CN1546651A - Bt solid bottom fermentation and continuous transplantation production method - Google Patents
Bt solid bottom fermentation and continuous transplantation production method Download PDFInfo
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- CN1546651A CN1546651A CNA2003101125218A CN200310112521A CN1546651A CN 1546651 A CN1546651 A CN 1546651A CN A2003101125218 A CNA2003101125218 A CN A2003101125218A CN 200310112521 A CN200310112521 A CN 200310112521A CN 1546651 A CN1546651 A CN 1546651A
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Abstract
The invention discloses a Bacillus thuringiensis solid bottom fermentation and continuous transplantation production method which comprises, (1) mixing the liquid state bacterial with solid state culture medium, storing in fermentation apparatus for fermentation, (2) treating the fermentate as bacterial to carry out continuous transfer and second fermentation. The invention realizes increased production efficiency.
Description
Technical field
The present invention relates to a kind of microorganism solid fermentation method, solid-state submerged fermentation of particularly a kind of bacillus thuringiensis (Bacillus thuringiensis) and the production method of transferring continuously.
Background technology
At present, the Bacillus thuringiensis solid state fermentation adopts tray formula, storehouse formula tray, box, bucket formula more, form is different, the stone operation is based on manual operations, time-consuming easy pollution, layer thickness, purifying air and wind supply quantity, warm and humid control do not reach standardization and stdn requirement, produce and often be subjected to region, weather, environmental influence and restriction, particularly production in summer temperature humidity is difficult to control, some medium component is difficult for pulverizing in the crushing process, cause the high waste of production energy consumption big, unstable product quality.
Summary of the invention
The production method that purpose of the present invention just provides the solid-state submerged fermentation of a kind of bacillus thuringiensis and transfers continuously is to realize with solid medium scale operation bacillus thuringiensis.
The present invention can realize in the following manner: (1) liquid bacterial classification is with after solid medium mixes, place the fermentation container fermentation, substratum thickness 300-800mm, fermentation container is in the fermenting house of sealing, room temp 26-55 ℃, humidity 35-95%, leavening temperature 28-32 ℃, humidity 35-85%, blast 112mmH
2O, air quantity 10588-12000m
3/ h, fermentation time 48-55 hour; (2) fermented product is transferred continuously as bacterial classification after again the fermentation, be when fermenting when the fermented product thalline quantity of logarithmic phase reaches 20,000,000,000/mg, extracting fermented product by fermentation container is transferred to respectively in 5-10 the fermentation container as bacterial classification and ferments, leavening temperature 28-32 ℃, humidity 35-85%, blast 112mmH
2O, air quantity 10588-12000m
3/ h fermentation time 48-55 hour, then,,, stops to shift continuously connecing as bacterial classification switching fermentation 3-5 time when finding that decline or phage appear in thalline with fermented product.
Spore content alive of the present invention can reach 400-500 hundred million/gram, and it is individual big, even to mix the spore crystal, and the toxicity evaluation of dry powder is stabilized in 16000IU/mg.The present invention has improved production efficiency greatly, has stable processing technique, is difficult for microbiological contamination, output height, the measured advantage of matter, reduces the making of first class inoculum, has reduced the phage contamination of heavy, thereby has reduced cost.It is fit to the solid state fermentation of aerobic microbiological very much.
Embodiment
Below in conjunction with embodiment, the present invention is carried out more detailed description.
Embodiment one
The solid-state submerged fermentation process of bacillus thuringiensis: the liquid amplification culture of bacterial classification, medium sterilization inoculation, solid-state submerged fermentation, oven dry, pulverizing.
1, fermentation container culture medium prescription and compound method: 665 kilograms in wheat bran, 14 kilograms of fish meal, 14 kilograms of yeast powders, the substratum mixing of materials is mixed thoroughly together, 7 kilograms of lime are put into 735 kg of water, get its supernatant liquor and substratum compound and stir and get final product.
2, medium sterilization: substratum is placed 121 ℃ of high-temperature sterilizations in the Sterilization Kettle, 35 minutes time.
3, inoculation and seed dressing: sand pipe bacterial classification inoculation, the breeding of eggplant bottle, the liquid cultivation of seeding tank, the inoculation of substratum normal temperature is also mixed thoroughly in still, then sub-material.
4, solid-state submerged fermentation:
4.1 fermentation container is arranged on 30 ℃ of the interior indoor temperature of fermenting house, humidity 65% is respectively spread the aseptic wet cloth of one deck up and down and is used for filtrated air and moisture-heat preservation in the fermentation container;
4.2 postvaccinal substratum is sent to fermentation container, paves thickness 450mm through tripper;
4.3 during fermentation, fermentation container adopts microcomputer frequency modulation control, 30 ℃ of temperature, humidity 85%, blast 112mmH
2O, air quantity 10588m
3/ h, wind direction is advanced with the long axis direction of fermentation container, under the effect of wake turbulence plate, wind direction is transferred vertically upward, evenly penetrate substratum, through lag period, logarithmic phase, stationary phase, the fermentation in 51 hours of ripening stage four-stage, fermented product parasporal crystal toxicity evaluation 〉=16000IU/mg, toxin protein 〉=2.0.
5, with dryer fermented product moisture is dried to below 7%, make product fineness reach 200 orders through the micronizer mill pulverizing.
