CN101863685A - Method for making high-concentration bacterial powder for microbial fertilizers - Google Patents
Method for making high-concentration bacterial powder for microbial fertilizers Download PDFInfo
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- CN101863685A CN101863685A CN201010192330A CN201010192330A CN101863685A CN 101863685 A CN101863685 A CN 101863685A CN 201010192330 A CN201010192330 A CN 201010192330A CN 201010192330 A CN201010192330 A CN 201010192330A CN 101863685 A CN101863685 A CN 101863685A
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Abstract
The invention provides a method for making high-concentration bacterial powder for microbial fertilizers, which is characterized in that mature and commercialized Bacillus mucilaginosus and Bacillus laterosporus are used for producing the sporeformer powder for composite microbial fertilizers. The Bacillus mucilaginosus and Bacillus laterosporus are subjected to fermentation culture, the culture solution of the Bacillus mucilaginosus and Bacillus laterosporus is obtained, the fermentation liquor is centrifuged at a high speed, concentrated and mixed and dried by spraying and thus the sporeformer powder for composite microbial fertilizers is obtained. The sporeformer powder provided by the invention is low in production and investment cost, low in technical process requirements, flexible in production, convenient and quick in use, high in biological stability, high in concentration and convenient to transport and can be stored for a long time.
Description
Technical field
The present invention relates to a kind of method of making high-concentration bacterial powder for microbial fertilizers.
Background technology
Bacillusmusilaginosiengineering (Bacillus mucilaginosus) is a kind of bacterium that can decompose silicate minerals, and more domestic scholars also call silicate bacteria to it.The morphological specificity of this bacterium is: the thick length of thalline is shaft-like, and the thick-walled pod film is arranged; Gemma is big, the sporangiocyst wall thickness, and ripe sporangiocyst does not expand; Bacterium colony is circular on silicate bacteria special culture media flat board, water white transparency, and there is cloud point at the middle part after 5-6 days, and the edge is transparent, smooth surface, thickness, elasticity is big, can pull into thread.
The key property of bacillusmusilaginosiengineering is the function with trace elements such as the invalid potassium, phosphorus and the magnesium that transform in the soil, iron, molybdenums, improve plant potassium, phosphorus and trophic level that some is micro-, produce several physiological active substances simultaneously, promote growth and development of plants, improve quality, strengthen the cold-resistant drought-enduring of plant and the effect of resisting disease and pest.At present, in the bio-feritlizer of the conversion that promotes the invalid phosphorus potassium of soil, the supply that increases soil phosphorus potassium, raising crop yield, this bacterioid is best bacterial classification.
Bacillus laterosporus (Bacillus laterosporus) belongs to bacillus, and Gram-positive can be changed into feminine gender, and gemma is oval, adnation, middle life or nearly middle giving birth to, and sporangiocyst expands, and free gemma is on one side than the other side thick (canoe shape).
Bacillus laterosporus can promote plant root growth, strengthens the root system receptivity, thereby improves crop yield; Suppress the inside and outside pathogenic bacteria breeding of plant materials, alleviate disease and pest, reduce pesticide residue; The improvement chessom solves the soil compaction phenomenon, thereby activating soil improves utilization rate of fertilizer; Strengthen plant metabolism, promote photosynthesis and strengthen the blade protection film, the opposing pathogenic bacteria; Strengthen photosynthesis, improve chemical fertilizer utilization ratio, reduce nitrate content.Solidify some heavy metals, reduce heavy metal content in the plant materials.
At present, it is low to contain bacteria concentration for microbial fertilizer, uses inconveniently, transport defectives such as not convenient, thus how to obtain high-concentration bacterial powder become how to obtain efficiently, one of the important technology of microbial fertilizer easily.
Summary of the invention
The objective of the invention is by design adopt set of parameter rationally, process stabilizing, industrialization degree is high and meet the production unit and the producing and manufacturing technique of biological inoculum characteristics, utilizes sophisticated, commercial bacillusmusilaginosiengineering, bacillus laterosporus to produce composite bio-fertilizer sporeformer powder.Sporeformer powder production cost of investment provided by the invention is low; Technical matters requires low; Produce flexible, convenient to use; Bacterium powder biologically stable is good, concentration height, convenient transportation, storage tolerance.
