CN112920982B - Paenibacillus spray drying protective agent and spray drying method using protective agent - Google Patents
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Abstract
The invention relates to a paenibacillus spray drying protective agent and a spray drying method using the protective agent, wherein the paenibacillus spray drying protective agent uses sodium carboxymethyl cellulose, bean pulp powder, rice hull powder and molasses as raw materials, the raw materials are widely available and low in price, and the production cost is low; furthermore, after the bean pulp powder and molasses in the spray drying protective agent are applied to soil, the effect of promoting the paenibacillus to proliferate at plant roots is exerted, and meanwhile, the bean pulp powder has the effect of enhancing the fertility of the soil in the use process, so that the paenibacillus spray drying protective agent has great market competitiveness.
Description
Technical Field
The invention belongs to the technical field of microbial agents, and particularly relates to a bacillus-like spray drying protective agent and a spray drying method using the protective agent.
Background
In recent years, chemical fertilizers play a vital role in agricultural production, but people gradually find that the chemical fertilizers have serious nutrient loss and high loss, and the utilization rate of the chemical fertilizers is lower and lower. The long-term application of chemical fertilizer easily causes harm to soil structure and physicochemical properties: the content of As in the soil can reach 50-60mg/kg after long-term application of ammonium nitrate, ammonium phosphate and compound fertilizer; accelerating soil acidification; resulting in soil nutrient imbalance; the soil aggregate structure is destroyed, so that the soil is seriously hardened, and the agricultural cultivation value is seriously even lost.
With the increasing prominence of resource and environmental problems, people are paying attention to ecological environment and food safety problems, and microbial fertilizers are used as important supplements of traditional fertilizers and are widely applied to ecological agriculture.
Agricultural Paenibacillus (such as Paenibacillus polymyxa, paenibacillus azoxypegani, paenibacillus macerans, paenibacillus lautus, etc.) is an important strain for producing microbial fertilizers, and the microbial agents thereof have been widely applied to agricultural production. Spray drying is the most economical and convenient method of concentrating microbial fermentation broth to make dry microbial inoculum, but the microbial is subjected to high temperature and drying pressure during spray drying, and the recovery rate of viable bacteria is generally low. Therefore, people generally use high polymer materials such as starch, skimmed milk powder, cyclodextrin, trehalose, amino acid, polyamino acid and the like as a protective agent for microbial spray drying, so as to achieve the aim of reducing the reduction or loss of microbial activity in the spray drying process. Patent CN201210190725 discloses that polyaspartic acid is used as a spray drying protective agent, and although the recovery rate of viable bacteria is improved, the cost is high, and the spray drying protective agent is not suitable for agricultural production, so that development of a spray drying protective agent with high recovery rate of viable bacteria and low cost is an urgent need for agricultural production.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a paenibacillus spray drying protective agent and a preparation method thereof. When the spray drying protective agent is used for the spray drying of paenibacillus, the activity loss of agricultural paenibacillus in the spray drying process can be effectively reduced, and the yield of the dried living bacteria is improved.
The technical scheme adopted by the invention is as follows:
a bacillus-like spray-drying protectant comprising the following components: sodium carboxymethyl cellulose, soybean meal, rice hull powder and molasses.
Further preferably, the paenibacillus spray drying protective agent comprises the following raw materials in percentage by mass: 2-5% of sodium carboxymethyl cellulose, 1-10% of soybean meal powder, 0.5-5% of rice hull powder and 0.2-1% of molasses.
The mass percentages of the raw materials are respectively as follows: sodium carboxymethylcellulose 2%, soybean meal 8%, rice hull powder 3% and molasses 0.5%.
The paenibacillus is agricultural paenibacillus.
The agricultural paenibacillus is one or more of paenibacillus polymyxa, nitrogen fixation paenibacillus, paenibacillus macerans and paenibacillus lautus.
The spray drying method using the paenibacillus spray drying protective agent comprises the following steps:
(1) Liquid fermentation is carried out on paenibacillus until the sporulation period, and concentrated solution of paenibacillus is obtained after concentration;
(2) And (3) sequentially adding sodium carboxymethylcellulose, bean pulp powder, rice hull powder and molasses into the concentrated solution of the paenibacillus in the step (1), fully and uniformly stirring, and then carrying out spray drying.
In the step (1), the formula of the liquid fermentation medium is as follows: 10g/L of glucose, 15g/L of sucrose, 15g/L of corn steep liquor, 5g/L of yeast powder, 20mg/L of ferrous sulfate, 5g/L of magnesium sulfate, 0.5g/L of potassium dihydrogen phosphate and 2.5g/L of disodium hydrogen phosphate.
