CN101669523A - Production process of bacillus thuringiensis suspension concentrates - Google Patents
Production process of bacillus thuringiensis suspension concentrates Download PDFInfo
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- 238000004519 manufacturing process Methods 0.000 title claims abstract description 18
- 239000004546 suspension concentrate Substances 0.000 title claims abstract description 16
- 241000193388 Bacillus thuringiensis Species 0.000 title claims abstract description 13
- 229940097012 bacillus thuringiensis Drugs 0.000 title claims abstract description 13
- 238000000855 fermentation Methods 0.000 claims abstract description 41
- 230000004151 fermentation Effects 0.000 claims abstract description 41
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 19
- 239000000843 powder Substances 0.000 claims abstract description 18
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 16
- 239000002689 soil Substances 0.000 claims abstract description 10
- 239000008223 sterile water Substances 0.000 claims abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 9
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 claims abstract description 9
- 235000010234 sodium benzoate Nutrition 0.000 claims abstract description 9
- 229920001285 xanthan gum Polymers 0.000 claims abstract description 9
- 229920005551 calcium lignosulfonate Polymers 0.000 claims abstract description 8
- 239000011780 sodium chloride Substances 0.000 claims abstract description 8
- 238000002156 mixing Methods 0.000 claims abstract description 7
- 239000002245 particle Substances 0.000 claims abstract description 7
- 238000012216 screening Methods 0.000 claims abstract description 7
- 241000894006 Bacteria Species 0.000 claims description 17
- 239000000203 mixture Substances 0.000 claims description 16
- 239000000243 solution Substances 0.000 claims description 14
- 238000005516 engineering process Methods 0.000 claims description 10
- 238000002703 mutagenesis Methods 0.000 claims description 8
- 231100000350 mutagenesis Toxicity 0.000 claims description 8
- 239000002002 slurry Substances 0.000 claims description 8
- 229920001817 Agar Polymers 0.000 claims description 7
- 239000001888 Peptone Substances 0.000 claims description 7
- 108010080698 Peptones Proteins 0.000 claims description 7
- 239000008272 agar Substances 0.000 claims description 7
- 235000015278 beef Nutrition 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- 235000019319 peptone Nutrition 0.000 claims description 7
- 230000003068 static effect Effects 0.000 claims description 7
- 231100000331 toxic Toxicity 0.000 claims description 7
- 230000002588 toxic effect Effects 0.000 claims description 7
- 238000005303 weighing Methods 0.000 claims description 7
- 230000001580 bacterial effect Effects 0.000 claims description 5
- 238000010790 dilution Methods 0.000 claims description 5
- 239000012895 dilution Substances 0.000 claims description 5
- 239000012530 fluid Substances 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 5
- 238000010008 shearing Methods 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 5
- 241000255789 Bombyx mori Species 0.000 claims description 4
- 229920002261 Corn starch Polymers 0.000 claims description 4
- 235000019733 Fish meal Nutrition 0.000 claims description 4
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 4
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 4
- 229910017052 cobalt Inorganic materials 0.000 claims description 4
- 239000010941 cobalt Substances 0.000 claims description 4
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 claims description 4
- 239000012141 concentrate Substances 0.000 claims description 4
- 239000008120 corn starch Substances 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 239000004467 fishmeal Substances 0.000 claims description 4
- 230000005251 gamma ray Effects 0.000 claims description 4
- 235000012054 meals Nutrition 0.000 claims description 4
- 230000000394 mitotic effect Effects 0.000 claims description 4
- 238000009423 ventilation Methods 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 abstract description 6
- 238000000227 grinding Methods 0.000 abstract description 3
- 231100000419 toxicity Toxicity 0.000 abstract description 3
- 230000001988 toxicity Effects 0.000 abstract description 3
- 239000006185 dispersion Substances 0.000 abstract description 2
- 239000004299 sodium benzoate Substances 0.000 abstract description 2
- 238000005507 spraying Methods 0.000 abstract description 2
- 230000002349 favourable effect Effects 0.000 abstract 3
- MCMNRKCIXSYSNV-UHFFFAOYSA-N Zirconium dioxide Chemical compound O=[Zr]=O MCMNRKCIXSYSNV-UHFFFAOYSA-N 0.000 abstract 2
- 231100000674 Phytotoxicity Toxicity 0.000 abstract 1
- RYAGRZNBULDMBW-UHFFFAOYSA-L calcium;3-(2-hydroxy-3-methoxyphenyl)-2-[2-methoxy-4-(3-sulfonatopropyl)phenoxy]propane-1-sulfonate Chemical compound [Ca+2].COC1=CC=CC(CC(CS([O-])(=O)=O)OC=2C(=CC(CCCS([O-])(=O)=O)=CC=2)OC)=C1O RYAGRZNBULDMBW-UHFFFAOYSA-L 0.000 abstract 1
