CN106701090A - Bacillus subtilis-containing soil amendment - Google Patents
Bacillus subtilis-containing soil amendment Download PDFInfo
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- CN106701090A CN106701090A CN201510783692.6A CN201510783692A CN106701090A CN 106701090 A CN106701090 A CN 106701090A CN 201510783692 A CN201510783692 A CN 201510783692A CN 106701090 A CN106701090 A CN 106701090A
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Abstract
The invention provides a method used to produce a bacillus subtilis-containing bactericide or a bacillus subtilis-containing soil amendment. In the method, a mixture of starch, protein and syrup, yeast powder, corn steep liquor and bentonite are used as raw materials of a culture medium, and the method comprises the steps of selecting a bacterial species, preparing the culture medium, performing fermentation processing, performing vacuum concentration, preparing a liquidpreparation, performing spray drying and preparing the bactericide or the soil amendment in a form of a powder or in other forms. A culture solution prepared by the disclosed method can be used as the bactericide or the soil amendment. The liquid preparation or a powdery preparation can be produced and applied to soil and seeds to provide a good effect on controlling bacterial wilt.
Description
Technical field
The present invention relates to a kind of method, for producing bactericide or soil conditioner, containing bacillus subtilis, culture bacillus subtilis is trained liquid or a kind of bactericide of powdery or soil improvement dosage form a nutrient solution and with post processing particularly in a culture medium.As the bactericide function of the completion of the strength of the device by biology, also referred to as biological electric control.
Background technology
A kind of soil conditioner containing bacillus subtilis, a kind of method, for producing bactericide or soil conditioner, containing bacillus subtilis, it mainly realizes starch, protein, the mixture of syrup, dusty yeast, corn pulp, bentonite and is made up of the following steps as a culture medium：
Bacillus subtilis is the flexible fungi of certain spore and is wherein general isolation to be typically used for produce enzyme or antibiotic.Discriminatively, the present invention has relational implementation to produce such bacillus subtilis of bactericide or soil conditioner and open spore and the material isolation from the special metabolism cultivated as insecticide and can improve the soil bacteria clump for being beneficial to plant growth for plant disease (such as powdery mildew of pea).
The content of the invention
The present invention disclosed herein provides a kind of method, for producing bactericide or soil conditioner, containing the bacillus subtilis for relating to the special kind bacillus subtilis of culture one, it is isolated in a fermentation tank for the culture medium for especially illustrating and under a predetermined operating condition until sporulation, vacuum, it is used to the nutrient solution of moistureproof synthesis and suitably prepares the nutrient solution of concentration accumulating the bactericide or soil conditioner of liquid.Spray drying can then be performed to the nutrient solution of above-mentioned concentration, and the powder tank of synthesis is the entrance powdered bacteriocide or soil conditioner for suitably preparing.Additionally, prepared by bactericide or soil conditioner can be implemented liquid by deep processing or powdery.
