CN103012009A - Organic acid plant soil conditioner and preparation method thereof - Google Patents
Organic acid plant soil conditioner and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a preparation method of an organic acid plant soil conditioner. The preparation method comprises the following steps of: carrying out filtering as well as evaporation and concentration on yellow water, a by-product of white wine fermentation; fully mixing and dissolving glucoses, (NH4)2SO4, KH2PO4, MgSO4.7H2O, MnSO4.H2O, NaCl and CaCO3 in the processed yellow water, and then carrying out high-temperature sterilization on the obtained solution; and feeding fermentation mother liquor subjected to sterilization to a mixed strain fermentation liquor to be subjected to fermentation for 24 to 48 hours, and bottling the obtained solution by an aseptic filling machine, wherein the mixed strain fermentation liquor is prepared by mixing rhodop seudomonas palustris accounting for 2wt%-5wt% of the fermentation mother liquor, pseudomonas fluorescens accounting for 2wt%-5wt% of the liquor, azotobacter chroococcum accounting for 2wt%-5wt% of the liquor, bacillus megatheriums accounting for 5wt%-10wt% of the liquor and bacillus mucilaginosus accounting for 5wt%-10wt% of the liquor.
Description
Technical field
The invention relates to a kind of organic acid plant soil conditioning agent and preparation method thereof, belong to the microbial organic fertilizer technical field.
Background technology
In the liquor fermentation process, water content produces a large amount of free-waters at 52～55% the wine unstrained spirits that enters to store after microbial metabolism.The gradually sedimentation of water that is not utilized by Institute of Micro-biology in these water and the wine unstrained spirits, with the acid in the wine unstrained spirits, Zulkovsky starch, yeast leachable, reducing sugar, tannin, alcohol and the stripping of fragrance precursor material, be deposited at last pond bottom, cellar for storing things and form brown color, be the liquid of flco shape, this liquid is called as yellow water.According to surveying and determination, the pH of yellow water is 3.0～3.5, be rich in the organic acid substances such as acetic acid, butyric acid, lactic acid, caproic acid and other alcohol, aldehyde material in the yellow water, also contain the liquor flavor materials such as ethyl acetate, ethyl lactate, ethyl hexanoate, and the organic matters such as the autolyzate of a small amount of remaining starch, residual sugar and alcohol, soil ulmin and yeast thalline, anerobe.Its COD, BOD are considerably beyond wastewater discharge standard.At present, to yellow water utilize approach less, utilization ratio is lower.Mainly be further distill yellow water wine, return cellar for storing things fermentation, mix poor unstrained spirits and return cellar for storing things fermentation etc. with the wine tail.Developing new yellow water and utilize approach, improve distillery waste discharging water quality, few effluent, turn waste into wealth, is a problem of being badly in need of solution.
Development and popularization along with the microbial organic fertilizer technology, some industry and agriculture byproduct constantly are exploited, if can be utilized effectively, it is a kind of biological organic fertilizer raw material of high-quality that the wine brewing byproduct yellow water that is rich in organic substance equally also be can yet be regarded as.
Summary of the invention
The object of the invention is to, for the problems referred to above, provide a kind of method of utilizing yellow water to produce organic acid plant soil conditioning agent.
For achieving the above object, the present invention is by the following technical solutions:
A kind of preparation method of organic acid plant soil conditioning agent is characterized in that,
(1) liquor fermentation by product yellow water is filtered, evaporation concentration processes;
(2) be disposed rear adding glucose, (NH of yellow water
4)
2SO
4, KH
2PO
4, MgSO
47H
2O, MnSO
4H
2O, NaCl and CaCO
3Fully carry out high-temperature sterilization behind the mixed dissolution;
(3) the fermentation mother liquor access fermented by mixed bacterium liquid fermentation 24～48h after sterilization finishes bottles by aseptic filler;
Described hybrid strain fermentating liquid is comprised of Rhodopseudomonas palustris (Rhodopseudomonaspalustris), Pseudomonas fluorescens (Pseudomonas fluorescens), blown-ball Azotobacter (Azotobacterchroococcum), bacillus megaterium (Bacillus megaterium) and bacillusmusilaginosiengineering (Bacillus mucilaginosus) bacterium liquid;
The inoculum size of Rhodopseudomonas palustris, Pseudomonas fluorescens, blown-ball Azotobacter, bacillus megaterium and bacillusmusilaginosiengineering is respectively 2～5%, 2～5%, 2～5%, 5～10%, 5～10% of fermentation mother liquor weight in the described fermented by mixed bacterium liquid.
