CN100367859C - Method for preparing bacillus thuringiensis microbiological pesticide by tumeric hydrolysis waste liquor - Google Patents
Method for preparing bacillus thuringiensis microbiological pesticide by tumeric hydrolysis waste liquor Download PDFInfo
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Abstract
The present invention relates to a method for preparing bacillus thuringiensis microbial insecticide from yellow ginger hydrolysis waste water. The method comprises the following steps of the pretreatment of waste water, strain domestication, medium component supplement, seed tank fermentation, fermentation tank fermentation and fermentation water post treatment, and the detailed method is given in a specification. The present invention has the advantages that the extraction process and the extraction efficiency of the original yellow ginger saponin are not influenced, and the microbial insecticide with good insecticidal effect and harmless environment safety is obtained, so that the present invention realizes the resource treatment of the yellow ginger hydrolysis waste water, and also reduces the production cost of the microbial insecticide.
Description
Technical field
The present invention relates to the recycling of yellow ginger waste hydrolyzed liquid, specifically, be that the waste hydrolyzed liquid that utilizes yellow ginger processing to extract in the saponin process prepares bacillus thuringiensis (Bacillus thuringiensis, be called for short bacillus thuringiensis,Bt or Bt) method of microorganism insecticide, belong to field of environment engineering technology.
Background technology
Yellow ginger (Dioscorea zingiberensis C.H.Wright) has another name called dioscorea zingiberensis wright, duration and degree of heating root, is the perennial voluble herb plant of Dioscoreaceae, is the distinctive medicine source plant resource of China.Yellow ginger also is the highest plant of China's Chinese yam saponin content, and the good reputation of " medicinal gold " is arranged, and its root-like stock contains the Chinese yam saponin of 1.1-16.15%, is the primary raw material of synthesizing steroid hormone medicine.
The production technology that saponin is extracted in yellow ginger processing mainly contains: (1) pre-fermentation-acid-hydrolysis method, technological process is: yellow ginger → pulverizing → pre-fermentation → sulphuric acid hydrolysis → hydrolysis residue extracts saponin, waste hydrolyzed liquid (is that yellow ginger is after acid hydrolysis is handled, hydrolysate is again through the liquid part of Separation of Solid and Liquid gained, and is corresponding with hydrolysis residue) discharging; (2) enzymatic hydrolysis-acid-hydrolysis method, technological process is: yellow ginger → wet-milling → amylase liquefaction, carbohydrase saccharification → filtrate acid hydrolysis → hydrolysis residue extract saponin, the waste hydrolyzed liquid discharging.Except that saponin, yellow ginger also contains the starch of 40-50%, the cellulose of 45-50%, 5% protein and other mineral element.Existing production technology is only extracted about 2% saponin wherein, and remains the form loss of beneficiating ingredients such as 98% starch, cellulose with waste hydrolyzed liquid and waste residue, and not only contaminated environment is also wasted resource.The sugar content of for example typical yellow ginger waste hydrolyzed liquid is 2-6%, SO
4 2-Be about 2-4%, acidity big (pH＜0), also contain minor N, P and trace element in addition.
Patent CN1124776A discloses a kind of method of extracting alcohol from the waste liquid of yellow ginger or earthworm production saponin, this method adds the alkaline matter neutralization with yellow ginger or earthworm hydrolyzed waste, add nitrogenous source and phosphorus source and carry out alcoholic fermentation, then distillation extraction alcohol.Patent ZL00131274.X discloses the technology of extracting in a kind of yellow ginger waste hydrolyzed liquid with refining glucose, glucose is extracted and made with extra care to this technology with the yellow ginger waste hydrolyzed liquid through processing steps such as acid hydrolysis, neutralization, desalination, decolourings under the prerequisite that does not influence the saponin extraction.Above-mentioned two patents have been carried out recycling to the sugar in the yellow ginger waste hydrolyzed liquid (starch hydrolysate), and the former focuses on conversion of glucose production alcohol, and the latter directly extracts and refining glucose, and both all are useful extensions of yellow ginger industrial chain.Yet above-mentioned two methods but lack engineering example, and main cause is that economic feasibility is not strong, and two methods all need comparatively expensive extraction and subtractive process, and the price of alcohol that obtains or glucose is comparatively cheap.
