical, not spraying pesticide.
Investigate the disease incidences such as the big brown spot of grape, yellowtop, black rot, cracked fruit and weight method measure vintage,
Spectrophotometry grape soluble sugar content.
The alga fertilizer of table 1 handles grape effect.
| Processing | Foliar fertilizer | Fertilization mode | The incidences of disease such as cracked fruit（%） | Vintage（Thousand grams/acre） | Soluble sugar content（Gram/100 grams） |
| 1 | Embodiment 1 | Each 1 time of seedling, Berry Growth | 5d | 2800a | 17.48c |
| 2 | Embodiment 2 | Each 1 time of seedling, Berry Growth | 7d | 2950a | 18.12b |
| 3 | Embodiment 3 | Each 1 time of seedling, Berry Growth | 7d | 2980a | 19.33a |
| 4 | Comparative example 1 | Each 1 time of seedling, Berry Growth | 35b | 2010c | 14.61e |
| 5 | Comparative example 2 | Each 1 time of seedling, Berry Growth | 41a | 2080c | 14.75e |
| 6 | Comparative example 3 | Each 1 time of seedling, Berry Growth | 30c | 2520b | 15.80d |
Field rice manure trial.
Test site：The horizontal Jiang Zhen in Jiangxi Province Ji'an City Jian County.Belong to middle subtropical zone hills mountain city monsoon moist climate, weather
Gently, sunshine is enriched, and makes a clear distinction between the four seasons, and summer in winter length, spring and autumn is short, and abundant rainfall, frost-free period is longer, average temperature of the whole year 17.5-
18.6℃。
For trying soil：Red soil, nutrient content are：The g kg of organic matter 9.60, the full mg/kg of nitrogen 186.25 are quick-acting
The mg/kg of phosphorus 5.89, the mg/kg of available potassium 45.66, soil pH 5.8.
Experimental cultivar：Y two excellent 1.
Experimental design：This experiment sets 6 processing altogether, and specific processing is shown in Table 1, and often processing is repeated 3 times, random district's groups arrangement,
Experimental plot area is 2000 square metres.
Test and carried out in 2016, cell sets section ridging, conventional to plant specification, 1500 grams of foliar fertilizers of Seedling Stage, converts
150 kilograms of water, stirs, and foliage-spray is once；Ear growth period, 3000 grams of foliar fertilizers, be watered 150 kilograms, stir
Foliage-spray is once.Base manure and each processing field management method are identical, not spraying pesticide.
Investigate rice blast, the sheath and culm blight of rice, the sheath and culm blight of rice, etc. disease incidence and weight method measure rice yield.
The alga fertilizer of table 2 handles rice effect.
| Processing | Foliar fertilizer | Fertilization mode | The incidences of disease such as rice blast（%） | Rice yield（Thousand grams/acre） |
| 1 | Embodiment 1 | Each 1 time of seedling, heading growth | 7c | 800a |
| 2 | Embodiment 2 | Each 1 time of seedling, heading growth | 7c | 820b |
| 3 | Embodiment 3 | Each 1 time of seedling, heading growth | 8c | 930a |
| 4 | Comparative example 1 | Each 1 time of seedling, heading growth | 21b | 500d |
| 5 | Comparative example 2 | Each 1 time of seedling, heading growth | 22b | 680c |
| 6 | Comparative example 3 | Each 1 time of seedling, heading growth | 29a | 770c |
Crop field tomato manure trial.
Test site：The horizontal Jiang Zhen in Jiangxi Province Ji'an City Jian County.Belong to middle subtropical zone hills mountain city monsoon moist climate, weather
Gently, sunshine is enriched, and makes a clear distinction between the four seasons, and summer in winter length, spring and autumn is short, and abundant rainfall, frost-free period is longer, average temperature of the whole year 17.5-
18.6℃。
For trying soil：Red soil, nutrient content are：The g kg of organic matter 9.80, the full mg/kg of nitrogen 186.25 are quick-acting
The mg/kg of phosphorus 5.89, the mg/kg of available potassium 45.66, soil pH 5.8.
Experimental cultivar：Rosy clouds powder tomato.
Experimental design：This experiment sets 6 processing altogether, and specific processing is shown in Table 1, and often processing is repeated 3 times, random district's groups arrangement,
Experimental plot area is 667 square metres.
