CN102992915A - Method for preparing environment-friendly and high-efficiency biological organic fertilizer by using distilled spirit vinasse - Google Patents
Method for preparing environment-friendly and high-efficiency biological organic fertilizer by using distilled spirit vinasse Download PDFInfo
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Abstract
The invention provides a method for preparing environment-friendly and high-efficiency biological organic fertilizer by using distilled spirit vinasse, comprising the following steps of: puffing fermentation substrate by a threaded rod type puffing machine; transporting into a wet material mixing machine, cooling and adding microelement mineral salt, and inoculating bacterium liquid in proportion when the temperature is reduced to 30-35DEG C, completely mixing; and filling the inoculated and mixed fermentation substrate into a novel silica gel breathable membrane fermentation bag, packaging and sealing by an automatic metering and packing system, and fermenting for 5-7 at constant temperature in a fermentation room to obtain a finished product. The fermentation substrate is prepared by evenly mixing 50-60 parts by weight of distilled spirit vinasse, 5-10 parts by weight of humic acid, 5-15 parts by weight of sunflower husk powder, 5-10 parts by weight of ammonium sulfate, 5-10 parts by weight of potassium chloride and 5-10 parts by weight of corn steep liquor, wherein the water content of the fermentation substrate is 30-40 wt%; and the mixed bacterium liquid comprises bacterium liquid of streptomyces jingyangensis, clostridium butyricum, pseudomonas mendocina, azotobacter chroococcum and rhizopus oryzae.
Description
Technical field
The invention relates to a kind of method of utilizing distillers ' grains to produce the environment-friendly high-efficiency bio-organic fertilizer, belong to microorganism compound organic fertilizer technical field.
Background technology
Chinese liquor distiller grains is the byproduct of wine-making industry.According to statistics, China produces Chinese liquor distiller grains per year and reaches tens million of tons, and amount is concentrated greatly, if untimely the processing, will be putrid and deteriorated, not only wasted valuable resource, also can the severe contamination surrounding environment.Therefore, the comprehensive utilization of vinasse is of great significance development of resources and the environment protection tool of China.At present China's research has in this regard obtained certain achievement, for example, utilizes distillers ' grains to produce glycerine, cultivate edible mushrooms, extract aminoacids complex and trace element, extract phytic acid and phytic acid ca, make vinegar, extract protein, produce amylase and cellulase, utilize the vinasse anaerobically fermenting to reclaim biogas, produce the SCP feed, produce craboraffin, make the dyeing reductive agent with the lime fermentation, be used for former wine and produce, external application is used for the treatment of sacroiliitis.Recently have report to produce aminobutyric acid take vinasse as raw material with short lactobacillus, the liquid vinasse also can be used for cultivating Bacillus thuringiensis, produce biogas, recovery waste heat.Utilize the liquor-making byproduct yellow water to improve vinosity etc., but this is wherein except being used as feed, additive method all can not fundamentally solve the final whereabouts problem of vinasse, and might produce more poor slag, in addition vinasse complicated component, water content is high, and difficult storage has again the fermentation weighting materials such as rice husk that are difficult for utilization, so still there is certain difficulty in the utilization of Chinese liquor distiller grains.Therefore, Comprehensive utilizes vinasse, prevents from polluting, avoids waste, turning waste into wealth becomes the emphasis that we study present stage.
Summary of the invention
The object of the invention is to, adapt to above-mentioned form, a kind of method of utilizing distillers ' grains to produce the environment-friendly high-efficiency bio-organic fertilizer is provided.
For achieving the above object, the present invention is by the following technical solutions:
A kind of method of utilizing distillers ' grains to produce the environment-friendly high-efficiency bio-organic fertilizer is characterized in that,
(1) fermentation substrate is carried out puffing by the screw bulking machine, and then be transported to and lower the temperature in the wet feed mixing machine and add micro-mineral salt, treat to inoculate in proportion when temperature drops to 30～35 ℃ mixed bacteria liquid and fully mix;
(2) fermentation substrate of inoculating and mixing is loaded in the silica gel respiratory membrane fermentation bag by the automatic gauge packaging system and packs, seals, and constant temperature bottom fermentation 5～7d in proving room is described environment-friendly high-efficiency bio-organic fertilizer finished product;
Described fermentation substrate be with: distillers ' grains 50～60 weight parts, humic acids 5～10 weight parts, Sunflower shell powder 5～15 weight parts, ammonium sulfate 5～10 weight parts, Repone K 5～10 weight parts, corn steep liquor 5～10 weight parts mix the fermentation substrate of making, and the fermentation substrate moisture content is 30～40wt%;
Described mixed bacteria liquid comprises the bacterium liquid of Jingyang streptomycete, clostridium butylicum, pseudomonas mendocina, blown-ball Azotobacter and Rhizopus oryzae.
Aforesaid method, preferably, in the step (1), described fermentation substrate carries out extruding puffing by the saturation steam of twin-screw extruder through 0.3～0.4MPa under 140～150 ℃ of conditions.
Aforesaid method, preferably, in the step (1), described fermentation substrate is transported to the wet feed mixing machine after puffing, passes into pressurized air and makes the fermentation substrate temperature be down to 30～35 ℃, adds simultaneously described micro-mineral salt: MnSO
4H
2O0.05～0.1%, CaCO
31～2%, MgSO
47H
2O0.1～0.2%, ZnSO
40.05～0.1% and FeSO
40.1～0.2%, fully mix, described percentage ratio accounts for the per-cent of fermentation substrate weight for each micro-mineral salt.