The present invention is achieved in that
The technical process of gel-shaped bacillus fermentation
The bacillusmusilaginosiengineering kind is seeded on the suitable slant medium, treat that growth fully after, place the refrigerator preservation about 4 ℃, manufacture slant strains.The bacterial classification that takes a morsel is forwarded on the Kolle flask slant medium, carries out enlarged culturing (30 ℃ constant temperature culture 2-5 days), treat growth fully after, place the refrigerator preservation about 4 ℃, make the Kolle flask slant strains.Bacterial classification in 30 ℃ of constant temperature culture 2-5 days, is added the washing of 40-50ml sterilized water, make the gemma suspension.To cultivate in the 500L seeding tank based on 0.11MPa, real jar of sterilization of vapor pressure 30 minutes in seeding tank, cultivated 15-20h with the gemma suspension inoculation for 30 ℃, tank pressure 0.05MPa, and ventilation is than 1: 1V/Vm, yeast culture is to the logarithmic growth later stage.With 5m
3Cultivate based on 0.11MPa real jar of sterilization of vapor pressure 30 minutes in the fermentor tank.Thalline and nutrient solution are transferred to 5m in the lump
3In the fermentor tank, cultivate 30-40h for 30 ℃, tank pressure 0.05MPa, ventilation is than 1: after 0.7-1V/Vm, thalline form gemma more than 90%, bacillus exfoliation 50-60%, fermentation ends finally obtains the fermented liquid of gemma content at 6-7 hundred million/gram.
Bacillus laterosporus zymotechnique flow process
The bacillus laterosporus kind is seeded on the suitable slant medium, treat that growth fully after, place the refrigerator preservation about 4 ℃, manufacture slant strains.The bacterial classification that takes a morsel is forwarded on the Kolle flask slant medium, carries out enlarged culturing (34-35 ℃ constant temperature culture 2-5 days), treat growth fully after, place the refrigerator preservation about 4 ℃, make the Kolle flask slant strains.Bacterial classification in 34-35 ℃ of constant temperature culture 2-5 days, is added the washing of 40-50ml sterilized water, make the gemma suspension.To cultivate in the 500L seeding tank based on 0.11MPa, real jar of sterilization of vapor pressure 30 minutes in seeding tank, cultivated 10-15h with the gemma suspension inoculation for 34-36 ℃, tank pressure 0.05MPa, and ventilation is than 1: 1V/Vm, yeast culture is to the logarithmic growth later stage.With 5m
3Cultivate based on 0.11MPa real jar of sterilization of vapor pressure 30 minutes in the fermentor tank.Thalline and nutrient solution are transferred to 5m in the lump
3In the fermentor tank, cultivate 18-25h for 34-36 ℃, tank pressure 0.05MPa, ventilation is than 1: 0.7-1V/Vm, thalline form gemma more than 90%, fermentation ends, the final fermented liquid that gets gemma content at 12-14 hundred million/gram.
Above-mentioned two kinds of fermented liquids through high speed centrifugation concentrate, mixing, spraying drying make the bacterium powder.
Preferably, in the above-mentioned high-concentration bacterial powder making processes, gel-shaped bacillus and bacillus laterosporus bacterial classification are respectively the quality strains that the inventor filters out from soil voluntarily.
Preferably, in the above-mentioned high-concentration bacterial powder making processes, fermention medium was consistent with the seed tank culture base during bacillusmusilaginosiengineering was cultivated, and it consists of: starch 5 grams, bean powder 3 grams, sugar 1 gram, CaCO
31 gram, sulphur ammonium 0.5 gram, 2 liters of oil, 1 liter in water, Ph7.0.Slant medium is consistent with the Kolle flask substratum, is the silicate bacteria substratum, and it consists of: sucrose (C
12H
22O
11) 5.0g, Sodium phosphate dibasic (Na
2HPO
412H
2O) 2.0g, sal epsom (MgSO
47H
2O) 0.5g, lime carbonate (CaCO
3) 0.1g, iron trichloride (FeCl
36H
2O) 0.005g, agar 18.0g, distilled water 1000mL, pH7.5～8.0.