A kind of soil conditioner containing bacillus subtilis, a kind of method, for producing bactericide or soil conditioner, containing bacillus subtilis, it mainly realizes starch, protein, the mixture of syrup, dusty yeast, corn pulp, bentonite and is made up of the following steps as a culture medium：
1. a kind of soil conditioner containing bacillus subtilis, a kind of method, for producing bactericide or soil conditioner, containing bacillus subtilis, it mainly realizes starch, protein, the mixture of syrup, dusty yeast, corn pulp, bentonite and is made up of the following steps as a culture medium：
A. species of bacterium：
An appropriate kind bacillus subtilis is selected to isolate its production for being used for bactericide or soil conditioner;
B. fermentation tank：
The fermentation tank for realizing the type of agitation inflation or some other type for equally providing hyperoxia intensity cultivate bacillus subtilis during a fermentation process for liquid, and it is used to support enough oxygen to aerobic bacillus subtilis;
C. culture medium：
Culture medium is prepared, the optimum combination scope with starch, 5 to 40 g/L of its change, protein, 3 to 24 g/L of its change, syrup, 1 to 8 g/L of its change, dusty yeast, 1.5 to 12.0 g/L of its change, corn pulp, its change, 3 to 35 g/L and bentonite, its change, 1 to 20g/L；
D. fermentation process：
One fermentation process of nutrient solution of operation, the species and culture medium that it contains the bacterium pH6.0-8.0 and do saturation at 25 ~ 30 DEG C more than 30%;
E. concentration：
Preferably operated less than below 0.5 atm and under 55 DEG C by the nutrient solution of the method being concentrated in vacuo;More preferably under 0.1 atm and under 38 DEG C, most preferably under 0.01 atm and under 30 DEG C;
Liquid preparation：
The water content for increasing the nutrient solution that an appropriate quantity for appropriate additive is concentrated to adjustment reaches a spore density with a predetermined product standard, and to obtain a liquid preparation, it can be employed as bactericide or soil conditioner;
G. it is spray-dried：
Fully make the culture fluid dewatering by using the concentration for thering is a spray dryer for the outlet air temperature of its nozzle to be obtained due to above-mentioned vacuum concentration under 75 DEG C, to ensure the activity of spore;
H. prepared by powdery：With
Increase an appropriate quantity for appropriate carrier, auxiliary or diluent enter the powder obtained in spray drying and reach spore density with a predetermined product standard, prepared with obtaining a powdery, it can be employed as bactericide or soil conditioner;
1. the other forms for preparing：
Treatment liquid or powdery prepare, to obtain the other form of preparation, such as lozenge or bentonite.
2. as quoted in claim 1, wherein the fermentation tank that the type for stirring inflation is used to cultivate bacillus subtilis can possess ability, it rises to 60000 to a kind of method from 60
Rise (60 tons) changes.
3. a kind of method in claim 1 as quoted, wherein including that liquid at least preserve and acid or alkali is used to adjust pH value fluctuation for the appropriate additive of liquid preparation.
4. a kind of method is in claim 3 as quoted, wherein the liquid preparation containing bacillus subtilis recognizes to increase appropriate water and acid or alkali liquid enters the nutrient solution concentrated to adjusting pH value to 7.0+-0.5
And spore density is adjusted to more than 10< 9 >/ml .
5. as quoted in claim 1, wherein carrier, the auxiliary material prepared for powdery or diluent include at least talcum powder to a kind of method, and bentonite simultaneously disperses surfactant.
6. a kind of method in claim 5 as quoting, wherein prepared by the powdery containing bacillus subtilis recognize that increase bentonitic appropriate quantity solves to adjust spore density to more than 10 into concentrate and spraying dried culture
< 9 >The method of/g.
7. as being quoted in claim 1, wherein bactericide or soil conditioner can be applied to plantation to a kind of method, sow and provide the excellent effect of the control in bacterial wilt.
Bacillus subtilis is the flexible fungi of certain spore and is wherein general isolation to be typically used for produce enzyme or antibiotic.The present invention has the such bacillus subtilis and open spore of realizing production bactericide or soil conditioner and the material isolation from the special metabolism cultivated as insecticide and can to improve the soil bacteria clump for being beneficial to plant growth for plant disease (such as powdery mildew of pea).
Brief description of the drawings
The present invention and use, the preferred pattern of further object and its advantage, will be best understood when being combined reading with accompanying drawing with reference to the detailed description below an illustrative embodiment, wherein：
Fig. 1 is a brief flow chart, and it illustrates the method or soil conditioner for producing bactericide, contains bacillus subtilis of the invention.
Specific embodiment
The present invention provides a kind of method, for producing bactericide or soil conditioner, contain bacillus subtilis, its mixture for mainly realizing starch, protein, syrup, dusty yeast, corn pulp, bentonite are constituted as a culture medium and by the following steps, and it can be seen with Fig. 1.
1. species one of bacterium：
The appropriate bacillus subtilis of selection isolates the production for bactericide or soil conditioner.
2. fermentation tank B：
When bacillus subtilis is aerobic, growth needs the thin spore oxygen of macrophage and in the disclosed methods wherein, fermentation process for cultivating the liquid of bacillus subtilis is preferably carried out in fermentation tank B, the type or some other type of its agitation inflation, equally provides hyperoxia intensity.In the present invention, large scale industry company from from 60 rise to 60000 liters (60 tons) stir together inflation fermentation tank example be application culture bacillus subtilis, and fermentation tank, with other capacity ranges, is also suitable.