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- 239000000230 xanthan gum Substances 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 16
- 239000000047 product Substances 0.000 description 7
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- 230000009969 flowable effect Effects 0.000 description 4
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- 230000032696 parturition Effects 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 238000009736 wetting Methods 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
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- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
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- YDEXUEFDPVHGHE-GGMCWBHBSA-L disodium;(2r)-3-(2-hydroxy-3-methoxyphenyl)-2-[2-methoxy-4-(3-sulfonatopropyl)phenoxy]propane-1-sulfonate Chemical compound [Na+].[Na+].COC1=CC=CC(C[C@H](CS([O-])(=O)=O)OC=2C(=CC(CCCS([O-])(=O)=O)=CC=2)OC)=C1O YDEXUEFDPVHGHE-GGMCWBHBSA-L 0.000 description 1
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- 235000013311 vegetables Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to a production process of bacillus thuringiensis suspension concentrates. The production process comprises the following steps: preparing a solution A by using xanthan gum, sodium chloride and the like; transferring a Bt LX-7 high virulent strain which is separated from soil onto a slant culture medium, and carrying out single colony screening on the Bt LX-7 high virulent strain; selecting a fermentation strain to be fermented; screening and filtering the fermentation liquid, and concentrating the fermentation strain by a centrifuge to obtain strain pulp; directly carrying out spray drying on the strain pulp to obtain Bt LX-7 raw powder, or fully mixing the strain pulp with sodium benzoate, calcium lignosulfonate and sterile water to obtain a solution B; and mixing the solution A and the solution B, and placing the mixing solution and zirconia into a high speed grinding cylinder to obtain the suspension concentrates with the particle size of 0.5-5 microns and thesuspensibility above 90 percent. The production process has high fermentation potency and low production cost. The suspension concentrates has favorable stability, high efficacy, favorable safety, aswell as favorable dispersion and suspensibility in water, is suitable for various spraying modes, is firmly adhered to the surface of a plant, resists rain wash, does not contain organic solvent, isnot inflammable and does not have phytotoxicity, irritation and toxicity caused by the organic solvent.
Description
Technical field
The present invention relates to a kind of production technology of bacillus thuringiensis suspension concentrates.
Background technology
Bacillus thuringiensis is a kind of Gram-positive bacillus, belong to bacillus, its thalline is a rod-short, give birth to flagellum, single giving birth to or the formation short chain, it produces the desinsection parasporal crystal albumen that is called delta-endotoxin in the gemma forming process, these albumen have very high insecticidal activity, it is the most well-known kind in the bacterium class biopesticide, can be from insect, soil, stock and dust, separate in sewage and vegetation etc. and obtain, its preparation is that purposes is the widest in the world at present, the microorganism insecticide of output maximum, account for more than 95% of microorganism insecticide total amount, because of its to pest efficient, to the person poultry safety, environmentally friendly and worldwide be widely used, control efficiency is good, prevention and treatment range is wide, can be widely used in farming, woods, really, vegetables and city trees and shrubs plant insect, the control of storage insect and sanitary insect pest etc., insect are difficult for producing resistance, and production technology is simple, main production raw material are agricultural byproducts, and production and use cost are low.
Suspension concentrates is that the former medicine of solid disperses, is suspended in the aqueous media that contains multiple auxiliary agent with fine droplet, utilizes wet method to carry out the sticky flowable suspension that ultramicro grinding is made.This preparation does not contain organic solvent, and is nonflammable, and safety is good, the poisoning, excitant and the toxicity that do not have organic solvent to cause.Compare with wetting powder, characteristics such as it has, and the powder diameter is little, no dust pollution, penetration are strong, drug effect height have the advantage of wetting powder and two kinds of formulations of missible oil concurrently, can mix use with water.Manufacturing is than missible oil, wetting powder difficulty, the cost height, and the angle of some developed countries from chemistry security is used at first carried out this respect work in the world, and China still is in the starting conceptual phase.
