Composite microbial deodorant and preparation method thereof

Abstract

The invention belongs to the technical field of microbial deodorants, and particularly relates to a composite microbial deodorant and a preparation method thereof. The preparation method comprises the following steps: preparing a bacillus subtilis seed solution, preparing a streptococcus thermophilus seed solution, preparing a streptomyces griseus seed solution, performing primary fermentation culture, modifying sepiolite and performing secondary fermentation culture. The sepiolite modification step comprises the following steps: putting sepiolite into a sodium hydroxide solution, raising the temperature to 60-64 DEG C, carrying out heat preservation stirring for 2.8-3.2 hours, after the heat preservation stirring is finished, putting the sepiolite into an ethanol solution, adding sodium dodecyl benzene sulfonate and polyethylene glycol 1000, carrying out heat preservation stirring at 38-42 DEG C for 26-33 minutes, after the heat preservation stirring is finished, adding kH550, raising the temperature to 58-62 DEG C, and carrying out stirring reaction for 1.8-2.2 hours; adding glutamic acid and chitosan oligosaccharide, and continuously stirring to react for 1.8-2.2 hours, so as to obtain modified sepiolite; the deodorant prepared by the invention is good in deodorization performance, good in long-term effect and excellent in stability.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial deodorants, and in particular relates to a composite microbial deodorant and a preparation method thereof. Background Art

[0002] With the rapid development of social economy, odor, as one of the environmental hazards, directly affects people's quality of life and endangers human health; The sources of foul odors are mainly divided into domestic sources and industrial sources; among them, domestic sources mainly refer to the foul odors produced in daily life, such as the foul gases produced in household toilets, public toilets, ditches, etc., which bring troubles to the lives of surrounding residents; industrial sources refer to the foul odors produced in industrial production, including farms, pharmaceutical factories, chemical plants and food processing plants.

[0003] Foul odors mainly include the following categories: one is sulfur-containing compounds, such as hydrogen sulfide, sulfur dioxide, thiols and thioethers; two is nitrogen-containing compounds, such as ammonia, indole and amines; three is halogens and their derivatives, such as halogenated hydrocarbons and chlorine; four is oxygen-containing organic compounds, such as organic acids, phenols, aldehydes, ketones, etc.; five is hydrocarbons, such as alkanes, alkenes, alkynes and aromatic hydrocarbons.

[0004] The removal of odor mainly includes physical, chemical and biological methods. The physical method includes methods such as absorbents to absorb odorous gases and mechanical ventilation to reduce the concentration of odorous gases, but the deodorization effect is not obvious. The chemical method mainly uses oxidants such as potassium permanganate, hydrogen peroxide and sulfur dioxide, and reducing agents such as sodium borohydride and sodium sulfite to remove odorous substances, but it has the problems of poor persistence and serious environmental pollution. The biological method utilizes the physiological metabolic activities of microorganisms to degrade malodorous substances and convert them into odorless substances. It has the advantages of high efficiency, safety, non-toxicity, and will not cause secondary pollution. It has become the main method for odor prevention and control research at home and abroad. However, it has defects such as single effect, poor adaptability and poor long-term effectiveness.

[0005] CN115353212A discloses a composite microbial deodorant containing medical stone powder and a preparation method thereof, and specifically discloses that the composite microbial deodorant includes the following components: a composite bacterial strain, a bacterial strain growth promoter, medical stone powder and water; the patent realizes the function of microbial deodorization through a composite bacterial strain composed of a combination of multiple professional deodorizing functional bacteria, and provides sufficient influence on the composite bacterial strain through the bacterial strain growth promoter, so that the deodorization of the composite bacterial strain is fast and the deodorization effect is good; at the same time, by adding medical stone powder with water purification function, based on the powerful adsorption performance and excellent dissolution performance of medical stone, on the one hand, the deodorization performance is improved, and on the other hand, mineral nutrition is provided for the composite bacterial strain, thereby further improving the deodorization efficiency and deodorization function of the composite microbial deodorant.

[0006] However, the composite microbial deodorant obtained by this method has good deodorizing performance only in a short period of time, has poor long-term effectiveness, and is greatly affected by ultraviolet radiation. In an environment with high ultraviolet intensity, the deodorizing performance is significantly reduced. Summary of the invention

[0007] In order to solve the technical problems existing in the prior art, the present invention provides a composite microbial deodorant and a preparation method thereof, which has good deodorizing performance, good long-term effect and excellent resistance to ultraviolet radiation.

[0008] In view of the above technical problems, the present invention adopts the following technical solutions: A method for preparing a composite microbial deodorant includes the steps of preparing a Bacillus subtilis seed solution, preparing a thermophilic Streptococcus seed solution, preparing a Streptomyces griseus seed solution, primary fermentation culture, sepiolite modification, and secondary fermentation culture, which are specifically as follows: 1. Preparation of Bacillus subtilis Seed Solution Placing Bacillus subtilis in a shake flask culture medium for shake flask culture, controlling the culture temperature to be 36-38° C., the culture time to be 16-20 hours, and the rotation speed to be 175-185 rpm, obtaining a shake flask culture solution after the culture is completed, and then inoculating the shake flask culture solution into a seed culture medium at an inoculum amount of 1.8-2.2% for seed culture, the culture temperature to be 36-38° C., and the culture time to be 18-22 hours, to obtain a Bacillus subtilis seed solution; The Bacillus subtilis is purchased from the market; The shake flask culture medium is composed of the following ingredients: 4.8-5.2 g / L beef extract, 4.4-4.8 g / L peptone, 3.4-3.6 g / L sodium chloride, 3.2-3.5 g / L glucose, and the pH value is controlled to be 7.0-7.2; The seed culture medium is composed of the following ingredients: 11.8-12.3 g / L of glucose, 3.3-3.5 g / L of peptone, 0.38-0.42 g / L of magnesium sulfate heptahydrate, 2.6-2.8 g / L of ammonium sulfate, and 4.8-5.2 g / L of sodium chloride.