Bacillus thuringiensis is a class can produce parasporal crystal (being insecticidal crystal protein) in metabolic process a Gram-positive bacteria, be the present microorganism insecticide of output maximum (accounting for more than 90% of microorganism insecticide total amount) in the world, be widely used in control farming, woods and fruit tree pest insect, storage insect and sanitary insect pest.Compare with chemical insecticide, it is strong that the Bt insecticide has selectivity, to the person poultry harmless, and advantages of environment protection.Bt insecticide main flow production technology is a liquid deep layer fermenting at present, mainly with various industrial and agricultural products or agricultural byproducts, as glucose, peptone, analysis for soybean powder, corn starch, wheat bran etc. is raw material, and cost of material too high (accounting for the 20-35% of total cost of production) is the key constraints that the Bt insecticide is difficult to popularize.Therefore, the agriculture and industry waste that how to adopt low cost, locality to get is a raw material, reduces the fermenting and producing cost, and is significant for applying of this ecological harmless type biopesticide of Bt insecticide.
Through domestic and foreign literature and patent retrieval, do not see the report that utilizes the yellow ginger waste hydrolyzed liquid to prepare the bacillus thuringiensis microbiological insecticide for fermenting raw materials so far as yet.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art part, and provide a kind of yellow ginger waste hydrolyzed liquid to prepare the method for bacillus thuringiensis microbiological insecticide.Keeping original saponin extraction process and do not influencing on the basis of saponin extraction efficiency, by fermentation technique the yellow ginger waste hydrolyzed liquid is prepared into the higher microorganism insecticide of added value, both can solve the problem of environmental pollution of yellow ginger waste hydrolyzed liquid, realized its recycling and final disposal again, turned waste into wealth.
The product that provides said method to make is provided another object of the present invention.Product can effectively be killed the Lepidoptera crop pests, and is environmentally safe harmless, and product quality meets " bacillus thuringiensis wetting powder " national standard (GB/T 19567.3-2004).
The present invention seeks to and can realize in the following way: the yellow ginger waste hydrolyzed liquid that provides prepares the method for microorganism insecticide, this method step is in the following order carried out: the preliminary treatment of waste liquid, strain domestication, medium component replenish, seeding tank fermentation, ferment tank and zymotic fluid post processing, and concrete grammar is as follows: wherein: L represents 1 liter of yellow ginger waste hydrolyzed liquid; % representation quality percentage, the umber of the inoculation of medium seed of promptly per 100 parts of quality;
1) preliminary treatment of waste liquid: in yellow ginger waste hydrolyzed liquid (solid content 2-5%), add the calcic alkaline matter of 10-80g/L, be used for and waste liquor PH, and make SO in the waste liquid
4 2-With Ca
2+Reaction generates CaSO
4Precipitation.Utilize gravity settling separation or centrifugal separation with CaSO
4Sediment is removed.Reaction back waste liquor PH is increased to 7.0-8.5, free SO4
2-Content is reduced to below the 5000mg/L.Used calcic alkaline matter refers to Ca (OH)
2, CaCO
3, CaO, wherein Ca (OH) preferably
2, Ca (OH)
2Optimum amount be 60g/L.
