CN104336076B - A kind of preparation technology of bacillus thuringiensis active bacteria formulation - Google Patents
A kind of preparation technology of bacillus thuringiensis active bacteria formulation Download PDFInfo
- Publication number
- CN104336076B CN104336076B CN201410436991.8A CN201410436991A CN104336076B CN 104336076 B CN104336076 B CN 104336076B CN 201410436991 A CN201410436991 A CN 201410436991A CN 104336076 B CN104336076 B CN 104336076B
- Authority
- CN
- China
- Prior art keywords
- medium
- fermentation
- bacillus thuringiensis
- culture
- glucose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
Access in fermentation medium and cultivate after I and II seed expands culture the present invention relates to a kind of preparation technology bacillus thuringiensis of bacillus thuringiensis active bacteria formulation, add the quantity and conversion ratio of carbon source increase bacillus by feed supplement stream during the fermentation, specific yield is significantly improved, reduces unit cost；Terminate fermentation, using vacuum film inspissator concentrated broth, soluble synergistic matter, growth-promoting material, Substance in zymotic fluid can be reclaimed effectively；The multistage cold nebulization fluidization drying apparatus drying-granulating of concentrated broth, sort particle, cyclone separator trapping fine powder returns to be granulated again, is obtained every gram of bacterium powder biologicall test and is not less than 80000IU/mg to bollworm activity, insecticidal crystal protein content is not less than 13% bacillus thuringiensis class product.Vent gas treatment, qualified discharge.The present invention can at utmost retain active principle in zymotic fluid, and product quality is high, utilization rate of equipment and installations is high, relatively good, the waste gas that bioactivity is kept, and discharged wastewater met the national standard can large-scale industrialized production.
Description
Technical field
The present invention relates to a kind of preparation technology of bacillus thuringiensis active bacteria formulation.
Background technology
Bacillus thuringiensis (Bacillus thuringiensis, abbreviation Bt) is that a kind of important Insect Pathogenic is thin
Bacterium, and the important member of countries in the world research and development microbial insecticide.The major advantage of Bt insecticides：1. pair people, animal are pacified
Entirely, not pollution of ecological environment；2. a pair target pest has specific toxic action, to the Non-target pests such as silkworm and natural enemy without
Poison；3. prevention effect is good, prevention and treatment range is wide, can be widely used for the preventing and treating of the insects such as agriculture, woods, fruit and vegetable and urban park plant, evil
Worm is not likely to produce resistance；User will not be caused to be poisoned 4. easy to use, during use；5. simple production process, main production is former
Material is agricultural byproducts, and production is low with use cost.
Bt insecticides can be used for preventing and treating cotton and other crops, such as：Vegetables, fruit tree, tobacco, forest and city trees and shrubs are planted
Thing endanger serious and refractory insect preventing and treating (including：The bollworm of Lepidoptera, diamondback moth, beet armyworm, prodenia litura, dish
Blue or green worm, the colorado potato bug of dendrolimus punctatus and coleoptera, daikon leaf beetle, willow herb are chrysomelid etc.), application is quite varied.It is international
Existing more than ten of the kind of upper Bt insecticides, its merchandized handling are concentrated mainly on the countries such as the U.S., France, Belgium.City of China
Demand is also very big, only with 70,000,000 mu of cottons, 1.3 hundred million mu of vegetables, 20,000,000 mu of potatos 50% areal calculation, press
Every mu uses 0.5 kilogram of Bt pulvis, needs ten thousand tons of pulvis about 7.5-8.0 years, equivalent newborn ten thousand tons of suspension 50-60, additional other agricultures,
The preventing and treating of the insects such as woods, fruit tree.Current demand of the domestic Bt insecticides in biological control of insect pests accounts for 90%, but annual production only accounts for
The 1% of agricultural chemicals total amount, determine that biological pesticide develops into 60% requirement and differed greatly from " world environments and development conference "；Cause
This, the exploitation of China's Bt insecticides has wide market prospects.
The common production method of sporeine preparation is using related bacillus thuringiensis production bacterium at present
Kind, batch fermentation culture, when to it, most or all form gemma, carried out by the way of plate-frame filtering or centrifugation dense
Contracting, is dried after adding auxiliary agent or spray drying obtains the active bacteria formulation of bacillus thuringiensis.There is following for these methods
Deficiency：
1st, fermentation tank coefficient is 60%-70%, and utilization rate of equipment and installations is not high；
2nd, due to the batch fermentation of use, initial medium excessive concentration of fermenting is caused, lag phase is longer after inoculation, fermentation
Byproduct in process thing forms more, and fermented product concentration does not increase, and it is larger to ultimately result in fermentation costs；
3rd, the flocculation/plate-frame filtering or centrifugation technique that the concentration of zymotic fluid typically uses, both technologies typically all can
Substantial amounts of waste water is produced, subsequent treatment cost is higher；
4th, it can also cause the active ingredient inside fermentation supernatant to lose using flocculation/plate-frame filtering or centrifugation technique, make
Into the decline of product effect；
5th, the inlet temperature that spray drying is typically set is 180 DEG C to 200 DEG C, and outlet temperature is 80 DEG C to 100 DEG C, at this
Under the higher operating temperature of sample, the bacterium number of some kinds has a certain degree of decline, directly affects the quality of product.