Aforesaid method, preferably, in the described mixed bacteria liquid, the inoculum size of Jingyang streptomycete, clostridium butylicum, pseudomonas mendocina, blown-ball Azotobacter and Rhizopus oryzae is respectively 2～5%, 5～10%, 5～10%, 2～5% and 5～10% of fermentation substrate weight percent; Fully mix 2～5min after the fermentation substrate inoculation.
Aforesaid method, preferably, in the step (2), after described fermentation substrate inoculation and the mixing, be loaded in the silica gel respiratory membrane fermentation bag by the automatic gauge packaging system and pack, to seal, packaged material is 30～35 ℃ of constant temperature bottom fermentation 5～7d in proving room, are described environment-friendly high-efficiency bio-organic fertilizer finished product.
Aforesaid method, preferably, described silica gel respiratory membrane fermentation bag, the bag sealing two ends is circular acerous design.
Aforesaid method, preferably, described wet feed mixing machine, twin-screw extruder and automatic gauge packaging system are the 316L stainless steel.
Aforesaid method, preferably, described bacterial classification is made by the following method:
(1) preparation method of described Jingyang streptomycete bacterium liquid is:
Preparation Jingyang streptomycete first order seed substratum: KNO
31g, NaCl0.5g, K
2HPO
40.5g, FeSO
47H
2O0.01g, MgSO
47H
2O0.5g, Zulkovsky starch 20g, distilled water 1000mL, pH7.2～7.4 are at 115～121 ℃ of lower sterilization 20min～30min of temperature;
Preparation Jingyang streptomycete secondary seed medium and fermention medium: glucose 20kg, soybean cake powder 20kg, NaCl3kg, CaCO
31kg, sterile pure water 1000L, pH7.2～7.4 are at 115～121 ℃ of lower sterilization 20～30min of temperature;
Jingyang streptomycete first order seed is cultivated: dress Jingyang streptomycete seed substratum 300mL in the 1000mL triangular flask, it is 1.0～2.0 * 10 that concentration is made on access streptomyces species inclined-plane, Jingyang
7Spore suspension 25～50mL of individual/mL, 28～30 ℃ of temperature, rotating speed 150～170r/min, shaking culture 24～48h;
Jingyang streptomycete secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through Jingyang streptomycete first order seed is inoculated in the streptomycete secondary seed tank of Jingyang according to 5～10% weight ratio, be under the condition of 1:1～1.5, with 100～120r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
Jingyang streptomycete liquid fermentation and culture: the above-mentioned bacterium liquid of cultivating through Jingyang streptomycete secondary seed is inoculated in the streptomycete fermentation tank of Jingyang according to 5～10% weight ratio, be under 1:1～1.5 conditions, with 100～120r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
(2) preparation method of described clostridium butylicum bacterium liquid is:
Preparation clostridium butylicum first order seed substratum: Tryptones 20g, beef extract 10g, yeast extract paste 6g, glucose 4g, dipotassium hydrogen phosphate 2g, potassium primary phosphate 1g, sal epsom 0.4g, calcium chloride 0.2g, ferrous sulfate 0.1g, cysteine hydrochloride 0.5g, distilled water 1000mL, pH7.2～7.4 are at 115～121 ℃ of lower sterilization 20min～30min of temperature;
Preparation clostridium butylicum secondary seed medium and fermention medium: glucose 10kg, Tryptones 10kg, yeast extract 5kg, beef extract 3kg, K
2HPO
42kg, MgSO
47H
2O0.5kg, MnSO
4H
2O0.2kg, NaHCO1kg, CaCO
31kg, distilled water 1000L mixes, and pH6.8～7.0 are at 115～121 ℃ of lower sterilization 20min～30min of temperature;
The clostridium butylicum first order seed is cultivated: dress clostridium butylicum first order seed substratum 900mL in the 1000mL triangular flask, the clostridium butylicum strain inclined plane is inoculated in the clostridium butylicum first order seed substratum in the ratio of ring/50～100mL, and 35～37 ℃ of temperature leave standstill cultivates 24h～48h;
The clostridium butylicum secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through the clostridium butylicum first order seed is inoculated in the clostridium butylicum secondary seed tank according to 5～10% weight ratio, and 35～37 ℃ of temperature leave standstill cultivates 24h～48h;
The clostridium butylicum liquid fermentation and culture: the above-mentioned bacterium liquid of cultivating through the clostridium butylicum secondary seed is inoculated in the clostridium butylicum fermentor tank according to 5～10% weight ratio, and 35～37 ℃ of temperature leave standstill cultivates 24h～48h;
(3) preparation method of described pseudomonas mendocina bacterium liquid is:
Preparation pseudomonas mendocina first order seed substratum: peptone 10g, beef extract 10g, NaCl5g, glucose 10g, distilled water 1000mL, pH6.8～7.0 are at 115～121 ℃ of lower sterilization 20min～30min of temperature;
Preparation pseudomonas mendocina secondary seed medium and fermention medium: peptone 10kg, yeast soak powder 10kg, glucose 10kg, NaCl2kg, distilled water 1000L, and pH6.8～7.0 are at 115～121 ℃ of lower sterilization 20min～30min of temperature;
The pseudomonas mendocina first order seed is cultivated: dress pseudomonas mendocina first order seed substratum 500mL in the 1000mL triangular flask, the pseudomonas mendocina strain inclined plane is inoculated in the pseudomonas mendocina first order seed substratum 28～30 ℃ of temperature, rotating speed 130～150r/min, shaking culture 24～48h in the ratio of ring/50～100mL;
The pseudomonas mendocina secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through the pseudomonas mendocina first order seed is inoculated in the pseudomonas mendocina secondary seed tank according to 5～10% weight ratio, be under the condition of 1:1～1.2, with 130～150r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
