CN112852669A - Composite microbial deodorant and preparation method thereof - Google Patents
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Abstract
The invention discloses a composite microbial deodorant and a preparation method thereof. In the invention, the screened strain seed liquid which is subjected to propagation culture is added into a fermentation tank for culture; in the step S13, the total amount of the inoculated seed liquid is about 1-5% of the mother liquid, fermentation culture is carried out, and the fermentation production can be stopped when the number of bacteria is over 1 multiplied by 107/ml order of magnitude; adding plant aromatic essential oil and activated carbon adsorption particles into the fermentation liquor obtained in the step S12, stirring and mixing uniformly, and then sealing and filling the solution in the fermentation tank to obtain the microbial deodorant; the microbial antibacterial deodorization shows attractive application prospects for air resistance/sterilization and organic matter purification. The microbial deodorization technology not only has strong broad-spectrum sterilization performance, but also can purify organic matters in the air, such as ethylene, and the like, and has the characteristics of high efficiency, low cost and no harm to the environment and human bodies, thereby showing important application prospects in sterilization and purification.
Description
Technical Field
The invention belongs to the technical field of deodorizers, and particularly relates to a compound microbial deodorizer and a preparation method thereof.
Background
Every family has more or less domestic garbage every day, and the garbage can release a large amount of harmful malodorous gases such as ammonia, hydrogen sulfide and the like when being not processed and stacked in the environment in time, thereby seriously polluting the atmosphere; the garbage contains pathogenic microorganisms, a large amount of acidic and alkaline organic pollutants are generated in the stacking and putrefaction process, heavy metals in the garbage are dissolved out to form a pollution source integrating organic substances, the heavy metals and the pathogenic microorganisms, and otherwise, a penetrating fluid generated by rainwater is poured can cause serious pollution to surface water and underground water. The garbage contains a plurality of pathogenic microorganisms, and is a breeding ground for mosquitoes, flies, cockroaches and mice, which inevitably endanger the health of vast citizens. The natural plant deodorant is prepared by extracting natural sterilization and deodorization factors from plants by adopting equipment and a process with completely independent intellectual property rights. No chemical substance is added, no toxic or side effect is caused to human bodies and livestock, and the use is safe. Has antibacterial, bactericidal and deodorant effects, and has good decomposition and removal effects on ammonia, hydrogen sulfide and other malodors.
However, the common deodorant can generate unpleasant odor during the reaction process, thereby causing troubles to operators.
Disclosure of Invention
The invention aims to: in order to solve the problems, a compound microbial deodorant and a preparation method thereof are provided.
The technical scheme adopted by the invention is as follows: a compound microorganism deodorant and a preparation method thereof are provided, wherein the compound microorganism deodorant and the preparation method thereof comprise the following steps:
s1, collecting and culturing original strains, namely collecting the original strains from mountains of natural forests in mountainous areas of Qinling mountains in Shaanxi province or original forest soil;
s2, strain separation is carried out from the strains collected in the step S1;
s3, weighing 10g of soil from the sample, and filling the soil into a triangular flask filled with 250mL of sterile water; vibrating for 20min to make the microorganisms in the soil fully and uniformly distributed in the liquid;
s4, bottling aerobic bacteria and anaerobic bacteria in an aerobic triangular bottle; taking out 0.5mL of original solution from the triangular flask, and performing a series of dilutions according to ten times of dilutions;