Vacuum film evaporation device is the general name of a kind of evaporator, feature be feed liquid along heating tube wall in membranaceous flowing and
Conducted heat and evaporated, advantage is heat transfer efficiency height, and evaporation rate is fast, and residence time of material is short, therefore is particularly suitable for thermal sensitivity
The evaporation of material.Such evaporator includes MVR (mechanical steam recompression) evaporator, falling film type concentration evaporator and board-like dense
Contracting evaporator etc..Usually it is applied to active Chinese drug component material to concentrate, food additives concentration, antibiotic concentration, enzyme preparation concentration
Deng field, but there is presently no in terms of active bacteria formulation concentration.
Multistage cold nebulization fluidized drying is granulated compared to conventional spray drying tower, employs a variety of advanced technologies, than
As used multiple spray gun structures in tower, spray gun is using high pressure nozzle, and the droplet dispersion effect of ejection is good, and multiple nozzles spray
The droplet gone out mutually collides, and easily forms bulky grain；Without forming then being trapped by cyclone separator for particle, tower is then back to
Push up agglomeration again and granulation that range of atomization carries out powder；And bottom of towe has fluid bed to carry out redrying to the particle of formation,
Further reduce the water content of particle；System requirements charging has very high solid content (10%-25%), need not so bond
Agent just can directly form particle；Due to being multilevel drying system, and material moisture is not high, so the operation temperature of the equipment
It is low compared to conventional spray drying tower, the control of general air temperature at 130 DEG C -180 DEG C, outlet temperature control 60 DEG C -80 DEG C it
Between, so the system possesses the ability for drying heat sensitive material well, the technology is applied in the drying of active bacteria formulation, can
The advantages that bioactivity being effectively maintained in product, obtains the product of high quality, and environmental protection, and process is few.
Fermentation and drying-granulating can produce substantial amounts of waste gas, and volatile organic matter (VOC), hydrogen sulfide, ammonia are often contained in the inside
The various pollutants such as gas, thio-alcohol, while with unpleasant foul smell, past fermentation enterprise typically directly discharges it or letter
It is single with being discharged after Water spray, two kinds of processing modes can cause the serious pollution to environment.It is direct using suitable chemical reagent
Contaminants associated in oxidation gaseous effluent or to decompose pollutant therein with physical method be relatively good processing method, can solve
Foul smell in removing exhaust gas, reduces environmental pollution.
The content of the invention
The purpose of the present invention is to be directed to above-mentioned present situation, it is desirable to provide a kind of utilization rate of equipment and installations is high, and product quality improves, biology
Relatively good, the waste gas that activity is kept, discharged wastewater met the national standard, can large-scale industrialized production, product quality is more stable, yield
It is higher, the preparation technology for the bacillus thuringiensis active bacteria formulation that production cost substantially reduces.