Aforesaid method, preferably, liquor fermentation by product yellow water described in the step (1) adopts many deep bed filter to filter, and removes wherein larger impurity, and then is that total solids content is 15% liquid by the evaporation concentration tank with the fermentation mother liquor evaporation concentration.
Aforesaid method, preferably, described glucose, (NH
4)
2SO
4, KH
2PO
4, MgSO
47H
2O, MnSO
4H
2O, NaCl and CaCO
3Add in the fermentation mother liquor by 1～2%, 0.5～1%, 0.01～0.02%, 0.01～0.02%, 0.001～0.005%, 0.01～0.02% and 1～2% weight ratio respectively;
Described high-temperature sterilization is boiling sterilization 20～30min under 115 ℃～121 ℃, 0.07MPa～0.11MPa condition.
Aforesaid method, preferably, described Rhodopseudomonas palustris bacterium liquid is made by the following method:
Preparation seed culture medium: K
2HPO
40.78g, KH
2PO
40.5g, MgSO
47H
2O0.2g, NaCl0.5g, CaCl
20.05g, (NH
4)
2SO
40.05g, yeast extract paste 2.5g, sodium-acetate 1.65g, Sodium Tetraborate 0.38g, ammonium molybdate 0.22g, distilled water 1000mL, pH 6.7～7.2, at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress seed culture medium 500mL in the 1000mL triangular flask, and in the ratio access seed culture medium of strain inclined plane in ring/50～100mL, 25～30 ℃ of temperature, rotating speed 100～120r/min, shaking culture 24～48h;
Secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the secondary seed tank according to 5～10% weight ratio, be under the condition of 1:0.5～1, with 80～100r/min mechanical stirring in 25～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h.
Aforesaid method, preferably, described Pseudomonas fluorescens bacterium liquid is made by the following method:
The preparation seed culture medium: peptone 5g, beef extract 3g, NaCl 5g, distilled water 1000mL mixes, and pH6.8～7.0 are at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress seed culture medium 500mL in the 1000mL triangular flask, and in the ratio access seed culture medium of strain inclined plane in ring/50～100mL, 28～30 ℃ of temperature, rotating speed 100～120r/min, shaking culture 24～48h;
Secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the 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.
Aforesaid method, preferably, described blown-ball Azotobacter bacterium liquid is made by the following method:
Preparation 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, pH 7.0～7.2, at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress seed culture medium 500mL in the 1000mL triangular flask, inoculate in the first order seed substratum 28～30 ℃ of temperature, rotating speed 100～120r/min, shaking culture 24～48h with strain inclined plane in the ratio of ring/50～100mL;
Secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the 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.
Aforesaid method, preferably, described bacillus megaterium bacterium liquid is made by the following method:
Preparation first order seed substratum: peptone 10.0g, beef extract 3.0g, NaCl 5.0g, glucose 5.0g, distilled water 1000mL, pH 6.8～7.0, at 115～121 ℃ of lower sterilization 20～30min of temperature;
Preparation secondary seed medium: Semen Maydis powder 10kg, analysis for soybean powder 10kg, K
2HPO
41.5kg, MgSO
47H
2O 1.5kg, CaCO
31.5kg, distilled water 1000L, pH 6.8～7.0, at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress first order seed substratum 500mL in the 1000mL triangular flask, and in the ratio access first order seed substratum of strain inclined plane in ring/50～100mL, 28～30 ℃ of temperature, rotating speed 130～150r/min, shaking culture 24～48h;
Secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the 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.