Embodiment two
The solid-state deep layer switching of bacillus thuringiensis fermenting process: medium sterilization, solid-state submerged fermentation thing are transferred as bacterial classification and are fermented, dry, pulverize.
1, fermentation container culture medium prescription and compound method: prepare burden in embodiment one culture medium prescription ratio, wheat bran, fish meal, yeast powder, lime respectively multiply by 5 times, add up to 3500 kilograms, the substratum mixing of materials is mixed thoroughly together, 35 kilograms of lime are put into 3675 kg of water, get its supernatant liquor and substratum compound and stir and get final product.
2, medium sterilization: substratum is placed 121 ℃ of high-temperature sterilizations in the Sterilization Kettle, 35 minutes time.
3, the solid state fermentation thing is transferred as bacterial classification and is fermented:
3.1 in the fermenting house in each fermentation container up and down all the aseptic wet cloth of place mat one deck be used for filtrated air and moisture-heat preservation; Send into fermentation container behind the medium sterilization, pave, thickness 600mm by tripper;
Extract with material moving machine to the fermented product of logarithmic phase thalline quantity 20,000,000,000/mg 3.2 will ferment subsequently and to be transferred to respectively in 5 fermentation containers as bacterial classification;
3.3 fermenting process is with embodiment 1.
Embodiment three
The switching fermentation more continuously of the solid-state deep layer of bacillus thuringiensis: medium sterilization, the continuous switching of solid state bacterial are fermented, dry, are pulverized.
1. culture medium prescription and compound method: culture medium prescription, ratio, quantity and compound method are with embodiment two (1);
2, medium sterilization: substratum is placed 121 ℃ of high-temperature sterilizations in the Sterilization Kettle, 35 minutes time.
3, solid state bacterial switching fermentation more continuously:
3.1 send into fermentation container behind the medium sterilization, pave by tripper, thickness 800mm,
Fermented to extract with material moving machine and be transferred to respectively in 10 fermentation containers 3.2 will execute example two (3.2) again, evenly fermented again after the seed dressing as bacterial classification to the fermented product of logarithmic phase thalline quantity 20,000,000,000/mg;
3.3 fermenting process is with embodiment one (4.3);
4. oven dry is pulverized with embodiment two (5);
Embodiment one, two, three forms the solid-state deep layer of complete bacillus thuringiensis and transfers continuously the fermentative production cycle, whole process only needs five days, feeds intake 17200 kilograms, compares production cycle shortening 1.6 days with common solid-state submerged fermentation, output improves 3.2 times, and the product toxicity evaluation improves 30%.
Bacillus thuringiensis solid state fermentation thing give birth to be surveyed through quality inspection, dissolve filler and standardization after, as the wettable powder commodity selling, this product can be mixed soil and use as pesticide-clay mixture, available other solubility preparations dissolve into formulations such as suspension agent, can directly convert water and spray.Its insecticidal effect, plate-out ability, wettability etc. all have good performance.Because the product substratum is made up of grain by-product, can lands after the dispenser and be fertilizer, the economic benefits and social benefits function of have desinsection, topdressing.
Claims (2)
1, the solid-state submerged fermentation of a kind of bacillus thuringiensis and the production method of switching continuously is characterized in that:
(1) liquid bacterial classification places the fermentation container fermentation with after solid medium mixes, substratum thickness 300-800mm, fermentation container in the fermenting house of sealing, room temp 26-55 ℃, humidity 35-95%, leavening temperature 28-32 ℃, humidity 35-85%, blast 112mmH
2O, air quantity 10588-12000m
3/ h, fermentation time 48-55 hour;
(2) fermented product is transferred continuously as bacterial classification after again the fermentation.
2, production method as claimed in claim 1, fermentation again after it is characterized in that fermented product transferred continuously as bacterial classification, be when fermenting when the fermented product thalline quantity of logarithmic phase reaches 20,000,000,000/mg, extracting fermented product by fermentation container is transferred to respectively in 5-10 the fermentation container as bacterial classification and ferments, leavening temperature 28-32 ℃, humidity 35-85%, blast 112mmH
2O, air quantity 10588-12000m
3/ h fermentation time 48-55 hour, then,,, stops to shift continuously connecing as bacterial classification switching fermentation 3-5 time when finding that decline or phage appear in thalline with fermented product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2003101125218A CN1546651A (en) | 2003-12-08 | 2003-12-08 | Bt solid bottom fermentation and continuous transplantation production method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2003101125218A CN1546651A (en) | 2003-12-08 | 2003-12-08 | Bt solid bottom fermentation and continuous transplantation production method |
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| CN1546651A true CN1546651A (en) | 2004-11-17 |
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| CNA2003101125218A Pending CN1546651A (en) | 2003-12-08 | 2003-12-08 | Bt solid bottom fermentation and continuous transplantation production method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120046377A1 (en) * | 2009-04-30 | 2012-02-23 | Siegwerk Benelux Sa | Photoinitiators |
-
2003
- 2003-12-08 CN CNA2003101125218A patent/CN1546651A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120046377A1 (en) * | 2009-04-30 | 2012-02-23 | Siegwerk Benelux Sa | Photoinitiators |
| US8598249B2 (en) * | 2009-04-30 | 2013-12-03 | Siegwerk Druckfarben Ag & Co. Kgaa | Photoinitiators |
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