Though China Bt output is bigger at present, quality is unsatisfactory, and the highest suspending agent of tiring of its finished product only is 8000IU/ul, pulvis only is 50000IU/mg, apart from international most advanced level certain gap is arranged still, and formulation is on the low side, can not satisfies the demand of market (particularly international market).In order to remedy the vacancy of product, must improved and perfect the preparation height product of tiring aspect bacterial classification, zymotechnique and the formulation.
Summary of the invention
The objective of the invention is at above-mentioned present situation, aim to provide a kind of fermentation titer height, the production technology of the bacillus thuringiensis suspension concentrates that production cost is low.
The implementation of the object of the invention is, the production technology of bacillus thuringiensis suspension concentrates, and concrete steps are as follows:
1) get 8 gram xanthans and mix well, slowly be poured over then in the 250ml water in the stirring, soaked 2 ± 1 hours, continue to be stirred to dissolving fully, obtain A liquid with 75 ± 5 gram sodium chloride,
2) the Bt LX-7 supper toxic strain that separates from soil, separating method be, gets the 1g soil sample, adds the 99ml sterile water and fully vibrate, and is diluted to 10 then
-7, getting back three dilution factors and be coated with flat board respectively, picking list bacterium colony inserts the inclined-plane behind 28 ℃ of cultivation 1-2d, shake flask fermentation,
3) strain separated is transferred to after the 60 gamma-ray and mutagenesis mutagenesis of 1.2kgy cobalt and carries out single bacterium colony screening on the slant medium, select the height of tiring, sync rates height, the antiphagin ability is strong, fermentation period is short, mitotic stability is good bacterial strain as fermentation strain in enormous quantities
Slant medium: peptone 1 ± 0.2%, beef extract 0.5 ± 0.2%, agar 2 ± 0.5%, pH6.8-7.2 sterilized 30 ± 5 minutes for 121 ± 1 ℃,
4) adopt one grade fermemtation, directly with the spore inoculating of fermentation strain in fermentation tank, 31 ± 1 ℃ of cultivation temperature, ventilation is than 1: 1,220 ± 10 rev/mins of speeds of agitator, pH6.8-7.2, fermentation time 30-35 hour.
Fermentation medium: corn starch 3.0%, dregs of beans 2.5%, fish meal 2.0%, dried silkworm chrysalis meal 1.5%, pH7.5 sterilized 30 minutes for 121 ℃,
Zymotic fluid is transferred pH to 5.0, filter through 100 mesh sieves, centrifuge concentrates for 3 times and obtains the bacterium slurry, and bacterium slurry or direct atomized drying obtain the former powder of Bt LX-7, or enter next step,
5) take by weighing the former powder of 220 gram Bt LX-7,3.5 ± 1 gram Sodium Benzoates, 8 ± 1 gram calcium lignosulfonates, add the 500ml sterile water, fully mix, obtain B liquid,
6) solution A and solution B mixing back are put into the static abrading cylinder that has chuck with 200 ± 20 gram zirconias, the medium of high-speed motion applies shearing force and impulsive force to material, obtains the bacillus thuringiensis suspension concentrates of particle diameter at 0.5～5 μ m.
Technology of the present invention, the fermentation titer height, production cost is low; Producing only needs a sand mill, and equipment is simple; No dust pollution in transportation and the use, noresidue.The strong suspending agent insecticidal crystal protein content that the present invention produces is that 2.4%.54 ℃ of two weeks of storage also are 1.6, and room temperature is placed and observed in three months, and no layering, flatulence, smelly phenomenon are shaken gently, keep even flowable, good stability.
Strong suspending agent is applicable to various spray patterns, also can be used for ultra low volume spraying, has good dispersiveness and suspension in water; And do not contain organic solvent, nonflammable, safety is good, the poisoning, excitant and the toxicity that do not have organic solvent to cause.It is more firm that strong suspending agent invests plant surface, resistance of rainwater washing against, and drug effect is higher.
The present invention is applicable to the production technology of the bacillus thuringiensis suspension concentrates of producing 8000-32000IU/mg.
Embodiment
The concrete steps of production technology of the present invention are as follows:
1) get 8 gram xanthans and mix well, slowly be poured over then in the 250ml water in the stirring, soaked 2 ± 1 hours, continue to be stirred to dissolving fully, obtain A liquid with 75 ± 5 gram sodium chloride,
2) the Bt LX-7 supper toxic strain that separates from soil, separating method be, gets the 1g soil sample, adds the 99ml sterile water and fully vibrate, and is diluted to 10 then
-7, getting back three dilution factors and be coated with flat board respectively, picking list bacterium colony inserts the inclined-plane behind 28 ℃ of cultivation 1-2d, shake flask fermentation, it is standby to choose the supper toxic strain preservation after giving birth to survey.