The implementation of the object of the invention is a kind of preparation technology of bacillus thuringiensis active bacteria formulation, specific steps
It is as follows：
1) first order seed culture：2000ml primary-seed mediums are housed, 115 DEG C -121 DEG C damp and hot in 5000ml triangular flasks
Sterilize 15-30min, take 10ml bacillus thuringiensis viable bacteria freeze pipe strain be inoculated in primary-seed medium, 30 DEG C-
35 DEG C of 8~10h of culture；The raw material and dosage of first order seed culture used medium be：Glucose 0.5%-3%, beef extract
0.5%-3%, peptone 0.5%-3%, sodium chloride 0.3-1%, medium pH 6.5-7.5；
2) secondary seed culture：150L or 300L secondary seed mediums are housed in 400L fermentation tanks, 115 DEG C -121 DEG C
Moist heat sterilization 15-30min, 3000ml first order seeds are inoculated in secondary seed medium；30 DEG C -35 DEG C culture 8~
10h；The raw material and dosage of secondary seed culture used medium be：Glucose 0.5%-3%, yeast extract 0.5%-3%,
Peptone 0.5%-3%, dipotassium hydrogen phosphate 0.02%-0.05%, sodium chloride 0.5%-1%, magnesium sulfate 0.02%-0.1%, training
Support base pH and be adjusted to 6.5-7.5；
3) ferment：In 5m3Or 40m34000L or 30m is housed in fermentation tank3Bacillus thuringiensis viable bacteria fermentation culture medium,
115 DEG C of -121 DEG C of moist heat sterilization 15-30min, bacillus thuringiensis viable bacteria fermentation is inoculated in by 150L or 300L secondary seeds
In culture medium, control tank pressure 0.02Mpa-0.05Mpa, in 28 DEG C of -35 DEG C of culture 30-36h；Ferment used medium raw material and
Dosage is：Glucose 0.5%-3%, dregs of beans 1%-4%, corn steep liquor 1%-3%, dusty yeast 1%-3%, peptone 1%-3%,
Dipotassium hydrogen phosphate 0.01%-0.05%, magnesium sulfate 0.1%-0.2%, manganese sulfate 0.002%, medium pH 6.5-7.5；
4) feed supplement：When fermentation to 8-12h starts stream plus the grape being configured to by glucose 20%-60% water 40%-80%
Sugared supplemented medium, ferment to 24-28 hours and terminate；
5) when being more than or equal to 90% to spore forming rate, fermentation is terminated；
6) enter inspissator when zymotic fluid being heated into 35 DEG C -60 DEG C, carried out in the case where vacuum is -70kpa~-95kpa
Concentration, when being concentrated into solid content and reaching 10%-25%, start to discharge；
7) granulation is dried with multistage cold nebulization fluidization drying apparatus in concentrated broth, and particle meets by sorting
Particle requirement directly goes out tower, obtains and is not less than 80000IU/mg to bollworm activity, and insecticidal crystal protein content is not less than 13%
Bacillus thuringiensis product；180 DEG C of the EAT, 60 DEG C of outlet temperature；
8) vent gas treatment：De-tasted using chemical oxidation, plasma irradiation or ultraviolet irradiation, the processing of ammonia nitrogen removal or depriving hydrogen sulphide
After discharge.
The invention has the advantages that：
1st, by the method for feed supplement, specific yield is significantly improved, reduces unit cost, fermentation tank coefficient is by original
First 60%-70% brings up to 80%-90%, and utilization rate of equipment and installations improves, and single tank yield greatly promotes；By fed-batch fermentation, reach
To high density fermentation, biomass is improved, zymotic fluid active constituent content improves 10-20%, higher than batch fermentation；
2nd, using multistage fermentation technique, it can be used on various scale fermentation tanks, shorten the fermentation period of bacillus,
Reduce fermentation microbiological contamination risk；
3rd, using the direct concentrated broth of cryogenic vacuum thin film concentrator, compared to using traditional flocculation/plate-frame filtering or
The condensing mode of person's centrifugation, soluble synergistic matter, growth-promoting material, Substance etc. are effectively returned in zymotic fluid
Receive；
4th, using low temperature multistage drying and granulating new technology, production technology is spray-dried compared to conventional centrifugal, import wind-warm syndrome and
It is low to export wind-warm syndrome, preferably remains bioactivity, and disposably obtain water dispersion granule product rather than powder product；
5th, not having to addition auxiliary agent after fermenting can dry or be spray-dried：
6th, caused condensed water in low temperature concentration process, biochemical oxygen demand (BOD) (BOD values) is extremely low, typically below 80, reaches
Qualified discharge standard；And traditional flocculation/plate-frame filtering or the method for concentration of centrifugation, a large amount of biochemical oxygen demand (BOD) (BOD can be produced
Value) very high waste water, it is necessary to carry out the processing of complexity after be only possible to reach discharge standard；
7th, fermentation tail gas, drying and granulating tail gas are collected by collection system, are irradiated through chemical oxidation, plasma, be ultraviolet
Can qualified discharge after irradiation etc. is de-tasted, ammonia nitrogen removal, depriving hydrogen sulphide vent gas treatment mode are handled.
Embodiment
The present invention takes bacillus thuringiensis seed to freeze pipe strain, is inoculated in 121 DEG C of moist heat sterilization 30min one-level kind
In sub- culture medium, first order seed culture is carried out.First order seed is inoculated in through 121 DEG C of sterilizing 30min, secondary seed medium
Secondary seed culture is carried out in fermentation tank.Fermentation, feed supplement during the fermentation, terminate fermentation.
I and II seed culture medium, fermentation medium, after moist heat sterilization, culture medium will be rapidly decreased to supplemented medium
30 DEG C or so.
Using vacuum film inspissator concentrated broth, inspissator is that MVR inspissators, falling film evaporator or plate evaporation are dense
Contracting device.Using MVR inspissators, the indirect steam come out from vaporization chamber compresses through compressor, and pressure and temp is increased to 100 DEG C -115
DEG C, heating chamber is delivered to when heating steam uses, and heating zymotic fluid to feeding temperature, heating steam is condensed into water in itself.