Test and carried out in 2016, cell sets section ridging, tomato 500 plants/acre of specification of plantation, 500 grams of Seedling Stage
Foliar fertilizer, 100 kilograms are watered, is stirred, foliage-spray is once；Bear fruit growth period, 1000 grams of foliar fertilizers, be watered 100 kilograms,
Foliage-spray stir once.Base manure and each processing field management method are identical, not spraying pesticide.
Investigate the disease incidences such as tomato droop, sclerotiniose, verticillium wilt and weight method measure tomato production.
The alga fertilizer of table 3 handles tomato effect.
| Processing | Foliar fertilizer | Fertilization mode | The incidences of disease such as droop（%） | Tomato production（Thousand grams/acre） |
| 1 | Embodiment 1 | Each 1 time of seedling, Berry Growth | 8c | 4550a |
| 2 | Embodiment 2 | Each 1 time of seedling, Berry Growth | 8c | 4430a |
| 3 | Embodiment 3 | Each 1 time of seedling, Berry Growth | 6c | 4560a |
| 4 | Comparative example 1 | Each 1 time of seedling, Berry Growth | 26b | 3580c |
| 5 | Comparative example 2 | Each 1 time of seedling, Berry Growth | 26b | 3550c |
| 6 | Comparative example 3 | Each 1 time of seedling, Berry Growth | 37a | 3790b |
Different fertilization is can be seen that to the disease such as grape fruit split incidence of disease, grape from table 1,2 and 3 result of the tests
The influence of yield and soluble sugar content reaches significant difference level, and soluble sugar content height is related to grape quality.It is solvable
Property sugared content is higher, and grape quality is better.Different fertilization is to the disease such as rice blast incidence of disease and tomato droop
Influence etc. the disease incidence of disease, rice and tomato production reaches significant difference level.Crop field grape, rice and tomato fertilizer efficiency
It is all highest that the alga fertilizer that test example 2 is prepared, which applies rear grape, rice and tomato production, grape, rice and west
The red persimmon incidence of disease is all minimum.It can be drawn from result of the test, alga fertilizer of the present invention can promote grape, rice and tomato life
It is long, effectively prevent and treat grape, rice and tomato disease.
Claims (7)
- A kind of 1. alga fertilizer preparation method using undaria pinnitafida as raw material, it is characterized in that according to the following steps：(1) undaria pinnitafida pre-processes：Undaria pinnitafida is cleaned with running water；Soaked 54 hours, changed with running water again within during which every 6 hours Water 1 time；Undaria pinnitafida after immersion is shredded with knife, and broken undaria pinnitafida crushes through colloid mill；Rear skirt band puree is crushed with distilled water with body Product ratio 1:2 ~ 4 are mixed, and suspension is handled through high pressure homogenizer；Homogenizer condition of work：60 ~ 80 MPas of pressure, cycle-index 3 ~ 5 It is secondary, 1000 ls/h of flow, room temperature condition；(2) first step enzyme digestion reaction：Undaria pinnitafida homogenate and distilled water are with mass ratio 1 after pretreatment:2 ~ 5 mix, and are placed in continuous Flow in stirred tank, through pueraria root residue immobilization, immobilized cellulase is placed in be turned cellulase together with agitating shaft Dynamic wire netting inframe, according to undaria pinnitafida and cellulase mass ratio 5 ~ 20:1 addition immobilized cellulase, adjusts pH to 5.0 ~ 6.0,160 ~ 320 revs/min of speed of agitator, after stirring 3 ~ 9 minutes, speed of agitator is adjusted to 40 ~ 120 revs/min, temperature control System is at 40 ~ 60 DEG C；After immobilized cellulase Biocatalytic Conversion 2 ~ 4 hours, start continuous feed and discharging, before reaction terminates Stop charging and discharging within 1 ~ 2 hour；Second step enzyme digestion reaction：Discharging enters in another continuous flowing stirred tank in step (2), alkali protease warp Pueraria root residue immobilization, immobilization alkali protease is placed in the wire netting inframe rotated together with agitating shaft, dense by alkali protease 12000 ~ 60000 mg/litres addition immobilization alkali protease is spent, adjusts pH to 6.0 ~ 7.5, by 170 ~ 340 revs/min of stirrings After 3 ~ 8 minutes, speed of agitator is adjusted to 60 ~ 120 revs/min, and temperature control is at 45 ~ 65 DEG C；After reaction starting 2 ~ 4 hours, open Beginning continuous feed and discharging, reaction terminate to stop charging and discharging in first 1 ~ 2 hour；(4) under the enzymolysis liquid room temperature condition that step (3) obtains, 6000 ~ 12000 revs/min, centrifuge 3 ~ 6 minutes, obtain supernatant Liquid and undaria pinnitafida residue, supernatant falling film evaporation technology are concentrated, 10 ~ 20 ls/h of evaporation capacity, 45 ~ 65 DEG C of temperature, pressure 0.02 ~ 0.08 MPa of power, cocnentration factor 2.1 ~ 3.5, concentrate obtain liquid seaweed extract after ultrafiltration；(5) the undaria pinnitafida residue in step (4) and cake of camellia oleifera seeds, pericarp, waste dish, brewage waste residue, chicken manure with mass ratio 1:3~5: 4~8:5~7:2~4:4 ~ 6 heap fermentations, 12 ~ 18 cubic metres of volume, 40 ~ 70 centimetres of height, 45 ~ 65 DEG C of temperature, by translating into hair Ferment material controls temperature, fermentation time 30 ~ 50 days；Fermentation ends obtain high-quality solid organic fertilizer.