3) strain separated is transferred to after the 60 gamma-ray and mutagenesis mutagenesis of 1.2kgy cobalt and carries out single bacterium colony screening on the slant medium, select the height of tiring, sync rates height, the antiphagin ability is strong, fermentation period is short, mitotic stability is good bacterial strain as fermentation strain in enormous quantities
Slant medium: peptone 1 ± 0.2%, beef extract 0.5 ± 0.2%, agar 2 ± 0.5%, pH6.8-7.2 sterilized 30 ± 5 minutes for 121 ± 1 ℃,
4) adopt one grade fermemtation, directly with the spore inoculating of fermentation strain in fermentation tank, 31 ± 1 ℃ of cultivation temperature, ventilation is than 1: 1,220 ± 10 rev/mins of speeds of agitator, pH6.8-7.2, fermentation time 30-35 hour, fermentation level can reach 6000IU/ul.
Fermentation medium: corn starch 3.0%, dregs of beans 2.5%, fish meal 2.0%, dried silkworm chrysalis meal 1.5%, pH7.5 sterilized 30 minutes for 121 ℃,
Zymotic fluid is transferred pH to 5.0, filter through 100 mesh sieves, centrifuge concentrates for 3 times and obtains the bacterium slurry, and bacterium slurry or direct atomized drying obtain the former powder of Bt LX-7, or enter next step.
Zymotic fluid is through above-mentioned postprocessing working procedures, and former powder is tired and improved greatly, and tiring reaches 80000IU/mg.Former powder water imbibition is poor, has solved the problem of mobile difference in the strong suspending agent preparation process.
5) take by weighing the former powder of 220 gram Bt LX-7,3.5 ± 1 gram Sodium Benzoates, 8 ± 1 gram calcium lignosulfonates, add the 500ml sterile water, fully mix, obtain B liquid,
6) solution A and solution B mixing back are put into the static abrading cylinder that has chuck with 200 ± 20 gram zirconias, the medium of high-speed motion applies shearing force and impulsive force to material, obtains particle diameter at the acquisition particle diameter of 0.5～5 μ m bacillus thuringiensis suspension concentrates at 0.5～5 μ m.
Utilize that the suitable sodium lignin sulfonate of orthogonal experiment screening is made dispersant, xanthans is made preservative as suspending agent, Sodium Benzoate, add sterile water and the former medicine of a certain amount of high-load, fully mix, constant volume, medium by high-speed motion applies shearing force and impulsive force to material, thereby obtain optimum dispersion, grinding effect, finally obtain even matter, exquisiteness, the toughness high-concentration suspension liquid formulation of particle diameter at 0.5～5 μ m, suspensibility is more than 90%, keep even flowable through storing, heat storage stability is good.
The applicant has made the every performance detection of suspension concentrates, and concrete outcome is as follows:
Biologicall test and insecticidal crystal protein assay: with the cotton bollworm is that biologicall test is tired and is that 17672IU/mg, insecticidal crystal protein content are 2.4%. for the examination worm
Heat storage stability detects: sample is loaded in the vial seals, store a week, two weeks in 54 ℃ of incubators respectively, carry out biologicall test and insecticidal crystal protein assay respectively,
| (IU/mg) tires | Crystalline protein content (%) | |
| Room temperature is placed | ??17672 | ??2.4 |
| 54 ℃ of heat one weeks of storage | ??13846 | ??2.2 |
| 54 ℃ of heat two weeks of storage | ??86342 | ??1.6 |
Room temperature is placed The apparent phenomenon: room temperature is placed and was observed in three months, and no layering, flatulence, smelly phenomenon are shaken gently, keep even flowable.
Enumerate specific embodiments of the invention below:
Example 1,1) get 8 gram xanthans and mix well, slowly be poured over then in the 250ml water in the stirring, soaked 2 hours, continue to be stirred to dissolving fully, obtain A liquid with 75 gram sodium chloride,
2) the Bt LX-7 supper toxic strain that separates from soil, separating method be, gets the 1g soil sample, adds the 99ml sterile water and fully vibrate, and is diluted to 10 then
-7, getting back three dilution factors and be coated with flat board respectively, picking list bacterium colony inserts the inclined-plane behind 28 ℃ of cultivation 2d, shake flask fermentation, it is standby to choose the supper toxic strain preservation after giving birth to survey.