Aforesaid method, preferably, described bacillusmusilaginosiengineering bacterium liquid is made by the following method:
Configuration seed culture medium: K
2HPO
40.2g, MgSO
47H
2O0.2g, CaCO
35g, NaCl0.2g, sucrose 10g, CaSO
42H
2O0.1g, distilled water 1000mL, pH 6.8～7.0, at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress seed culture medium 500mL in the 1000mL triangular flask, inoculate in the first order seed substratum 25～28 ℃ of temperature, rotating speed 130～150r/min, shaking culture 24～48h with strain inclined plane in the ratio of ring/50～100mL;
Secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the 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 25～28 ℃ of temperature, ventilation volume ratio, cultivate 24～48h.
Aforesaid method, preferably, after fermentation mother liquor described in the step (3) sterilization, the inoculation, be under the condition of 1:0.5～1 in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h with 100～120r/min mechanical stirring, then bottle by aseptic filler.
The organic acid plant soil conditioning agent for preparing of method as mentioned above.
According to the organic acid plant soil conditioning agent that the method for the invention prepares, total solid in this product 〉=17.00%, calcium of organic acid content 〉=1.00%, organic acid total amount 〉=30.00g/L, N+P+K+Mg+Mn+S 〉=11.00g/L, Rhodopseudomonas palustris (CFU/g) 〉=2.50 * 10
8, Pseudomonas fluorescens 〉=1.00 * 10
9, blown-ball Azotobacter (CFU/g) 〉=2.00 * 10
8, bacillus megaterium (CFU/g) 〉=1.55 * 10
9, bacillusmusilaginosiengineering (CFU/g) 〉=1.20 * 10
9
Beneficial effect of the present invention is:
The present invention selects the biological organic fertilizer of excellent property to produce bacterial classification, adopts advanced microbic liquid deep layer fermentation technology, with liquor fermentation by product yellow water as fermentation mother liquor, after filtration, add certain proportion glucose, (NH after the concentration
4)
2SO
4, KH
2PO
4, MgSO
47H
2O, MnSO
4H
2O, NaCl and CaCO
3Fully carry out high-temperature sterilization behind the mixed dissolution, then inoculate the mixed strains that is formed by Rhodopseudomonas palustris (Rhodopseudomonaspalustris), Pseudomonas fluorescens (Pseudomonas fluorescens), blown-ball Azotobacter (Azotobacterchroococcum), bacillus megaterium (Bacillus megaterium) and bacillusmusilaginosiengineering (Bacillusmucilaginosus), through deep layer liquid state fermentation, can make a kind of plant soil conditioning agent that is rich in the necessary organic trace element of organic acid, beneficial microorganism and plant-growth.
Contain abundant organic acid and residual sugar in the raw materials used yellow water of the present invention, can provide good nutritional condition for the microorganism growth breeding.Select good microbial organic fertilizer to produce bacterium, adopt advanced microorganism liquid state fermentation technology, organic matter remaining in the yellow water and carbohydrate are carried out submerged fermentation, add during the fermentation the necessary trace element of some plant-growths, decompose and organic chelated effect generation biological organic matter by the thalline fermentation, can make farm crop be easier to absorb and utilize.Simultaneously, a large amount of beneficial microorganisms and abundant microbial metabolites and organic acid have also been produced by microbial fermentation.Make the biological organic fertilizer of yellow water by fermentation production can be balanced, coordinate growing and stimulating root growth of farm crop, improve soil salt alkalescence, increase crop yield and soil fertility, improve crop quality; The immunizing power of crop be can strengthen again, disease-resistant, drought resisting, water conservation, cold tolerance improved.Also reduce simultaneously the discharging of waste water, wine by product processed is effectively reclaimed and utilized, turned waste into wealth, reduced environmental pollution, had good ecological benefits, economic benefit and DEVELOPMENT PROSPECT.
More particularly, the preparation method of organic acid plant soil conditioning agent provided by the invention has the following advantages:
1, the calcium of organic acid that is rich in the product of the present invention can be by biological complexing, replacement(metathesis)reaction, and unnecessary Na on the soil granular is removed in displacement
+, the difficult P that utilizes in the activation salt affected soil
5+, Fe2
+, Ca
2+, Mg
2+Plasma and trace element change it into and can utilize state by plant absorbing, remove the plant physiology nutritional deficiency symptom.Pass through simultaneously Na
+Reduce activation Ca
2+, Mg
2+After the plasma, soil water conducting energy (HC) is increased, make soil moisture easier to be mobile, thereby saline alkali and the crop root environment of soil have effectively been improved, promote root growth, guarantee that the crop seedling is neat, seedling strong, make plant and improve output in the saltings growth.
2, the organic acid that is rich in of product of the present invention, but efficient neutralization and alleviate soil alkaline.
3, the beneficial microorganism in the product of the present invention can produce glucide, account for 0.1% of the soil organism, with plant mucilage, mineral idiosome and organic colloid combine, can improve soil aggregate, strengthen the physicals of soil and the loss of minimizing soil particle, under certain conditions, can also participate in soil ulmin and form.Can improve soil physical property so use microbial fertilizer, be conducive to increase soil fertility.
4, product of the present invention can replace chemical fertilizer and agricultural chemicals, relaxes or the minimizing pollution of agricultural products, and can improve the quality of agricultural-food.
5, the beneficial microbe colony that is rich in the product of the present invention can promote soil material to transform, and improves Soil structure, increases soil fertility, and promotes plant growth.By the microorganism nitrogen fixation, can improve the soil nitrogen level, can effectively improve some organic composition, sulfide and ammonia-state nitrogen in the soil by its Metabolic activity, and promote the conversion of noxious pollutant such as agricultural chemicals etc.Can promote simultaneously beneficial microorganism propagation, make it jointly to participate in the material cycle of soil ecology.
6, the beneficial microbe colony in the product of the present invention can strengthen crop disease-resistant diseases prevention ability, can promote the beneficial microorganism breedings such as actinomycetes, suppresses the pernicious bacteria growths such as filamentous fungus, thereby effectively suppresses some generation of planting disease and spread.
7, the microorganism in the product of the present invention has the effect of phosphorus decomposing, potassium decomposing, fixed nitrogen, can improve utilization rate of fertilizer more than 10%～30%.A large amount of productions of this fertilizer and promoting the use of can be saved the resources such as manure resources and associated coal, Sweet natural gas, oil, and alleviating energy crisis is had certain help.
8, the product of using the inventive method and making can reduce peasant's production cost, improve crop yield, increases farmers' income.
9, in the organic acid plant soil conditioning agent provided by the invention, total solid 〉=17.00%, calcium of organic acid content 〉=1.00%, organic acid total amount 〉=30.00g/L, N+P+K+Mg+Mn+S 〉=11.00g/L, Rhodopseudomonas palustris (CFU/g) 〉=2.50 * 10
8, Pseudomonas fluorescens 〉=1.00 * 10
9, blown-ball Azotobacter (CFU/g) 〉=2.00 * 10
8, bacillus megaterium (CFU/g) 〉=1.55 * 10
9, bacillusmusilaginosiengineering (CFU/g) 〉=1.20 * 10
9
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 Rhodopseudomonas palustris (Rhodopseudomonaspalustris) in following examples of the present invention, Pseudomonas fluorescens (Pseudomonas fluorescens), blown-ball Azotobacter (Azotobacterchroococcum), bacillus megaterium (Bacillus megaterium) and bacillusmusilaginosiengineering (Bacillusmucilaginosus) are to buy respectively in Chinese industrial microbial preservation administrative center (CICC), the wild-type strain of China common micro-organisms culture presevation administrative center (CGMCC) and Chinese agriculture microbial strains preservation administrative center (ACCC).Preserving number is respectively: ACCC 10649, CICC 21461, CGMCC 1.175, CICC 21694, CICC 23575.