Preferably, in the above-mentioned high-concentration bacterial powder making processes, fermention medium was consistent with the seed tank culture base during bacillus laterosporus was cultivated, and it consists of: soybean cake powder 4 grams, Semen Maydis powder 4 grams, sweet potato powder 4 grams, CaCO
32 grams, 1 liter in water, Ph nature.Slant medium is consistent with the Kolle flask substratum, is nutrient agar medium, and it consists of: peptone 10.0g, extractum carnis 3.0g, sodium-chlor (NaCl) 5.0g, agar 18.0g, distilled water 1000mL, pH7.2～7.5.
Preferably, in the above-mentioned high-concentration bacterial powder making processes, during high speed centrifugation, with fermented liquid disc centrifuge (7302 rev/mins), concrete controlled variable: 3 seconds sealing time, sealing cycle 1 minute, 1 second deslagging time, in 10 minutes deslagging cycles, the 3 seconds water time of sealing concentrates, and obtains concentrating 10 times of bacterium liquid.
Preferably, in the above-mentioned high-concentration bacterial powder making processes, during mixing, with the concentrated solutions of two kinds of bacterium mixed according to 1: 1.
Preferably, in the above-mentioned high-concentration bacterial powder making processes, during spraying drying, the bacterium mud interpolation that mixes is carried out spraying drying by 1/0.3-0.4CaCO3, when inlet temperature reaches 140-150 ℃, begin spraying, control flows into the speed of centrifugal pan slurries, makes the temperature in the drying tower middle level of spraying remain on 60-65 ℃.
Side spore, the potassium sporeformer powder that finally reaches production by above flow process has bacterial classification survival rate height, viable bacteria content height (〉=200 hundred million/gram) reaches hundred million of bacillusmusilaginosiengineering 70-80/gram, hundred million of bacillus laterosporus 120-130/restrain.
Description of drawings
Accompanying drawing 1 microbial bacteria powder technological process of production figure.
Embodiment
1, gel-shaped bacillus fermentation technical process
The bacillusmusilaginosiengineering kind is seeded on the suitable slant medium, treat that growth fully after, place the refrigerator preservation about 4 ℃, manufacture slant strains.The bacterial classification that takes a morsel is forwarded on the Kolle flask slant medium, carries out enlarged culturing (30 ℃ constant temperature culture 2-5 days), treat growth fully after, place the refrigerator preservation about 4 ℃, make the Kolle flask slant strains.Bacterial classification in 30 ℃ of constant temperature culture 2-5 days, is added the washing of 40-50ml sterilized water, make the gemma suspension.To cultivate in the 500L seeding tank based on 0.11MPa, real jar of sterilization of vapor pressure 30 minutes in seeding tank, cultivated 15-20h with the gemma suspension inoculation for 30 ℃, tank pressure 0.05MPa, and ventilation is than 1: 1V/Vm, yeast culture is to the logarithmic growth later stage.With 5m
3Cultivate based on 0.11MPa real jar of sterilization of vapor pressure 30 minutes in the fermentor tank.Thalline and nutrient solution are transferred to 5m in the lump
3In the fermentor tank, cultivate 30-40h for 30 ℃, tank pressure 0.05MPa, ventilation is than 1: after 0.7-1V/Vm, thalline form gemma more than 90%, bacillus exfoliation 50-60%, fermentation ends finally obtains the fermented liquid of gemma content at 6-7 hundred million/gram.
Wherein slant medium is consistent with the Kolle flask substratum, is the silicate bacteria substratum, and it consists of: sucrose (C
12H
22O
11) 5.0g, Sodium phosphate dibasic (Na
2HPO
412H
2O) 2.0g, sal epsom (MgSO
47H
2O) 0.5g, lime carbonate (CaCO
3) 0.1g, iron trichloride (FeCl
36H
2O) 0.005g, agar 18.0g, distilled water 1000mL, pH7.5～8.0.