3) strain separated is transferred to after the 60 gamma-ray and mutagenesis mutagenesis of 1.2kgy cobalt and carries out single bacterium colony screening on the slant medium, select the height of tiring, sync rates height, the antiphagin ability is strong, fermentation period is short, mitotic stability is good bacterial strain as fermentation strain in enormous quantities
Slant medium: peptone 1%, beef extract 0.5%, agar 2%, pH7.0 sterilized 30 minutes for 121 ℃,
4) adopt one grade fermemtation, directly with the spore inoculating of fermentation strain in fermentation tank, 31 ℃ of cultivation temperature, ventilation is than 1: 1,220 rev/mins of speeds of agitator, pH7.0, fermentation time 33 hours, fermentation level can reach 6000IU/ul.
Fermentation medium: corn starch 3.0%, dregs of beans 2.5%, fish meal 2.0%, dried silkworm chrysalis meal 1.5%, pH7.5 sterilized 30 minutes for 121 ℃,
Zymotic fluid is transferred pH to 5.0, filter through 100 mesh sieves, centrifuge concentrates for 3 times and obtains the bacterium slurry, and bacterium slurry or direct atomized drying obtain the former powder of Bt LX-7,
5) take by weighing the former powder of 220 gram Bt LX-7,3.5 gram Sodium Benzoates, 8 gram calcium lignosulfonates, add the 500ml sterile water, fully mix, obtain B liquid,
6) solution A and solution B mixing back are put into the static abrading cylinder that has chuck with 200 gram zirconias, the medium of high-speed motion applies shearing force and impulsive force to material, obtains the bacillus thuringiensis suspension concentrates of particle diameter at 0.5～5 μ m.
It is as follows give birth to survey the result: the product 18100IU/mg that tires, crystalline content 2.2%.
Example 2, with example 1, different is, gets 8 gram xanthans and restrains sodium chloride with 80 and mix well, and slowly is poured over then in the 250ml water in the stirring, soaked 1 hour,
Separate Bt LX-7 supper toxic strain, get back three dilution factors and be coated with flat board, cultivate 1d for 28 ℃,
Slant medium: peptone 0.8%, beef extract 0.6%, agar 1.8%, pH6.8 sterilized 38 minutes for 120 ℃,
One grade fermemtation, 30 ℃ of fermentation tank culture temperature, 210 rev/mins of speeds of agitator, pH6.8, fermentation time 32 hours,
Take by weighing the former powder of 220 gram Bt LX-7,3.0 gram Sodium Benzoates, 8.5 gram calcium lignosulfonates,
Solution A and solution B are mixed the back and are put into the static abrading cylinder that has chuck with 180 gram zirconias.
It is as follows give birth to survey the result: the product 17800IU/mg that tires, crystalline content 2.1%.
Example 3, with example 1, different is, get 8 gram xanthans and restrain sodium chloride with 70 and mix well,
Slant medium: peptone 1.2%, beef extract 0.7%, agar 2.5%, pH7.2 sterilized 35 minutes for 122 ℃,
One grade fermemtation, 30 ℃ of fermentation tank culture temperature, 230 rev/mins of speeds of agitator, pH7.2, fermentation time 35 hours.
Take by weighing the former powder of 220 gram Bt LX-7,4.5 gram Sodium Benzoates, 9 gram calcium lignosulfonates,
Solution A and solution B are mixed the back and are put into the static abrading cylinder that has chuck with 220 gram zirconias.
It is as follows give birth to survey the result: the product 17980IU/mg that tires, crystalline content 2.45%.
Example 4, with example 1, different is, get 8 gram xanthans and restrain sodium chloride with 78 and mix well,
Slant medium: peptone 1.1%, beef extract 0.4%, agar 2.2%, pH7.1 sterilized 36 minutes for 120 ℃,
One grade fermemtation, 30 ℃ of fermentation tank culture temperature, 225 rev/mins of speeds of agitator, pH6.9, fermentation time 34 hours.
Take by weighing the former powder of 220 gram Bt LX-7,3.6 gram Sodium Benzoates, 7 gram calcium lignosulfonates,
Solution A and solution B are mixed the back and are put into the static abrading cylinder that has chuck with 210 gram zirconias.
It is as follows give birth to survey the result: the product 17850IU/mg that tires, crystalline content 2.48%.