3. culture medium C：
The carbon-nitrogen ratio and the concentration of other micronutrients of culture medium C applied in the present invention are determined after optimization, the yield of its balanced growth for being used to advance bacillus subtilis and spore and its metabolic material.Optimal culture medium C has following proportional composition：5 grams of starch：3 grams of protein：1 gram of syrup：1.5 grams of dusty yeasts：0.5 gram of corn pulp：0.4 gram of bentonite.
Desirable culture medium C is to cultivate bacillus subtilis to change 5 to 40 range of compositions preparations of the starch of g/L, protein, its change 3 to 24
G/L, syrup, 1 to 8 g/L of its change
, dusty yeast, its change 1.5 to 12.0
G/L, if culture medium possesses the concentration lower than combinations of the above thing scope, corn pulp, its change 0.5 to 4.0 g/L and bentonite, its change 1.5 to 2.0g/L, 10 to 80 milligrams/L of retarded growth can be induced on bacillus subtilis, and if culture medium possesses the concentration higher than combinations of the above thing scope, growth inhibition can be induced on bacillus subtilis.
Preferable range of compositions relative to the culture medium C of the rapid growth for promoting bacillus subtilis contains starch, 15 to 30 g/L of its change, protein, 9 to 18 g/L of its change, syrup, 3 to 6 g/L of its change, dusty yeast, 4.5 to 9.0 g/L of its change, corn pulp, its change 1.5 to 3.0 g/L and bentonite, its change 1.2 to 5.5g/L.
The most desirable range of compositions of the culture medium C of the rapid growth of bacillus subtilis is promoted to contain from 20-25 with respect to the present invention
G/L different starch, protein, it changes from 12-15 g/L, syrup, 4 to 5 g/L of its change, dusty yeast, 6.0 to 7.5 g/L of its change, corn pulp, its change 2.0 to 2.5 g/L and bentonite, its change 1.2 to 5.5g/L.
Suitable starch in the present invention can turn into corn flour, wheaten starch, rice starch or bentonite.Suitable protein in the present invention can turn into soybean protein, cruel element or whey protein.No significant difference of influence is known that realizes various thering is a little starch or protein.Dusty yeast should be the necessary composition of culture medium C because it raises money growth factor, to help bacillus subtilis, the material of its production spore and metabolism.The bacillus subtilis cultivated in the culture medium lacked for dusty yeast has the development of difference and is therefore inappropriate to manufacture bactericide or soil conditioner.Although syrup, dusty yeast and corn pulp inherently contain small micronutrient, the supplement of an inorganic salts is to be necessary to culture medium C of the invention.
4. fermentation process D
In the present invention, experimental design, with confirm Bacillus globigii spores and metabolism material maximum production together with optimal culture medium C can by fermentation process method obtain under a specific operating condition perform, as will below by explanation.
Preferably, culture medium C captures the ability of at most 85% fermentation tank B to accommodate the foam on the surface for being formed at culture medium C and to prevent culture medium C from overflowing during stirring.Culture medium C is sterilized capacity of the change by fermentation tank B within the whole unusual time at 121 DEG C with saturated vapor.But, it is to avoid result from the possible denaturation of the excessively culture medium C of long-term heating, period in the possible cycle should be minimized it is most short-term between.During being proposed, in 121 DEG C of sterilized culture C of the invention, it was one 50 liters of fermentation tank B than 20 minutes, for the amount of 600 liters of fermentation tanks and than 40 minutes was 3 tons of amounts of fermentation tank than 30 minutes.During the moist heat sterilization of culture medium C, relevant program, such as steam are introduced, and heating, the temperature of maintenance, temperature quenching, internal water cooling, inflation, Valve controlling can be automatically carried out by a control computer and belong to fermentation tank B.
In the present invention, the Bacillus globigii spores for being taken out without thermal shock activation at 4 DEG C from a culture dish are cultured into the use shaking flask method or fermentation method performed in a small fermentation tank B, and it is used to then be transferred into a big fermenting case.