Multistage cold nebulization fluidized drying is granulated：Concentrated broth is dried with multistage cold nebulization fluidization drying apparatus
It is granulated, particle meets particle requirement and directly go out tower, obtain and be not less than 80000IU/mg to bollworm activity, desinsection is brilliant by sorting
Body protein content is not less than 13% bacillus thuringiensis product；130 DEG C -180 DEG C of the EAT, outlet temperature 60
℃-80℃；
The present invention is described in detail with specific embodiment below：
Embodiment 1：(with 5m3Exemplified by fermentation tank)
1) first order seed culture：2000ml primary-seed mediums, 121 DEG C of moist heat sterilizations are housed in 5000ml triangular flasks
15min, take 10ml bacillus thuringiensis viable bacteria to freeze pipe strain and be inoculated in primary-seed medium, 14h is cultivated at 30 DEG C,
On inspection, strain has no microbiological contamination, and thalli morphology is preferable, can carry out next step operation；First order seed culture used medium
Raw material and dosage are：Glucose 0.5%, beef extract 3%, peptone 1.5%, sodium chloride 0.5%, medium pH 7.5；
2) secondary seed culture：150L secondary seed mediums, 121 DEG C of moist heat sterilizations are housed in 400L fermentation tanks
15min, 3000ml first order seeds are inoculated in secondary seed medium；14h is cultivated at 30 DEG C, on inspection, strain has no dye
Bacterium, thalli morphology is preferable, can carry out next step operation；The raw material and dosage of secondary seed culture used medium be：Grape
Sugar 0.5%, yeast extract 0.5%, peptone 3%, dipotassium hydrogen phosphate 0.02%, medium pH is adjusted to 7.5；
3) ferment：In 5m34000L bacillus thuringiensis viable bacteria fermentation culture mediums are housed, 121 DEG C damp and hot to go out in fermentation tank
Bacterium 15min, 150L secondary seeds are inoculated in bacillus thuringiensis viable bacteria fermentation culture medium, control tank pressure 0.02Mpa-
0.05Mpa, 36h is cultivated at 30 DEG C.Ferment used medium raw material and dosage be：Glucose 1%, dregs of beans 1%, corn steep liquor
1%, peptone 3%, dipotassium hydrogen phosphate 0.05%, magnesium sulfate 0.1%, manganese sulfate 0.002%, medium pH 7.5；
4) feed supplement：Start stream plus the glucose supplemented medium being configured to by glucose 60%, water 40% to 8h when fermenting,
Fermentation terminated to 28 hours；
5) when gemma and crystal have 10% separation, fermentation is terminated；
6) enter inspissator when zymotic fluid being heated into 35 DEG C, concentrated in the case where vacuum is -95kpa, be concentrated into and contain
Gu amount reaches 10%-20%, start to discharge；
7) granulation is dried with multistage cold nebulization fluidization drying apparatus in concentrated broth, and particle meets by sorting
Particle requirement directly goes out tower, obtains and is not less than 80000IU/mg to bollworm activity, and insecticidal crystal protein content is not less than 13%
Bacillus thuringiensis product；130 DEG C of the EAT, 60 DEG C of outlet temperature；
8) vent gas treatment：Drying and granulating tail gas is collected by collection system, then by chemical scrubber, with chemical oxidation
Solution-air contact occurs for agent such as hydrogen peroxide, sodium hypochlorite, and the malodorous elements during this in gas phase are transferred to liquid phase, and are changed
Learn it is oxidizing, so as to reach the purpose of de-tasting, ammonia nitrogen removal, depriving hydrogen sulphide, then qualified discharge in a organized way.
Embodiment 2, with embodiment 1, unlike,
1) raw material of first order seed culture used medium and dosage are：Glucose 1%, beef extract 0.5%, peptone
1.5%, sodium chloride 0.5%, medium pH 7.5；115 DEG C of moist heat sterilization 30min, 35 DEG C of culture 8h after inoculation.
2) raw material of secondary seed culture used medium and dosage are：Glucose 3%, yeast extract 1.5%, albumen
Peptone 3%, dipotassium hydrogen phosphate 0.05%, medium pH is adjusted to 7.5；115 DEG C of moist heat sterilization 30min, 35 DEG C of culture 8h after inoculation.