2) strain domestication: the method domestication bacillus thuringiensis,Bt fermented bacterium of taking in beef-protein medium, progressively to improve yellow ginger waste hydrolyzed liquid concentration.The waste hydrolyzed liquid that in medium, progressively adds 1/8 times of original content, inoculation Bt bacterial strain in solid inclined-plane that adds variable concentrations yellow ginger waste hydrolyzed liquid or flat board, aerobic cultivation is 72 hours under 30 ℃, get normal colony inoculation then in the medium that contains 1/4 concentration waste hydrolyzed liquid, cultivated 72 hours at similarity condition, as stated above the concentration of yellow ginger waste hydrolyzed liquid is increased to 1/2 and original content then, carries out successive transfer culture.Said process can carry out tens of times repeatedly, and tiring approaching or reach starting strain until the desinsection of the Bt of gained naturalized strain stops when the desinsection of conventional medium is tired.Used Bt starting strain is meant the main production bacterial strain of the existing use of Bt insecticide producer, and as BtHD-1,7216 etc., microbial preservation mechanism is by directly buying at home.The composition of used beef-protein medium is: beef extract 5g/L, peptone 10g/L, agar 15 g/L; PH7.2-7.5, running water 1000mL, 15 pounds of pressure and 121 ℃ of following sterilizations 30 minutes.The form of medium is solid inclined-plane or flat board, and slant medium is simultaneously also as the storage medium of domesticated strain.
3) medium component replenishes: to adding the nitrogenous source of 10-30g/L, the mineral salt of 0.2-1.5g/L, the defoamer of 1-5g/L through pretreated yellow ginger waste hydrolyzed liquid, sterilized 30-60 minute through 15 pounds of pressure and 121 ℃ of temperature in seeding tank or fermentation tank in the back that stirs, be cooled to 30-35 ℃ standby.Used nitrogenous source is meant big dregs of beans, peanut meal, rapeseed dregs, fish meal, corn steep liquor, dusty yeast and peptone etc., and preferably big dregs of beans, optimum amount are 20g/L; The mixture that described mineral salt is made up of dipotassium hydrogen phosphate, magnesium sulfate, three kinds of materials of manganese sulphate, the mass ratio of three kinds of materials is 20-100: 10-30: 1-5 in the mixture.Used defoamer is meant bubble enemy, polypropylene glycol, soya-bean oil, peanut wet goods, and wherein preferably bubble opposes, and bubble enemy's optimum amount is 1.5g/L.
4) seeding tank fermentation: bottle bacterium liquid that shakes to culture medium inoculated Bt naturalized strain, carry out fermented and cultured.Fermentation condition is: inoculum concentration 1-3%, jar warm 28-32 ℃, tank pressure 0.02-0.06MPa, throughput 1: 1.0-1.5 (V/V/ branch), mixing speed 220-260 rev/min, fermentation time 8-12 hour.
5) ferment tank: the inoculum concentration of pressing 8-15% with the seed liquor culture transferring to fermentation tank.Condition of culture is: jar warm 28-32 ℃, tank pressure 0.02-0.06MPa, throughput 1: 1.0-1.5 (VN/ branch), mixing speed 180-220 rev/min, fermentation time 30-42 hour.Put a jar standard: parasporal crystal comes off 20%, zymotic fluid pH is 8.5-9.0.
6) zymotic fluid post processing: zymotic fluid adds dispersant and the 1-4g/L emulsifier of 20-50g/L, stirs, and sends into the spray drying tower drying, EAT 150-220 ℃, outlet temperature 60-70 ℃, is lower than 4%, packing to moisture content.Described dispersant is meant kaolin, diatomite, bentonite or precipitated calcium carbonate, kaolin preferably wherein, and optimum amount is 40g/L.Described emulsifier is meant sodium lignosulfonate, Tween-80 or TX-10, sodium lignosulfonate preferably wherein, and the optimum amount of sodium lignosulfonate is 2g/L.
Know-why of the present invention is: sugar and a spot of protein, phosphorus and the trace element contained with the yellow ginger waste hydrolyzed liquid are main nutrient matter, replenish partial nitrogen source and mineral element simultaneously, under the condition of keeping appropriate pH, temperature and throughput, bacillus thuringiensis,Bt is grown and propagation, form insecticide active substances such as gemma and insecticidal crystal protein, zymotic fluid is made the microorganism insecticide product through concentrate drying.
The present invention has the following advantages and good effect compared to existing technology:
(1) solved the disposal problem of outlet of high concentration yellow ginger waste hydrolyzed liquid, both prevented the loss of utility and pollution, realized comprehensive utilization of resources again, played the effect that turns waste into wealth environment.