Pseudomonas mendocina liquid fermentation and culture: the above-mentioned bacterium liquid of cultivating through the pseudomonas mendocina secondary seed is inoculated in the pseudomonas mendocina fermentor tank according to 5～10% weight ratio, be under the condition of 1:1～1.2, with 130～150r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
(4) preparation method of described blown-ball Azotobacter bacterium liquid is:
Preparation blown-ball Azotobacter seed culture medium: K
2HPO
40.8g, KH
2PO
40.2g, MgSO
47H
2O0.2g,
NaCl0.5g, CaCl
20.1g, N.F,USP MANNITOL 20g, yeast extract paste 0.5g, FeCl
30.005g, Na
2MoO
4.2H
2O0.002g, distilled water 1000mL, pH7.0～7.2 are at 115～121 ℃ of lower sterilization 20～30min of temperature;
The blown-ball Azotobacter first order seed is cultivated: dress blown-ball Azotobacter seed culture medium 500mL in the 1000mL triangular flask, the blown-ball Azotobacter strain inclined plane is inoculated in the blown-ball Azotobacter first order seed substratum 28～30 ℃ of temperature, rotating speed 100～120r/min, shaking culture 24～48h in the ratio of ring/50～100mL;
The blown-ball Azotobacter secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through the blown-ball Azotobacter first order seed is inoculated in the blown-ball Azotobacter secondary seed tank according to 5～10% weight ratio, be under the condition of 1:0.5～1, with 100～120r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
Blown-ball Azotobacter liquid fermentation and culture: the above-mentioned bacterium liquid of cultivating through the blown-ball Azotobacter secondary seed is inoculated in the blown-ball Azotobacter fermentor tank according to 5～10% weight ratio, be under the condition of 1:0.5～1, with 100～120r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
(5) preparation method of described Rhizopus oryzae liquid is:
Preparation Rhizopus oryzae first order seed substratum: glucose sugar 20g, potato leaching liquid 1000mL, the pH nature is at 115～121 ℃ of lower sterilization 20～30min of temperature;
Wherein, the preparation method of potato leaching liquid is: remove the potato 200g of skin, be cut into small pieces, add water 1000mL and boil 30min elimination potato ball, filtrate is complemented to 1000mL;
Preparation Rhizopus oryzae secondary seed medium and fermention medium: yam starch 100kg, (NH
4)
2SO
41.5kg, MgSO
47H
2O0.25kg, KH
2PO
40.3kg, CaCO
360kg, ZnSO
47H
2O0.05kg sterile pure water 1000L, the pH nature is at 115～121 ℃ of lower sterilization 20～30min of temperature;
The Rhizopus oryzae first order seed is cultivated: dress Rhizopus oryzae seed culture medium 250mL in the 1000mL triangular flask, it is 2.5～4.0 * 10 that concentration is made on access Rhizopus oryzae bacterial classification inclined-plane
7Spore suspension 10～30mL of individual/mL, 25～28 ℃ of temperature, rotating speed 150～180r/min, shaking culture 24～48h;
The Rhizopus oryzae secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through the Rhizopus oryzae first order seed is inoculated in the Rhizopus oryzae secondary seed tank according to 5～10% weight ratio, be under 1:1～1.5 conditions, with 100～120r/min mechanical stirring in 25～28 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
Rhizopus oryzae liquid fermentation and culture: the above-mentioned bacterium liquid of cultivating through the Rhizopus oryzae secondary seed is inoculated in the Rhizopus oryzae fermentor tank according to 5～10% weight ratio, in 25～28 ℃ of temperature, ventilation volume ratio be under 1:1～1.5 conditions, 100～120r/min mechanical stirring, cultivate 24～48h.
The environment-friendly high-efficiency bio-organic fertilizer for preparing of method as mentioned above.
According to the environment-friendly high-efficiency bio-organic fertilizer that method as mentioned above prepares, humic acids content 〉=6.55%, biochemical humic acid content 〉=18.50%, N+P
2O
5+ K 〉=7.11%, Ca+Zn+Mn+Mg+Fe+S 〉=2.00%, beneficial microorganism total count (CFU/g) 〉=3.50 * 10
9, miscellaneous bacteria rate≤10.00%, organic acid total amount 〉=7.32g/kg.
Beneficial effect of the present invention is:
The present invention is that to select distillers ' grains, Sunflower shell powder, ammonium sulfate, Repone K, humic acids and corn steep liquor be raw material, adds in proportion MnSO after puffing
4H
2O, CaCO
3, MgSO
47H
2O, ZnSO
4And FeSO
4Then inoculate the mixed bacteria liquid that Jingyang streptomycete (Streptomycesjingyangensis), clostridium butylicum (Clostridium butyricum), pseudomonas mendocina (Pseudomonas mendocina), blown-ball Azotobacter (Azotobacter chroococcum) and Rhizopus oryzae (Rhizopus oryzae) form, the complex microorganism solid organic fertilizer product that makes by efficient respiratory membrane process for solid state fermentation.The advantage of the technology of the present invention is that the fermentation substrate raw material is after high temperature, high pressure swelling are processed, nutritive substance is decomposed and release fully, volatile hazardous and noxious substances and antinutritional factor are effectively removed, robust fibre is become digestible small molecule structure by physical decomposition, the gelatinization of starch height, the utilization ratio of the nutritive substances such as fat, protein improves greatly, and pathogenic bacterium and miscellaneous bacteria be by effectively deactivation and inhibition, thereby provides good nutritional condition for the beneficial microorganism amount reproduction.The fermentation substrate raw material has generated a large amount of biochemical humic acids and organic acid after the effective microorganisms fermentation simultaneously, and ventilation property and water-retentivity, the activating soil nutrient that improves salting of soil, raising soil played vital role.And micro-stable being chelated to that the effect of process microorganism makes humic acids, organic acid add with external source effectively prevented antagonistic action between each element, can obviously improve plant to the absorption rate of trace element.
More particularly, the technology of a kind of distillers ' grains production of the present invention environment-friendly high-efficiency bio-organic fertilizer has the following advantages:
1, the present invention makes effectively rehabilitating soil ecology of product, make soil reduction process in antioxidant status, effectively replenish the required organic matter of plant growth, probiotics and various trace elements, farm crop surprising energy for growth in benign state is given full play in secretion and synthetic various organic acids, amino acid, enzyme, active hormones, antioxidative enzyme etc.
2, in the inventive method, the fermentation substrate raw material is after high temperature, high pressure swelling are processed, nutritive substance is decomposed and release fully, volatile hazardous and noxious substances and antinutritional factor are effectively removed, the gelatinization of starch height, the utilization ratio of the nutritive substances such as fat, protein improves greatly, pathogenic bacterium and effectively deactivation and the inhibition of miscellaneous bacteria quilt, thereby for beneficial microorganism is grown, breeding provides good nutritional condition.
3, product of the present invention has slowly-releasing, long-acting characteristics.Therefore, use composite microbiological fertilizer, can not get instant result as using the chemical fertilizer effect, also can not run off in a large number because of eluviation, cause the pollution of water source and soil.
4, product of the present invention concentrates on fertilizer, chemical fertilizer and microbial fertilizer all over the body, use conveniently, overcome the shortcoming that soil physical and chemical property is degenerated and agricultural product quality descends that simple chemical fertilizer causes to soil, improved Soil structure, increase soil fertility, the needed nutrition of balanced crop growth has strengthened the crop anti-adversity energy.
5, the present invention makes product has activation soil moisture conservation and improving effect to alkaline earth, black earth.Can form regenerative system, phosphorus decomposing, potassium decomposing, fixed nitrogen compile the energy three-dimensional, and improve the acid, alkali of soil, sticking, sand and easy easily drought, the undesirable property such as harden of waterlogging, promote granuleization, greatly improve water conservation and the permeability of soil.
6, the present invention makes product and can prevent and treat physiological disease; Fertilizer damage is removed in the absorption of balance nitrogen, phosphorus, potassium.
7, rural economy prosperity and Sustainable development are guaranteed in the low input of the inventive method, high repayment.
8, adopt in the environment-friendly high-efficiency bio-organic fertilizer of the inventive method production humic acids content 〉=6.55%, biochemical humic acid content 〉=18.50%, N+P
2O
5+ K 〉=7.11%, Ca+Zn+Mn+Mg+Fe+S 〉=2.00%, beneficial microorganism total count (CFU/g) 〉=3.50 * 10
9, miscellaneous bacteria rate≤10.00%, organic acid total amount 〉=7.32g/kg.
Description of drawings
Fig. 1 is the schematic flow sheet of the inventive method.
Embodiment
Below describe technology of the present invention and characteristics in detail by specific embodiment, but these embodiment limit protection scope of the present invention.
Used Jingyang streptomycete (Streptomycesjingyangensis) in following examples of the present invention, clostridium butylicum (Clostridium butyricum), pseudomonas mendocina (Pseudomonas mendocina), blown-ball Azotobacter (Azotobacter chroococcum) and Rhizopus oryzae (Rhizopus oryzae) bacterial classification are the wild-type strains of buying respectively in Chinese industrial microbial strains preservation administrative center (CICC) and Chinese common micro-organisms culture presevation administrative center (CGMCC).Preserving number is respectively: ACCC40126, CICC20036, ACCC01078, CICC10309, CGMCC3.1267.
Embodiment 1
Referring to Fig. 1, prepare in accordance with the following methods the bio-organic fertilizer of present embodiment:
1. accurately take by weighing humic acids 100kg, distillers ' grains 600kg, Sunflower shell powder 100kg, ammonium sulfate 100kg, Repone K 50kg, corn steep liquor 50kg mixes and makes fermentation substrate, and the fermentation substrate moisture content is 30%.
2. the fermentation substrate raw material proportions afterwards by the saturation steam of twin-screw extruder through 0.4MPa, carries out extruding puffing under 150 ℃ of conditions.
3. fermentation substrate is transported to the wet feed mixing machine after puffing, passes into pressurized air and makes the fermentation substrate temperature be down to 35 ℃, adds simultaneously MnSO
4H
2O0.05%, CaCO
32%, MgSO
47H
2O0.2%, ZnSO
40.05% and FeSO
40.1%(is the per-cent that accounts for fermentation substrate weight) fully mix.
4. the inoculum size with Jingyang streptomycete, clostridium butylicum, pseudomonas mendocina, blown-ball Azotobacter and Rhizopus oryzae is respectively 5%, 10%, 10%, 5% and 10% of fermentation substrate weight; Fully mix 4min after the fermentation substrate inoculation.
5. after fermentation substrate inoculation and the mixing, be loaded in the silica gel respiratory membrane fermentation bag by the automatic gauge packaging system and pack, to seal, do not need air in the bag emptyingly in the wrapping process, packaged material 35 ℃ of condition bottom fermentation 7d in proving room are finished product.
6. Jingyang streptomycete (Streptomycesjingyangensis), clostridium butylicum (Clostridium butyricum), pseudomonas mendocina (Pseudomonas mendocina), blown-ball Azotobacter (Azotobacter chroococcum) and Rhizopus oryzae (Rhizopus oryzae) being increased respectively by the following method bacterium cultivates and liquid state fermentation:
(1) Jingyang streptomycete (Streptomycesjingyangensis)
A. first order seed substratum: KNO
31g, NaCl0.5g, K
2HPO
40.5g, FeSO
47H
2O0.01g, MgSO
47H
2O0.5g, Zulkovsky starch 20g, distilled water 1000mL, pH7.2 is at 121 ℃ of lower sterilization 30min of temperature.
B. secondary seed medium and fermention medium: glucose 20kg, soybean cake powder 20kg, NaCl3kg, CaCO
31kg, sterile pure water 1000L, pH7.2 is at 121 ℃ of lower sterilization 30min of temperature.
C. culture condition:
First order seed is cultivated: dress first order seed substratum 300mL in the 1000mL triangular flask, the spore suspension (2.0 * 10 that the access strain inclined plane is made
7Individual/mL) 30mL, 30 ℃ of temperature, rotating speed 170r/min, shaking culture 24h.
Secondary seed is cultivated: the 10L automated seed canned secondary seed medium 5L that ferments, the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the secondary seed tank according to 10% weight ratio, and 30 ℃ of temperature, ventilation volume ratio be 1:1.5, mechanical stirring 100r/min, cultivate 24h.
Liquid fermentation and culture: the 100L automated seed canned fermention medium 50L that ferments, the above-mentioned bacterium liquid of cultivating through secondary seed is inoculated in the fermentor tank according to 10% weight ratio, and 30 ℃ of temperature, ventilation volume ratio be 1:1.5, mechanical stirring 100r/min, cultivate 24h.
(2) clostridium butylicum (Clostridium butyricum)
A. prepare the first order seed substratum: Tryptones 20g, beef extract 10g, yeast extract paste 6g, glucose 4g, dipotassium hydrogen phosphate 2g, potassium primary phosphate 1g, sal epsom 0.4g, calcium chloride 0.2g, ferrous sulfate 0.1g, cysteine hydrochloride 0.5g, distilled water 1000mL, pH7.2 is at 121 ℃ of lower sterilization 30min of temperature.
B. prepare secondary seed medium and fermention medium: glucose 10kg, Tryptones 10kg, yeast extract 5kg, beef extract 3kg, K
2HPO
42kg, MgSO
47H
2O0.5kg, MnSO
4H
2O0.2kg, NaHCO1kg, CaCO
31kg, distilled water 1000L mixes, and pH6.8 is at 121 ℃ of lower sterilization 30min of temperature.
C. culture condition:
First order seed is cultivated: dress first order seed substratum 900mL in the 1000mL triangular flask, strain inclined plane to be inoculated in the first order seed substratum in the ratio of ring/100mL, and 37 ℃ of temperature leave standstill cultivation 24h.
Secondary seed is cultivated: the 20L automated seed canned secondary seed medium 10L that ferments, the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the secondary seed tank according to 10% weight ratio, and 37 ℃ of temperature leave standstill cultivates 24h.
Liquid fermentation and culture: the 200L automated seed canned fermention medium 100L that ferments, the above-mentioned bacterium liquid of cultivating through secondary seed is inoculated in the fermentor tank according to 10% weight ratio, 37 ℃ of temperature leave standstill cultivates 24h.
(3) pseudomonas mendocina (Pseudomonas mendocina)
A. prepare the first order seed substratum: peptone 10g, beef extract 10g, NaCl5g, glucose 10g, distilled water 1000mL, pH7.0 is at 121 ℃ of lower sterilization 30min of temperature.
B. prepare secondary seed medium and fermention medium: peptone 10kg, yeast soak powder 10kg, glucose 10kg, NaCl2kg, distilled water 1000L, and pH7.0 is at 121 ℃ of lower sterilization 30min of temperature.
C. culture condition:
First order seed is cultivated: dress first order seed substratum 500mL in the 1000mL triangular flask, inoculate strain inclined plane in the first order seed substratum in the ratio of ring/100mL 28 ℃ of temperature, rotating speed 130r/min, shaking culture 24h.
Secondary seed is cultivated: the 20L automated seed canned secondary seed medium 10L that ferments, the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the secondary seed tank according to 10% weight ratio, and 28 ℃ of temperature, ventilation volume ratio be 1:1, mechanical stirring 130r/min, cultivate 24h.
Liquid fermentation and culture: the 200L automated seed canned fermention medium 100L that ferments, the above-mentioned bacterium liquid of cultivating through secondary seed is inoculated in the fermentor tank according to 10% weight ratio, and 28 ℃ of temperature, ventilation volume ratio be 1:1, mechanical stirring 130r/min, cultivate 24h.
(4) blown-ball Azotobacter (Azotobacter chroococcum)
A. prepare seed culture medium: K
2HPO
40.8g, KH
2PO
40.2g, MgSO
47H
2O0.2g, NaCl0.5g, CaCl
20.1g, N.F,USP MANNITOL 20g, yeast extract paste 0.5g, FeCl
30.005g, Na
2MoO
4.2H
2O0.002g, distilled water 1000mL, pH7.0 is at 121 ℃ of lower sterilization 30min of temperature.
B. culture condition:
First order seed is cultivated: dress seed culture medium 500mL in the 1000mL triangular flask, inoculate strain inclined plane in the first order seed substratum in the ratio of ring/50mL 28 ℃ of temperature, rotating speed 120r/min, shaking culture 24h.
Secondary seed is cultivated: the 10L automated seed canned seed culture medium 5L that ferments, the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the secondary seed tank according to 10% weight ratio, and 28 ℃ of temperature, ventilation volume ratio be 1:1, mechanical stirring 120r/min, cultivate 24h.
Liquid fermentation and culture: the 100L automated seed canned seed culture medium 50L that ferments, the above-mentioned bacterium liquid of cultivating through secondary seed is inoculated in the fermentor tank according to 10% weight ratio, and 28 ℃ of temperature, ventilation volume ratio be 1:1, mechanical stirring 120r/min, cultivate 24h.
(5) Rhizopus oryzae (Rhizopus oryzae)
A. prepare the first order seed substratum: glucose sugar 20g, potato leaching liquid 1000mL, the pH nature is at 121 ℃ of lower sterilization 30min of temperature.
The preparation method of potato leaching liquid: remove the potato 200g of skin, be cut into small pieces, add water 1000mL and boil 30min elimination potato ball, filtrate is complemented to 1000mL.
B. prepare secondary seed medium and fermention medium: yam starch 100kg, (NH
4)
2SO
41.5kg, MgSO
47H
2O0.25kg, KH
2PO
40.3kg, CaCO
360kg, ZnSO
47H
2O0.05kg sterile pure water 1000L, the pH nature is at 121 ℃ of lower sterilization 30min of temperature.
C. culture condition:
First order seed is cultivated: dress first order seed substratum 250mL in the 1000mL triangular flask, the spore suspension (3.0 * 10 that the access strain inclined plane is made
7Individual/mL) 25mL, 28 ℃ of temperature, rotating speed 180r/min, shaking culture 24h.
Secondary seed is cultivated: the 20L automated seed canned secondary seed medium 10L that ferments, the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the secondary seed tank according to 10% weight ratio, and 28 ℃ of temperature, ventilation volume ratio be 1:1.5, mechanical stirring 100r/min, cultivate 24h.
Liquid fermentation and culture: the 200L automated seed canned fermention medium 100L that ferments, the above-mentioned bacterium liquid of cultivating through secondary seed is inoculated in the fermentor tank according to 10% weight ratio, and 28 ℃ of temperature, ventilation volume ratio be 1:1.5, mechanical stirring 100r/min, cultivate 24h.
7. the environment-friendly high-efficiency bio-organic fertilizer for preparing according to above method, humic acids content 〉=10.05%, biochemical humic acid content 〉=20.11%, N+P in this product
2O
5+ K 〉=7.85%, Ca+Zn+Mn+Mg+Fe+S 〉=2.50%, beneficial microorganism total count (CFU/g) 〉=8.00 * 10
9, miscellaneous bacteria rate≤10.00%, organic acid total amount 〉=10.10g/kg.
Embodiment 2
Referring to Fig. 1, prepare in accordance with the following methods the bio-organic fertilizer of present embodiment:
1. accurately take by weighing humic acids 50kg, distillers ' grains 500kg, Sunflower shell powder 150kg, ammonium sulfate 100kg, Repone K 100kg, corn steep liquor 100kg mixes and makes fermentation substrate, and the fermentation substrate moisture content is 30%.
2. the fermentation substrate raw material proportions afterwards by the saturation steam of twin-screw extruder through 0.4MPa, carries out extruding puffing under 150 ℃ of conditions.
3. fermentation substrate is transported to the wet feed mixing machine after puffing, passes into pressurized air and makes the fermentation substrate temperature be down to 35 ℃, adds simultaneously MnSO
4H
2O0.05%, CaCO
31%, MgSO
47H
2O0.1%, ZnSO
40.1% and FeSO
40.1% fully mixes.
4. the inoculum size with Jingyang streptomycete, clostridium butylicum, pseudomonas mendocina, blown-ball Azotobacter and Rhizopus oryzae is respectively 5%, 10%, 10%, 5% and 10% of fermentation substrate weight; Fully mix 4min after the fermentation substrate inoculation.
5. after fermentation substrate inoculation and the mixing, be loaded in the silica gel respiratory membrane fermentation bag by the automatic gauge packaging system and pack, to seal, do not need air in the bag emptyingly in the wrapping process, packaged material 35 ℃ of condition bottom fermentation 7d in proving room are finished product.
6. Jingyang streptomycete (Streptomycesjingyangensis), clostridium butylicum (Clostridium butyricum), pseudomonas mendocina (Pseudomonas mendocina), blown-ball Azotobacter (Azotobacter chroococcum) and Rhizopus oryzae (Rhizopus oryzae) being increased bacterium by method as described in Example 1 respectively cultivates and liquid state fermentation.
7. the environment-friendly high-efficiency bio-organic fertilizer for preparing according to above method, humic acids content 〉=6.55%, biochemical humic acid content 〉=18.50%, N+P in this product
2O
5+ K 〉=7.11%, Ca+Zn+Mn+Mg+Fe+S 〉=2.00%, beneficial microorganism total count (CFU/g) 〉=3.50 * 10
9, miscellaneous bacteria rate≤10.00%, organic acid total amount 〉=7.32g/kg.