3) raw material of fermentation used medium and dosage are：Glucose 1%, dregs of beans 4%, corn steep liquor 1%, peptone 3%,
Dipotassium hydrogen phosphate 0.05%, magnesium sulfate 0.1%, manganese sulfate 0.002%, medium pH 6.5；115 DEG C of moist heat sterilization 30min,
35 DEG C of culture 8h after inoculation.
4) feed supplement：When fermentation to 12h starts stream plus the glucose feed-batch culture being configured to by glucose 50%, water 50%
Base, ferment to 25 hours and terminate；
6) enter vacuum film inspissator when zymotic fluid being heated into 60 DEG C using falling film evaporator, vacuum for-
It is evaporated under 70kpa, then sequentially enters two effects and triple effect evaporator, when being concentrated into solid content and reaching 15%-25%, is started
Discharging.
7) granulation is dried with multistage cold nebulization fluidization drying apparatus in concentrated broth, and particle meets by sorting
Particle requirement directly goes out tower, obtains and is not less than 80000IU/mg to bollworm activity, and insecticidal crystal protein content is not less than 13%
Bacillus thuringiensis product；180 DEG C of the EAT, 80 DEG C of outlet temperature；
The made bacillus thuringiensis dry powder active bacteria formulation of the present embodiment is not less than 80000IU/mg to bollworm activity,
Insecticidal crystal protein content is not less than 13%.
Embodiment 3, with embodiment 1, unlike,
1) raw material of first order seed culture used medium and dosage are：Glucose 1.5%, beef extract 3%, peptone
0.5%, sodium chloride 1%, medium pH 7.0；115 DEG C of moist heat sterilization 30min, 35 DEG C of culture 8h after inoculation.
2) raw material of secondary seed culture used medium and dosage are：Glucose 2.5%, yeast extract 0.5%, egg
White peptone 3%, dipotassium hydrogen phosphate 0.05%, medium pH is adjusted to 7.0；115 DEG C of moist heat sterilization 30min, 35 DEG C of culture 8h after inoculation.
3) raw material of fermentation used medium and dosage are：Glucose 1%, dregs of beans 4%, corn steep liquor 3%, peptone 3%,
Dipotassium hydrogen phosphate 0.05%, magnesium sulfate 0.2%, manganese sulfate 0.002%, medium pH 7.0；115 DEG C of moist heat sterilization 30min,
35 DEG C of culture 30h after inoculation.
4) feed supplement：When fermentation to 10h starts stream plus the glucose feed-batch culture being configured to by glucose 40%, water 60%
Base, ferment to 27 hours and terminate；
6) plate evaporation inspissator concentrated broth is used：Enter plate evaporation inspissator when zymotic fluid is heated into 50 DEG C
Concentrated, concentration is evaporated under vacuum -86kp.Then when zymotic fluid, which is concentrated into solid content, reaches 15%-25%,
Start to discharge.
7) granulation is dried with multistage cold nebulization fluidization drying apparatus in concentrated broth, and particle meets by sorting
Particle requirement directly goes out tower, obtains and is not less than 80000IU/mg to bollworm activity, and insecticidal crystal protein content is not less than 13%
Bacillus thuringiensis product；140 DEG C of the EAT, 70 DEG C of outlet temperature；
The made bacillus thuringiensis dry powder active bacteria formulation of the present embodiment is not less than 80000IU/mg to bollworm activity,
Insecticidal crystal protein content is not less than 13%.
Embodiment 4：(with 40m3Exemplified by fermentation tank), with embodiment 1, the difference is that
1) raw material of first order seed culture used medium and dosage are：Glucose 2.5%, beef extract 3%, peptone
0.5%, sodium chloride 0.5%, medium pH 7.5；
2)) secondary seed culture：300L secondary seed mediums are housed in 400L fermentation tanks, used in secondary seed culture
The raw material and dosage of culture medium be：Glucose 2.5%, yeast extract 3%, peptone 0.5%, dipotassium hydrogen phosphate 0.05%,
Medium pH is adjusted to 7.5；
3) ferment：In 40m330m is housed in fermentation tank3Bacillus thuringiensis fermentation medium, access 300L two level kinds
Son.Ferment used medium raw material and dosage be：Glucose 3%, dregs of beans 4%, corn steep liquor 1%, peptone 1%, phosphoric acid hydrogen
Dipotassium 0.05%, magnesium sulfate 0.1%, manganese sulfate 0.002%, medium pH 7.5；
4) feed supplement：Start stream plus the glucose supplemented medium being configured to by glucose 20%, water 80% to 9h when fermenting,
Fermentation terminated to 27 hours；
7) granulation is dried with multistage cold nebulization fluidization drying apparatus in concentrated broth, and particle meets by sorting
Particle requirement directly goes out tower, obtains and is not less than 80000IU/mg to bollworm activity, and insecticidal crystal protein content is not less than 13%
Bacillus thuringiensis product；150 DEG C of the EAT, 70 DEG C of outlet temperature；
The made bacillus thuringiensis dry powder active bacteria formulation of the present embodiment is not less than 80000IU/mg to bollworm activity,
Insecticidal crystal protein content is not less than 13%.