Embodiment 1
Referring to Fig. 1, prepare in accordance with the following methods the organic acid plant soil conditioning agent of present embodiment:
1. take by weighing liquor fermentation by product yellow water 1000L, adopt many deep bed filter to filter, remove wherein larger impurity.And then be that total solids content is 15% liquid by the evaporation concentration tank with the fermentation mother liquor evaporation concentration.
2. after yellow water is disposed, with glucose, (NH
4)
2SO
4, KH
2PO
4, MgSO
47H
2O, MnSO
4H
2O, NaCl and CaCO
3Add in the fermentation mother liquor by 1%, 1%, 0.02%, 0.02%, 0.005%, 0.02% and 2% weight ratio respectively, then boiling sterilization 30min under 121 ℃, 0.11MPa condition.
3. Rhodopseudomonas palustris (Rhodopseudomonas palustris), Pseudomonas fluorescens (Pseudomonas fluorescens), blown-ball Azotobacter (Azotobacter chroococcum), bacillus megaterium (Bacillus megaterium) and bacillusmusilaginosiengineering (Bacillus mucilaginosus) being increased respectively by the following method bacterium cultivates:
(1) Rhodopseudomonas palustris (Rhodopseudomonas palustris)
A. prepare seed culture medium: K
2HPO
40.78g, KH
2PO
40.5g, MgSO
47H
2O0.2g,
NaCl 0.5g, CaCl
20.05g, (NH
4)
2SO
40.05g, yeast extract paste 2.5g, sodium-acetate 1.65g, Sodium Tetraborate 0.38g, ammonium molybdate 0.22g, distilled water 1000mL, pH 6.7, 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, and in the ratio access first order seed substratum of strain inclined plane in ring/50mL, 30 ℃ of temperature, rotating speed 100r/min, shaking culture 24h.
Secondary seed is cultivated: the 100L automated seed canned seed culture medium 50L 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, 30 ℃ of temperature, ventilation volume ratio 1:0.5, mechanical stirring 100r/min, cultivates 24h.
(2) Pseudomonas fluorescens (Pseudomonas fluorescens)
A. prepare seed culture medium: peptone 5g, beef extract 3g, NaCl 5g, distilled water 1000mL mixes, and 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, and in the ratio access first order seed substratum of strain inclined plane in ring/50mL, 30 ℃ of temperature, rotating speed 100r/min, shaking culture 24h.
Secondary seed is cultivated: the 100L automated seed canned seed culture medium 50L 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, 30 ℃ of temperature, ventilation volume ratio 1:0.5, mechanical stirring 100/min, cultivates 24h.
(3) blown-ball Azotobacter (Azotobacter chroococcum)
A. prepare seed culture medium: K
2HPO
40.8g, KH
2PO
40.2g, MgSO
47H
2O 0.2g,
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, pH 7.2, 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, and in the ratio access first order seed substratum of strain inclined plane in ring/50mL, 30 ℃ of temperature, rotating speed 120r/min, shaking culture 24h.
Secondary seed is cultivated: the 100L automated seed canned seed culture medium 50L 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, 30 ℃ of temperature, ventilation volume ratio 1:1, mechanical stirring 120r/min, cultivates 24h.
(4) bacillus megaterium (Bacillus megaterium)
A. prepare the first order seed substratum: peptone 10.0g, beef extract 3.0g, NaCl 5.0g, glucose 5.0g, distilled water 1000mL, pH 7.0, at 121 ℃ of lower sterilization 30min of temperature.
B. prepare secondary seed medium: Semen Maydis powder 10kg, analysis for soybean powder 10kg, K
2HPO
41.5kg, MgSO
47H
2O1.5kg, CaCO
31.5kg, distilled water 1000L, pH 7.0, 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, and in the ratio access first order seed substratum of strain inclined plane in ring/100mL, 30 ℃ of temperature, rotating speed 150r/min, shaking culture 24h.
Secondary seed is cultivated: the 200L automated seed canned secondary seed medium 100L 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, 30 ℃ of temperature, ventilation volume ratio 1:1, mechanical stirring 150r/min, cultivates 24h.