[0009] 2. Preparation of Streptococcus thermophilus seed solution Placing the thermophilic Streptococcus in a shake flask culture medium for shake flask culture, controlling the culture temperature to be 30-33° C., the culture time to be 15-17 hours, and the rotation speed to be 210-230 rpm, obtaining a shake flask culture solution after the culture is completed, and then inoculating the shake flask culture solution into a seed culture medium at an inoculum amount of 2.4-2.6% for seed culture, the culture temperature to be 35-37° C., and the culture time to be 20-23 hours, to obtain a thermophilic Streptococcus seed solution; The thermophilic streptococcus is purchased from the market; The shake flask culture medium is composed of the following ingredients: 9.8-10.2 g / L peptone, 4.8-5.2 g / L sodium chloride, 11.8-12.3 g / L agar, and 3.4-3.6 g / L beef extract; The seed culture medium is composed of the following components: 8.4-8.7 g / L glucose, 9.8-10.3 g / L peptone, 9.7-10.2 g / L beef extract, 0.08-0.12 g / L manganese sulfate, 0.27-0.29 g / L magnesium sulfate heptahydrate, 1.9-2.2 g / L diammonium citrate, 4.2-4.4 g / L sodium acetate, and 18-22 mg / L cobalt chloride hexahydrate.

[0010] 3. Preparation of Streptomyces griseus Seed Solution The Streptomyces griseus is placed in a shake flask culture medium for shake flask culture, the culture temperature is controlled to be 30-32° C., the culture time is 13.8-14.2 hours, and the rotation speed is 165-175 rpm. After the culture is completed, the shake flask culture solution is inoculated into a seed culture medium at an inoculum amount of 1.8-2.2% for seed culture, the culture temperature is 28-32° C., and the culture time is 46-50 hours to obtain a Streptomyces griseus seed solution; The shake flask culture medium is composed of the following ingredients: 9.8-10.2 g / L peptone, 4.8-5.2 g / L yeast powder, and 7.4-7.6 g / L sodium chloride; The seed culture medium is composed of the following ingredients: 19.5-20.6 g / L of glucose, 4.7-5.3 g / L of yeast powder, 5.0-5.4 g / L of peptone, 6.2-6.5 g / L of potassium dihydrogen phosphate, 9.8-10.2 mg / L of ammonium molybdate, 0.24-0.26 g / L of proline, and 14.0-16.0 g / L of polyethylene glycol 600.

[0011] 4. Primary fermentation culture 8-12% volume of Bacillus subtilis seed liquid, 8-12% volume of thermophilic Streptococcus seed liquid, and 8-12% volume of Streptomyces griseus seed liquid are respectively taken and inoculated into a fermentation medium for fermentation culture at a fermentation temperature of 24-26° C. for a fermentation time of 4.5-5.5 days. After the fermentation is completed, a primary fermentation culture liquid is obtained; The fermentation medium is composed of the following ingredients: 16.5-17.0 g / L of glucose, 12.2-12.7 g / L of corn flour, 6.0-6.2 g / L of soybean flour, 0.53-0.58 g / L of potassium dihydrogen phosphate, 0.24-0.28 g / L of manganese sulfate, 3.6-3.8 g / L of calcium chloride, 2.3-2.7 g / L of sodium nitrate, 0.7-0.9 g / L of glycine, 3.2-3.6 g / L of L-glutamine, and 1.3-1.5 g / L of Tween 80.

[0012] 5. Modification of sepiolite The sepiolite is placed in a sodium hydroxide solution, the temperature is raised to 60-64°C, and the mixture is stirred for 2.8-3.2 hours. After the stirring is completed, the mixture is placed in an ethanol solution, sodium dodecylbenzene sulfonate and polyethylene glycol 1000 are added, and the mixture is stirred for 26-33 minutes at 38-42°C. After the stirring is completed, kH550 is added, the temperature is raised to 58-62°C, and the mixture is stirred for 1.8-2.2 hours. Then, glutamic acid and chitosan oligosaccharide are added, and the mixture is stirred for 1.8-2.2 hours to obtain a modified sepiolite. The particle size of the sepiolite is 110-130nm; The mass ratio of the sepiolite, sodium hydroxide solution, ethanol solution, sodium dodecylbenzene sulfonate, polyethylene glycol 1000, kH550, glutamic acid and chitosan oligosaccharide is 10.0-10.5:68-72:98-104:0.8-1.2:0.6-0.8:1.0-1.3:0.4-0.6:0.5-0.8; The mass concentration of the sodium hydroxide solution is 40-43%; The mass concentration of the ethanol solution is 48-52%.