- A kind of 2. alga fertilizer preparation method using undaria pinnitafida as raw material according to claim 1, it is characterized in that in step (2) Continuous feed and discharge method are taken, speed is consistent, and is 4 ~ 6 kgs/hr.
- A kind of 3. alga fertilizer preparation method using undaria pinnitafida as raw material according to claim 1, it is characterized in that in step (2) The pueraria root residue preprocess method is：The distilled water flushing 3 ~ 5 times of pueraria root residue caused by kudzu-vine root powder production process, is placed in beaker In, pueraria root residue is with 40% ~ 50% Eusol with mass ratio 1:1 ~ 3 mixes, and vibrates in shaking table, is shaken by 180 ~ 360 revs/min After swinging 2 ~ 6 minutes, adjustment of rotational speed is 40 ~ 120 revs/min, and temperature control duration of oscillation 36 ~ 54 hours, vibrates at 20 ~ 40 DEG C Pueraria root residue distilled water flushing 3 ~ 5 times after end.
- A kind of 4. alga fertilizer preparation method using undaria pinnitafida as raw material according to claim 1, it is characterized in that in step (2) The cellulase immobilization method is：Cellulase is dissolved in 0.01 ~ 0.04 mol/L phosphate buffer, pH of cushioning fluid For 5.0 ~ 6.0, enzyme concentration is 60 ~ 90 g/l；Pueraria root residue and cellulase solution are with mass ratio 1 after pretreatment:2 ~ 5 mix, Vibrated in shaking table, after being vibrated 2 ~ 6 minutes by 180 ~ 360 revs/min, adjustment of rotational speed is 40 ~ 120 revs/min, and temperature control is 20 ~ 40 DEG C, duration of oscillation 36 ~ 54 hours, vibration secures the pueraria root residue of cellulase distilled water flushing 3 ~ 5 times after terminating.
- A kind of 5. alga fertilizer preparation method using undaria pinnitafida as raw material according to claim 1, it is characterized in that in step (3) Continuous feed and discharge method are taken, speed is consistent, and is 5 ~ 7 kgs/hr.
- A kind of 6. alga fertilizer preparation method using undaria pinnitafida as raw material according to claim 1, it is characterized in that in step (3) Described basic protein enzyme immobilization method is as follows：Alkali protease is dissolved in 0.01 ~ 0.04 mol/L phosphate buffer, delays Fliud flushing pH value is 6.5 ~ 7.5, and enzyme concentration is 70 ~ 100 g/l；Pueraria root residue and alkaline protease solution are with mass ratio after pretreatment 1:2 ~ 4 mix, and are vibrated in shaking table, and after being vibrated 3 ~ 6 minutes by 190 ~ 380 revs/min, adjustment of rotational speed is 60 ~ 120 revs/min, For temperature control at 30 ~ 40 DEG C, duration of oscillation 54 ~ 72 hours, vibration secures the pueraria root residue distillation of alkali protease after terminating Water rinses 3 ~ 5 times.
- A kind of 7. alga fertilizer preparation method using undaria pinnitafida as raw material according to claim 1, it is characterized in that in step (5) Described pericarp is banana skin, watermelon peel or apple skin, and waste dish is Chinese cabbage, romaine lettuce or lettuce waste dish, and it is white wine to brewage waste residue Brewage waste residue.
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