9) vent gas treatment：Drying and granulating tail gas is collected by collection system, then rich by plasma reaction area, the area
Containing high chemically active material, such as high energy electron, ion, free radical and excited state molecule etc., polluter in waste gas with
These high energy capacity materials produce reaction, are decomposed in very short time, so as to reach de-taste, the mesh such as ammonia nitrogen removal, depriving hydrogen sulphide
, then qualified discharge in a organized way.
Embodiment 5：(with 40m3Exemplified by fermentation tank) with embodiment 1, the difference is that,
1) raw material of first order seed culture used medium and dosage are：Glucose 2.5%, beef extract 2.6%, peptone
0.8%, sodium chloride 0.5%, medium pH 7.5；
2) secondary seed culture：300L secondary seed mediums are housed in 400L fermentation tanks, used in secondary seed culture
The raw material and dosage of culture medium be：Glucose 2.5%, yeast extract 3%, peptone 1.5%, dipotassium hydrogen phosphate 0.02%,
Medium pH is adjusted to 7.5；
3) ferment：In 40m330m is housed in fermentation tank3Bacillus thuringiensis fermentation medium, access 300L two level kinds
Son.Ferment used medium raw material and dosage be：Glucose 2%, dregs of beans 3%, corn steep liquor 2%, peptone 1%, phosphoric acid hydrogen
Dipotassium 0.1%, magnesium sulfate 0.1%, manganese sulfate 0.002%, medium pH 7.5；
4) feed supplement：When fermentation to 10h starts stream plus the glucose feed-batch culture being configured to by glucose 20%, water 80%
Base, ferment to 26 hours and terminate；
6) enter vacuum film inspissator when zymotic fluid being heated into 45 DEG C using falling film evaporator, vacuum for-
It is evaporated under 88kpa, then sequentially enters two effects and triple effect evaporator, when being concentrated into solid content and reaching 15%-25%, is started
Discharging.
The made bacillus thuringiensis dry powder active bacteria formulation of the present embodiment is not less than 80000IU/mg to bollworm activity,
Insecticidal crystal protein content is not less than 13%.
9) vent gas treatment：Fermentation tail gas, drying and granulating tail gas are collected by collection system, then pass through high energy UV ultraviolets
Reaction zone, the oxygen molecule in the air in the area, which can be decomposed, produces free oxygen, and free oxygen is unstable, and that generation is combined with oxygen molecule is smelly
Oxygen.Pre-Ozonation on Organic Matter has extremely strong oxidation, can be produced with the organic pollution in waste gas and decompose oxidation reaction, make its drop
Solution changes into low molecular compound, water and carbon dioxide, so as to reach the purpose of de-tasting, ammonia nitrogen removal, depriving hydrogen sulphide, then there is group
Knit qualified discharge.
Embodiment 6：(with 40m3Exemplified by fermentation tank), with embodiment 1, the difference is that,
1) raw material of first order seed culture used medium and dosage are：Glucose 2.5%, beef extract 2%, peptone
0.8%, sodium chloride 0.5%, medium pH 7.5；
2) 300L secondary seed mediums, 121 DEG C of moist heat sterilization 30min, by 3000ml mono- are housed in 400L fermentation tanks
Level seed is inoculated in secondary seed medium；The raw material and dosage of secondary seed culture used medium be：Glucose 2%,
Yeast extract 2.5%, peptone 1.5%, dipotassium hydrogen phosphate 0.02%, medium pH is adjusted to 7.5.
3) ferment：In 40m330m is housed in fermentation tank3Bacillus coagulans fermentation medium, sterilize 30min's through 121 DEG C
Bacillus thuringiensis fermentation medium, access 300L secondary seeds, control tank pressure 0.02Mpa-0.05Mpa, in 30 DEG C of cultures
35h；Tank is put when sporulation has 90%.Ferment used medium raw material and dosage be：Glucose 2%, dregs of beans 3% are beautiful
Rice ＆ peanut milk 2%, peptone 2%, dipotassium hydrogen phosphate 0.05%, magnesium sulfate 0.2%, manganese sulfate 0.002%, medium pH 7.5.
5) fermentation is terminated：When being more than or equal to 90% to spore forming rate, terminate fermentation；
6) plate evaporation inspissator concentrated broth is used：Enter plate evaporation inspissator when zymotic fluid is heated into 45 DEG C
Concentrated, be evaporated under vacuum -88kp.When being concentrated into solid content and reaching 15%-25%, start to discharge.
The made bacillus thuringiensis dry powder active bacteria formulation of the present embodiment is not less than 80000IU/mg to bollworm activity,
Insecticidal crystal protein content is not less than 13%.