[0013] 6. Secondary fermentation culture Adding modified sepiolite to a primary fermentation culture solution, performing secondary fermentation culture, the fermentation time is 11.5-12.5 hours, the fermentation temperature is 24-26° C., after the fermentation culture is completed, adding modified bamboo fiber, and then performing ultrasonic treatment, the ultrasonic time is 14-16 minutes, the ultrasonic power is 245-255W, the ultrasonic frequency is 34-37kHz, after the ultrasonic treatment is completed, stirring the reaction at 34-36° C. for 2.8-3.2 hours, and drying at 36-38° C. to obtain a composite microbial deodorant; The fermentation medium of the secondary fermentation culture is completely the same as that of the primary fermentation culture medium; The mass ratio of the primary fermentation culture liquid, the modified sepiolite, and the modified bamboo fiber is 98-102:11.7-12.3:3.5-3.7; The preparation method of the modified bamboo fiber is as follows: 4.8-5.2g of bamboo fiber is placed in 98-102g of deionized water, and then 9.7-10.3g of sodium periodate is added for stirring, and after stirring evenly, the mixture is reacted in a dark environment at room temperature for 7.5-8.5h, and then 3.6-4.0g of ethylene glycol is added to terminate the reaction, and the mixture is filtered, washed, and dried to obtain the modified bamboo fiber; The mass ratio of the bamboo fiber, deionized water, sodium periodate and ethylene glycol is 4.8-5.2:98-102:9.7-10.3:3.6-4.0.

[0014] A composite microbial deodorant is prepared by the above-mentioned preparation method.

[0015] Compared with the prior art, the present invention has achieved the following beneficial effects: 1. The present invention adopts specific strains for compounding, and then performs shake flask culture and seed culture respectively, and then performs mixed fermentation culture. In the mixed fermentation culture process, specific glycine, L-glutamine, Tween 80 and other components are added, wherein glycine and L-glutamine can provide more nutrition and energy, promote the growth and metabolism of strains, and regulate the balance of the internal and external environment of cells. Tween 80, as a surfactant, can change the permeability of cell membranes, thereby promoting cell reproduction and growth. In the secondary fermentation culture process, the same culture medium components as those in the primary fermentation culture process are used, and modified sepiolite is added, which has good fixation. It can be fermented and cultured together with microorganisms to improve the fixation performance of microorganisms. It will not cause damage, and ensure the activity of microorganisms. In addition, the modified sepiolite has more hydroxyl and amino groups, and its amino group can be cross-linked with the aldehyde group of bamboo fiber to form a stable network structure, thereby improving the stability of the deodorant, enhancing the persistence, and achieving effective removal of odor. 2. The deodorant prepared by the method of the present invention, in stinky water with an ammonia concentration of 250 mg / L and a hydrogen sulfide concentration of 190 mg / L, after standing for 6.0 hours, the removal rate of ammonia is 84.3-85.5%, and the removal rate of hydrogen sulfide is 86.1-87.3%; after standing for 12.0 hours, the removal rate of ammonia is 89.5-90.4%, and the removal rate of hydrogen sulfide is 92.1-92.7%; after standing for 48.0 hours, the removal rate of ammonia is 89.9-91.1%, and the removal rate of hydrogen sulfide is 92.5-93.2%; after standing for 168.0 hours, the removal rate of ammonia is 85.4-87.4%, and the removal rate of hydrogen sulfide is 86.8-88.3%; 3. The deodorant prepared by the method of the present invention has a UV intensity of 400W / cm 2 After being irradiated under ultraviolet light for 24.0 hours, the samples were placed in smelly water with an ammonia concentration of 250 mg / L and a hydrogen sulfide concentration of 190 mg / L for 12.0 hours. The removal rates of ammonia and hydrogen sulfide were 85.5-88.3% and 87.6-89.5%. DETAILED DESCRIPTION

[0016] In order to more clearly understand the technical features, purposes and effects of the present invention, specific embodiments of the present invention are now described.

[0017] Example 1 1. Preparation of Bacillus subtilis Seed Solution The Bacillus subtilis was placed in a shake flask culture medium for shake flask culture, the culture temperature was controlled to be 37° C., the culture time was 18 hours, the rotation speed was 180 rpm, and after the culture was completed, a shake flask culture solution was obtained, and then the shake flask culture solution was inoculated into a seed culture medium at an inoculum amount of 2.0% for seed culture, the culture temperature was 37° C., and the culture time was 20 hours, to obtain a Bacillus subtilis seed solution; The Bacillus subtilis is purchased from the market; The shake flask culture medium is composed of the following ingredients: 5.0 g / L beef extract, 4.6 g / L peptone, 3.5 g / L sodium chloride, 3.4 g / L glucose, and the pH value is controlled to be 7.1; The seed culture medium is composed of the following ingredients: 12.0 g / L glucose, 3.4 g / L peptone, 0.40 g / L magnesium sulfate heptahydrate, 2.7 g / L ammonium sulfate, and 5.0 g / L sodium chloride.

[0018] 2. Preparation of Streptococcus thermophilus seed solution The thermophilic Streptococcus was placed in a shake flask culture medium for shake flask culture, the culture temperature was controlled to be 32° C., the culture time was 16 h, the rotation speed was 220 rpm, and after the culture was completed, a shake flask culture solution was obtained, and then the shake flask culture solution was inoculated into a seed culture medium at an inoculum amount of 2.5% for seed culture, the culture temperature was 36° C., and the culture time was 22 h, to obtain a thermophilic Streptococcus seed solution; The thermophilic streptococcus is purchased from the market; The shake flask culture medium is composed of the following ingredients: 10.0 g / L peptone, 5.0 g / L sodium chloride, 12.0 g / L agar, and 3.5 g / L beef extract; The seed culture medium is composed of the following ingredients: 8.5 g / L glucose, 10.0 g / L peptone, 10.0 g / L beef extract, 0.10 g / L manganese sulfate, 0.28 g / L magnesium sulfate heptahydrate, 2.0 g / L diammonium citrate, 4.3 g / L sodium acetate, and 20 mg / L cobalt chloride hexahydrate.

[0019] 3. Preparation of Streptomyces griseus Seed Solution The Streptomyces griseus was placed in a shake flask medium for shake flask culture, the culture temperature was controlled to be 31° C., the culture time was 14.0 h, and the rotation speed was 170 rpm. After the culture was completed, the shake flask culture solution was inoculated into a seed culture medium at an inoculum amount of 2.0% for seed culture, the culture temperature was 30° C., and the culture time was 48 h, to obtain a Streptomyces griseus seed solution; The shake flask culture medium is composed of the following ingredients: 10.0 g / L peptone, 5.0 g / L yeast powder, and 7.5 g / L sodium chloride; The seed culture medium consists of the following ingredients: 20.0 g / L glucose, 5.0 g / L yeast powder, 5.2 g / L peptone, 6.3 g / L potassium dihydrogen phosphate, 10.0 mg / L ammonium molybdate, 0.25 g / L proline, and 15.0 g / L polyethylene glycol 600.

[0020] 4. Primary fermentation culture 10% volume of Bacillus subtilis seed liquid, 10% volume of thermophilic Streptococcus seed liquid, and 10% volume of Streptomyces griseus seed liquid were respectively taken and inoculated into the fermentation medium for fermentation culture at a fermentation temperature of 25° C. for 5 days. After the fermentation was completed, a primary fermentation culture liquid was obtained; The fermentation medium is composed of the following ingredients: 16.7 g / L glucose, 12.5 g / L corn flour, 6.1 g / L soybean flour, 0.55 g / L potassium dihydrogen phosphate, 0.26 g / L manganese sulfate, 3.7 g / L calcium chloride, 2.5 g / L sodium nitrate, 0.8 g / L glycine, 3.4 g / L L-glutamine, and 1.4 g / L Tween 80.

[0021] 5. Modification of sepiolite 10.2 g of sepiolite was placed in 70 g of 42 wt% sodium hydroxide solution, the temperature was raised to 62°C, and the mixture was stirred for 3.0 h. After the stirring was completed, the mixture was placed in 100 g of 50 wt% ethanol solution, 1.0 g of sodium dodecylbenzene sulfonate and 0.7 g of polyethylene glycol 1000 were added, and the mixture was stirred for 30 min at 40°C. After the stirring was completed, 1.2 g of kH550 was added, the temperature was raised to 60°C, and the mixture was stirred for 2.0 h. Then, 0.5 g of glutamic acid and 0.7 g of chitosan oligosaccharide were added, and the mixture was stirred for 2.0 h to obtain modified sepiolite. The particle size of the sepiolite is 120 nm.

[0022] 6. Secondary fermentation culture 12.0 g of modified sepiolite was added to 100 g of the primary fermentation culture solution, and a secondary fermentation culture was carried out. The fermentation time was 12.0 h, and the fermentation temperature was 25° C. After the fermentation culture was completed, 3.6 g of modified bamboo fiber was added, and then ultrasonic treatment was carried out. The ultrasonic time was 15 min, the ultrasonic power was 250 W, and the ultrasonic frequency was 36 kHz. After the ultrasonic treatment was completed, the reaction was stirred at 35° C. for 3.0 h, and the mixture was dried at 37° C. to obtain a composite microbial deodorant; The fermentation medium of the secondary fermentation culture is completely the same as that of the primary fermentation culture medium; The preparation method of the modified bamboo fiber is as follows: 5.0g of bamboo fiber is placed in 100g of deionized water, and then 10.0g of sodium periodate is added for stirring. After stirring evenly, the mixture is reacted in a dark environment at room temperature for 8.0h, and then 3.8g of ethylene glycol is added to terminate the reaction. The mixture is filtered, washed, and dried to obtain the modified bamboo fiber.

[0023] Example 2 1. Preparation of Bacillus subtilis Seed Solution Bacillus subtilis was placed in a shake flask culture medium for shake flask culture, the culture temperature was controlled to be 36° C., the culture time was 16 h, the rotation speed was 175 rpm, and after the culture was completed, a shake flask culture solution was obtained, and then the shake flask culture solution was inoculated into a seed culture medium at an inoculum amount of 1.8% for seed culture, the culture temperature was 36° C., and the culture time was 18 h, to obtain a Bacillus subtilis seed solution; The Bacillus subtilis is purchased from the market; The shake flask culture medium is composed of the following ingredients: 4.8 g / L beef extract, 4.4 g / L peptone, 3.4 g / L sodium chloride, and 3.2 g / L glucose, and the pH value is controlled to be 7.0; The seed culture medium is composed of the following ingredients: 11.8 g / L glucose, 3.3 g / L peptone, 0.38 g / L magnesium sulfate heptahydrate, 2.6 g / L ammonium sulfate, and 4.8 g / L sodium chloride.

[0024] 2. Preparation of Streptococcus thermophilus seed solution The thermophilic Streptococcus was placed in a shake flask culture medium for shake flask culture, the culture temperature was controlled to be 30° C., the culture time was 15 h, the rotation speed was 210 rpm, and after the culture was completed, a shake flask culture solution was obtained, and then the shake flask culture solution was inoculated into a seed culture medium at an inoculum amount of 2.4% for seed culture, the culture temperature was 35° C., and the culture time was 20 h, to obtain a thermophilic Streptococcus seed solution; The thermophilic streptococcus is purchased from the market; The shake flask culture medium is composed of the following ingredients: 9.8 g / L peptone, 4.8 g / L sodium chloride, 11.8 g / L agar, and 3.4 g / L beef extract; The seed culture medium is composed of the following ingredients: 8.4 g / L glucose, 9.8 g / L peptone, 9.7 g / L beef extract, 0.08 g / L manganese sulfate, 0.27 g / L magnesium sulfate heptahydrate, 1.9 g / L diammonium citrate, 4.2 g / L sodium acetate, and 18 mg / L cobalt chloride hexahydrate.

[0025] 3. Preparation of Streptomyces griseus Seed Solution The Streptomyces griseus was placed in a shake flask medium for shake flask culture, the culture temperature was controlled to be 30°C, the culture time was 13.8 hours, and the rotation speed was 165 rpm. After the culture was completed, the shake flask culture solution was inoculated into a seed culture medium at an inoculum amount of 1.8% for seed culture, the culture temperature was 28°C, and the culture time was 46 hours, to obtain a Streptomyces griseus seed solution; The shake flask culture medium is composed of the following ingredients: 9.8 g / L peptone, 4.8 g / L yeast powder, and 7.4 g / L sodium chloride; The seed culture medium is composed of the following ingredients: 19.5 g / L glucose, 4.7 g / L yeast powder, 5.0 g / L peptone, 6.2 g / L potassium dihydrogen phosphate, 9.8 mg / L ammonium molybdate, 0.24 g / L proline, and 14.0 g / L polyethylene glycol 600.

[0026] 4. Primary fermentation culture 8% volume of Bacillus subtilis seed liquid, 8% volume of thermophilic Streptococcus seed liquid, and 8% volume of Streptomyces griseus seed liquid were respectively inoculated into the fermentation medium for fermentation culture at a fermentation temperature of 24° C. for a fermentation time of 4.5 days. After the fermentation was completed, a primary fermentation culture liquid was obtained; The fermentation medium is composed of the following ingredients: 16.5 g / L glucose, 12.2 g / L corn flour, 6.0 g / L soybean flour, 0.53 g / L potassium dihydrogen phosphate, 0.24 g / L manganese sulfate, 3.6 g / L calcium chloride, 2.3 g / L sodium nitrate, 0.7 g / L glycine, 3.2 g / L L-glutamine, and 1.3 g / L Tween 80.

[0027] 5. Modification of sepiolite 10.0 g of sepiolite was placed in 68 g of 40 wt% sodium hydroxide solution, the temperature was raised to 60°C, and the mixture was stirred for 2.8 h. After the stirring was completed, the mixture was placed in 98 g of 48 wt% ethanol solution, 0.8 g of sodium dodecylbenzene sulfonate and 0.6 g of polyethylene glycol 1000 were added, and the mixture was stirred for 26 min at 38°C. After the stirring was completed, 1.0 g of kH550 was added, the temperature was raised to 58°C, and the mixture was stirred for 1.8 h. Then, 0.4 g of glutamic acid and 0.5 g of chitosan oligosaccharide were added, and the mixture was stirred for 1.80 h to obtain modified sepiolite. The particle size of the sepiolite is 110 nm.

[0028] 6. Secondary fermentation culture 11.7 g of modified sepiolite was added to 98 g of the primary fermentation culture solution, and a secondary fermentation culture was carried out. The fermentation time was 11.5 h, and the fermentation temperature was 24° C. After the fermentation culture was completed, 3.5 g of modified bamboo fiber was added, and then ultrasonic treatment was carried out. The ultrasonic time was 14 min, the ultrasonic power was 245 W, and the ultrasonic frequency was 34 kHz. After the ultrasonic treatment was completed, the reaction was stirred at 34° C. for 2.8 h, and dried at 36° C. to obtain a composite microbial deodorant; The fermentation medium of the secondary fermentation culture is completely the same as that of the primary fermentation culture medium; The preparation method of the modified bamboo fiber is as follows: 4.8g of bamboo fiber is placed in 98g of deionized water, and then 9.7g of sodium periodate is added for stirring. After stirring evenly, the mixture is reacted in a dark environment at room temperature for 7.5h, and then 3.6g of ethylene glycol is added to terminate the reaction. The modified bamboo fiber is obtained by filtering, washing, and drying.

[0029] Example 3 1. Preparation of Bacillus subtilis Seed Solution Bacillus subtilis was placed in a shake flask culture medium for shake flask culture, the culture temperature was controlled to be 38° C., the culture time was 20 h, the rotation speed was 185 rpm, and after the culture was completed, a shake flask culture solution was obtained, and then the shake flask culture solution was inoculated into a seed culture medium at an inoculum amount of 2.2% for seed culture, the culture temperature was 38° C., and the culture time was 22 h, to obtain a Bacillus subtilis seed solution; The Bacillus subtilis is purchased from the market; The shake flask culture medium is composed of the following ingredients: 5.2 g / L beef extract, 4.8 g / L peptone, 3.6 g / L sodium chloride, 3.5 g / L glucose, and the pH value is controlled to be 7.2; The seed culture medium is composed of the following ingredients: 12.3 g / L glucose, 3.5 g / L peptone, 0.42 g / L magnesium sulfate heptahydrate, 2.8 g / L ammonium sulfate, and 5.2 g / L sodium chloride.

[0030] 2. Preparation of Streptococcus thermophilus seed solution The thermophilic Streptococcus was placed in a shake flask culture medium for shake flask culture, the culture temperature was controlled to be 33° C., the culture time was 17 h, the rotation speed was 230 rpm, and after the culture was completed, a shake flask culture solution was obtained, and then the shake flask culture solution was inoculated into a seed culture medium at an inoculum amount of 26% for seed culture, the culture temperature was 37° C., and the culture time was 23 h, to obtain a thermophilic Streptococcus seed solution; The thermophilic streptococcus is purchased from the market; The shake flask culture medium is composed of the following ingredients: 10.2 g / L peptone, 5.2 g / L sodium chloride, 12.3 g / L agar, and 3.6 g / L beef extract; The seed culture medium is composed of the following ingredients: 8.7 g / L glucose, 10.3 g / L peptone, 10.2 g / L beef extract, 0.12 g / L manganese sulfate, 0.29 g / L magnesium sulfate heptahydrate, 2.2 g / L diammonium citrate, 4.4 g / L sodium acetate, and 22 mg / L cobalt chloride hexahydrate.