(2) existing bacillus thuringiensis microbiological insecticide is a fermentation raw material with industrial and agricultural products such as glucose, peptone, analysis for soybean powder, corn starch, wheat bran or agricultural byproducts mainly, and cost of material is too high to be its key constraints that is difficult to popularize.Bacillus thuringiensis,Bt mainly relies in the yellow ginger waste hydrolyzed liquid utilized composition (sugar, protein and the mineral salt etc.) growth and propagation of self among the present invention, the composition that adds is less, and can significantly reduce the production cost that has reduced microorganism insecticide, can promote being applicable of this ecological harmless biopesticide of bacillus thuringiensis,Bt.
(3) with prior art---the yellow ginger waste hydrolyzed liquid extracts glucose or the fermentation preparing alcohol is compared, and the present invention does not need expensive complicated extraction refining step, and therefore the added value of products obtained therefrom has the better economic feasibility than glucose and alcohol height.
(4) the present invention does not change existing yellow ginger Chinese yam saponin extraction process, does not influence extraction efficiency yet.
Description of drawings
Fig. 1 is a process chart of the present invention.
Embodiment
Below in conjunction with accompanying drawing, enumerate 6 embodiment, the present invention is further specified, but the present invention is not only limited to these embodiment.
Embodiment 1
(1) yellow ginger waste hydrolyzed liquid
Get the waste hydrolyzed liquid of Shiyan City, Hubei Province yellow ginger processing factory, this factory adopts pre-fermentation-acid-hydrolysis method to extract saponin, and technological process is: yellow ginger → pulverizing → pre-fermentation → sulphuric acid hydrolysis → hydrolysis residue extracts saponin, the waste hydrolyzed liquid discharging.Content of reducing sugar is that 20.1g/L, crude protein content are 2.2g/L, sulfate radical content 38.9g/L in the waste liquid by analysis, pH＜0.
(2) yellow ginger waste hydrolyzed liquid preliminary treatment
In the yellow ginger waste hydrolyzed liquid, add the Ca (OH) of 54.0g/L
2Powder fully stirs, and surveying waste liquor PH is 7.50, free SO
4 2-Concentration is 3200mg/L.Gravitational settling 3 hours is with CaSO
4Sediment is removed.
(3) Bt strain domestication
Adopting the BtHD-1 that produces bacterial strain commonly used of microorganism insecticide factory is starting strain.The composition of beef extract-peptone solid culture medium is: beef extract 5g/L, peptone 10g/L, agar 15g/L; Running water 1000mL, pH 7.2-7.5,15 pounds of pressure and 121 ℃ of following sterilizations 30 minutes.Get 8 times of yellow ginger waste hydrolyzed liquid 1000mL of dilution, replace the running water of medium, other components unchanged is made the solid plate that contains 1/8 times of original content waste hydrolyzed liquid, and inoculation Bt HD-1 starting strain is cultivated 72 hours to bacterium colony occurring down at 30 ℃.With crystal gemma difference decoration method microexamination strain morphology, get normal colony inoculation in the solid plate that contains 1/4 concentration waste hydrolyzed liquid, under similarity condition, cultivated 72 hours, and as stated above the concentration of waste liquid in the solid plate was increased to 1/2 and original content then, carry out successive transfer culture.Said process can carry out 12-15 time repeatedly, and the desinsection of the Bt naturalized strain of gained is tired and starting strain basic identical at conventional medium as a result, and obtained strains is a domesticated strain.Standby with the slant medium preservation.
(4) medium component replenishes
To through Ca (OH)
2Add the big dregs of beans of 15g/L, the dipotassium hydrogen phosphate of 0.5g/L, the magnesium sulfate of 0.2g/L, the manganese sulphate of 0.02g/L and the bubble enemy of 2g/L in the yellow ginger waste hydrolyzed liquid of powder pre-treating.Respectively in seeding tank and fermentation tank through 15 pounds of pressure and the sterilization of 121 ℃ of temperature 60 minutes, be cooled to 30 ℃ standby.
Other defoamer is polypropylene glycol, soya-bean oil, peanut oil, replaces the bubble enemy respectively, and effect is identical.
(5) seeding tank fermentation
Inoculation 1% above-mentioned Bt naturalized strain shake a bottle bacterium liquid fermented and cultured.Condition of culture is: jar 30 ℃ of temperature, tank pressure 0.03MPa, throughput 1: 1.0 (V/V/ branch), 240 rev/mins of mixing speeds, fermentation time 12 hours.
(6) ferment tank
The seed liquor of inoculation 10% is to fermentation tank.Condition of culture is: jar 30 ℃ of temperature, tank pressure 0.03MPa, throughput 1: 1.2 (V/V/ branch), 180 rev/mins of mixing speeds, fermentation time 32 hours.
(7) zymotic fluid post processing
Zymotic fluid adds the dispersant kaolin of 40g/L and the emulsifier sodium lignosulfonate of 2g/L, stirs, and sends into the spray drying tower drying, 150 ℃ of EATs, and 60 ℃ of outlet temperatures are lower than 4%, packing to moisture content.
Other dispersant is diatomite, bentonite or precipitated calcium carbonate, and effect is identical.
Other emulsifier is Tween-80 or TX-10, replaces sodium lignosulfonate, and effect is identical.
Embodiment 2
(1) yellow ginger waste hydrolyzed liquid
With embodiment 1.
(2) yellow ginger waste hydrolyzed liquid preliminary treatment
With embodiment 1.
(3) Bt strain domestication
With embodiment 1.
(4) medium component replenishes
In pretreated yellow ginger waste hydrolyzed liquid, add the peanut meal of 20g/L, the dipotassium hydrogen phosphate of 0.75g/L, the magnesium sulfate of 0.3g/L, the manganese sulphate of 0.03g/L and the bubble enemy of 2.5g/L.Respectively in seeding tank and fermentation tank through 15 pounds of pressure and the sterilization of 121 ℃ of temperature 60 minutes, be cooled to 35 ℃ standby.
(5) seeding tank fermentation
Inoculation 1.5% above-mentioned Bt naturalized strain shake a bottle bacterium liquid fermented and cultured.Condition of culture is: jar 30 ℃ of temperature, tank pressure 0.02MPa, throughput 1: 1.2 (V/V/ branch), 250 rev/mins of mixing speeds, fermentation time 10 hours.
(6) ferment tank
The seed liquor of inoculation 8% is to fermentation tank.Condition of culture is: jar 30 ℃ of temperature, tank pressure 0.02MPa, throughput 1: 1.1 (V/V/ branch), 200 rev/mins of mixing speeds, fermentation time 36 hours.
(7) zymotic fluid post processing
Zymotic fluid adds diatomite and the 1.5g/L Tween-80 of 30g/L, stirs, and sends into the spray drying tower drying, 220 ℃ of EATs, and 70 ℃ of outlet temperatures are lower than 4%, packing to moisture content.
Embodiment 3
(1) yellow ginger waste hydrolyzed liquid
With embodiment 1.
(2) yellow ginger waste hydrolyzed liquid preliminary treatment
Slowly add the CaO powder of 41.0g/L in the yellow ginger waste hydrolyzed liquid, fully stir, surveying waste liquor PH is 7.55, free 8O
4 2-Concentration is 2600mg/L.Gravitational settling 3 hours is with CaSO
4Sediment is removed.
(3) Bt strain domestication
With embodiment 1.
(4) medium component replenishes
To in pretreated yellow ginger waste hydrolyzed liquid, adding the fish meal of 15g/L and the soya-bean oil of 5.0g/L.Do not add mineral salt, respectively in seeding tank and fermentation tank through 15 pounds of pressure and the sterilization of 121 ℃ of temperature 60 minutes, be cooled to 35 ℃ standby.
(5) seeding tank fermentation
Inoculation 1.5% above-mentioned Bt naturalized strain shake a bottle bacterium liquid fermented and cultured.Condition of culture is: jar 32 ℃ of temperature, tank pressure 0.02MPa, throughput 1: 1.5 (V/V/ branch), 260 rev/mins of mixing speeds, fermentation time 8 hours.
(6) ferment tank
The seed liquor of inoculation 10% is to fermentation tank.Condition of culture is: jar 32 ℃ of temperature, tank pressure 0.02MPa, throughput 1: 1.5 (V/V/ branch), 220 rev/mins of mixing speeds, fermentation time 32 hours.
(7) zymotic fluid post processing
Zymotic fluid adds bentonite and the 2.5g/LTX-10 of 30g/L, stirs, and sends into the spray drying tower drying, 200 ℃ of EATs, and 70 ℃ of outlet temperatures are lower than 4%, packing to moisture content.
Embodiment 4
(1) yellow ginger waste hydrolyzed liquid
Get the waste hydrolyzed liquid of Hubei Province merit Xi Xian yellow ginger processing factory, this factory adopts enzymatic hydrolysis-acid-hydrolysis method, and technological process is: yellow ginger → wet-milling → amylase liquefaction, carbohydrase saccharification → filtrate acid hydrolysis → hydrolysis residue extract saponin, the waste hydrolyzed liquid discharging.Content of reducing sugar is that 32.5g/L, crude protein content are 3.1g/L, sulfate radical content 29.3g/L in the waste liquid by analysis, pH＜0.
(2) yellow ginger waste hydrolyzed liquid preliminary treatment
In the yellow ginger waste hydrolyzed liquid, add the Ca (OH) of 40.0g/L
2Powder fully stirs, and surveying waste liquor PH is 7.42, free SO
4 2-Concentration is 4600mg/L.With conventional centrifugal separation with CaSO
4Sediment is removed.
(3) Bt strain domestication
Adopting production bacterial strain Bt7216 commonly used is starting strain.Domesticating method is with embodiment 1.
(4) medium component replenishes
In pretreated yellow ginger waste hydrolyzed liquid, add the big dregs of beans of 15g/L, the dipotassium hydrogen phosphate of 0.8g/L, the magnesium sulfate of 0.3g/L, the manganese sulphate of 0.04g/L and the bubble enemy of 2g/L.Respectively in seeding tank and fermentation tank through 15 pounds of pressure and the sterilization of 121 ℃ of temperature 60 minutes, be cooled to 30 ℃ standby.
(5) seeding tank fermentation
Inoculation 1% above-mentioned Bt naturalized strain shake a bottle bacterium liquid fermented and cultured.Condition of culture is: jar 28 ℃ of temperature, tank pressure 0.02MPa, throughput 1: 1.0 (V/V/ branch), 220 rev/mins of mixing speeds, fermentation time 12 hours.
(6) ferment tank
Inoculum concentration with 8% moves into fermentation tank with seed liquor.Condition of culture is: jar 28 ℃ of temperature, tank pressure 0.02MPa, throughput 1: 1.2 (V/V/ branch), 200 rev/mins of mixing speeds, fermentation time 42 hours.
(7) zymotic fluid post processing
Zymotic fluid adds the kaolin of 50g/L and the Tween-80 of 2.0g/L, stirs, and sends into the spray drying tower drying, 160 ℃ of EATs, and 65 ℃ of outlet temperatures are lower than 4%, packing to moisture content.
Embodiment 5
(1) yellow ginger waste hydrolyzed liquid
With embodiment 4.
(2) yellow ginger waste hydrolyzed liquid preliminary treatment
Slowly add the CaO powder of 30g/L in the yellow ginger waste hydrolyzed liquid, fully stir, surveying waste liquor PH is 7.35, free SO
4 2-Concentration is 4800mg/L.Gravitational settling 3 hours is with CaSO
4Sediment is removed.
(3) Bt strain domestication
With embodiment 4.
(4) medium component replenishes
To corn steep liquor, the dipotassium hydrogen phosphate of 0.25g/L, the magnesium sulfate of 0.1g/L, the manganese sulphate of 0.01g/L and the peanut oil of 5g/L through adding 30g/L in the pretreated yellow ginger waste hydrolyzed liquid.Respectively in seeding tank and fermentation tank through 15 pounds of pressure and the sterilization of 121 ℃ of temperature 60 minutes, be cooled to 32 ℃ standby.
(5) seeding tank fermentation
Inoculation 2% above-mentioned Bt naturalized strain shake a bottle bacterium liquid fermented and cultured.Condition of culture is: jar 30 ℃ of temperature, tank pressure 0.02MPa, throughput 1: 1.2 (V/V/ branch), 240 rev/mins of mixing speeds, fermentation time 10 hours.
(6) ferment tank
Inoculum concentration with 10% moves into fermentation tank with seed liquor.Condition of culture is: jar 30 ℃ of temperature, tank pressure 0.02MPa, throughput 1: 1.2 (V/V/ branch), 200 rev/mins of mixing speeds, fermentation time 38 hours.
(7) zymotic fluid post processing
Zymotic fluid adds the diatomite of 25g/L and the sodium lignosulfonate of 1.0g/L, stirs, and sends into the spray drying tower drying, 180 ℃ of EATs, and 65 ℃ of outlet temperatures are lower than 4%, packing to moisture content.
Embodiment 6
(1) yellow ginger waste hydrolyzed liquid
With embodiment 4.
(2) yellow ginger waste hydrolyzed liquid preliminary treatment
With embodiment 4.
(3) Bt strain domestication
With embodiment 4.
(4) medium component replenishes
To opposing through the dusty yeast that adds 20g/L in the pretreated yellow ginger waste hydrolyzed liquid, the dipotassium hydrogen phosphate of 0.5g/L, the magnesium sulfate of 0.2g/L, the manganese sulphate of 0.02g/L and the bubble of 1g/L.Respectively in seeding tank and fermentation tank through 15 pounds of pressure and the sterilization of 121 ℃ of temperature 60 minutes, be cooled to 35 ℃ standby.
(5) seeding tank fermentation
Inoculation 3% above-mentioned Bt naturalized strain shake a bottle bacterium liquid fermented and cultured.Condition of culture is: jar 32 ℃ of temperature, tank pressure 0.02MPa, throughput 1: 1.5 (V/V/ branch), 250 rev/mins of mixing speeds, fermentation time 8 hours.
(6) ferment tank
Inoculum concentration with 12% moves into fermentation tank with seed liquor.Condition of culture is: jar 32 ℃ of temperature, tank pressure 0.02MPa, throughput 1: 1.5 (V/V/ branch), 210 rev/mins of mixing speeds, fermentation time 32 hours.
(7) ferment liquid post processing
Zymotic fluid adds the precipitated calcium carbonate of 50g/L and the sodium lignosulfonate of 2.0g/L, stirs, and sends into the spray drying tower drying, 220 ℃ of EATs, and 70 ℃ of outlet temperatures are lower than 4%, packing to moisture content.
Claims (8)
1. the yellow ginger waste hydrolyzed liquid prepares the method for bacillus thuringiensis microbiological insecticide, its characteristic is: this method step is in the following order carried out: the preliminary treatment of waste liquid, strain domestication, medium component replenish, seeding tank fermentation, ferment tank and zymotic fluid post processing, and concrete grammar is as follows:
Wherein: L represents 1 liter of yellow ginger waste hydrolyzed liquid;
1) preliminary treatment of waste liquid: add the calcic alkaline matter of 10-80g/L in solid content is the yellow ginger waste hydrolyzed liquid of 2-5%, fully stirring reaction is removed CaSO
4Sediment; Reaction back waste liquor PH is 7.0-8.5, free 5O
4 2-Content is reduced to 500-5000mg/L;
2) strain domestication: the method domestication Bt fermented bacterium of taking in the beef extract-peptone solid culture medium, progressively to improve yellow ginger waste hydrolyzed liquid concentration, inoculation Bt bacterial strain in solid inclined-plane that adds variable concentrations yellow ginger waste hydrolyzed liquid or flat board, cultivated 72 hours down, carry out repeatedly successively for 30 ℃;
3) medium component replenishes: to adding the nitrogenous source of 10-30g/L, the mineral salt of 0.2-1.5g/L, the defoamer of 1-5g/L through pretreated yellow ginger waste hydrolyzed liquid; Sterilized 30-60 minute through 15 pounds of pressure and 121 temperature in seeding tank or fermentation tank in the back that stirs, be cooled to 30-35 ℃ standby;
4) seeding tank fermentation: bottle bacterium liquid that shakes to culture medium inoculated Bt naturalized strain, carry out fermented and cultured; Fermentation condition is: inoculum concentration 1-3%, warm 28-32 ℃ in jar, tank pressure 0.02-0.06MPa, throughput 1: 1.0-1.5V/V/ branch, mixing speed 220-260 rev/min, fermentation time 8-12 hour;
5) ferment tank: to fermentation tank, fermentation condition is the inoculum concentration of pressing 8-15% with the seed liquor culture transferring: jar warm 28-32 ℃, tank pressure 0.02-0.06MPa, throughput 1: 1.0-1.5V/V/ branch, mixing speed 180-220 rev/min, fermentation time 30-42 hour;
6) zymotic fluid post processing: zymotic fluid adds the dispersant of 20-50g/L and the emulsifier of 1-4g/L, stirs, and sends into the spray drying tower drying, and EAT 150-220 ℃, outlet temperature 60-70 ℃, packing.
2. prepare the method for bacillus thuringiensis microbiological insecticide according to the described yellow ginger waste hydrolyzed liquid of claim 1, it is characterized in that: used calcic alkaline matter is Ca (OH) 1)
2, CaCO
3Or CaO.
3. prepare the method for bacillus thuringiensis microbiological insecticide according to the described yellow ginger waste hydrolyzed liquid of claim 1, it is characterized in that: used CaSO 1)
4Sediment removal method is to utilize gravity settling separation or centrifugal separation.
4. prepare the method for bacillus thuringiensis microbiological insecticide according to the described yellow ginger waste hydrolyzed liquid of claim 1, it is characterized in that: used nitrogenous source is big dregs of beans, peanut meal, rapeseed dregs, fish meal, corn steep liquor, dusty yeast or peptone 3).
5. the method for preparing the bacillus thuringiensis microbiological insecticide according to the described yellow ginger waste hydrolyzed liquid of claim 1, it is characterized in that: the mixture that used mineral salt is made up of dipotassium hydrogen phosphate, magnesium sulfate, three kinds of materials of manganese sulphate 3), three's mass ratio is 20-100: 10-30: 1-5 in the mixture.
6. prepare the method for bacillus thuringiensis microbiological insecticide according to the described yellow ginger waste hydrolyzed liquid of claim 1, it is characterized in that: used defoamer is bubble enemy, polypropylene glycol, soya-bean oil or peanut oil 3).
7. prepare the method for bacillus thuringiensis microbiological insecticide according to the described yellow ginger waste hydrolyzed liquid of claim 1, it is characterized in that: used described dispersant is kaolin, diatomite, bentonite or precipitated calcium carbonate 6).
8. prepare the method for bacillus thuringiensis microbiological insecticide according to the described yellow ginger waste hydrolyzed liquid of claim 1, it is characterized in that: the emulsifier 6) is sodium lignosulfonate, Tween-80 or TX-10.
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| CN100409751C (en) * | 2006-09-07 | 2008-08-13 | 广东省生态环境与土壤研究所 | Microorganism mosquitocide prepared by using urban sludge and preparation method thereof |
| CN101988043B (en) * | 2009-08-05 | 2013-03-27 | 北京大学 | Bacillus thuringiensis microbial insecticide, preparation method and special culture medium thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1246284A (en) * | 1999-09-14 | 2000-03-08 | 中国科学院生态环境研究中心 | Process for preparing microbe insecticide with high-concentration waste organic water in fermentation industry |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1246284A (en) * | 1999-09-14 | 2000-03-08 | 中国科学院生态环境研究中心 | Process for preparing microbe insecticide with high-concentration waste organic water in fermentation industry |
Non-Patent Citations (1)
| Title |
|---|
| 以淀粉废液为原料生产生物农药. 王丽芳.微生物学杂志，第8卷第4期. 1988 * |
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