9) fermentation tail gas, drying and granulating tail gas are collected by collection system, should then by high energy UV ultraviolet reaction zones
Oxygen molecule in the air in area, which can be decomposed, produces free oxygen, and unstable combined with oxygen molecule of free oxygen produces ozone.Ozone pair
Organic matter has extremely strong oxidation, can with waste gas organic pollution produce decompose oxidation reaction, make its Degradation and Transformation into
Low molecular compound, water and carbon dioxide, so as to reach the purpose of de-tasting, ammonia nitrogen removal, depriving hydrogen sulphide, then row up to standard in a organized way
Put.
Claims (4)
1. a kind of preparation technology of bacillus thuringiensis active bacteria formulation, it is characterised in that comprise the following steps that：
1) first order seed culture：2000ml primary-seed mediums, 115 DEG C of -121 DEG C of moist heat sterilizations are housed in 5000ml triangular flasks
15-30min, take 10ml bacillus thuringiensis viable bacteria to freeze pipe strain and be inoculated in primary-seed medium, at 30 DEG C -35 DEG C
Cultivate 8~14h；The raw material and dosage of first order seed culture used medium be：Glucose 0.5%-2.5%, beef extract
0.5%-3%, peptone 0.5%-1.5%, sodium chloride 0.5-1%, medium pH 7.0-7.5；
2) secondary seed culture：150L or 300L secondary seed mediums are housed in 400L fermentation tanks, 115 DEG C -121 DEG C damp and hot
Sterilized 15-30min, and 3000ml first order seeds are inoculated in secondary seed medium；In 30 DEG C of -35 DEG C of 8~14h of culture；Two
Level seed culture used medium raw material and dosage be：Glucose 0.5%-2.5%, yeast extract 0.5%-3%, albumen
Peptone 0.5%-3%, dipotassium hydrogen phosphate 0.02%-0.05%, medium pH are adjusted to 7.0-7.5；
3) ferment：In 5m3Or 40m34000L or 30m is housed in fermentation tank3Bacillus thuringiensis viable bacteria fermentation culture medium, 115
DEG C -121 DEG C of moist heat sterilization 15-30min, bacillus thuringiensis viable bacteria fermentation culture is inoculated in by 150L or 300L secondary seeds
In base, control tank pressure 0.02Mpa-0.05Mpa, in 30 DEG C of -35 DEG C of culture 30-36h；The raw material and dosage of fermentation used medium
For：Glucose 1%-3%, dregs of beans 1%-4%, corn steep liquor 1%-3%, peptone 1%-3%, dipotassium hydrogen phosphate 0.05%-
0.1%, magnesium sulfate 0.1%-0.2%, manganese sulfate 0.002%, medium pH 6.5-7.5；
4) feed supplement：When the glucose that fermentation to 8-12h starts stream plus is configured to by glucose 20%-60%, water 40%-80% is mended
Expect culture medium, ferment to 25-28 hours and terminate；
5) when spore forming rate is more than or equal to 90%, fermentation is terminated；
6) enter inspissator when zymotic fluid being heated into 35 DEG C -60 DEG C, concentrated in the case where vacuum is -70kpa~-95kpa,
When being concentrated into solid content and reaching 10%-25%, start to discharge；
7) granulation is dried with multistage cold nebulization fluidization drying apparatus in concentrated broth, and particle meets particle by sorting
It is required that directly going out tower, obtain and 80000IU/mg is not less than to bollworm activity, insecticidal crystal protein content is not less than 13% Su Yun
Golden bacillus product；130-180 DEG C of the EAT, 60-80 DEG C of outlet temperature；
8) vent gas treatment：De-tasted using chemical oxidation, plasma irradiation or ultraviolet irradiation, ammonia nitrogen removal or depriving hydrogen sulphide processing heel row
Put.
2. the preparation technology of a kind of bacillus thuringiensis active bacteria formulation according to claim 1, it is characterised in that one, two
Level seed culture medium, fermentation medium, for supplemented medium after moist heat sterilization, culture medium will be rapidly decreased to 30 DEG C.
A kind of 3. preparation technology of bacillus thuringiensis active bacteria formulation according to claim 1, it is characterised in that concentration
Device is MVR inspissators, falling film evaporator or plate evaporation inspissator.