In the step (1), the specific conditions of the liquid fermentation are as follows:
the culture temperature is 28-33 ℃, the initial pH is 7.0, the aeration ratio is l 0.6, the stirring speed is 120r/min in the early stage and 220r/min in the later stage, and after fermentation is carried out for 18-24 hours, the fermentation is finished when the dissolved oxygen is increased to more than 70%.
In the step (1), the ceramic membrane is adopted for concentration for 2 to 5 times, and then the concentrated solution of the paenibacillus is obtained.
In the step (2), the spray drying is carried out in a spray drying tower, the temperature of an air inlet is 140-150 ℃, the temperature of an air outlet is 50-85 ℃, and the pressure of a spray head is 140-170kPa;
the water content after spray drying was below 7%.
The beneficial effects of the invention are as follows:
the paenibacillus spray drying protective agent takes sodium carboxymethyl cellulose, bean pulp powder, rice hull powder and molasses as raw materials, and the raw materials have wide sources and wide pricesThe method has the advantages that the method is low in production cost, when the prepared spray-drying protective agent is used for the spray-drying of paenibacillus, the activity loss of agricultural paenibacillus in the spray-drying process can be effectively reduced, the yield of the dried live bacteria is improved, and the data show that the average yield of the agricultural paenibacillus bacterial agent produced by the method is above 68 percent, and the number of the live bacteria is 2 multiplied by 10 10 The content of the organic fertilizer is more than one per gram, and the requirements of producing the compound microbial fertilizer or the microbial organic fertilizer are met. Furthermore, after the bean pulp powder and molasses in the spray drying protective agent are applied to soil, the effect of promoting the paenibacillus to proliferate at plant roots is exerted, and meanwhile, the bean pulp powder has the effect of enhancing the fertility of the soil in the use process, so that the paenibacillus spray drying protective agent has great market competitiveness.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It will be apparent that the embodiments described are only some, but not all, embodiments of the invention. All other embodiments, based on the examples herein, which are within the scope of the invention as defined by the claims, will be within the scope of the invention as defined by the claims.
Example 1
The embodiment provides a nitrogen-fixing paenibacillus spray drying protective agent, which takes the volume of concentrated liquid of nitrogen-fixing paenibacillus as a reference, and the mass ratio of the added raw materials is as follows: sodium carboxymethylcellulose 2% (w/v), soybean meal 2% (w/v), rice hull 1% (w/v), and molasses 0.3% (w/v).
The spray drying method using the nitrogen-fixing paenibacillus spray drying protective agent comprises the following steps:
(1) Performing liquid fermentation on the nitrogen-fixing paenibacillus to a spore formation period, and concentrating by 3 times by adopting a 100nm ceramic membrane to obtain a concentrated solution of the nitrogen-fixing paenibacillus;
the formula of the culture medium for liquid fermentation is as follows: 10g/L of glucose, 15g/L of sucrose, 15g/L of corn steep liquor, 5g/L of yeast powder, 20mg/L of ferrous sulfate, 5g/L of magnesium sulfate, 0.5g/L of potassium dihydrogen phosphate and 2.5g/L of disodium hydrogen phosphate; the specific conditions for carrying out the liquid fermentation are as follows: the culture temperature is 28-33 ℃, the initial pH is 7.0, the aeration ratio is l, the stirring speed is 120r/min in the early stage, 220r/min in the later stage, and after fermentation for 24 hours, the fermentation is finished when the dissolved oxygen rises to more than 70%;
(2) Sequentially adding sodium carboxymethylcellulose, soybean meal powder, rice hull powder and molasses into the concentrated solution of the nitrogen-fixing paenibacillus in the step (1), fully and uniformly stirring, and then spray-drying in a spray-drying tower, wherein the temperature of an air inlet is 145 ℃, the temperature of an air outlet is 70 ℃, the pressure of a spray nozzle is 160kPa, and the water content after spray-drying is lower than 7%.
Comparative example 1
The comparative example differs from example 1 only in the kind and composition of the raw materials of the spray-drying protecting agent, specifically: based on the volume of the concentrated solution of the nitrogen-fixing paenibacillus, the adding mass ratio of the raw materials is as follows: 5% (w/v) of light calcium carbonate and 3% (w/v) of polyaspartic acid.
Example 2
The embodiment provides a paenibacillus polymyxa spray drying protective agent, which takes the volume of concentrated solution of paenibacillus polymyxa as a reference, and the added mass percentages of the raw materials are as follows: sodium carboxymethylcellulose 3%, soybean meal 2%, rice hull powder 3% and molasses 0.3%.
The spray drying method using the paenibacillus polymyxa spray drying protective agent comprises the following steps:
(1) Performing liquid fermentation on Paenibacillus polymyxa until a spore formation period, and concentrating by 2 times with a 100nm ceramic membrane to obtain Paenibacillus polymyxa concentrated solution;
the formula of the culture medium for liquid fermentation is as follows: 10g/L of glucose, 15g/L of sucrose, 15g/L of corn steep liquor, 5g/L of yeast powder, 20mg/L of ferrous sulfate, 5g/L of magnesium sulfate, 0.5g/L of potassium dihydrogen phosphate and 2.5g/L of disodium hydrogen phosphate; the specific conditions for carrying out the liquid fermentation are as follows: the culture temperature is 28-33 ℃, the initial pH is 7.0, the aeration ratio is l, the stirring speed is 120r/min in the early stage, 220r/min in the later stage, and the fermentation is finished when the dissolved oxygen rises to more than 70% after 18h of fermentation;
(2) Sequentially adding sodium carboxymethylcellulose, soybean meal powder, rice hull powder and molasses into the concentrated solution of the paenibacillus polymyxa in the step (1), fully and uniformly stirring, and then spray-drying in a spray-drying tower, wherein the temperature of an air inlet is 145 ℃, the temperature of an air outlet is 70 ℃, the pressure of a spray nozzle is 160kPa, and the water content after spray-drying is lower than 7%.
Comparative example 2
The comparative example differs from example 2 only in the kind and composition of the raw materials of the spray-drying protecting agent, specifically: based on the volume of the concentrated solution of the paenibacillus polymyxa, the mass ratio of the added raw materials is as follows: 5% (w/v) of light calcium carbonate and 3% (w/v) of poly-gamma-glutamic acid.
Example 3
The embodiment provides a bacillus lautus spray drying protective agent, which takes the volume of concentrated liquid of bacillus lautus as a reference, and the added mass percentages of the raw materials are as follows: sodium carboxymethylcellulose 4%, soybean meal 6%, rice hull powder 4% and molasses 1%.
A spray drying method using the protective agent for spray drying of the bacillus lautus, comprising the following steps:
(1) Liquid fermentation is carried out on the Paenibacillus lautus until the spore formation period, and a 100nm ceramic film is adopted for concentration for 5 times, so as to obtain concentrated solution of Paenibacillus polymyxa;
the formula of the culture medium for liquid fermentation is as follows: 10g/L of glucose, 15g/L of sucrose, 15g/L of corn steep liquor, 5g/L of yeast powder, 20mg/L of ferrous sulfate, 5g/L of magnesium sulfate, 0.5g/L of potassium dihydrogen phosphate and 2.5g/L of disodium hydrogen phosphate; the specific conditions for carrying out the liquid fermentation are as follows: the culture temperature is 28-33 ℃, the initial pH is 7.0, the aeration ratio is l 0.6, the stirring speed is 120r/min in the early stage and 220r/min in the later stage, after fermentation is carried out for 20h, when the dissolved oxygen is raised to more than 70%, the fermentation is finished;
(2) Sequentially adding sodium carboxymethylcellulose, soybean meal powder, rice hull powder and molasses into the concentrated solution of the bacillus lautus in the step (1), fully and uniformly stirring, and then spray-drying in a spray-drying tower, wherein the temperature of an air inlet is 150 ℃, the temperature of an air outlet is 75 ℃, the pressure of a spray nozzle is 160kPa, and the water content after spray-drying is lower than 7%.
Comparative example 3
The comparative example differs from example 3 only in the kind and composition of the raw materials of the spray-drying protecting agent, specifically: based on the volume of the concentrated solution of the bacillus lautus, the adding mass ratio of the raw materials is as follows: 5% (w/v) of light calcium carbonate, 5% (w/v) of polyaspartic acid, 3% (w/v) of poly gamma glutamic acid and 3% (w/v) of maltodextrin.
Example 4
The embodiment provides a paenibacillus macerans spray drying protective agent, which takes the volume of concentrated liquid of paenibacillus macerans as a reference, and the added mass percentages of the raw materials are as follows: sodium carboxymethylcellulose 4%, soybean meal 6%, rice hull powder 4% and molasses 1%.
The spray drying method using the Paenibacillus macerans spray drying protective agent comprises the following steps:
(1) Liquid fermentation is carried out on Paenibacillus macerans until a spore formation period, and a 100nm ceramic membrane is adopted for concentration for 2 times, so that concentrated liquid of Paenibacillus macerans is obtained;
the formula of the culture medium for liquid fermentation is as follows: 10g/L of glucose, 15g/L of sucrose, 15g/L of corn steep liquor, 5g/L of yeast powder, 20mg/L of ferrous sulfate, 5g/L of magnesium sulfate, 0.5g/L of potassium dihydrogen phosphate and 2.5g/L of disodium hydrogen phosphate; the specific conditions for carrying out the liquid fermentation are as follows: the culture temperature is 28-33 ℃, the initial pH is 7.0, the aeration ratio is l, the stirring speed is 120r/min in the early stage, 220r/min in the later stage, and after fermentation for 22h, the fermentation is finished when the dissolved oxygen rises to more than 70%;
(2) And (3) sequentially adding sodium carboxymethylcellulose, soybean meal powder, rice hull powder and molasses into the concentrated solution of the Paenibacillus macerans in the step (1), fully and uniformly stirring, and then performing spray drying in a spray drying tower, wherein the temperature of an air inlet is 140 ℃, the temperature of an air outlet is 60 ℃, the pressure of a spray nozzle is 170kPa, and the water content after spray drying is lower than 7%.
Comparative example 4
The comparative example differs from example 4 only in the kind and composition of the raw materials of the spray-drying protecting agent, specifically: based on the volume of the concentrated solution of the Paenibacillus macerans, the mass ratio of the added raw materials is as follows: 5% (w/v) of light calcium carbonate, 5% (w/v) of polyaspartic acid, 3% (w/v) of poly gamma glutamic acid and 5% (w/v) of maltodextrin.
Example 5
The embodiment provides a nitrogen-fixing paenibacillus spray drying protective agent, which takes the volume of concentrated liquid of nitrogen-fixing paenibacillus as a reference, and the mass ratio of the added raw materials is as follows: sodium carboxymethylcellulose 5% (w/v), soybean meal 10% (w/v), rice hull 5% (w/v), molasses 0.5% (w/v).
The spray drying method using the nitrogen-fixing paenibacillus spray drying protective agent comprises the following steps:
(1) Performing liquid fermentation on the nitrogen-fixing paenibacillus to a spore formation period, and concentrating by 2 times by adopting a 100nm ceramic membrane to obtain a concentrated solution of the nitrogen-fixing paenibacillus;
the formula of the culture medium for liquid fermentation is as follows: 10g/L of glucose, 15g/L of sucrose, 15g/L of corn steep liquor, 5g/L of yeast powder, 20mg/L of ferrous sulfate, 5g/L of magnesium sulfate, 0.5g/L of potassium dihydrogen phosphate and 2.5g/L of disodium hydrogen phosphate; the specific conditions for carrying out the liquid fermentation are as follows: the culture temperature is 28-33 ℃, the initial pH is 7.0, the aeration ratio is l, the stirring speed is 120r/min in the early stage, 220r/min in the later stage, and after fermentation for 24 hours, the fermentation is finished when the dissolved oxygen rises to more than 70%;
(2) Sequentially adding sodium carboxymethylcellulose, soybean meal powder, rice hull powder and molasses into the concentrated solution of the nitrogen-fixing paenibacillus in the step (1), fully and uniformly stirring, and then spray-drying in a spray-drying tower, wherein the temperature of an air inlet is 145 ℃, the temperature of an air outlet is 50 ℃, the pressure of a spray nozzle is 160kPa, and the water content after spray-drying is lower than 7%.
Example 6
The embodiment provides a nitrogen-fixing paenibacillus spray drying protective agent, which takes the volume of concentrated liquid of nitrogen-fixing paenibacillus as a reference, and the mass ratio of the added raw materials is as follows: sodium carboxymethylcellulose 5% (w/v), soybean meal 1% (w/v), rice hull 0.5% (w/v), molasses 0.2% (w/v).
The spray drying method using the nitrogen-fixing paenibacillus spray drying protective agent comprises the following steps:
(1) Performing liquid fermentation on the nitrogen-fixing paenibacillus to a spore formation period, and concentrating by 2 times by adopting a 100nm ceramic membrane to obtain a concentrated solution of the nitrogen-fixing paenibacillus;
the formula of the culture medium for liquid fermentation is as follows: 10g/L of glucose, 15g/L of sucrose, 15g/L of corn steep liquor, 5g/L of yeast powder, 20mg/L of ferrous sulfate, 5g/L of magnesium sulfate, 0.5g/L of potassium dihydrogen phosphate and 2.5g/L of disodium hydrogen phosphate; the specific conditions for carrying out the liquid fermentation are as follows: the culture temperature is 28-33 ℃, the initial pH is 7.0, the aeration ratio is l, the stirring speed is 120r/min in the early stage, 220r/min in the later stage, and the fermentation is finished when the dissolved oxygen rises to more than 70% after 18h of fermentation;
(2) Sequentially adding sodium carboxymethylcellulose, soybean meal powder, rice hull powder and molasses into the concentrated solution of the nitrogen-fixing paenibacillus in the step (1), fully and uniformly stirring, and then spray-drying in a spray-drying tower, wherein the temperature of an air inlet is 145 ℃, the temperature of an air outlet is 85 ℃, the pressure of a spray nozzle is 140kPa, and the water content after spray-drying is lower than 7%.
Experimental example
The results of spray drying and then examining the yields of viable bacteria in the above examples and comparative examples are shown in Table 1.
TABLE 1 yields of viable bacteria after spray drying of different protectants
As can be seen from Table 1, when the protective agent produced by the method of the invention is used for spray drying of agricultural paenibacillus bacterial agent, the yield of viable bacteria is more than 68 percent on average, and the viable bacteria number is 2 multiplied by 10 10 The number of the catalyst is more than one per gram,meets the requirements of producing compound microbial fertilizer or microbial organic fertilizer.
The foregoing is merely illustrative of the present invention, and the present invention is not limited thereto, and any person skilled in the art will readily recognize that variations or substitutions are within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (8)
1. The bacillus-like spray drying protective agent is characterized by comprising sodium carboxymethyl cellulose, soybean meal powder, rice hull powder and molasses;
based on the volume of the concentrated solution of the paenibacillus, the adding mass percentages of the raw materials are as follows: 2-5% of sodium carboxymethyl cellulose, 1-10% of soybean meal powder, 0.5-5% of rice hull powder and 0.2-1% of molasses.
2. The paenibacillus spray drying protective agent according to claim 1, wherein the mass percentages of the raw materials are respectively: sodium carboxymethylcellulose 2%, soybean meal 8%, rice hull powder 3% and molasses 0.5%.
3. The paenibacillus spray drying protective agent according to claim 1, wherein the paenibacillus is one or more of paenibacillus polymyxa, paenibacillus azotemmae, paenibacillus macerans and paenibacillus lautus.
4. A spray drying method using the paenibacillus spray drying protective agent according to any one of claims 1 to 3, characterized by comprising the steps of:
(1) Liquid fermentation is carried out on paenibacillus until the sporulation period, and concentrated solution of paenibacillus is obtained after concentration;
(2) And (3) sequentially adding sodium carboxymethylcellulose, bean pulp powder, rice hull powder and molasses into the concentrated solution of the paenibacillus in the step (1), fully and uniformly stirring, and then carrying out spray drying.
5. The method of claim 4, wherein in step (1), the liquid fermentation medium is formulated as follows: 10g/L of glucose, 15g/L of sucrose, 15g/L of corn steep liquor, 5g/L of yeast powder, 20mg/L of ferrous sulfate, 5g/L of magnesium sulfate, 0.5g/L of potassium dihydrogen phosphate and 2.5g/L of disodium hydrogen phosphate.
6. The method according to claim 4, wherein in step (1), the specific conditions of the liquid fermentation are as follows:
the culture temperature is 28-33 ℃, the initial pH is 7.0, and the aeration ratio is l:0.6, the earlier stage of stirring speed is 120r/min, the later stage is 220r/min, and after fermentation is carried out for 18-24 hours, when dissolved oxygen rises to more than 70%, the fermentation is finished.
7. The method according to claim 4, wherein in the step (1), the concentration is carried out 2 to 5 times by using a ceramic membrane to obtain a concentrated liquid of Paenibacillus.
8. The method according to claim 4, wherein in the step (2), the spray drying is performed in a spray drying tower, the temperature of an air inlet is 140-150 ℃, the temperature of an air outlet is 50-85 ℃, and the pressure of a spray nozzle is 140-170kPa; the water content after spray drying was below 7%.
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