Claims (1)
1, the production technology of bacillus thuringiensis suspension concentrates is characterized in that concrete steps are as follows:
1) get 8 gram xanthans and mix well, slowly be poured over then in the 250ml water in the stirring, soaked 2 ± 1 hours, continue to be stirred to dissolving fully, obtain A liquid with 75 ± 5 gram sodium chloride,
2) the Bt LX-7 supper toxic strain that separates from soil, separating method be, gets the 1g soil sample, adds the 99ml sterile water and fully vibrate, and is diluted to 10 then
-7, getting back three dilution factors and be coated with flat board respectively, picking list bacterium colony inserts the inclined-plane behind 28 ℃ of cultivation 1-2d, shake flask fermentation,
3) strain separated is transferred to after the 60 gamma-ray and mutagenesis mutagenesis of 1.2kgy cobalt and carries out single bacterium colony screening on the slant medium, select the height of tiring, sync rates height, the antiphagin ability is strong, fermentation period is short, mitotic stability is good bacterial strain as fermentation strain in enormous quantities
Slant medium: peptone 1 ± 0.2%, beef extract 0.5 ± 0.2%, agar 2 ± 0.5%, pH6.8-7.2 sterilized 30 ± 5 minutes for 121 ± 1 ℃,
4) adopt one grade fermemtation, directly with the spore inoculating of fermentation strain in fermentation tank, 31 ± 1 ℃ of cultivation temperature, ventilation is than 1: 1,220 ± 10 rev/mins of speeds of agitator, pH6.8-7.2 fermented 30-35 hour.
Fermentation medium: corn starch 3.0%, dregs of beans 2.5%, fish meal 2.0%, dried silkworm chrysalis meal 1.5%, pH7.5 sterilized 30 minutes for 121 ℃,
Zymotic fluid is transferred pH to 5.0, filter through 100 mesh sieves, centrifuge concentrates for 3 times and obtains the bacterium slurry, and bacterium slurry or direct atomized drying obtain the former powder of Bt LX-7, or enter next step,
5) take by weighing the former powder of 220 gram Bt LX-7,3.5 ± 1 gram Sodium Benzoates, 8 ± 1 gram calcium lignosulfonates, add the 500ml sterile water, fully mix, obtain B liquid,
6) solution A and solution B mixing back are put into the static abrading cylinder that has chuck with 200 ± 20 gram zirconias, the medium of high-speed motion applies shearing force and impulsive force to material, obtains the bacillus thuringiensis suspension concentrates of particle diameter at 0.5～5 μ m.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102910981A (en) * | 2012-10-16 | 2013-02-06 | 南通派斯第农药化工股份有限公司 | 8,000 UI/mg bacillus thuringiensis SC and preparation method thereof |
| CN109022283A (en) * | 2018-08-14 | 2018-12-18 | 华侨大学 | A kind of method of stable liquid photosynthetic bacteria active bacteria formulation |
| CN109593680A (en) * | 2018-12-26 | 2019-04-09 | 武汉科诺生物科技股份有限公司 | A kind of Dipel liquid fermentation medium and its bacterium powder and oil suspending agent |
| CN112501083A (en) * | 2020-12-22 | 2021-03-16 | 武汉科诺生物科技股份有限公司 | Bacillus thuringiensis liquid fermentation medium and fermentation culture method |
-
2009
- 2009-08-19 CN CN200910063658A patent/CN101669523A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102910981A (en) * | 2012-10-16 | 2013-02-06 | 南通派斯第农药化工股份有限公司 | 8,000 UI/mg bacillus thuringiensis SC and preparation method thereof |
| CN109022283A (en) * | 2018-08-14 | 2018-12-18 | 华侨大学 | A kind of method of stable liquid photosynthetic bacteria active bacteria formulation |
| CN109593680A (en) * | 2018-12-26 | 2019-04-09 | 武汉科诺生物科技股份有限公司 | A kind of Dipel liquid fermentation medium and its bacterium powder and oil suspending agent |
| CN109593680B (en) * | 2018-12-26 | 2022-02-22 | 武汉科诺生物科技股份有限公司 | Bacillus thuringiensis liquid fermentation medium and bacterial powder and oil suspension agent thereof |
| CN112501083A (en) * | 2020-12-22 | 2021-03-16 | 武汉科诺生物科技股份有限公司 | Bacillus thuringiensis liquid fermentation medium and fermentation culture method |
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