One desirable operating temperature, for fermentation process D, in 23-45
Between DEG C, and the retarded growth of bacillus subtilis can occur beyond the scope in an operating temperature.At the same time, more preferably between 26-38 DEG C, most preferably the best operating temperature between 20-29 DEG C, for fermentation process D, is kept operating temperature during the sporulation stage between 28-36 DEG C.
Nutrient solution now refers to zymotic fluid and with adjustment with its pH value between 5.5-8.5, is used to be beneficial to fermentation process with aseptic sour and alkali liquid (such as sulfuric acid and NaOH).PH value outside this scope is proposed that the growth inhibition of bacillus subtilis.The pH value of zymotic fluid, for the rapid growth of bacillus subtilis, more preferably by scope 6.5 to 8.5 and most preferably by scope 6.5 to 7.5.During the growth course of bacillus subtilis, the pH value of zymotic fluid forces down the cycle to the centre of vegetative growth phase throughout the lag phase, it is required to the introduction of the liquid of the alkali of there, and cycle of the pH value throughout the centre of the vegetative growth phase to the sporulation stage recovers, the introduction of acid solution body is required to keep the pH value with the revised distance to there.
The dissolving oxygen saturation of zymotic fluid is preferably maintained more than 33% really not so growth inhibition, and the bacteriolysis of bacillus subtilis can be caused.Party saturation, for the rapid growth of bacillus subtilis, more preferably terminates to be 55% and most preferably more than 75%.During the growth course of bacillus subtilis, the rising in the cycle of the centre throughout vegetative growth phase that the cycle throughout the centre in lag phase to vegetative growth phase and the change to sporulation stage and pH value are coordinated is forced down really.Party saturation can be adjusted the saturation by changing mixing speed and/or the inflation mixing speed that the mixing speed of that repeating and/or the saturation of inflation influence amplification weaken in mode and/or inflation influence reduction simultaneously.
Party saturation for making zymotic fluid reaches maximum, and fermentation tank B is preferably operated in highest mixing speed, its ability for being susceptible to fermentation tank B.The mixing speed of application can be the 85% of the maximum limit mixing speed of the permission described in detail in the operation instructions of fermentation tank (if mixing speed is adjustable) or be probably a specific mixing speed, and it is preprogramed in fermentation tank B.For example；50 liters of fermentation tanks can be operated with 350 rev/min;500 liters of fermentation tanks can be operated with 120 rev/min;5 tons of fermentation tanks can be operated with 22 rev/min.The thin spore cluster that powerful shearing stress from the head-tail of connection unit separate together with the agitation of high speed can be constructed and enter individual unit, and without adverse effect to bacterial growth.
When fermentation tank B is operated in the mixing speed, the inflation together with filtration and aseptic gas tank is adjusted belonging to the saturation that fermentation tank B is preprogramed the minimum zymotic fluid together with by control computer.During the growth course of bacillus subtilis, inflation is increased for supporting party to be saturated to the cycle of the centre of vegetative growth phase in minimum value throughout the lag phase and being depressed the sporulation stage with the cycle coordinated throughout the centre of vegetative growth phase that changes of pH value.When big fermentation tank is implemented, if necessary complete air-breathing ability can be used wherein, be clipped in the middle the oxygen demand of the maximum bacterium of generation, its limitation for being used to overcome mixing speed with the cycle for catering to vegetative growth phase.
During fermentation process, when the protein included in culture medium is by stirring and inflating, the thin spore foam annihilator of macrophage is on the surface of zymotic fluid so as to occur, the automatic resistance foam device for loading aseptic defoamer starts to eliminate foam, and wherein defoamer can be material or bleeding agent based on silicon and suitably be diluted.
The point that fermentation process can wherein discharge free ripe spore after its dissolving is terminated.One sporulation ratio higher can still turn into time-consuming inapproachable therefore display fermentation tank unhelpful operation.The whole operation term of fermentation process can be varied from different size fermentation tank B.In the present invention, an operational term being proposed is that 60 liters of fermentation tank B are less than 20 hours;For 600 liters of fermentation tank B less than 23 hours;For 5 tons of fermentation tank B less than 22 hours and for 50 tons of fermentation tank B less than 28 hours..
