CN109504623B - Bacillus brevis thermophilus for producing amylase and application thereof - Google Patents
Bacillus brevis thermophilus for producing amylase and application thereof Download PDFInfo
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Abstract
The invention discloses a bacillus brevis thermophilus for producing amylase and application thereof, belonging to the technical field of environmental biology. The invention provides a thermophilic Brevibacillus brevis B205CCTCC No. M2018472 strain, which has good degradation capability of starch contained in kitchen waste, can be well symbiotic with other strains, has scientific and reasonable compounding proportion, uses the microbial inoculum to compost, has short time for the compost to reach high temperature, long duration time of the high temperature and high starch degradation rate, can effectively accelerate the composting reaction speed, can improve the harmless effect of the compost, and has wide application prospect.
Description
Technical Field
The invention relates to a bacillus brevis thermophilus for producing amylase and application thereof, belonging to the technical field of environmental biology.
Background
China has a large population, is a large country for generating garbage, and has more than two hundred million tons of urban garbage generation amount per year according to urban garbage statistical data issued by the Ministry of construction; still more than 1500 counties produce nearly 0.7 million tons of waste; in the aspect of garbage of villages and towns, accurate statistical data is not available for a while because the quantity of the villages and the towns is too dispersed. In general, the domestic garbage production amount in China is more than four hundred million tons. The most main constituent part of domestic garbage in China is kitchen garbage which is more than 60 percent, and some areas even reach 70 to 80 percent, while the main domestic garbage in Europe and America is paper, and the kitchen garbage only accounts for 25 percent. In addition, the water content of the kitchen waste is high, and the kitchen waste is easy to decay and deteriorate and pollutes the environment.
Aerobic composting refers to a process of stabilizing organic substances in organic wastes by means of the action of aerobic microorganisms under aerobic conditions, wherein certain heat is generated in the composting process, the high temperature duration is long, and harmless treatment can be realized without an external heating source; the capital investment is low, the management is easy, and the equipment is simple; the product is tasteless and odorless, has loose texture and low water content, is convenient to transport and process subsequent commercial fertilizers, and is one of important ways for realizing the recycling of the kitchen waste.
Rice and flour food residues are important components of the kitchen waste, and the screening of strains capable of efficiently degrading starch and the research and development of compound microbial agents containing different microbial strains and compound proportions have great significance for accelerating the composting process of the starch-containing kitchen waste.
Disclosure of Invention
The first purpose of the invention is to provide Bacillus brevis (Bacillus theroruber strain) B205 which is preserved in China center for type culture collection in 13.7.2018 with the preservation number of CCTCC No. M2018472 and the preservation address of China, Wuhan and Wuhan university. The fermentation culture conditions of the strain are as follows: the temperature is 30-60 ℃, the ventilation volume is 0.8-1.5vvm, the stirring speed is 200-.
The second purpose of the invention is to provide a microbial agent containing the Bacillus brevis B205.
In one embodiment of the invention, the microbial agent comprises B205B, B-tolerant L-lysine (Lysinibacillus boroniculolerans), L-lysine fusiform (Lysinibacillus fusiformis), B105B and T.viridans (Trichoderma atroviride).
In one embodiment of the invention, the Bacillus licheniformis (strain) B105 has been preserved in the China center for type culture Collection in 2018, 7 and 13 months, with the preservation number of CCTCC No: M2018470, and the preservation address of China, Wuhan university.
In one embodiment of the invention, the B-tolerant L.lysinibacillus, L.fusiformis and T.viridans are commercial strains including, but not limited to, Lysinibacillus boroniculatus ATCC BAA-1146, Lysinibacillus fusiformis ATCC 7055, Trichoderma atroviride IMI 206040.
In one embodiment of the invention, the ratio of the viable count of the Bacillus brevis B205, the B-resistant lysine bacillus, the L-lysine bacillus fusiformis, the B105 and the T.viridis in the composite microbial agent is 390-460:5-12:0.5-1.5:160-240: 0.6-1.6.
In one embodiment of the present invention, the total viable bacteria count of the complex microbial inoculant is 1.0 × 109-1.0×1010CFU/mL。
In one embodiment of the present invention, the total viable bacteria count of the complex microbial inoculant is 1.0 × 109-1.0×1010CFU/g。
The third purpose of the invention is to provide a method for preparing the compound microbial agent, which comprises the following steps: respectively inoculating bacillus brevis B205, boron-resistant lysine bacillus, lysine bacillus fusiformis, bacillus licheniformis B105 and trichoderma atroviride into a culture medium to obtain fermentation liquor of the bacillus brevis B205, the boron-resistant lysine bacillus, the lysine bacillus fusiformis, the bacillus licheniformis B105 and the trichoderma atroviride, mixing the obtained fermentation liquor according to the proportion of 390 plus 460:5-12:0.5-1.5:160 plus 240:0.6-1.6, and adding a carrier auxiliary material to obtain the liquid composite microbial agent containing the bacillus brevis B205.
In one embodiment of the present invention, the carrier auxiliary material includes one or more of lignite, zeolite, and stone powder.
In one embodiment of the present invention, the preparation method of the complex microbial agent comprises: mixing the bacillus brevis B205, the boron-resistant lysine bacillus, the lysine bacillus fusiformis, the bacillus licheniformis B105 and the trichoderma atroviride bacterial liquid according to a certain proportion, adding one or more of crushed straws, carbonized straws and bran, and drying by hot air to obtain the solid compound microbial agent containing the bacillus brevis B205.
In the present inventionIn one embodiment of the invention, the bacterial liquid is prepared by respectively inoculating bacillus brevis B205, lysine bacillus boron-resistant, lysine bacillus fusiformis and bacillus licheniformis B105 into a nutrient broth culture medium after high-temperature sterilization, wherein the inoculation amount is 3-10% (v/v) of the volume of a fermentation culture medium, and standing and culturing for 20-30 h; the culture temperature of the B105 of the B-resistant lysine bacillus, the lysine bacillus fusiformis and the Bacillus licheniformis is 28-37 ℃; the culture temperature of the bacillus brevis B205 is 30-60 ℃; when the number of viable bacteria is greater than or equal to 1 × 108And (3) obtaining the fermentation liquor of the bacillus brevis thermophilus B205, the lysine bacillus boragenes, the lysine bacillus fusiformis and the bacillus licheniformis B105 when the concentration is CFU/mL.
In one embodiment of the invention, the nutrient broth is a nutrient broth after autoclaving.
In one embodiment of the invention, the ingredients of the nutrient broth medium comprise glucose 15.0g, yeast extract 5.0g, calcium chloride (with two crystal waters) 0.2g, water 1000 mL.
In one embodiment of the invention, the fermentation liquor for obtaining trichoderma atroviride is prepared by respectively inoculating trichoderma atroviride into a potato glucose culture medium subjected to high-temperature sterilization, wherein the inoculation amount is 3% -10% (v/v) of the volume of the fermentation culture medium, standing and culturing for 20-30h, and the culture temperature is 20-30 ℃; when the number of viable bacteria is greater than or equal to 1 × 108And at CFU/mL, obtaining the fermentation liquor of the trichoderma atroviride.
In one embodiment of the present invention, the potato dextrose culture medium is sterilized at high temperature.
In one embodiment of the invention, the ingredients of the potato dextrose medium comprise 6.0g of potato extract powder, 20.0g of glucose and 1000mL of water.
The fourth purpose of the invention is to provide the application of the compound microbial agent.
In one embodiment of the invention, the application comprises mixed composting of kitchen waste, sewage treatment excess sludge of food plants and straws.
In one embodiment of the invention, the application comprises the following steps:
step (1): and (3) mixing the compound microbial agent and the compost raw materials according to the weight ratio of 0.1-0.5: mixing at a ratio of 100;
step (2): composting the mixture, measuring the temperature of the pile body, and turning the pile according to the temperature change.
In one embodiment of the present invention, the step (2) composts the mixture, measures the temperature of the compost at 8:00 am and 6:00 pm, turns the compost once a day when the temperature is higher than 55 ℃, starts to decrease as the composting time increases, stops turning when the temperature is lower than 40 ℃, and ends the composting.
The invention also claims the application of the bacillus brevis thermophilus in the fields of food, biology, chemical industry and environment.
The invention has the beneficial effects that:
(1) the invention discloses a bacillus brevis B205 capable of efficiently degrading starch in kitchen waste. The amylase activity of the strain is 3712.6U/mL after the strain is cultured for 26h, and the strain has better starch degradation capability.
(2) The compost composite microbial agent provided by the invention can be used for mixing and composting kitchen waste, food plant sewage treatment residual sludge and straw, and better realizes resource utilization of mixed waste.
(3) After the compost composite microbial agent provided by the invention is composted for 24 hours, the degradation rate of starch in compost raw materials can reach 94.9% -99.8%, and the degradation rate of starch in a control experiment is 68.7%.
(4) The microbial agent provided by the invention has obvious advantages in composting and can effectively shorten the temperature rise time of the compost, so that the time required by the compost to reach the high temperature of 55 ℃ is shortened from 60 hours in a control experiment to 12-28 hours.
(5) The microbial agent provided by the invention has obvious advantages in composting and can effectively improve the highest temperature of a compost, so that the highest temperature of the compost is improved to 61-68 ℃ from 57 ℃ of a control experiment.
(6) The microbial agent provided by the invention has obvious advantages in composting and can effectively prolong the high-temperature duration of a compost, the duration of the high temperature of the compost is 59-88h at 55 ℃, and the duration of a control experiment is 10 h.
(7) The microbial agent provided by the invention has obvious advantages in composting, can effectively improve the composting speed and the stabilization effect, and can reduce the carbon-nitrogen ratio of a mixed material to 11.8-16.9 from 22.7 of a control experiment after 24 hours of composting.
(8) The microbial agent provided by the invention has obvious advantages in composting and can improve the harmless effect of the composting, the number of faecal coliform and the death rate of roundworm eggs are respectively 52-78/g and 96.2% -97.9% when the composting is finished, and the number of faecal coliform and the death rate of roundworm eggs are respectively 126/g and 77.2% in comparison experiments.
Biological material preservation
Bacillus brevis (Bacillus theroruber strain) B205, which has been preserved in China center for type culture Collection in 2018, 7.13.M, with the preservation number of CCTCC No. M2018472, and the preservation address of China, Wuhan university.
Bacillus licheniformis (Bacillus licheniformis strain) B105 has been preserved in China center for type culture Collection in 2018, 7-13.7.M, with the preservation number of CCTCC No: M2018470, and the preservation address of China, Wuhan university.
Detailed Description
The detection method is referred to the quality standard of the biological organic fertilizer in China (NY 884-.
The detection method of the starch content comprises the following steps: the detection is carried out by an acid hydrolysis method, namely after fat and soluble saccharides are removed, the starch is hydrolyzed into monosaccharide with reducibility by acid, and then the monosaccharide is converted into the starch by measuring reducing sugar. The degradation rate of the starch is R (%):
R＝[(m1-m2)/m1/*100％
wherein m is1Is the mass (mg) of starch in the starting material, m2Is the mass (mg) of starch in the material after a certain period of composting.
The method for detecting the faecal coliform population comprises the following steps: the detection method (GB/T19524.1-2004) for the number of faecal coliform bacteria in the fertilizer is adopted for detection.
The detection method of the death rate of the roundworm eggs comprises the following steps: the detection method (GB/T19524.2-2004) for the number of faecal coliform bacteria in the fertilizer is adopted for detection.
In a specific embodiment, the B.borotolerant Lysinibacillus, L.fusiformis and T.viridans may be selected from the commercial strains Lysinibacillus boroniculatus ATCC BAA-1146, Lysinibacillus fusiformis ATCC 7055, Trichoderma atroviride IMI 206040, but are not limited thereto.
Example 1 screening of Bacillus brevis B205
(1) Kitchen waste, food plant sewage treatment excess sludge and corn stalks are mixed according to the weight ratio of 0.85: 0.85: 0.8, inoculating 5% (w/w) of rotten straw stalks in a certain area of Wuxi city of Jiangsu province, wherein the water content of the mixed material is 55%, the C/N ratio is 25, composting after uniform mixing, measuring the temperature of a compost at 8:00 am and 6:00 pm every day, turning the compost once every day when the temperature is higher than 55 ℃, beginning to reduce the temperature of the compost along with the extension of composting time, stopping turning the compost when the temperature is lower than 40 ℃, and sampling;
(2) weighing 10g of compost sample, putting the compost sample into a triangular flask filled with 90mL of sterilized water, and shaking the mixture for 1h by a 120r/min shaking table to obtain 10-1Diluting, and sequentially making into 10-4、10-5And 10-6Respectively sucking 100 mu L of the three compost sample diluents, uniformly coating the three compost sample diluents on nutrient agar culture media, Gao's first agar culture media and potato agar culture medium plates, respectively putting the three compost sample diluents into an incubator at 45 ℃ for inverted culture for 1-7 days, and then repeatedly scribing and separating until pure strains are obtained;
(3) and (2) directly dripping a proper amount of 0.02mol/L iodine solution on the colony surface of each pure strain, if the colony surface is a colorless transparent ring, the strain is an amylase producing strain and can degrade starch, the amylase producing capability of the strain can be judged according to the size of the transparent ring, the larger the transparent ring is, the stronger the starch degrading capability is, and finally, the strain which produces the amylase and has the strongest starch degrading capability is obtained by screening.
Example 2 identification of Bacillus brevis B205 thermophilus
After being cultured on a nutrient agar medium plate at 45 ℃ overnight, the culture medium plate forms white, oval, smooth-surfaced and irregular-edge colonies, and the colonies are observed to be gram-positive, rod-shaped, spore-producing and large-amount of exopolysaccharide by an optical microscope.
Taxonomic molecular identification is carried out by 16SrDNA sequencing, the sequencing result (1469bp) is subjected to Blast comparison in NCBI, and the similarity of the 16SrDNA base sequence and the Bacillus brevis (Brevibacillus thermophilus strain) is the highest and is 99%.
By combining the morphological characteristics, physiological and biochemical characteristics and hereditary characteristics, the amylase producing strain can be identified to be the Bacillus brevis thermophilus (Brevibacillus thermophilus strain), the strain is named as the Bacillus brevis thermophilus B205 and is preserved in the China center for type culture collection in 7-13 months in 2018, and the strain preservation number is CCTCC No. M2018472.
The amylase producing ability of the strain is verified. Inoculating the strain to a nutrient agar culture medium plate, culturing at 45 ℃ for 26h, then transferring to a nutrient broth culture medium, culturing at 45 ℃ for 26h, centrifuging the fermentation liquor, and determining the activity of amylase by using a DNS method, wherein the activity of the amylase is determined to be 3712.6U/mL.
Kitchen waste, food plant sewage treatment excess sludge and corn stalks are pressed according to the weight ratio of 0.75: 0.75: 1, mixing the materials according to a proportion, wherein the water content of the mixed materials is 55 percent, the C/N ratio is 30, uniformly mixing 0.3 percent (by mass) of fermentation liquor of Geobacillus stearothermophilus B205 with the materials, composting, measuring the temperature of two heaps at 8:00 am and 6:00 pm every day, turning the heaps once a day when the temperature is higher than 55 ℃, beginning to reduce the temperature of the heaps along with the extension of composting time, stopping turning the heaps when the temperature is lower than 40 ℃, and sampling, wherein the result shows that the starch content in the raw materials is reduced by 72.2 percent after 24 hours of composting.
Example 3 activation and culture of Bacillus brevis B205 species
The Bacillus brevis B205CCTCC No. M2018472 is inoculated on a nutrient agar culture medium plate and cultured for 22h at 45 ℃. The nutrient agar culture medium comprises glucose 15.0g, yeast extract 5.0g, and calcium chloride (with two crystal water) 0.2 g.
Inoculating the activated strain into a 250mL shaking flask filled with 100mL liquid culture medium, and performing shaking culture at 45 ℃ at 500r/min for 22 h. The shake flask culture medium consists of 15.0g of glucose, 5.0g of yeast extract, 0.2g of calcium chloride (with two crystal waters), 1000mL of water and pH 7.0.
Inoculating the shake flask culture solution into a 3L fermentation tank according to the volume ratio of 3-10%, stirring at the rotation speed of 200-600r/min, ventilation of 0.8-1.5vvm and dissolved oxygen of more than 30%, and performing fermentation culture at 30-60 ℃ for 20-30 h. The culture medium of the fermentation tank comprises 15.0g of glucose, 5.0g of yeast extract, 0.2g of calcium chloride (with two crystal waters), 2.0g of potassium dihydrogen phosphate and 5.0g of sodium chloride, and the pH value is 7.0.
Example 4 preparation of liquid Complex microbial Agents containing Bacillus brevis B205 thermophilus
The compost composite microbial agent is prepared by compounding fermentation liquor of Bacillus brevis B205CCTCC No. M2018472 and fermentation liquor of other strains according to different viable count ratios, and the preparation method comprises the following steps:
(1) the fermentation broth of the bacillus brevis B205, the boron-resistant lysine bacillus, the fusiform lysine bacillus and the bacillus licheniformis B105 is prepared by respectively inoculating the bacillus brevis B205, the boron-resistant lysine bacillus, the fusiform lysine bacillus and the bacillus licheniformis B105 into a nutrient broth culture medium after high-temperature sterilization, wherein the inoculation amount is 3-10% (v/v) of the volume of the fermentation culture medium, and culturing for 20-30 h; the culture temperature of the B105 of the B-resistant lysine bacillus, the lysine bacillus fusiformis and the Bacillus licheniformis is 28-37 ℃; the culture temperature of the bacillus brevis B205 is 30-60 ℃; when the number of viable bacteria is greater than or equal to 1 × 108Obtaining fermentation liquor of the bacillus brevis B205, the lysine bacillus boresii, the lysine bacillus fusiformis and the bacillus licheniformis B105 when the concentration is CFU/mL;
(2) the fermentation liquid of Trichoderma atroviride is prepared by inoculating Trichoderma atroviride into potato glucose agar culture medium sterilized at high temperature, wherein the inoculation amount is 3% -10% (v/v) of the volume of the fermentation culture medium, and culturing for 20-30h at 20-30 deg.C(ii) a When the number of viable bacteria is greater than or equal to 1 × 108And at CFU/mL, obtaining the fermentation liquor of the trichoderma atroviride.
(3) Mixing the obtained fermentation liquor of the Bacillus brevis B205, the boron-resistant lysine bacillus, the lysine bacillus fusiformis, the Bacillus licheniformis B105 and the trichoderma atroviride according to a certain proportion, adding one or more of commercially available carrier lignite auxiliary materials, zeolite and stone powder which account for 0.1-10% of the mass of the fermentation liquor into the fermentation liquor of the Bacillus brevis B205, the boron-resistant lysine bacillus, the lysine bacillus fusiformis, the Bacillus licheniformis B105 and the trichoderma atroviride according to the ratio of the viable count of 390-9-1.0×1010CFU/mL liquid compound microbial agent.
Example 5 preparation of solid Complex microbial preparation containing Bacillus brevis B205 thermophilus
Mixing the obtained fermentation liquor of the bacillus brevis B205, the boron-resistant lysine bacillus, the lysine bacillus fusiformis, the bacillus licheniformis B105 and the trichoderma atroviride according to a certain proportion, adding one or more of commercially available crushed straws, carbonized straws and bran into the mixture of the bacillus brevis B205, the boron-resistant lysine bacillus, the lysine bacillus fusiformis, the bacillus licheniformis B105 and the trichoderma atroviride according to the viable count ratio of 390-460:5-12:0.5-1.5:160-240:0.6-1.6, and drying by hot air to obtain the solid composite microbial agent containing the bacillus brevis B205.
Example 6 application of composite microbial Agents for composting
Kitchen waste, food plant sewage treatment excess sludge and corn stalks are mixed according to the weight ratio of 0.85: 0.85: 0.8, mixing the mixture according to the mass ratio of 0.3: 100, the addition amount of the microbial inoculum is 0.3 percent, composting is carried out after uniform mixing, the temperature of two heaps is measured at 8:00 in the morning and 6:00 in the afternoon every day, when the temperature is higher than 55 ℃, the heaps are turned once every day, the temperature of the heaps begins to drop along with the extension of composting time, when the temperature is lower than 40 ℃, the turning is stopped, sampling is carried out, and various quality indexes of the compost are measured. The result shows that the time required for the pile body to reach the highest temperature is 21h, the highest temperature is 65 ℃, and the duration time of the pile body higher than 55 ℃ is 66 h; after being piled for 24 hours, the degradation rate of the starch is 97.8 percent; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 14.8, and the number of faecal coliform bacteria and the death rate of roundworm eggs are 73/g and 96.7 percent respectively.
Example 7 use of compost composite microbial Agents
The kitchen waste, the food plant sewage treatment residual sludge and the corn straws are added according to the proportion of 1.1: 1.1: 1, mixing the mixture according to the mass ratio of 0.5: 100, the addition amount of the microbial inoculum is 0.5 percent, composting is carried out after uniform mixing, the temperature of two heaps is measured at 8:00 in the morning and 6:00 in the afternoon every day, when the temperature is higher than 55 ℃, the heaps are turned once every day, the temperature of the heaps begins to drop along with the extension of composting time, when the temperature is lower than 40 ℃, the turning is stopped, sampling is carried out, and various quality indexes of the compost are measured. The result shows that the temperature of the pile body rises rapidly, the time required for reaching the highest temperature is 23 hours, the highest temperature is 63 ℃, and the duration time of the temperature higher than 55 ℃ is 63 hours; after being piled for 24 hours, the degradation rate of the starch is 96.4 percent; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 13.9, and the number of faecal coliform bacteria and the death rate of roundworm eggs are 69/g and 97.2 percent respectively.
Example 8 use of compost composite microbial Agents
The kitchen waste, the food plant sewage treatment residual sludge and the corn straws are added according to the proportion of 1.1: 1.1: 1, mixing the mixture according to the mass ratio of 0.5: 100, the addition amount of the microbial inoculum is 0.5 percent, composting is carried out after uniform mixing, the temperature of two heaps is measured at 8:00 in the morning and 6:00 in the afternoon every day, when the temperature is higher than 55 ℃, the heaps are turned once every day, the temperature of the heaps begins to drop along with the extension of composting time, when the temperature is lower than 40 ℃, the turning is stopped, sampling is carried out, and various quality indexes of the compost are measured. The result shows that the temperature of the pile body rises rapidly, the time required for reaching the highest temperature is 12 hours, the highest temperature is 68 ℃, and the duration time of the temperature higher than 55 ℃ is 88 hours; after being piled for 24 hours, the degradation rate of the starch is 99.8 percent; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 11.9, and the number of faecal coliform bacteria and the death rate of roundworm eggs are 52/g and 97.9 percent respectively.
Example 9 use of compost composite microbial Agents
The kitchen waste, the food plant sewage treatment residual sludge and the corn straws are added according to the proportion of 1.1: 1.1: 1, mixing the mixture according to the mass ratio of 0.1: 100, the addition amount of the microbial inoculum is 0.1 percent, composting is carried out after uniform mixing, the temperature of two heaps is measured at 8:00 in the morning and 6:00 in the afternoon every day, when the temperature is higher than 55 ℃, the heaps are turned once every day, the temperature of the heaps begins to drop along with the extension of composting time, when the temperature is lower than 40 ℃, the turning is stopped, sampling is carried out, and various quality indexes of the compost are measured. The result shows that the temperature of the pile body rises rapidly, the time required for reaching the highest temperature is 28 hours, the highest temperature is 68 ℃, and the duration time of the temperature higher than 55 ℃ is 59 hours; after being piled for 24 hours, the degradation rate of the starch is 94.9 percent; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 18.1, and the number of faecal coliform bacteria and the death rate of roundworm eggs are 78/g and 96.2 percent respectively.
Comparative example 1
Kitchen waste, food plant sewage treatment excess sludge and corn stalks are mixed according to the weight ratio of 0.85: 0.85: 0.8, wherein the water content of the mixed material is 55 percent, the C/N ratio is 30, and the liquid compound microbial agent without the bacillus brevis B205 is mixed with the mixed material according to the mass ratio of 0.3: 100, the addition amount of the microbial inoculum is 0.3 percent, wherein,
the ratio of the number of viable bacteria of the boron-resistant lysine bacillus, the fusiform lysine bacillus, the bacillus licheniformis B105 and the trichoderma aureoviride is 8:1:195:1.1, the materials are uniformly mixed and piled, the temperature of two piles is measured at 8:00 am and 6:00 pm every day, when the temperature is higher than 55 ℃, the piles are turned once every day, the temperature of the piles begins to decrease along with the prolonging of the piling time,
when the temperature is lower than 40 ℃, stopping turning the compost, sampling, and measuring various quality indexes of the compost. The result shows that the time required for the pile body to reach the highest temperature is 60 hours, the highest temperature is 57 ℃, and the duration time of the pile body higher than 55 ℃ is 10 hours; after the materials are stacked for 24 hours,
the starch degradation rate is 68.7%; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 22.7, and the number of faecal coliform bacteria and the death rate of roundworm eggs are 126/g and 77.2% respectively.
Comparative example 2
The specific implementation manner is the same as that of example 6, except that the ratio of viable count of Bacillus brevis B205, B-lysine resistant Bacillus, B-lysine fusiformis, B105 and Trichoderma atroviride is different, as shown in Table 1. The result shows that the time required for the pile body to reach the highest temperature is prolonged to 52-62 h; after being piled for 24 hours, the starch degradation rate is only 52.8-71.1%; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 19.4-23.2, and the number of faecal coliform and the death rate of ascaris ova are 490-870/g and 41.
TABLE 1
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. A Bacillus brevis (Bacillus theroruber) B205 is preserved in China center for type culture collection (CCTCC No: M2018472) at 13.7.2018 with the preservation address of university in Wuhan, Wuhan.
2. A microbial agent comprising the Bacillus brevis B205 as set forth in claim 1.
3. The microbial agent according to claim 2, comprising Bacillus brevis B205, B-tolerant lysine Bacillus (Lysinibacillus boroniculolerans), L-lysine Bacillus fusiformis (Lysinibacillus fusiformis), B105 and Trichoderma atroviride (Trichoderma atroviride); the Bacillus licheniformis (Bacillus licheniformis) B105 has been preserved in China center for type culture Collection in 2018, 7 and 13 months, with the preservation number of CCTCC No: M2018470, and the preservation address of China, Wuhan university.
4. The microbial inoculant according to claim 3, wherein the ratio of the viable count of B205, B105 and B105 is 390-460:5-12:0.5-1.5:160-240: 0.6-1.6; the total viable count of the microbial inoculum is 1.0 × 109-1.0×1010CFU/mL or CFU/g.
5. The microbial agent according to claim 4, which is prepared by the following steps: respectively inoculating bacillus brevis B205, boron-resistant lysine bacillus, lysine bacillus fusiformis, bacillus licheniformis B105 and trichoderma atroviride into a culture medium to obtain fermentation liquor of the bacillus brevis B205, the boron-resistant lysine bacillus, the lysine bacillus fusiformis, the bacillus licheniformis B105 and the trichoderma atroviride, mixing the obtained fermentation liquor in proportion, and adding a carrier auxiliary material to obtain a liquid composite microbial agent containing the bacillus brevis B205; the carrier auxiliary material comprises one or more of lignite, zeolite and stone powder.
6. The microbial agent according to claim 4, which is prepared by the following steps: mixing the bacillus brevis B205, the boron-resistant lysine bacillus, the lysine bacillus fusiformis, the bacillus licheniformis B105 and the trichoderma atroviride bacterial liquid in proportion, adding one or more of crushed straws, carbonized straws and bran, and drying by hot air to obtain the solid compound microbial agent containing the bacillus brevis B205.
7. The use of Bacillus brevis B205 as claimed in claim 1 for degrading starch in the fields of food, chemical industry and environment.
8. A method for promoting mixed composting of kitchen waste, excess sludge of sewage treatment of food factories and straws, which is characterized in that the composting is carried out by using a microbial agent containing the Bacillus brevis B205 as claimed in claim 1.
9. The method of claim 8, comprising the steps of:
step (1): and (3) mixing the microbial agent and the compost raw materials according to the ratio of 0.1-0.5: mixing at a ratio of 100;
step (2): composting the mixture, measuring the temperature of the pile body, and turning the pile according to the temperature change.
10. The method according to claim 9, wherein the step (2) comprises composting the mixture, measuring the temperature of the compost 1-2 times a day in the morning and afternoon, turning the compost once a day when the temperature is higher than 55 ℃, wherein the temperature of the compost starts to decrease with the increase of the composting time, and stopping turning the compost when the temperature is lower than 40 ℃ to finish the composting.
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