Claims (9)
1. a method of utilizing distillers ' grains to produce the environment-friendly high-efficiency bio-organic fertilizer is characterized in that,
(1) fermentation substrate is carried out puffing by the screw bulking machine, and then be transported to and lower the temperature in the wet feed mixing machine and add micro-mineral salt, treat to inoculate in proportion when temperature drops to 30～35 ℃ mixed bacteria liquid and fully mix;
(2) fermentation substrate of inoculating and mixing is loaded in the silica gel respiratory membrane fermentation bag by the automatic gauge packaging system and packs, seals, and constant temperature bottom fermentation 5～7d in proving room is described environment-friendly high-efficiency bio-organic fertilizer finished product;
Described fermentation substrate be with: distillers ' grains 50～60 weight parts, humic acids 5～10 weight parts, Sunflower shell powder 5～15 weight parts, ammonium sulfate 5～10 weight parts, Repone K 5～10 weight parts, corn steep liquor 5～10 weight parts mix the fermentation substrate of making, and the fermentation substrate moisture content is 30～40wt%;
Described mixed bacteria liquid comprises the bacterium liquid of Jingyang streptomycete, clostridium butylicum, pseudomonas mendocina, blown-ball Azotobacter and Rhizopus oryzae.
2. method according to claim 1 is characterized in that, in the step (1), described fermentation substrate carries out extruding puffing by the saturation steam of twin-screw extruder through 0.3～0.4MPa under 140～150 ℃ of conditions.
3. method according to claim 1 is characterized in that, in the step (1), described fermentation substrate is transported to the wet feed mixing machine after puffing, passes into pressurized air the fermentation substrate temperature is down to
30～35 ℃, add simultaneously described micro-mineral salt: MnSO
4H
2O0.05～0.1%, CaCO
31～2%, MgSO
47H
2O0.1～0.2%, ZnSO
40.05～0.1% and FeSO
40.1～0.2%, fully mix, described percentage ratio accounts for the per-cent of fermentation substrate weight for each micro-mineral salt.
4. method according to claim 1, it is characterized in that, in the described mixed bacteria liquid, the inoculum size of Jingyang streptomycete, clostridium butylicum, pseudomonas mendocina, blown-ball Azotobacter and Rhizopus oryzae is respectively 2～5%, 5～10%, 5～10%, 2～5% and 5～10% of fermentation substrate weight percent; Fully mix 2～5min after the fermentation substrate inoculation.
5. method according to claim 1, it is characterized in that, in the step (2), after described fermentation substrate inoculation and the mixing, be loaded in the silica gel respiratory membrane fermentation bag by the automatic gauge packaging system and pack, to seal, packaged material is 30～35 ℃ of constant temperature bottom fermentation 5～7d in proving room, are described environment-friendly high-efficiency bio-organic fertilizer finished product.
6. method according to claim 1 is characterized in that, described silica gel respiratory membrane fermentation bag, and the bag sealing two ends is circular acerous design.
7. method according to claim 1 is characterized in that, described wet feed mixing machine, twin-screw extruder and automatic gauge packaging system are the 316L stainless steel.
8. according to claims 1 described method, it is characterized in that described bacterial classification is made by the following method:
(1) preparation method of described Jingyang streptomycete bacterium liquid is:
Preparation Jingyang streptomycete first order seed substratum: KNO
31g, NaCl0.5g, K
2HPO
40.5g, FeSO
47H
2O0.01g, MgSO
47H
2O0.5g, Zulkovsky starch 20g, distilled water 1000mL, pH7.2～7.4 are at 115～121 ℃ of lower sterilization 20min～30min of temperature;
Preparation Jingyang streptomycete secondary seed medium and fermention medium: glucose 20kg, soybean cake powder 20kg, NaCl3kg, CaCO
31kg, sterile pure water 1000L, pH7.2～7.4 are at 115～121 ℃ of lower sterilization 20～30min of temperature;
Jingyang streptomycete first order seed is cultivated: dress Jingyang streptomycete seed substratum 300mL in the 1000mL triangular flask, it is 1.0～2.0 * 10 that concentration is made on access streptomyces species inclined-plane, Jingyang
7Spore suspension 25～50mL of individual/mL, 28～30 ℃ of temperature, rotating speed 150～170r/min, shaking culture 24～48h;
Jingyang streptomycete secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through Jingyang streptomycete first order seed is inoculated in the streptomycete secondary seed tank of Jingyang according to 5～10% weight ratio, be under the condition of 1:1～1.5, with 100～120r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
Jingyang streptomycete liquid fermentation and culture: the above-mentioned bacterium liquid of cultivating through Jingyang streptomycete secondary seed is inoculated in the streptomycete fermentation tank of Jingyang according to 5～10% weight ratio, be under 1:1～1.5 conditions, with 100～120r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
(2) preparation method of described clostridium butylicum bacterium liquid is:
Preparation clostridium butylicum first order seed substratum: Tryptones 20g, beef extract 10g, yeast extract paste 6g, glucose 4g, dipotassium hydrogen phosphate 2g, potassium primary phosphate 1g, sal epsom 0.4g, calcium chloride 0.2g, ferrous sulfate 0.1g, cysteine hydrochloride 0.5g, distilled water 1000mL, pH7.2～7.4 are at 115～121 ℃ of lower sterilization 20min～30min of temperature;
Preparation clostridium butylicum secondary seed medium and fermention medium: glucose 10kg, Tryptones 10kg, yeast extract 5kg, beef extract 3kg, K
2HPO
42kg, MgSO
47H
2O0.5kg, MnSO
4H
2O0.2kg, NaHCO1kg, CaCO
31kg, distilled water 1000L mixes, and pH6.8～7.0 are at 115～121 ℃ of lower sterilization 20min～30min of temperature;
The clostridium butylicum first order seed is cultivated: dress clostridium butylicum first order seed substratum 900mL in the 1000mL triangular flask, the clostridium butylicum strain inclined plane is inoculated in the clostridium butylicum first order seed substratum in the ratio of ring/50～100mL, and 35～37 ℃ of temperature leave standstill cultivates 24h～48h;
The clostridium butylicum secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through the clostridium butylicum first order seed is inoculated in the clostridium butylicum secondary seed tank according to 5～10% weight ratio, and 35～37 ℃ of temperature leave standstill cultivates 24h～48h;
The clostridium butylicum liquid fermentation and culture: the above-mentioned bacterium liquid of cultivating through the clostridium butylicum secondary seed is inoculated in the clostridium butylicum fermentor tank according to 5～10% weight ratio, and 35～37 ℃ of temperature leave standstill cultivates 24h～48h;
(3) preparation method of described pseudomonas mendocina bacterium liquid is:
Preparation pseudomonas mendocina first order seed substratum: peptone 10g, beef extract 10g, NaCl5g, glucose 10g, distilled water 1000mL, pH6.8～7.0 are at 115～121 ℃ of lower sterilization 20min～30min of temperature;
Preparation pseudomonas mendocina secondary seed medium and fermention medium: peptone 10kg, yeast soak powder 10kg, glucose 10kg, NaCl2kg, distilled water 1000L, and pH6.8～7.0 are at 115～121 ℃ of lower sterilization 20min～30min of temperature;
The pseudomonas mendocina first order seed is cultivated: dress pseudomonas mendocina first order seed substratum 500mL in the 1000mL triangular flask, the pseudomonas mendocina strain inclined plane is inoculated in the pseudomonas mendocina first order seed substratum 28～30 ℃ of temperature, rotating speed 130～150r/min, shaking culture 24～48h in the ratio of ring/50～100mL;
The pseudomonas mendocina secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through the pseudomonas mendocina first order seed is inoculated in the pseudomonas mendocina secondary seed tank according to 5～10% weight ratio, be under the condition of 1:1～1.2, with 130～150r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
Pseudomonas mendocina liquid fermentation and culture: the above-mentioned bacterium liquid of cultivating through the pseudomonas mendocina secondary seed is inoculated in the pseudomonas mendocina fermentor tank according to 5～10% weight ratio, be under the condition of 1:1～1.2, with 130～150r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
(4) preparation method of described blown-ball Azotobacter bacterium liquid is:
Preparation blown-ball Azotobacter seed culture medium: K
2HPO
40.8g, KH
2PO
40.2g, MgSO
47H
2O0.2g,
NaCl0.5g, CaCl
20.1g, N.F,USP MANNITOL 20g, yeast extract paste 0.5g, FeCl
30.005g, Na
2MoO
4.2H
2O0.002g, distilled water 1000mL, pH7.0～7.2 are at 115～121 ℃ of lower sterilization 20～30min of temperature;
The blown-ball Azotobacter first order seed is cultivated: dress blown-ball Azotobacter seed culture medium 500mL in the 1000mL triangular flask, the blown-ball Azotobacter strain inclined plane is inoculated in the blown-ball Azotobacter first order seed substratum 28～30 ℃ of temperature, rotating speed 100～120r/min, shaking culture 24～48h in the ratio of ring/50～100mL;
The blown-ball Azotobacter secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through the blown-ball Azotobacter first order seed is inoculated in the blown-ball Azotobacter secondary seed tank according to 5～10% weight ratio, be under the condition of 1:0.5～1, with 100～120r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
Blown-ball Azotobacter liquid fermentation and culture: the above-mentioned bacterium liquid of cultivating through the blown-ball Azotobacter secondary seed is inoculated in the blown-ball Azotobacter fermentor tank according to 5～10% weight ratio, be under the condition of 1:0.5～1, with 100～120r/min mechanical stirring in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
(5) preparation method of described Rhizopus oryzae liquid is:
Preparation Rhizopus oryzae first order seed substratum: glucose sugar 20g, potato leaching liquid 1000mL, the pH nature is at 115～121 ℃ of lower sterilization 20～30min of temperature;
Wherein, the preparation method of potato leaching liquid is: remove the potato 200g of skin, be cut into small pieces, add water 1000mL and boil 30min elimination potato ball, filtrate is complemented to 1000mL;
Preparation Rhizopus oryzae secondary seed medium and fermention medium: yam starch 100kg, (NH
4)
2SO
41.5kg, MgSO
47H
2O0.25kg, KH
2PO
40.3kg, CaCO
360kg, ZnSO
47H
2O0.05kg sterile pure water 1000L, the pH nature is at 115～121 ℃ of lower sterilization 20～30min of temperature;
The Rhizopus oryzae first order seed is cultivated: dress Rhizopus oryzae seed culture medium 250mL in the 1000mL triangular flask, it is 2.5～4.0 * 10 that concentration is made on access Rhizopus oryzae bacterial classification inclined-plane
7Spore suspension 10～30mL of individual/mL, 25～28 ℃ of temperature, rotating speed 150～180r/min, shaking culture 24～48h;
The Rhizopus oryzae secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through the Rhizopus oryzae first order seed is inoculated in the Rhizopus oryzae secondary seed tank according to 5～10% weight ratio, be under 1:1～1.5 conditions, with 100～120r/min mechanical stirring in 25～28 ℃ of temperature, ventilation volume ratio, cultivate 24～48h;
Rhizopus oryzae liquid fermentation and culture: the above-mentioned bacterium liquid of cultivating through the Rhizopus oryzae secondary seed is inoculated in the Rhizopus oryzae fermentor tank according to 5～10% weight ratio, in 25～28 ℃ of temperature, ventilation volume ratio be under 1:1～1.5 conditions, 100～120r/min mechanical stirring, cultivate 24～48h.
9. the environment-friendly high-efficiency bio-organic fertilizer for preparing according to each described method in the claim 1～8.
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