5. E is concentrated in vacuo：
The nutrient solution that the zymotic fluid for following fermentation process to complete synthesizes.Operation vacuum concentrator, vacuum is preferably with 0.5
atm;More preferably under 0.1 atm, most preferably 0.01
The boiling point of nutrient solution preferably under 55 DEG C, more preferably under 45 DEG C, the rising osmotic pressure of nutrient solution is most preferably caused under as 38 DEG C and promotes to keep new and extend it to shelf-life simultaneously under atm.
6. liquid preparation F：
The nutrient solution of the concentration obtained in step E can further be produced and be matched spore density with a predetermined product standard as the liquid preparation of the appropriate quantity by increasing to the appropriate additive for adjusting its water content.Suitable additive includes the liquid of at least preservative and acid or alkali, for adjusting pH value fluctuation.Therefore, liquid preparation can be employed as bactericide or soil conditioner.
7. G is spray-dried：
The nutrient solution of the concentration obtained by above-mentioned vacuum concentration E is fully dehydrated now by using a spray dryer.The outlet air temperature of nozzle should be by 75 DEG C of control, to ensure the activity of spore.
8. powdery prepares H：
The powder of the synthesis of acquisition can be further produced as one by adding the aid in or thin powdery preparation for being adapted to the appropriate quantity of the appropriate carrier of spore density with a predetermined product standard in step spray drying G.Suitable carrier adjuvant or diluent include at least wood powder, bentonite and dispersion surfactant.Therefore, powdery is prepared and can be employed as bactericide or soil conditioner.
9. the other forms of I are prepared：
Prepare other form, such as lozenge or gelatin, can by treatment liquid or powdery prepare realize.
Embodiment 1
Method for cultivating bacillus subtilis
Take from a Bacillus globigii spores for PDA disks at 5 DEG C to be activated by shaking flask method, it realizes a shaking flask, with bottom baffle, it is used to strengthen oxygen diffusion and stirring effect.Culture medium in shaking flask (hereinafter referred to as " culture medium of standard ") is by 20 ratios of g/l of corn flour, 12 g/l of soybean protein, 4 g/l of syrup, 6 g/l of dusty yeast, 2 g/l of corn pulp, bentonitic 3 g/l compositions.Culture medium can be preferably filled with about 1/5th of the ability of shaking flask;That is 100 ml culture mediums are used for one 500 ml shaking flasks.Shaking flask is put into an incubator in 31+-1 DEG C and 200 rev/min at that time.
When optical density (OD or absorptivity) is wherein 0.5, when bacillus subtilis in the medium comes to the point, nutrient solution is ready to be transferred to one 5 liters of fermentation tanks, and large-scale farming is used for containing 3.5 liters of culture mediums of standard.Enough quantity inoculations of bacillus subtilis enter fermentation tank, to obtain 10% rate of vaccination.5 liters of fermentation tanks are operated at 31+-1 DEG C at that time, and therefore the mixing speed of fermentation tank is that it is that, more than 1 liter, pH value 7.0+-0.1 and 70% does saturation to inflate per minute more than 600 rev/min.
When the OD of the bacillus subtilis in 5 liters of fermentation tanks reaches 0.5, to 50 liters of fermentation tanks, subsequent large-scale farming to be used for containing 35 liters of culture mediums of standard, nutrient solution is ready to further be transferred.50 liters of fermentation tanks are operated at 31+-1 DEG C at that time, and therefore the mixing speed of fermentation tank is that it is that, more than 5 liters, pH value 7.0+-0.1 and 70% does saturation to inflate per minute more than 300 rev/min.Fermentation process can be carried out until when the unit cultivated is produced and discharges the spore of maturation, nutrient in the medium exhausts (it spends about 20 hours after inoculation).Nutrient solution can further be processed with vacuum concentration.Alternatively, fermentation process can carry out providing product as culture for use in the further fermentation process in one 500 liters of fermentation tanks until the OD of the bacillus subtilis in 50 liters of fermentation tanks reaches 0.5.
When the OD of the bacillus subtilis in 50 liters of fermentation tanks reaches 0.5, nutrient solution is ready to be transferred into 500 liters of fermentation tanks, and advanced large-scale farming is used for containing 350 liters of culture mediums of standard.500 liters of fermentation tanks are operated at 31+-1 DEG C at that time, and therefore the mixing speed of fermentation tank is that it is that, more than 15 liters, pH value 7.0+-0.1 and 60% does saturation to inflate per minute more than 100 rev/min.Fermentation process can be carried out until when the unit cultivated is produced and discharges the spore of maturation, nutrient in the medium exhausts (it spends about 23 hours after inoculation).Nutrient solution can further be processed with vacuum concentration.Alternatively, fermentation process can carry out providing product as culture for use in the further fermentation process in one 5000 liters of (5 tons) fermentation tanks until the OD of the bacillus subtilis in 500 liters of fermentation tanks reaches 0.5.
When the OD of the bacillus subtilis in 500 liters of fermentation tanks reaches 0.5, nutrient solution is ready to be transferred into one 5000 liters of fermentation tanks, and advanced large-scale farming is used for containing 3500 liters of culture mediums of standard.5000 liters of fermentation tanks are at that time in 31+-1
[degree] C. is operated, and therefore the mixing speed of fermentation tank is that it is that, more than 30 liters, pH value 7.0+-0.1 and 50% does saturation to inflate per minute more than 50 rev/min.Fermentation process can be carried out until when the unit cultivated is produced and discharges the spore of maturation, nutrient in the medium exhausts (it spends about 25 hours after inoculation).Nutrient solution can further be processed with vacuum concentration.Alternatively, fermentation process can carry out providing product as culture for use in the further fermentation process in one 50000 liters of (50 tons) fermentation tanks until the OD of the bacillus subtilis in 5000 liters of fermentation tanks reaches 0.5.
When the OD of the bacillus subtilis in 5000 liters of fermentation tanks reaches 0.5, nutrient solution is ready to be transferred into 50000 liters of fermentation tanks, and advanced large-scale farming is used for containing 35000 liters of culture mediums of standard.50000
Rise fermentation tank to be operated at 31+-1 DEG C at that time, therefore the mixing speed of fermentation tank is that it is that, more than 50 liters, pH value 7.010.1 and 50% does saturation to inflate per minute more than 20 rev/min.Fermentation process can be carried out until when the unit cultivated is produced and discharges the spore of maturation, nutrient in the medium exhausts (it spends about 26 hours after inoculation).Nutrient solution can further be processed with vacuum concentration.
Embodiment 2
Enter bactericide or soil conditioner, the method containing bacillus subtilis for productive culture liquid
The all of batch of nutrient solution fermented in the fermentation tank as described in embodiment 1 is while the boiling point of nutrient solution is under 35 DEG C, be with a vacuum concentrator possibility of the vacuumizing under 0.1 atm keep nutrient can with nutrient solution together with concentrated and be mixed into organic fertilizer, it is used to make crop fertile suitable for domain variability.Meanwhile, it is such to arrange help, for avoiding environmental pollution, the unloading of its discarded nutrient solution resulted from together with the outside holding nutrient in factory.
The liquid of concentration equally previously discussed with by increasing appropriate water and acid or alkali liquid, with to adjustment to more than 10
< 9 >The liquid preparation of the spore density of/ml and the pH value in 7.0+-0.5 can be produced.
The nutrient solution of concentration can be prepared as powdery by using the spray dryer of the outlet air temperature for having the nozzle under 70 DEG C and alternatively be produced and be increased the appropriate quantity of clay powder, with to adjustment to more than 10
< 9 >/g 。
Embodiment 3
Field experiment contains bacillus subtilis in powdered bacteriocide or soil conditioner
The liquid preparation obtained in embodiment 2 is to use 600 times water-reducible.Being mixed has to 1000 liquid preparations for fertilizer grade amino-acid liquid of dilution and aside rest mixture 3 hours, and the activation of Bacillus globigii spores can strengthen.The mixture of dilution is fallen by the plant per about 500 ml around the root of tomato and enters soil, it is transplanted 10 days by once every 7 days, for at least 4 times, continuously help the preventing and treating of tomato bacterial wilt disease and contribute to vigorous growth, extend growth and the fructufy stage of plant.