[0031] 3. Preparation of Streptomyces griseus Seed Solution The Streptomyces griseus was placed in a shake flask culture medium for shake flask culture, the culture temperature was controlled to be 32°C, the culture time was 14.2 hours, and the rotation speed was 175 rpm. After the culture was completed, the shake flask culture solution was inoculated into a seed culture medium at an inoculum amount of 2.2% for seed culture, the culture temperature was 32°C, and the culture time was 50 hours, to obtain a Streptomyces griseus seed solution; The shake flask culture medium is composed of the following ingredients: 10.2 g / L peptone, 5.2 g / L yeast powder, and 7.6 g / L sodium chloride; The seed culture medium is composed of the following ingredients: 20.6 g / L glucose, 5.3 g / L yeast powder, 5.4 g / L peptone, 6.5 g / L potassium dihydrogen phosphate, 10.28 mg / L ammonium molybdate, 0.26 g / L proline, and 16.0 g / L polyethylene glycol 600.

[0032] 4. Primary fermentation culture 12% volume of Bacillus subtilis seed liquid, 12% volume of thermophilic Streptococcus seed liquid, and 12% volume of Streptomyces griseus seed liquid were respectively inoculated into the fermentation medium for fermentation culture at a fermentation temperature of 26° C. for 5.5 days. After the fermentation was completed, a primary fermentation culture liquid was obtained; The fermentation medium is composed of the following ingredients: 17.0 g / L glucose, 12.7 g / L corn flour, 6.2 g / L soybean flour, 0.58 g / L potassium dihydrogen phosphate, 0.28 g / L manganese sulfate, 3.8 g / L calcium chloride, 2.7 g / L sodium nitrate, 0.9 g / L glycine, 3.6 g / L L-glutamine, and 1.5 g / L Tween 80.

[0033] 5. Modification of sepiolite 10.5 g of sepiolite was placed in 72 g of 43 wt% sodium hydroxide solution, the temperature was raised to 64°C, and the mixture was stirred for 3.2 h. After the stirring was completed, the mixture was placed in 104 g of 52 wt% ethanol solution, 1.2 g of sodium dodecylbenzene sulfonate and 0.8 g of polyethylene glycol 1000 were added, and the mixture was stirred for 33 min at 42°C. After the stirring was completed, 1.3 g of kH550 was added, the temperature was raised to 62°C, and the mixture was stirred for 2.2 h. Then, 0.6 g of glutamic acid and 0.8 g of chitosan oligosaccharide were added, and the mixture was stirred for 2.2 h to obtain modified sepiolite. The particle size of the sepiolite is 130 nm.

[0034] 6. Secondary fermentation culture 12.3 g of modified sepiolite was added to 102 g of the primary fermentation culture solution, and a secondary fermentation culture was carried out. The fermentation time was 12.5 h, and the fermentation temperature was 26° C. After the fermentation culture was completed, 3.7 g of modified bamboo fiber was added, and then ultrasonic treatment was carried out. The ultrasonic time was 16 min, the ultrasonic power was 255 W, and the ultrasonic frequency was 37 kHz. After the ultrasonic treatment was completed, the reaction was stirred at 36° C. for 3.2 h, and dried at 38° C. to obtain a composite microbial deodorant; The fermentation medium of the secondary fermentation culture is completely the same as that of the primary fermentation culture medium; The preparation method of the modified bamboo fiber is as follows: 5.2g of bamboo fiber is placed in 102g of deionized water, and then 10.3g of sodium periodate is added for stirring. After stirring evenly, the mixture is reacted in a dark environment at room temperature for 8.5h, and then 4.0g of ethylene glycol is added to terminate the reaction. The modified bamboo fiber is obtained by filtering, washing, and drying.

[0035] Comparative Example 1 Based on Example 1, the changes are as follows: In the first fermentation culture and the second fermentation culture steps, glycine, L-glutamine, and Tween 80 in the fermentation medium are replaced with deionized water in equal amounts; The modification step of sepiolite is omitted, and an equal amount of sepiolite in the secondary fermentation culture step is replaced by sepiolite without any treatment; The rest of the operations are the same.

[0036] Comparative Example 2 Based on Example 1, the changes are as follows: The secondary fermentation culture step is as follows: after the modification of the sepiolite is completed, 12.0 g of the modified sepiolite is added to 100 g of the primary fermentation liquid, and the mixture is fermented at 35° C. for 3.0 h and dried at 37° C. to obtain a composite microbial deodorant; The rest of the operations are the same.

[0037] Performance Testing The composite microbial deodorants prepared in Examples 1-3 and Comparative Examples 1-2 were used in deodorization tests, as follows: 1. Basic deodorizing performance Take 5 portions of 1L of stinky water and place them in closed containers respectively. The main components of the stinky water are ammonia and hydrogen sulfide, the concentration of ammonia is 250mg / L, and the concentration of hydrogen sulfide is 190m / L. Then add 85g of the composite microbial deodorant prepared in Example 1-3 and Comparative Example 1-2 respectively, stir at room temperature for 10min, let it stand after stirring, test the concentrations of ammonia and hydrogen sulfide in the sewage after standing for different times, and calculate the ammonia removal rate and hydrogen sulfide removal rate respectively; the test results are the average of three parallel tests; The test results are as follows:

[0038] 2. UV resistance The composite microbial deodorant prepared in Example 1-3 and Comparative Example 1-2 was subjected to ultraviolet light with an intensity of 400 W / cm 2 Irradiate under ultraviolet light for 24.0 hours to obtain a composite microbial deodorant after irradiation; Take 5 portions of 1L of stinky water and place them in closed containers respectively. The main components of the stinky water are ammonia and hydrogen sulfide, the concentration of ammonia is 250mg / L, and the concentration of hydrogen sulfide is 190m / L. Then add 85g of the composite microbial deodorant irradiated by Example 1-3 and Comparative Example 1-2 respectively, stir at room temperature for 10.0min, let stand for 12.0h after stirring, test the ammonia and hydrogen sulfide concentrations in the sewage again, and calculate the ammonia removal rate and hydrogen sulfide removal rate after standing for 12.0h; the test results are the average of three parallel tests; The test results are as follows:

[0039] The invention adopts specific strains for compounding, and then respectively carries out shake flask culture and seed culture, and then carries out mixed fermentation culture. In the mixed fermentation culture process, specific components such as glycine, L-glutamine, Tween 80 and the like are added, wherein glycine and L-glutamine can provide more nutrition and energy, promote the growth and metabolism of strains, and regulate the balance of the internal and external environment of cells; Tween 80, as a surfactant, can change the permeability of cell membranes, thereby promoting the reproduction and growth of cells; in the secondary fermentation culture process, the same culture medium components as those in the primary fermentation culture process are adopted, and modified sepiolite is added, which has good fixation, and fermentation cultured with microorganisms can better improve the fixation performance of microorganisms, and will not cause damage, thereby ensuring the activity of microorganisms; and the modified sepiolite has more hydroxyl and amino groups, and the amino groups can be cross-linked with the aldehyde groups of bamboo fibers, thereby forming a stable network structure, thereby improving the stability of the deodorant, enhancing the lasting effect, and realizing the effective removal of odor.

[0040] In Comparative Example 1, the nutrients and surfactants in the fermentation medium were omitted, which affected the metabolism and reproduction of the microorganisms. In addition, the sepiolite was not modified and could not be cross-linked with the modified bamboo fiber, thereby affecting the stability and long-term effectiveness of the deodorant. In Comparative Example 2, the modified sepiolite was directly added to the primary fermentation culture solution for immobilization, which had poor immobilization performance for microorganisms. In addition, the modified bamboo fiber was omitted and cross-linking could not be achieved, which ultimately resulted in poor deodorization performance, long-term performance and stability of the deodorant.

[0041] Unless otherwise specified, all ratios and percentages described in the present invention are by mass ratios and percentages are by mass percentages.

[0042] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention is described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing a composite microbial deodorant, characterized in that: The method comprises the steps of preparing a seed solution of Bacillus subtilis, preparing a seed solution of Streptococcus thermophilus, preparing a seed solution of Streptomyces griseus, primary fermentation culture, modification of sepiolite and secondary fermentation culture; The sepiolite modification step comprises placing the sepiolite in a sodium hydroxide solution, raising the temperature to 60-64° C., keeping the temperature and stirring for 2.8-3.2 hours, and after the keeping temperature and stirring, placing the sepiolite in an ethanol solution, adding sodium dodecylbenzene sulfonate and polyethylene glycol 1000, keeping the temperature and stirring for 26-33 minutes at 38-42° C., and after the keeping temperature and stirring, adding kH550, raising the temperature to 58-62° C., stirring and reacting for 1.8-2.2 hours, and then adding glutamic acid and chitosan oligosaccharide, and continuing to stir and react for 1.8-2.2 hours to obtain the modified sepiolite; The secondary fermentation and culturing step comprises: adding modified sepiolite to the primary fermentation culture solution, carrying out secondary fermentation and culturing, wherein the fermentation time is 11.5-12.5 hours, the fermentation temperature is 24-26° C., after the fermentation and culturing is completed, adding modified bamboo fiber, and then carrying out ultrasonic treatment, wherein the ultrasonic treatment time is 14-16 minutes, the ultrasonic power is 245-255 W, and the ultrasonic frequency is 34-37 kHz. After the ultrasonic treatment is completed, stirring and reacting at 34-36° C. for 2.8-3.2 hours, and drying at 36-38° C., to obtain a composite microbial deodorant.

2. The method for preparing a composite microbial deodorant according to claim 1, characterized in that: The step of preparing the Bacillus subtilis seed liquid is as follows: placing the Bacillus subtilis in a shake flask culture medium for shake flask culture, controlling the culture temperature to be 36-38° C., the culture time to be 16-20 hours, and the rotation speed to be 175-185 rpm, obtaining a shake flask culture liquid after the culture is completed, and then inoculating the shake flask culture liquid into a seed culture medium at an inoculation amount of 1.8-2.2% for seed culture, the culture temperature to be 36-38° C., and the culture time to be 18-22 hours, to obtain the Bacillus subtilis seed liquid; The Bacillus subtilis is purchased from the market; The shake flask culture medium is composed of the following ingredients: 4.8-5.2 g / L beef extract, 4.4-4.8 g / L peptone, 3.4-3.6 g / L sodium chloride, 3.2-3.5 g / L glucose, and the pH value is controlled to be 7.0-7.2; The seed culture medium is composed of the following ingredients: 11.8-12.3 g / L of glucose, 3.3-3.5 g / L of peptone, 0.38-0.42 g / L of magnesium sulfate heptahydrate, 2.6-2.8 g / L of ammonium sulfate, and 4.8-5.2 g / L of sodium chloride.

3. The method for preparing a composite microbial deodorant according to claim 1, characterized in that: The step of preparing the thermophilic streptococcus seed liquid is as follows: placing the thermophilic streptococcus in a shake flask culture medium for shake flask culture, controlling the culture temperature to be 30-33° C., the culture time to be 15-17 hours, and the rotation speed to be 210-230 rpm, obtaining a shake flask culture liquid after the culture is completed, and then inoculating the shake flask culture liquid into a seed culture medium at an inoculation amount of 2.4-2.6% for seed culture, the culture temperature to be 35-37° C., and the culture time to be 20-23 hours, to obtain the thermophilic streptococcus seed liquid; The thermophilic streptococcus is purchased from the market; The shake flask culture medium is composed of the following ingredients: 9.8-10.2 g / L peptone, 4.8-5.2 g / L sodium chloride, 11.8-12.3 g / L agar, and 3.4-3.6 g / L beef extract; The seed culture medium is composed of the following components: 8.4-8.7 g / L glucose, 9.8-10.3 g / L peptone, 9.7-10.2 g / L beef extract, 0.08-0.12 g / L manganese sulfate, 0.27-0.29 g / L magnesium sulfate heptahydrate, 1.9-2.2 g / L diammonium citrate, 4.2-4.4 g / L sodium acetate, and 18-22 mg / L cobalt chloride hexahydrate.

4. The method for preparing a composite microbial deodorant according to claim 1, characterized in that: The step of preparing the griseus seed solution comprises placing the griseus in a shake flask culture medium for shake flask culture, controlling the culture temperature to be 30-32° C., the culture time to be 13.8-14.2 h, and the rotation speed to be 165-175 rpm, and after the culture is completed, inoculating the shake flask culture solution into the seed culture medium at an inoculum amount of 1.8-2.2% for seed culture, the culture temperature to be 28-32° C., and the culture time to be 46-50 h, to obtain the griseus seed solution; The shake flask culture medium is composed of the following ingredients: 9.8-10.2 g / L peptone, 4.8-5.2 g / L yeast powder, and 7.4-7.6 g / L sodium chloride; The seed culture medium is composed of the following ingredients: 19.5-20.6 g / L of glucose, 4.7-5.3 g / L of yeast powder, 5.0-5.4 g / L of peptone, 6.2-6.5 g / L of potassium dihydrogen phosphate, 9.8-10.2 mg / L of ammonium molybdate, 0.24-0.26 g / L of proline, and 14.0-16.0 g / L of polyethylene glycol 600.

5. The method for preparing a composite microbial deodorant according to claim 1, characterized in that: The primary fermentation culture step comprises taking 8-12% volume of Bacillus subtilis seed liquid, 8-12% volume of thermophilic Streptococcus seed liquid, and 8-12% volume of Streptomyces griseus seed liquid respectively, inoculating them into fermentation medium for fermentation culture, the fermentation temperature is 24-26° C., the fermentation time is 4.5-5.5 days, and after the fermentation is completed, a primary fermentation culture liquid is obtained.

6. The method for preparing a composite microbial deodorant according to claim 5, characterized in that: The fermentation medium is composed of the following ingredients: 16.5-17.0 g / L of glucose, 12.2-12.7 g / L of corn flour, 6.0-6.2 g / L of soybean flour, 0.53-0.58 g / L of potassium dihydrogen phosphate, 0.24-0.28 g / L of manganese sulfate, 3.6-3.8 g / L of calcium chloride, 2.3-2.7 g / L of sodium nitrate, 0.7-0.9 g / L of glycine, 3.2-3.6 g / L of L-glutamine, and 1.3-1.5 g / L of Tween 80.

7. The method for preparing a composite microbial deodorant according to claim 1, characterized in that: In the sepiolite modification step, the particle size of the sepiolite is 110-130 nm; The mass ratio of the sepiolite, sodium hydroxide solution, ethanol solution, sodium dodecylbenzene sulfonate, polyethylene glycol 1000, kH550, glutamic acid and chitosan oligosaccharide is 10.0-10.5:68-72:98-104:0.8-1.2:0.6-0.8:1.0-1.3:0.4-0.6:0.5-0.8; The mass concentration of the sodium hydroxide solution is 40-43%; The mass concentration of the ethanol solution is 48-52%.

8. The method for preparing a composite microbial deodorant according to claim 1, characterized in that: In the secondary fermentation culture step, the fermentation medium of the secondary fermentation culture is completely the same as the primary fermentation medium; The mass ratio of the primary fermentation culture solution, the modified sepiolite and the modified bamboo fiber is 98-102:11.7-12.3:3.5-3.

7.

9. The method for preparing a composite microbial deodorant according to claim 1, characterized in that: The preparation method of the modified bamboo fiber is as follows: 4.8-5.2g of bamboo fiber is placed in 98-102g of deionized water, and then 9.7-10.3g of sodium periodate is added for stirring, and after stirring evenly, the mixture is reacted in a dark environment at room temperature for 7.5-8.5h, and then 3.6-4.0g of ethylene glycol is added to terminate the reaction, and the mixture is filtered, washed, and dried to obtain the modified bamboo fiber; The mass ratio of the bamboo fiber, deionized water, sodium periodate and ethylene glycol is 4.8-5.2:98-102:9.7-10.3:3.6-4.

0.

10. The composite microbial deodorant prepared according to the preparation method according to any one of claims 1 to 9.

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