(5) bacillusmusilaginosiengineering (Bacillus mucilaginosus)
A. prepare seed culture medium: K
2HPO
40.2g, MgSO
47H
2O0.2g, CaCO
35g, NaCl0.2g, sucrose 10g, CaSO
42H
2O0.1g, 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, and in the ratio access first order seed substratum of strain inclined plane in ring/100mL, 25 ℃ of temperature, rotating speed 150r/min, shaking culture 24h.
Secondary seed is cultivated: the 200L automated seed canned seed culture medium 100L 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, 25 ℃ of temperature, ventilation volume ratio 1:1～1, mechanical stirring 150r/min, cultivates 24h.
4. the inoculum size of Rhodopseudomonas palustris, Pseudomonas fluorescens, blown-ball Azotobacter, bacillus megaterium and bacillusmusilaginosiengineering is respectively 5%, 5%, 5%, 10%, 10% of fermentation mother liquor weight percent in the fermented by mixed bacterium liquid.
5. after fermentation mother liquor sterilization, the inoculation, 30 ℃ of temperature, ventilation volume ratio 1:1, mechanical stirring 120r/min, cultivation 48h bottle by aseptic filler.
6. the organic acid plant soil conditioning agent for preparing according to above method, wherein total solid 〉=19.25%, calcium of organic acid content 〉=1.50%, organic acid total amount 〉=30.00g/L, N+P+K+Mg+Mn+S 〉=20.00g/L, Rhodopseudomonas palustris (CFU/g) 〉=5.00 * 10
8, Pseudomonas fluorescens 〉=1.50 * 10
9, blown-ball Azotobacter (CFU/g) 〉=3.00 * 10
8, bacillus megaterium (CFU/g) 〉=3.00 * 10
9, bacillusmusilaginosiengineering (CFU/g) 〉=2.50 * 10
9
Embodiment 2
Referring to Fig. 1, prepare in accordance with the following methods the organic acid plant soil conditioning agent of present embodiment:
1. take by weighing liquor fermentation by product yellow water 1000L, adopt many deep bed filter to filter, remove wherein larger impurity.And then be that total solids content is 20% liquid by the evaporation concentration tank with the fermentation mother liquor evaporation concentration.
2. after yellow water is disposed, with glucose, (NH
4)
2SO
4, KH
2PO
4, MgSO
47H
2O, MnSO
4H
2O, NaCl and CaCO
3Add in the fermentation mother liquor by 1.5%, 0.5%, 0.01%, 0.01%, 0.002%, 0.01% and 1.5% weight ratio respectively, then boiling sterilization 30min under 121 ℃, 0.11MPa condition.
3. Rhodopseudomonas palustris (Rhodopseudomonas palustris), Pseudomonas fluorescens (Pseudomonas florescens), blown-ball Azotobacter (Azotobacter chroococcum), bacillus megaterium (Bacillus megaterium) and bacillusmusilaginosiengineering (Bacillus mucilaginosus) being increased bacterium by method as described in Example 1 respectively cultivates.
4. in the fermented by mixed bacterium liquid, the inoculum size of Rhodopseudomonas palustris, Pseudomonas fluorescens, blown-ball Azotobacter, bacillus megaterium and bacillusmusilaginosiengineering is respectively 3%, 3%, 5%, 5%, 5% of fermentation mother liquor weight percent.
5. after fermentation mother liquor sterilization, the inoculation, 30 ℃ of temperature, ventilation volume ratio 1:1, mechanical stirring 120r/min, cultivation 48h bottle by aseptic filler.
6. the organic acid plant soil conditioning agent for preparing according to above method, wherein total solid 〉=17.00%, calcium of organic acid content 〉=1.00%, organic acid total amount 〉=30.00g/L, N+P+K+Mg+Mn+S 〉=11.00g/L, Rhodopseudomonas palustris (CFU/g) 〉=2.50 * 10
8, Pseudomonas fluorescens 〉=1.00 * 10
9, blown-ball Azotobacter (CFU/g) 〉=2.00 * 10
8, bacillus megaterium (CFU/g) 〉=1.55 * 10
9, bacillusmusilaginosiengineering (CFU/g) 〉=1.20 * 10
9
Claims (10)
1. the preparation method of an organic acid plant soil conditioning agent is characterized in that,
(1) liquor fermentation by product yellow water is filtered, evaporation concentration processes;
(2) be disposed rear adding glucose, (NH of yellow water
4)
2SO
4, KH
2PO
4, MgSO
47H
2O, MnSO
4H
2O, NaCl and CaCO
3Fully carry out high-temperature sterilization behind the mixed dissolution;
(3) the fermentation mother liquor access fermented by mixed bacterium liquid fermentation 24～48h after sterilization finishes bottles by aseptic filler;
Described hybrid strain fermentating liquid is comprised of Rhodopseudomonas palustris, Pseudomonas fluorescens, blown-ball Azotobacter, bacillus megaterium and bacillusmusilaginosiengineering bacterium liquid;
The inoculum size of Rhodopseudomonas palustris, Pseudomonas fluorescens, blown-ball Azotobacter, bacillus megaterium and bacillusmusilaginosiengineering is respectively 2～5%, 2～5%, 2～5%, 5～10%, 5～10% of fermentation mother liquor weight in the described fermented by mixed bacterium liquid.
2. method according to claim 1, it is characterized in that, liquor fermentation by product yellow water described in the step (1) adopts many deep bed filter to filter, remove wherein larger impurity, and then be that total solids content is 15% liquid by the evaporation concentration tank with the fermentation mother liquor evaporation concentration.
3. method according to claim 1 is characterized in that, described glucose, (NH
4)
2SO
4, KH
2PO
4, MgSO
47H
2O, MnSO
4H
2O, NaCl and CaCO
3Add in the fermentation mother liquor by 1～2%, 0.5～1%, 0.01～0.02%, 0.01～0.02%, 0.001～0.005%, 0.01～0.02% and 1～2% weight ratio respectively;
Described high-temperature sterilization is boiling sterilization 20～30min under 115 ℃～121 ℃, 0.07MPa～0.11MPa condition.
4. method according to claim 1 is characterized in that, described Rhodopseudomonas palustris bacterium liquid is made by the following method:
Preparation seed culture medium: K
2HPO
40.78g, KH
2PO
40.5g, MgSO
47H
2O 0.2g, NaCl0.5g, CaCl
20.05g, (NH
4)
2SO
40.05g, yeast extract paste 2.5g, sodium-acetate 1.65g, Sodium Tetraborate 0.38g, ammonium molybdate 0.22g, distilled water 1000mL, pH 6.7～7.2, at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress seed culture medium 500mL in the 1000mL triangular flask, and in the ratio access seed culture medium of strain inclined plane in ring/50～100mL, 25～30 ℃ of temperature, rotating speed 100～120r/min, shaking culture 24～48h;
Secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the secondary seed tank according to 5～10% weight ratio, be under the condition of 1:0.5～1, with 80～100r/min mechanical stirring in 25～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h.
5. method according to claim 1 is characterized in that, described Pseudomonas fluorescens bacterium liquid is made by the following method:
The preparation seed culture medium: peptone 5g, beef extract 3g, NaCl 5g, distilled water 1000mL mixes, and pH6.8～7.0 are at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress seed culture medium 500mL in the 1000mL triangular flask, and in the ratio access seed culture medium of strain inclined plane in ring/50～100mL, 28～30 ℃ of temperature, rotating speed 100～120r/min, shaking culture 24～48h;
Secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the 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.
6. method according to claim 1 is characterized in that, described blown-ball Azotobacter bacterium liquid is made by the following method:
Preparation 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, pH 7.0～7.2, at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress seed culture medium 500mL in the 1000mL triangular flask, inoculate in the first order seed substratum 28～30 ℃ of temperature, rotating speed 100～120r/min, shaking culture 24～48h with strain inclined plane in the ratio of ring/50～100mL;
Secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the 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.
7. method according to claim 1 is characterized in that, described bacillus megaterium bacterium liquid is made by the following method:
Preparation first order seed substratum: peptone 10.0g, beef extract 3.0g, NaCl 5.0g, glucose 5.0g, distilled water 1000mL, pH 6.8～7.0, at 115～121 ℃ of lower sterilization 20～30min of temperature;
Preparation secondary seed medium: Semen Maydis powder 10kg, analysis for soybean powder 10kg, K
2HPO
41.5kg, MgSO
47H
2O1.5kg, CaCO
31.5kg, distilled water 1000L, pH 6.8～7.0, at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress first order seed substratum 500mL in the 1000mL triangular flask, and in the ratio access first order seed substratum of strain inclined plane in ring/50～100mL, 28～30 ℃ of temperature, rotating speed 130～150r/min, shaking culture 24～48h;
Secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the 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.
8. method according to claim 1 is characterized in that, described bacillusmusilaginosiengineering bacterium liquid is made by the following method:
Configuration seed culture medium: K
2HPO
40.2g, MgSO
47H
2O0.2g, CaCO
35g, NaCl0.2g, sucrose 10g, CaSO
42H
2O0.1g, distilled water 1000mL, pH 6.8～7.0, at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress seed culture medium 500mL in the 1000mL triangular flask, inoculate in the first order seed substratum 25～28 ℃ of temperature, rotating speed 130～150r/min, shaking culture 24～48h with strain inclined plane in the ratio of ring/50～100mL;
Secondary seed is cultivated: the above-mentioned bacterium liquid of cultivating through first order seed is inoculated in the 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 25～28 ℃ of temperature, ventilation volume ratio, cultivate 24～48h.
9. method according to claim 1, it is characterized in that, after the sterilization of fermentation mother liquor described in the step (3), the inoculation, be under the condition of 1:0.5～1 in 28～30 ℃ of temperature, ventilation volume ratio, cultivate 24～48h with 100～120r/min mechanical stirring, then bottle by aseptic filler.
10. the organic acid plant soil conditioning agent for preparing according to each described method in the claim 1～9.
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| CN104609992A (en) * | 2014-12-31 | 2015-05-13 | 天津北洋百川生物技术有限公司 | Dedicated composite bacterial fertilizer for saline-alkali soil and preparation method thereof |
| CN106479907A (en) * | 2015-09-02 | 2017-03-08 | 上海师范大学 | A kind of culture medium improving composite bacteria fermentation Biomass and its using method |
| CN107986875A (en) * | 2017-12-20 | 2018-05-04 | 四川国科中农生物科技有限公司 | A kind of humic acid Water soluble fertilizer and its preparation process using yellow water as carrier |
| CN110128214A (en) * | 2019-06-19 | 2019-08-16 | 乐斯福(明光)有限公司 | A method of it is produced using yeast thick slurry waste liquid containing humic acid water-soluble fertilizer |
| CN110241041A (en) * | 2019-05-31 | 2019-09-17 | 南京工业大学 | Compound microbial preparation, preparation method and application thereof |
| CN111908979A (en) * | 2020-08-17 | 2020-11-10 | 江苏高生生物饲料有限公司 | Production method of water-soluble carbon energy fertilizer by taking white spirit yellow water as raw material |
| CN114702350A (en) * | 2022-04-02 | 2022-07-05 | 西华大学 | Preparation method and application of novel ecological modifier composition for alkaline soil |
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| CN104609992A (en) * | 2014-12-31 | 2015-05-13 | 天津北洋百川生物技术有限公司 | Dedicated composite bacterial fertilizer for saline-alkali soil and preparation method thereof |
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| CN110128214A (en) * | 2019-06-19 | 2019-08-16 | 乐斯福(明光)有限公司 | A method of it is produced using yeast thick slurry waste liquid containing humic acid water-soluble fertilizer |
| CN111908979A (en) * | 2020-08-17 | 2020-11-10 | 江苏高生生物饲料有限公司 | Production method of water-soluble carbon energy fertilizer by taking white spirit yellow water as raw material |
| CN114702350A (en) * | 2022-04-02 | 2022-07-05 | 西华大学 | Preparation method and application of novel ecological modifier composition for alkaline soil |
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