The bacillusmusilaginosiengineering fermention medium is consistent with the seed tank culture base, and it consists of: starch 5 grams, bean powder 3 grams, sugar 1 gram, CaCO
31 gram, sulphur ammonium 0.5 gram, 2 liters of oil, 1 liter in water, Ph7.0.
2, bacillus laterosporus zymotechnique flow process
The bacillus laterosporus kind is seeded on the suitable slant medium, treat that growth fully after, place the refrigerator preservation about 4 ℃, manufacture slant strains.The bacterial classification that takes a morsel is forwarded on the Kolle flask slant medium, carries out enlarged culturing (34-35 ℃ constant temperature culture 2-5 days), treat growth fully after, place the refrigerator preservation about 4 ℃, make the Kolle flask slant strains.Bacterial classification in 34-35 ℃ of constant temperature culture 2-5 days, is added the washing of 40-50ml sterilized water, make the gemma suspension.To cultivate in the 500L seeding tank based on 0.11MPa, real jar of sterilization of vapor pressure 30 minutes in seeding tank, cultivated 10-15h with the gemma suspension inoculation for 34-36 ℃, tank pressure 0.05MPa, and ventilation is than 1: 1V/Vm, yeast culture is to the logarithmic growth later stage.With 5m
3Cultivate based on 0.11MPa real jar of sterilization of vapor pressure 30 minutes in the fermentor tank.Thalline and nutrient solution are transferred to 5m in the lump
3In the fermentor tank, cultivate 18-25h for 34-36 ℃, tank pressure 0.05MPa, ventilation is than 1: 0.7-1V/Vm, thalline form gemma more than 90%, fermentation ends, the final fermented liquid that gets gemma content at 12-14 hundred million/gram.
Wherein slant medium is consistent with the Kolle flask substratum, is nutrient agar medium, and it consists of: peptone 10.0g, extractum carnis 3.0g, sodium-chlor (NaCl) 5.0g, agar 18.0g, distilled water 1000mL, pH7.2～7.5.
The bacillus laterosporus fermention medium is consistent with the seed tank culture base, and it consists of: soybean cake powder 4 grams, Semen Maydis powder 4 grams, sweet potato powder 4 grams, CaCO
32 grams, 1 liter in water, Ph nature.
3, high speed centrifugation concentrates
With fermented liquid disc centrifuge (7302 rev/mins), concrete controlled variable: 3 seconds sealing time, sealing cycle 1 minute, 1 second deslagging time, in 10 minutes deslagging cycles, the 3 seconds water time of sealing concentrates, and obtains concentrating 10 times of bacterium liquid.
4, mix
With the concentrated solutions of two kinds of bacterium mixed according to 1: 1.
5, spraying drying
The bacterium mud interpolation that mixes is carried out spraying drying by 1/0.3-0.4CaCO3, when inlet temperature reaches 140-150 ℃, begin spraying, control flows into the speed of centrifugal pan slurries, makes the temperature in the drying tower middle level of spraying remain on 60-65 ℃.
Side spore, the potassium sporeformer powder that finally reaches production by above flow process has bacterial classification survival rate height, viable bacteria content height (〉=200 hundred million/gram) reaches hundred million of bacillusmusilaginosiengineering 70-80/gram, hundred million of bacillus laterosporus 120-130/restrain.
Claims (7)
1. method of making high-concentration bacterial powder for microbial fertilizers, it is characterized in that utilizing sophisticated, produce composite bio-fertilizer sporeformer powder with commercial gel-shaped bacillus, bacillus laterosporus.
1.1 gel-shaped bacillus fermentation technical process
The bacillusmusilaginosiengineering kind is seeded on the suitable slant medium, treat that growth fully after, place the refrigerator preservation about 4 ℃, manufacture slant strains.The bacterial classification that takes a morsel is forwarded on the Kolle flask slant medium, carries out enlarged culturing (30 ℃ constant temperature culture 2-5 days), treat growth fully after, place the refrigerator preservation about 4 ℃, make the Kolle flask slant strains.Bacterial classification in 30 ℃ of constant temperature culture 2-5 days, is added the washing of 40-50ml sterilized water, make the gemma suspension.To cultivate in the 500L seeding tank based on 0.11MPa, real jar of sterilization of vapor pressure 30 minutes in seeding tank, cultivated 15-20h with the gemma suspension inoculation for 30 ℃, tank pressure 0.05MPa, and ventilation is than 1: 1V/Vm, yeast culture is to the logarithmic growth later stage.With 5m
3Cultivate based on 0.11MPa real jar of sterilization of vapor pressure 30 minutes in the fermentor tank.Thalline and nutrient solution are transferred to 5m in the lump
3In the fermentor tank, cultivate 30-40h for 30 ℃, tank pressure 0.05MPa, ventilation is than 1: after 0.7-1V/Vm, thalline form gemma more than 90%, bacillus exfoliation 50-60%, fermentation ends finally obtains the fermented liquid of gemma content at 6-7 hundred million/gram.
1.2 bacillus laterosporus zymotechnique flow process
The bacillus laterosporus kind is seeded on the suitable slant medium, treat that growth fully after, place the refrigerator preservation about 4 ℃, manufacture slant strains.The bacterial classification that takes a morsel is forwarded on the Kolle flask slant medium, carries out enlarged culturing (34-35 ℃ constant temperature culture 2-5 days), treat growth fully after, place the refrigerator preservation about 4 ℃, make the Kolle flask slant strains.Bacterial classification in 34-35 ℃ of constant temperature culture 2-5 days, is added the washing of 40-50ml sterilized water, make the gemma suspension.To cultivate in the 500L seeding tank based on 0.11MPa, real jar of sterilization of vapor pressure 30 minutes in seeding tank, cultivated 10-15h with the gemma suspension inoculation for 34-36 ℃, tank pressure 0.05MPa, and ventilation is than 1: 1V/Vm, yeast culture is to the logarithmic growth later stage.With 5m
3Cultivate based on 0.11MPa real jar of sterilization of vapor pressure 30 minutes in the fermentor tank.Thalline and nutrient solution are transferred to 5m in the lump
3In the fermentor tank, cultivate 18-25h for 34-36 ℃, tank pressure 0.05MPa, ventilation is than 1: 0.7-1V/Vm, thalline form gemma more than 90%, fermentation ends finally obtains the fermented liquid of gemma content at 12-14 hundred million/gram.
1.3 above-mentioned two kinds of fermented liquids through high speed centrifugation concentrate, mixing, spraying drying make the bacterium powder.
2. according to claim 1, it is characterized in that: gel-shaped bacillus bacterial classification and bacillus laterosporus bacterial classification are respectively the quality strains that the inventor filters out from soil voluntarily.
3. according to claim 1, it is characterized in that: fermentation tank culture medium was consistent with the seed tank culture base during bacillusmusilaginosiengineering was cultivated, and it consists of: starch 5 grams, bean powder 3 grams, sugar 1 gram, CaCO
31 gram, sulphur ammonium 0.5 gram, 2 liters of oil, 1 liter in water, Ph7.0.Slant medium is consistent with the Kolle flask substratum, is the silicate bacteria substratum, and it consists of: sucrose (C
12H
22O
11) 5.0g, Sodium phosphate dibasic (Na
2HPO
412H
2O) 2.0g, sal epsom (MgSO
47H
2O) 0.5g, lime carbonate (CaCO
3) 0.1g, iron trichloride (FeCl
36H
2O) 0.005g, agar 18.0g, distilled water 1000mL, pH7.5～8.0.
4. according to claim 1, it is characterized in that: fermentation tank culture medium was consistent with the seed tank culture base during bacillus laterosporus was cultivated, and it consists of: soybean cake powder 4 grams, Semen Maydis powder 4 grams, sweet potato powder 4 grams, CaCO
32 grams, 1 liter in water, Ph nature.Slant medium is consistent with the Kolle flask substratum, is nutrient agar medium, and it consists of: peptone 10.0g, extractum carnis 3.0g, sodium-chlor (NaCl) 5.0g, agar 18.0g, distilled water 1000mL, pH7.2～7.5.
5. according to claim 1, it is characterized in that: during high speed centrifugation, with fermented liquid disc centrifuge (7302 rev/mins), concrete controlled variable: 3 seconds sealing time, sealing cycle 1 minute, 1 second deslagging time, 10 minutes deslagging cycles, the 3 seconds water time of sealing concentrates, and obtains concentrating 10 times of bacterium liquid.
6. according to claim 1, it is characterized in that: during mixing, the concentrated solutions of two kinds of bacterium mixed according to 1: 1.
7. according to claim 1, it is characterized in that: during spraying drying, the bacterium mud interpolation that mixes is carried out spraying drying by 1/0.3-0.4CaCO3, when inlet temperature reaches 140-150 ℃, begin spraying, control flows into the speed of centrifugal pan slurries, makes the temperature in the drying tower middle level of spraying remain on 60-65 ℃.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104396486A (en) * | 2014-10-27 | 2015-03-11 | 山东省农作物种质资源中心 | Winter wheat planting method |
| CN104450600A (en) * | 2014-11-14 | 2015-03-25 | 武汉科诺生物科技股份有限公司 | Preparation method of bacillus mucilaginosus raw powder containing 40-100 billion live spores per gram |
| CN105850512A (en) * | 2016-06-03 | 2016-08-17 | 开县代祥食用菌种植厂 | Planting method for white mushrooms |
| CN108795805A (en) * | 2018-06-08 | 2018-11-13 | 山东宸青生物科技有限公司 | Brevibacillus laterosporus fermentation modification method |
| CN110066194A (en) * | 2019-04-02 | 2019-07-30 | 漳州三炬生物技术有限公司 | Novel more bacterium composite microbe fertilizers of one kind and its preparation method and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101643710A (en) * | 2009-08-25 | 2010-02-10 | 山东汇丰源农业科技发展有限公司 | Bacillus subtilis and application thereof |
| CN101665374A (en) * | 2009-09-23 | 2010-03-10 | 新疆农业科学院微生物应用研究所 | Functional microbial fertilizer production by utilizing agricultural wastes |
| CN101671205A (en) * | 2009-09-27 | 2010-03-17 | 上海创博生物技术有限公司 | Composite microbial preparation for promoting growth of rape and preparation method thereof |
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2010
- 2010-06-07 CN CN 201010192330 patent/CN101863685B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101643710A (en) * | 2009-08-25 | 2010-02-10 | 山东汇丰源农业科技发展有限公司 | Bacillus subtilis and application thereof |
| CN101665374A (en) * | 2009-09-23 | 2010-03-10 | 新疆农业科学院微生物应用研究所 | Functional microbial fertilizer production by utilizing agricultural wastes |
| CN101671205A (en) * | 2009-09-27 | 2010-03-17 | 上海创博生物技术有限公司 | Composite microbial preparation for promoting growth of rape and preparation method thereof |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104396486A (en) * | 2014-10-27 | 2015-03-11 | 山东省农作物种质资源中心 | Winter wheat planting method |
| CN104396486B (en) * | 2014-10-27 | 2016-06-22 | 山东省农作物种质资源中心 | A kind of implantation methods of winter wheat |
| CN104450600A (en) * | 2014-11-14 | 2015-03-25 | 武汉科诺生物科技股份有限公司 | Preparation method of bacillus mucilaginosus raw powder containing 40-100 billion live spores per gram |
| CN104450600B (en) * | 2014-11-14 | 2017-06-16 | 武汉科诺生物科技股份有限公司 | A kind of preparation method of 400 1,000 hundred million live spores per gram gel-shaped class bacillus original powder |
| CN105850512A (en) * | 2016-06-03 | 2016-08-17 | 开县代祥食用菌种植厂 | Planting method for white mushrooms |
| CN108795805A (en) * | 2018-06-08 | 2018-11-13 | 山东宸青生物科技有限公司 | Brevibacillus laterosporus fermentation modification method |
| CN110066194A (en) * | 2019-04-02 | 2019-07-30 | 漳州三炬生物技术有限公司 | Novel more bacterium composite microbe fertilizers of one kind and its preparation method and application |
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| CN101863685B (en) | 2013-06-12 |
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