Claims (7)
1.OnePlant the soil conditioner containing bacillus subtilis, a kind of method, for producing bactericide or soil conditioner, contains bacillus subtilis, it mainly realizes starch, protein, the mixture of syrup, dusty yeast, corn pulp, bentonite as a culture medium, and is made up of the following steps：
A. species of bacterium：
A kind of appropriate bacillus subtilis of selection isolates its production for being used for bactericide or soil conditioner;
B. fermentation tank：
The fermentation tank for realizing agitation type or some other type for equally providing hyperoxia intensity cultivate bacillus subtilis during a fermentation process for liquid, and it is used to support enough oxygen to aerobic bacillus subtilis
C. culture medium：
Culture medium is prepared, the optimum combination scope with starch, 5 to 40 g/L of its change, protein, 3 to 24 g/L of its change, syrup, 1 to 8 g/L of its change, dusty yeast, 1.5 to 12.0 g/L of its change, corn pulp, its change, 3 to 35 g/L and bentonite, its change, 1 to 20g/L；
D. fermentation process：
One fermentation process of nutrient solution of operation, the species and culture medium that it contains the bacterium pH6.0-8.0 and do saturation at 25 ~ 30 DEG C more than 30%;
E. concentration：
Preferably operated less than below 0.5 atm and under 55 DEG C by the nutrient solution of the method being concentrated in vacuo;More preferably under 0.1 atm and under 38 DEG C, most preferably under 0.01 atm and under 30 DEG C;
F liquid preparations：
The water content for increasing the nutrient solution that an appropriate quantity for appropriate additive is concentrated to adjustment reaches a spore density with a predetermined product standard, and to obtain a liquid preparation, it can be employed as bactericide or soil conditioner;
G. it is spray-dried：
Fully make the culture fluid dewatering by using the concentration for thering is a spray dryer for the outlet air temperature of its nozzle to be obtained due to above-mentioned vacuum concentration under 75 DEG C, to ensure the activity of spore;
H. prepared by powdery：
Increase an appropriate quantity for appropriate carrier, auxiliary or diluent enter the powder obtained in spray drying and reach spore density with a predetermined product standard, prepared with obtaining a powdery, it can be employed as bactericide or soil conditioner.
2. the other forms for preparing：
Treatment liquid or powdery prepare, to obtain the other form of preparation, such as lozenge or gelatin.
3. a kind of method in claim 1 as quoted, wherein the fermentation tank that the type for stirring inflation is used to cultivate bacillus subtilis can possess ability, it rises to 60000 liters of (60 tons) changes from 60.
4. a kind of method in claim 1 as quoted, wherein including that liquid at least preserve and acid or alkali is used to adjust pH value fluctuation for the appropriate additive of liquid preparation.
5. a kind of method in claim 3 as quoted, wherein the liquid preparation containing bacillus subtilis recognizes that increasing an appropriate water and the liquid of sour or alkali enters the nutrient solution concentrated to adjustment pH value to 7.0+-0.5 and adjust spore density to more than 10< 9 >/ml .
6. as quoted in claim 1, wherein carrier, the adjuvant prepared for powdery or diluent include at least gelatin to a kind of method, and bentonite simultaneously disperses surfactant.
7. a kind of method in claim 5 as quoting, wherein prepared by the powdery containing bacillus subtilis recognize that increase bentonitic appropriate quantity solves to adjust spore density to more than 10 into concentrate and spraying dried culture< 9 >The method of/g.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111149814A (en) * | 2020-01-14 | 2020-05-15 | 河南农贝得农业科技有限公司 | Agricultural microbial agent with insect expelling effect and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111149814A (en) * | 2020-01-14 | 2020-05-15 | 河南农贝得农业科技有限公司 | Agricultural microbial agent with insect expelling effect and preparation method thereof |
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Application publication date: 20170524 |