4. the preparation technology of a kind of bacillus thuringiensis active bacteria formulation according to claim 1, it is characterised in that use
MVR inspissators, the indirect steam come out from vaporization chamber compress through compressor, and temperature is increased to 100 DEG C -115 DEG C, delivers to heating chamber
When heating steam uses, heating zymotic fluid to feeding temperature, heating steam is condensed into water in itself.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410436991.8A CN104336076B (en) | 2014-08-29 | 2014-08-29 | A kind of preparation technology of bacillus thuringiensis active bacteria formulation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410436991.8A CN104336076B (en) | 2014-08-29 | 2014-08-29 | A kind of preparation technology of bacillus thuringiensis active bacteria formulation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104336076A CN104336076A (en) | 2015-02-11 |
| CN104336076B true CN104336076B (en) | 2017-12-08 |
Family
ID=52493158
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410436991.8A Active CN104336076B (en) | 2014-08-29 | 2014-08-29 | A kind of preparation technology of bacillus thuringiensis active bacteria formulation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104336076B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105284795A (en) * | 2015-10-19 | 2016-02-03 | 江西新龙生物科技股份有限公司 | Technology for preparing high-content bisultap granular through MVR technology |
| CN107090420B (en) * | 2017-06-07 | 2021-01-26 | 青岛农业大学 | Fermentation culture method of bacillus thuringiensis |
| CN107267424B (en) * | 2017-08-02 | 2018-03-27 | 上海乾界生物科技有限公司 | A kind of process for preparing microbial insecticide |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1366822A (en) * | 2001-07-18 | 2002-09-04 | 湖南师范大学 | Double engineering bacterium biological pesticide and its production method |
| CN102948621A (en) * | 2012-11-12 | 2013-03-06 | 东北农业大学 | Prebiotic peptide biological feed additive and preparation method and application thereof |
-
2014
- 2014-08-29 CN CN201410436991.8A patent/CN104336076B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1366822A (en) * | 2001-07-18 | 2002-09-04 | 湖南师范大学 | Double engineering bacterium biological pesticide and its production method |
| CN102948621A (en) * | 2012-11-12 | 2013-03-06 | 东北农业大学 | Prebiotic peptide biological feed additive and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 苏云金芽袍杆菌9803菌株工业发酵研究;孟世芬;《中国优秀硕士学位论文全文数据库（电子期刊） 农业科技辑》;20090615(第6期);第D046-18页；第1.4-1.5节、第2.2-2.8节、第3.1节、3.3节、3.6节、3.7.4节 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104336076A (en) | 2015-02-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102910988B (en) | Compound microorganism liquid bacterial fertilizer for preventing plant diseases and insect pests and preparation method of compound microorganism liquid bacterial fertilizer | |
| CN104232526A (en) | Process for preparing viable bacillus subtilis preparation | |
| CN101967455B (en) | Bacillus amyloliquefaciens EA19 for controlling wheat root diseases and preparation thereof | |
| CN104232530A (en) | Process for preparing viable bacillus licheniformis preparation | |
| WO2011032330A1 (en) | Antagonistic bacteria for preventing and treating bacterial wilt disease of continuously planted tobacco and microorganism organic fertilizer thereof | |
| CN108264401B (en) | Bio-organic fertilizer, preparation process and application method thereof | |
| CN104232528A (en) | Process for preparing viable bacillus amyloliquefaciens preparation | |
| CN111793498A (en) | Microbial agent and preparation method and application thereof | |
| CN104336076B (en) | A kind of preparation technology of bacillus thuringiensis active bacteria formulation | |
| CN112760248A (en) | Streptomyces lincolnensis capable of preventing and treating peach brown rot and application thereof | |
| CN104671853A (en) | Preparation method of compound photosynthetic bacterial fertilizer | |
| CN101352177A (en) | Compound successive crop-resistance micro-ecological formulation special for cotton and special bacterial strain and use thereof | |
| CN1186441C (en) | Microbial fertilizer bacterial agent for direct decomposing straw as fertilizer and its production process and application method | |
| CN104232525A (en) | Process for preparing viable bacillus coagulans preparation | |
| CN101215532B (en) | Bacillus megaterium and its application and application method in ferment bacteria | |
| CN104232527A (en) | Process for preparing viable bacillus megaterium preparation | |
| CN108707032A (en) | The biological organic fertilizer and preparation method thereof that microbial degradation house refuse obtains | |
| CN110200017B (en) | Compound microbial agent for preventing and treating root-knot nematode and preparation method thereof | |
| CN105948831B (en) | A method of biological and ecological methods to prevent plant disease, pests, and erosion fertilizer is produced using brewex's grains | |
| CN110122485A (en) | A kind of trichoderma, Trichoderma and preparation method thereof | |
| CN106399131B (en) | One plant of production dark purple mould and its application | |
| CN100596305C (en) | Atrazine herbicide degradation bacterium preparation making method | |
| CN104263675A (en) | Preparation technique of Bacillus polymyxa viable bacterium preparation | |
| CN111892440A (en) | Microbial fertilizer for preventing and treating potato scab and preparation method and use method thereof | |
| CN105733963A (en) | Composite fermentation straw degradation agent, preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |