CN112704083B - Paecilomyces lilacinus microcapsule suspending agent and preparation method thereof - Google Patents
Paecilomyces lilacinus microcapsule suspending agent and preparation method thereof Download PDFInfo
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Abstract
The invention provides a paecilomyces lilacinus microcapsule suspending agent which consists of an oil phase forming component, a water phase forming component and a capsule wall material; the components for forming the oil phase comprise paecilomyces lilacinus concentrated bacterial liquid and an auxiliary agent; the components for forming the water phase comprise an emulsifier and paecilomyces lilacinus supernatant, the capsule wall material is formed by polymerizing a capsule wall material prepolymer, and the capsule wall material prepolymer comprises isocyanate and organic amine. The auxiliary agent comprises one or more of an acid-base regulator, a dispersing agent, solvent oil, an antifreezing agent, a defoaming agent, a preservative and a viscosity regulator; the paecilomyces lilacinus concentrated bacterial liquid and the supernatant are obtained by separating paecilomyces lilacinus fermented bacterial liquid. The paecilomyces lilacinus microcapsule suspending agent provided by the invention contains metabolites and effective viable bacteria of paecilomyces lilacinus in a water phase and an oil phase respectively, and compared with conventional powder, granules and water dispersing agents, the microcapsule suspending agent has the advantages of longer shelf life and more stable performance.
Description
Technical Field
The invention belongs to the technical field of microbial agents, and particularly relates to a paecilomyces lilacinus biological agent and a preparation method thereof.
Background
Paecilomyces lilacinus (A)Paecilomyces lilacinus(Thorn.) Samson is a fungus belonging to the genus paecilomyces of the family hyphomycetaceae, belonging to the genus parasitic fungus, and produces various organic acids, enzymes, physiologically active substances through secretory metabolism. The paecilomyces lilacinus can parasitize eggs and also infect larvae and female worms, after spores of the paecilomyces lilacinus germinate, the produced hyphae can penetrate the egg shells, the larvae and the walls of female adults of the nematodes, the hyphae absorb nutrition in the nematode body and propagate, and the normal physiological metabolism of the eggs, the larvae and the female adults is damaged, so that the plant nematodes die.
The paecilomyces lilacinus can inhibit nematode infection and promote the growth of plant root system and plant nutritive organs (research and development of paecilomyces lilacinus, pancang, fine and special chemicals, 2003, 11 (6)), and application modes such as seed dressing before sowing, hole application during planting and the like are preferably adopted. Therefore, there is a need for a paecilomyces lilacinus preparation which is stable and easily dispersible. The dosage forms of the microbial agent mainly comprise powder, granules, suspending agents, suspending emulsions and the like, and for the microbial agent for seed dressing, the powder and the granules are not as convenient to use as liquid dosage forms; the suspending agent or the suspending emulsion in the common liquid microbial inoculum formulation has the defects of poor stability and dispersibility. Therefore, there is a need to develop a paecilomyces lilacinus microcapsule suspension agent to improve the stability and dispersibility of the liquid microbial inoculum.
The preparation method of the microcapsule suspending agent comprises an interfacial polymerization method, a complex coacervation method and an in-situ polymerization method. The encapsulating material adopted by the interfacial polymerization method has higher toxicity and is more suitable for non-biological materials; the capsule wall materials used by the complex coacervation method are gelatin, Arabic gum, sodium alginate and the like, are natural, non-toxic and good in biocompatibility, but the encapsulation efficiency of the micro-capsule is not high, and the stability of the micro-capsule is poor. The in-situ polymerization method for obtaining the capsule wall material by polymerization of the prepolymer has the advantages of simple and easily obtained raw materials, no toxicity and degradability of the capsule wall material; the microcapsule prepared by the method has high encapsulation efficiency, good stability, strong water penetration resistance and more uniform grain diameter.
Disclosure of Invention
Aiming at the defects in the field, the invention aims to provide a paecilomyces lilacinus microcapsule suspending agent to improve the application effect of paecilomyces lilacinus.
The second purpose of the invention is to provide a preparation method of the paecilomyces lilacinus microcapsule suspending agent.
The technical scheme for realizing the aim of the invention is as follows:
a Paecilomyces lilacinus microcapsule suspension comprises components for forming oil phase, components for forming water phase, and capsule wall material;
the components for forming the oil phase comprise paecilomyces lilacinus concentrated bacterial liquid and an auxiliary agent; the components for forming the water phase comprise an emulsifier, paecilomyces lilacinus supernatant; the capsule wall material is polymerized by a capsule wall material prepolymer, and the capsule wall material prepolymer comprises isocyanate and organic amine; the auxiliary agent comprises one or more of an acid-base regulator, a dispersing agent, solvent oil, an antifreezing agent, a defoaming agent, a preservative and a viscosity regulator;
the paecilomyces lilacinus concentrated bacterial liquid and the supernatant are obtained by separating paecilomyces lilacinus fermented bacterial liquid.
Specifically, the paecilomyces lilacinus concentrated bacterial liquid and the supernatant are obtained by centrifugally separating paecilomyces lilacinus zymocyte liquid.
Preferably, the concentration of the paecilomyces lilacinus concentrated bacterial liquid is 150-300 hundred million/mL; the mass ratio of the paecilomyces lilacinus concentrated bacterial liquid in the components for forming the oil phase is 1-50%.
Wherein the mass ratio of the isocyanate to the organic amine is 1: (1-3); the isocyanate is one or more of TDI (toluene diisocyanate), isophorone diisocyanate trimer (IPDI), Hexamethylene Diisocyanate (HDI), HDI biuret, HDI trimer, TDI trimer, dicyclohexylmethane diisocyanate (HMDI) and addition products thereof; the organic amine is Tetraethylenepentamine (TEPA) or triethylene tetramine (TETA).
The microcapsule prepared by taking isocyanate as a material has no inhibition effect on the growth of crop root systems; the organic amine is preferably triethylene tetramine (TETA), and the obtained microcapsule has smaller particle size.
Wherein the solvent oil is one or more of soybean oil, n-hexane, rosin-based solvent ND-45, benzene, toluene, xylene, aliphatic hydrocarbon solvent oil, cyclohexanone, methyl esterified vegetable oil, mineral oil, aromatic hydrocarbon solvent oil, dichloromethane, petroleum ether, pine nut base oil, No. 100 solvent oil, No. 150 solvent oil and No. 200 solvent oil;
the emulsifier is one or more of Sodium Dodecyl Sulfate (SDS), alkylphenol polyoxyethylene, ethylene oxide propylene oxide block copolyether, fatty alcohol polyoxyethylene, nonylphenol polyoxyethylene ether phosphate, styrene phenol formaldehyde resin polyoxyethylene ether phosphate, styrene phenol polyoxyethylene polyoxypropylene ether, alkyl naphthalene formaldehyde condensation sodium sulfonate, alkyl succinate sodium sulfonate, span and tween.
The dispersing agent is one or more of naphthalenesulfonate formaldehyde condensate, tristyrylphenol polyoxyethylene ether sulfate ammonium salt, fatty alcohol polyoxyethylene ether sodium sulfate, fatty alcohol polyoxyethylene ether phosphate, alkylphenol polyoxyethylene ether sodium sulfate, alkylphenol polyoxyethylene ether phosphate, EO-PO block copolymer, styrene phenol formaldehyde resin polyoxyethylene ether phosphate, styrylphenol polyoxyethylene polyoxypropylene ether, alkyl naphthalene formaldehyde condensate sodium sulfonate, polymeric carboxylic acid sodium, methylene naphthalene sulfonate, polycarboxylate, lignosulfonate, pyrrolidone, tea saponin and bisphenol A dispersing agent;
wherein the pH regulator is one or more of sulfuric acid, hydrochloric acid, phosphoric acid, acetic acid, oxalic acid, citric acid, ammonium sulfate, ammonium chloride, zinc sulfate, triethanolamine, diethylamine, triethylamine, ethylenediamine, sodium hydroxide, potassium hydroxide, sodium carbonate or potassium carbonate;
the viscosity regulator is one or more of polyethylene glycol, polyvinyl alcohol, magnesium aluminum silicate, bentonite, kaolin, diatomite, attapulgite, carboxymethyl cellulose, xanthan gum, carboxymethyl starch and chitosan.
The antifreeze agent comprises one or more of but is not limited to ethylene glycol, propylene glycol, glycerol, isopropanol or urea;
the antifoaming agent includes, but is not limited to, one or more of epoxidized soybean oil, epoxidized linseed oil, fatty acid, epoxidized ester, xylitol;
the preservatives include, but are not limited to, one or more of potassium sorbate, sodium benzoate, formaldehyde, and isothiazolinones.
The preparation method of the paecilomyces lilacinus microcapsule suspending agent comprises the following steps:
1) obtaining paecilomyces lilacinus fermentation liquor through liquid submerged fermentation, wherein the bacteria content in the fermentation liquor is 20-40 hundred million/mL,
2) obtaining concentrated bacterial liquid through centrifugation, wherein the concentration of the concentrated bacterial liquid is 150-300 hundred million/mL, collecting supernatant for later use,
3) mixing the concentrated solution with isocyanate, sequentially adding solvent oil and a dispersing agent, adding an auxiliary agent, adjusting the pH value to 5.5-8.5, and uniformly stirring to obtain an oil phase; mixing the supernatant with emulsifier and organic amine to obtain water phase, and mixing the oil phase and the water phase.
Wherein the formula of the culture medium for liquid submerged fermentation is as follows: 10 g of sucrose, 20 g of yeast powder, 4g of peptone and MgSO4·7H2O0.5 g, KH2PO40.5 g and 5g of bean cake powder. Mixing the above components, adding water to 1000 ml, adjusting pH to 5.6 with hydrochloric acid, and sterilizing to obtain fermentation culture medium.
In the preparation process, the pH value is controlled to be 5.5-8.5, so that the suitable living conditions of the paecilomyces lilacinus can be provided. Preferably, the pH value is controlled to be 6.5-8.0.
Wherein the mass ratio of the oil phase to the water phase is 1: (0.8 to 1.2). The proportion of the auxiliary agent in the total mass of the oil phase and the water phase can be 0.1-10%.
In the step 3), pouring the oil phase into the water phase under a shearing condition, wherein the shearing rate is preferably 1500-5000 r/min, so as to obtain a small microcapsule particle size; preferably, the shear rate is 2000-3000 r/min, 5 min.
The paecilomyces lilacinus culture step comprises the main steps of inoculating to a slant culture medium for culture, shaking culture, submerged fermentation in a fermentation tank and the like. The formulation of the culture medium for slant culture may employ those known in the art, for example: 200 g of potato, 20 g of glucose and 20 g of agar, mixing and dissolving the components, adding water to a constant volume of 1000 ml, adjusting the pH value to 5.6 by using hydrochloric acid, and sterilizing to obtain a slant culture medium;
the medium formulation for shake flask culture may be the same as that for liquid submerged culture.
Further preferably, the liquid submerged fermentation conditions are: taking the seed culture in the middle and later period of logarithmic growth, transferring the seed culture into a fermentation tank filled with a fermentation culture medium with the inoculation amount of 5-8% (V/V), keeping the tank temperature at 22-28 ℃, the tank pressure at 0.1-1.0 kg/cm, the stirring speed at 100-200rpm, the ventilation quantity at 1: 0.5-0.8, and the fermentation culture time at 60-96 hours, preferably 70-85 hours.
By adopting a submerged fermentation method, higher viable count can be obtained in shorter time, and the production efficiency is improved.
The mass portion of the invention can be gram, kilogram, ton or the self-defined mass unit for convenient operation. In the same formulation, the unit mass represented by "part" is the same. The formulations of the present invention may be scaled up or down, or made into liquid or solid media by means known in the art.
The invention has the beneficial effects that:
the paecilomyces lilacinus microcapsule suspending agent provided by the invention contains metabolites and effective viable bacteria (conidia) of paecilomyces lilacinus in water phase and oil phase, and compared with conventional powder, granules and water dispersing agent, the microcapsule suspending agent has more stable performance.
The suspension microcapsule suspending agent prepared by the preparation method has good compatibility of the oil phase and the water phase, and the oil phase plays a role in wrapping and isolating oxygen and water for conidiospores, so that the conidiospores are in a stable dormant state, the storage time of the paecilomyces lilacinus conidiospores is remarkably prolonged, and the control effect is greatly improved.
In the preparation method, the supernatant after the centrifugal fermentation liquid is adopted in the water phase, so that the water consumption is saved, the number of effective viable bacteria (conidia) in the product is increased, and the fermentation liquid also provides nutrient components for plant growth.
Detailed Description
The following examples are intended to illustrate the invention but should not be construed as limiting the scope thereof. In the examples, all the means used are conventional in the art unless otherwise specified.
In the examples, the parts are by mass unless otherwise specified. Unless otherwise specified, all ratios are mass ratios.
Paecilomyces lilacinus (A)Paecilomyces lilacinus) The preservation number of the strain is CGMCC number 11579, the preservation time is 2015, 10 months and 30 days, and the address is as follows: china general microbiological culture Collection center.
Example 1: preparation of paecilomyces lilacinus zymocyte liquid by liquid submerged fermentation
Preparation of a fermentation medium: weighing 10 g of sucrose, 20 g of yeast powder, 4g of peptone and MgSO4·7H2O0.5 g, KH2PO40.5 g and 5g of bean cake powder, adding water to 1000 ml, adjusting the pH value to 5.6 by hydrochloric acid, and sterilizing to obtain the fermentation medium. The culture medium for submerged fermentation in the fermentation tank is obtained by amplifying according to the formula. The volume of the medium in this example was 5000L.
Taking the seed culture in the middle and later logarithmic growth period, inoculating into a fermentation tank filled with fermentation medium at an inoculation amount of 8% (V/V), keeping the tank temperature at 25 deg.C and the tank pressure at 0.5kg/cm, stirring at 150rpm, ventilating at 1: 0.6, and fermenting for 60 hr.
The bacteria content in the zymophyte liquid is 20 hundred million/mL.
EXAMPLE 2 preparation of Paecilomyces lilacinus microcapsule suspension
The paecilomyces lilacinus microcapsule suspension provided by the embodiment consists of ingredients for forming an oil phase, ingredients for forming an aqueous phase and a capsule wall material; the components for forming the oil phase comprise paecilomyces lilacinus concentrated bacterial liquid and an auxiliary agent; the components for forming the water phase comprise an emulsifier, paecilomyces lilacinus supernatant; the capsule wall material is polymerized by a capsule wall material prepolymer, the capsule wall material prepolymer comprises isocyanate and organic amine, and the auxiliary agent comprises an acid-base regulator, a dispersant and solvent oil; the paecilomyces lilacinus concentrated bacterial liquid and the supernatant are obtained by separating paecilomyces lilacinus fermented bacterial liquid.
The preparation method of the paecilomyces lilacinus microcapsule suspending agent comprises the following steps:
1) obtaining paecilomyces lilacinus zymocyte liquid through a liquid submerged fermentation process, wherein the bacterium content in the zymocyte liquid is 20-40 hundred million/mL,
2) obtaining concentrated bacterial liquid through centrifugation, wherein the concentration of the concentrated bacterial liquid is 150-300 hundred million/mL, pouring out supernatant for later use,
3) and mixing the concentrated solution and the TDI trimer, sequentially adding a dispersing agent and solvent oil, adding an acid-base regulator to regulate the pH value to 5.5-8.5, and uniformly stirring to obtain an oil phase.
4) Mixing the supernatant with emulsifier and triethylene tetramine, stirring to obtain water phase, and pouring oil phase into water phase under shearing condition (shearing rate of 3000r/min, 5 min) to obtain Paecilomyces lilacinus microcapsule suspension.
In this example, the bacteria content in the fermentation broth was 20 hundred million/mL, and the centrifugation conditions were: 10000rpm for 15 minutes, and the concentration of the obtained concentrated bacterial liquid is 150 hundred million/mL. The dispersant is Sokalan CP (polycarboxylate) and the solvent oil is soybean oil.
Wherein the dosage ratio of the TDI tripolymer, the triethylene tetramine and the supernatant fluid is 1 g: 1.5 g: 100mL, 1% (mass ratio) SDS was added to the supernatant. The mass ratio of the concentrated solution, the dispersing agent and the solvent oil in the step 3) is 2: 35, adjusting the pH value to 7.5 by using an acid-base regulator ethylenediamine;
in the step 4), the water phase and the oil phase are mixed according to the mass ratio of 1: 1.
Table 1: example 2 Paecilomyces lilacinus microcapsule suspension test results
Example 3: preparation of paecilomyces lilacinus zymocyte liquid by liquid submerged fermentation
The fermentation medium was prepared as in example 1.
The seed culture in the middle and later logarithmic growth stage (the culture method is the same as that in examples 1 and 2) is inoculated into a fermentation tank filled with a fermentation medium in an inoculation amount of 8% (V/V), the temperature of the tank is kept at 25 ℃, the pressure of the tank is kept at 0.5kg/cm, the stirring speed is 150rpm, the ventilation rate is 1: 0.6, and the fermentation culture time is 72 hours.
The bacteria content in the zymophyte liquid is 30 hundred million/mL.
EXAMPLE 4 preparation of Paecilomyces lilacinus microcapsule suspension
1) Obtaining paecilomyces lilacinus zymocyte liquid through a liquid submerged fermentation process, wherein the bacterium content in the zymocyte liquid is 20-40 hundred million/mL,
2) obtaining concentrated bacterial liquid through centrifugation, wherein the concentration of the concentrated bacterial liquid is 150-300 hundred million/mL, pouring out supernatant for later use,
3) and mixing the concentrated solution and the TDI trimer, sequentially adding a dispersing agent and solvent oil, adding an acid-base regulator to regulate the pH value to 5.5-8.5, and uniformly stirring to obtain an oil phase.
4) Mixing the supernatant with emulsifier and triethylene tetramine, stirring to obtain water phase, and pouring oil phase into water phase under shearing condition (shearing rate of 3000r/min, 5 min).
In this example, the bacteria content in the fermentation broth was 30 hundred million/mL, and the centrifugation conditions were: 10000rpm for 15 minutes, and the concentration of the obtained concentrated bacterial liquid is 200 hundred million/mL.
The composition ratios were the same as in example 2.
Table 2: example 4 Paecilomyces lilacinus microcapsule suspension test results
Example 5: preparation of paecilomyces lilacinus zymocyte liquid by liquid submerged fermentation
The fermentation medium was prepared as in example 1.
Taking the seed culture in the middle and later logarithmic growth period, inoculating into a fermentation tank filled with fermentation medium at an inoculation amount of 8% (V/V), keeping the tank temperature at 25 deg.C and the tank pressure at 0.5kg/cm, stirring at 150rpm, ventilating at 1: 0.6, and fermenting for 84 hr. The bacteria content in the zymophyte liquid is 40 hundred million/mL.
EXAMPLE 6 preparation of Paecilomyces lilacinus microcapsule suspension
1) Obtaining paecilomyces lilacinus zymocyte liquid through a liquid submerged fermentation process, wherein the bacterium content in the zymocyte liquid is 20-40 hundred million/mL,
2) obtaining concentrated bacteria liquid through centrifugation, wherein the concentration of the concentrated bacteria liquid is 150-300 hundred million/mL, pouring out supernatant for later use,
3) mixing the concentrated solution with TDI trimer, sequentially adding a dispersant and solvent oil, adding an acid-base regulator to regulate the pH value to 5.5-8.5, and uniformly stirring to obtain an oil phase; the dispersant is Sokalan CP (polycarboxylate) and the solvent oil is soybean oil.
4) Mixing the supernatant with emulsifier and triethylene tetramine, stirring to obtain water phase, and pouring the oil phase into the water phase under shearing condition. (shear rate 3000r/min, 5 min).
In this example, the bacteria content in the fermentation broth was 40 hundred million/mL, and the centrifugation conditions were: 10000rpm for 15 minutes, and the concentration of the obtained concentrated bacterial liquid is 300 hundred million/mL.
The composition ratios were the same as in example 2.
Table 3: example 6 Paecilomyces lilacinus microcapsule suspension test results
Example 7: preparation of paecilomyces lilacinus zymocyte liquid by liquid submerged fermentation
The fermentation medium was prepared as in example 1.
Taking the seed culture in the middle and later logarithmic growth period, inoculating into a fermentation tank filled with fermentation medium at an inoculation amount of 8% (V/V), maintaining the tank temperature at 25 deg.C and the tank pressure at 0.5kg/cm, stirring at 150rpm, ventilating at 1: 0.6, and fermenting for 90 hr. The bacteria content in the zymophyte liquid is 30 hundred million/mL.
In production, the longer fermentation time can not obtain more viable bacteria. Therefore, the fermentation time is preferably 70 to 85 hours.
Test examples
This test example compares the shelf life of different forms of paecilomyces lilacinus. The preparation formulations are respectively as follows:
paecilomyces lilacinus water dispersion agent: preparing concentrated bacterial liquid obtained by centrifugal separation of the zymophyte liquid obtained in the embodiment 5 into bacterial powder, adding an auxiliary agent, and preparing water dispersion agent by a conventional method;
paecilomyces lilacinus powder: the concentrated bacterial liquid obtained by centrifugal separation of the fermented bacterial liquid obtained in example 5 was made into bacterial powder, and then added with an auxiliary agent to make into powder by a conventional method.
Paecilomyces lilacinus microcapsule suspension of example 6.
The preparation is prepared, viable bacteria count is carried out by using a dilution coating flat plate method, and viable bacteria count is carried out by using a dilution coating flat plate method after sampling for 3 months, 6 months and 12 months. The results are shown in Table 4.
Table 4: viable bacteria count results
| Dosage forms | Counting for 3 months | Counting for 6 months | 12 month count |
| Water dispersion agent | 12 hundred million/mL | Can not detect out | Can not detect out |
| Powder preparation | 25 hundred million/g | 12 hundred million/g | Can not detect out |
| Microcapsule suspension | 37 hundred million/mL | 26 hundred million/mL | 20 hundred million/mL |
Note: the initial concentration of each of the three dosage forms was 40 hundred million/mL (or 40 hundred million/gram).
The number of viable bacteria on the flat plate is only 30% of the initial count of the water dispersant at 3 months, the number of viable bacteria on the flat plate is only about 60% of the initial count of the water dispersant at 3 months, the number of viable bacteria on the flat plate is 20-30% of the initial count at 6 months, and the number of viable bacteria cannot be detected at 12 months; the viable count of the microcapsule suspending agent on the flat plate is about 90 percent of the initial count in 3 months, the viable count on the flat plate is 60-70 percent of the initial count in 6 months, and the viable count is about 50 percent of the initial count in 12 months. The suspension microcapsule suspending agent prepared by the method obviously prolongs the storage time of the paecilomyces lilacinus conidia, and greatly improves the prevention and treatment effect.
Although the present invention has been described in detail hereinabove, it will be apparent to those skilled in the art that modifications and improvements can be made thereto based on the present invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (6)
1. A paecilomyces lilacinus microcapsule suspension is characterized by consisting of ingredients for forming an oil phase, ingredients for forming a water phase and a capsule wall material;
the component for forming the oil phase is paecilomyces lilacinus (A)Paecilomyces lilacinus) Concentrated bacterial liquid and an auxiliary agent; the components for forming the water phase consist of an emulsifier and paecilomyces lilacinus supernatant; the capsule wall material is formed by polymerizing a capsule wall material prepolymer, the capsule wall material prepolymer comprises TDI tripolymer and triethylene tetramine, and the mass ratio of the TDI tripolymer to the triethylene tetramine is 1: (1-3), wherein the auxiliary agent consists of an acid-base regulator, a dispersing agent and solvent oil;
the paecilomyces lilacinus concentrated bacterial liquid and the supernatant are obtained by separating paecilomyces lilacinus fermented bacterial liquid;
the paecilomyces lilacinus microcapsule suspending agent is prepared by the following steps:
1) obtaining paecilomyces lilacinus fermentation liquor through liquid submerged fermentation, wherein the bacteria content in the fermentation liquor is 20-40 hundred million/mL, and the conditions of the liquid submerged fermentation are as follows:
taking the seed culture in the middle and later period of logarithmic growth, transferring the seed culture into a fermentation tank filled with a fermentation culture medium in an inoculation amount of 5-8% (V/V), keeping the tank temperature at 22-28 ℃ and the tank pressure at 0.1-1.0 kg/cm2Stirring the mixtureThe speed is 100-200rpm, the ventilation quantity is 1: 0.5-0.8, and the fermentation culture time is 70-85 hours;
2) obtaining concentrated bacterial liquid through centrifugation, wherein the concentration of the concentrated bacterial liquid is 150-300 hundred million/mL, collecting supernatant for later use,
3) mixing the concentrated solution with TDI trimer, sequentially adding solvent oil and a dispersant, adding an acid-base regulator, regulating the pH value to 5.5-8.5, uniformly stirring to obtain an oil phase, mixing the supernatant with an emulsifier and triethylene tetramine to obtain a water phase, pouring the oil phase into the water phase under a shearing condition, wherein the shearing rate is 2000-3000 r/min, and the shearing time is 5 min.
2. The paecilomyces lilacinus microcapsule suspension according to claim 1, wherein the concentration of the paecilomyces lilacinus concentrated bacterial liquid is 150-300 hundred million/mL; the mass content of the paecilomyces lilacinus concentrated bacterial liquid in the components for forming the oil phase is 1-50%.
3. The paecilomyces lilacinus microcapsule suspension according to claim 1, wherein the solvent oil is one or more of soybean oil, n-hexane, rosin-based solvent ND-45, benzene, toluene, xylene, aliphatic hydrocarbon solvent oil, cyclohexanone, methyl esterified vegetable oil, mineral oil, aromatic hydrocarbon solvent oil, dichloromethane, petroleum ether, pine seed base oil, solvent oil No. 100, solvent oil No. 150 or solvent oil No. 200;
the emulsifier is one or more of lauryl sodium sulfate, alkylphenol polyoxyethylene, ethylene oxide propylene oxide block copolyether, fatty alcohol polyoxyethylene, nonylphenol polyoxyethylene phosphate, styrene phenol formaldehyde resin, polyoxyethylene phosphate, styrene phenol polyoxyethylene polyoxypropylene ether, alkyl naphthalene formaldehyde condensate sodium sulfonate, alkyl succinate sodium sulfonate, span and tween.
4. A paecilomyces lilacinus microcapsule suspension according to claim 1,
the dispersing agent is one or more of naphthalenesulfonate formaldehyde condensate, tristyrylphenol polyoxyethylene ether sulfate ammonium salt, fatty alcohol polyoxyethylene ether sodium sulfate, fatty alcohol polyoxyethylene ether phosphate, alkylphenol polyoxyethylene ether sodium sulfate, alkylphenol polyoxyethylene ether phosphate, EO-PO block copolymer, styrene phenol formaldehyde resin polyoxyethylene ether phosphate, styrylphenol polyoxyethylene polyoxypropylene ether, alkyl naphthalene formaldehyde condensate sodium sulfonate, polymeric sodium carboxylate, methylene naphthalene sulfonate, polycarboxylate, lignosulfonate and tea saponin;
the acid-base regulator is one or more of sulfuric acid, hydrochloric acid, phosphoric acid, acetic acid, oxalic acid, citric acid, ammonium sulfate, ammonium chloride, triethanolamine, diethylamine, triethylamine, ethylenediamine, sodium hydroxide, potassium hydroxide, sodium carbonate or potassium carbonate.
5. The paecilomyces lilacinus microcapsule suspension according to any one of claims 1 to 4, wherein the formula of the culture medium for liquid submerged fermentation is as follows: 10 g of sucrose, 20 g of yeast powder, 4g of peptone and MgSO4·7H2O0.5 g, KH2PO40.5 g and 5g of bean cake powder; mixing the above components, adding water to 1000 ml, adjusting pH to 5.6 with hydrochloric acid, and sterilizing to obtain fermentation culture medium.
6. The paecilomyces lilacinus microcapsule suspension according to any one of claims 1 to 4, wherein the mass ratio of the oil phase to the water phase is 1: (0.8 to 1.2).
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