s5, inoculating 0.5mL of the diluted solution from each dilution in a test tube containing 4.5mL of the culture medium; respectively dripping 0.2mL of liquid from the first 4 dilutions into an improved Gauss culture medium plate and a PDA plate, and uniformly pushing by using a glass scraper; culturing and separating medium temperature bacteria, actinomycetes, fungi and yeast at 30 ℃, and culturing and separating high temperature resistant or thermophilic microorganisms at 50 ℃; the variety of media is numerous and the choice of which medium is the key to the success of the fermentation. Whether the composition and the proportion of the culture medium are proper or not has great influence on the growth of microorganisms, the selection of processes, the quality and the yield of products and the like. Different microorganisms are suitable for different culture media, the production application (or action) is different, and the selection standard of the culture media is different, for example, a spore culture medium is a common solid culture medium for breeding spores of strains, the requirement of the culture medium is that the strains can grow rapidly to produce more high-quality spores, and the culture medium is not easy to cause variation of the strains. Therefore, the basic preparation requirements of the spore culture medium are that the nutrition is not too rich (especially organic nitrogen source), otherwise, spores are not easy to generate; the concentration of the inorganic salt used is appropriate, otherwise the spore amount and color, etc. are affected. The seed culture medium is used for spore germination and strain growth and propagation, and has rich and complete nutrients easy to be absorbed and utilized by thallus. The contents of nitrogen source and vitamins are suitably high, but the total concentration is preferably slightly diluted so as to facilitate the reproduction of the bacteria. The fermentation medium is a medium for the growth, propagation and synthesis of the bacteria. It not only makes the seed grow quickly after inoculation to reach a certain thallus concentration, but also makes the growing bacteria synthesize the required product quickly. Therefore, the composition of the fermentation medium contains specific elements, preconditions, promoters and the like required for the product in addition to elements and compounds necessary for the growth of the microorganism;
s6, adding the collected sample into fresh pig manure according to the inoculation amount of 30 percent, and repeatedly enriching until the pig manure is odorless; adding odorless pig manure serving as an initial strain into simulated organic garbage (the inoculation amount is 30%), performing acclimation culture, and sampling from the treated simulated garbage;
s7, separating medium temperature bacteria, actinomycetes, fungi and yeast at 25 deg.C by streaking or serial dilution method with beef extract peptone, Gao 'S I, Gao' S II, and martin as culture medium, and separating high temperature resistant or thermophilic microorganisms at 50 deg.C; the simulated garbage comprises 50 percent of vegetables, 15 to 25 percent of rice, 5 to 25 percent of meat and 10 percent of rice hull as a loosening agent;
s8, weighing 1g of sample, adding 5ml of sterile water, fully oscillating and scattering to prepare a suspension liquid, then putting the suspension liquid and liquid culture into a large test tube of 40x180mm according to a ratio of 1: 5, adding a layer of sterilized liquid paraffin on the liquid surface to form anaerobism, placing the test tube at a temperature of 28-30 ℃ and under an illumination intensity of 2000Lux for anaerobic culture, wherein the culture solution is obviously dark red after three days; then, the culture solution is used for agar plate streaking separation and purification;
s9 taking the culture solution from the step S8 and adding filter sterilized NaHCO into the culture solution3(ii) a 5.0/50mL water;
s10, using an epi-plane of sterilized liquid paraffin on the surface of the culture medium under the same culture conditions, wherein a red microcolony appears after three days, a single colony grows well after six days, and the single colony is picked and separated repeatedly until a pure single colony is obtained;
s11: taking the strain with good deodorization performance, then carrying out propagation culture on the strain, and after culturing a mother solution, starting large-scale preparation;
s12: taking a fermentation tank, adding a 95% glucose solution into the fermentation tank, starting a high-temperature disinfection function inside the fermentation tank, and disinfecting and sterilizing the inside of the fermentation tank; then the strain seed liquid which is screened out in the step S11 and is subjected to propagation culture is added into a fermentation tank for culture;
s13: adding plant aromatic essential oil and activated carbon adsorption particles into the fermentation liquor obtained in the step S12, stirring and mixing uniformly, and then sealing and filling the solution in the fermentation tank to obtain the microbial deodorant
In a preferred embodiment, in step S1, the sampling should be performed according to the screening day, distribution of microorganisms, the main characteristics of the strains and their ecological relationship, etc., to determine the specific time, environment and target. It is observed that many original forests in the mountainous area of the Qin mountains in Shaanxi province and mountains rich in vegetation are full of fallen leaves, thick fallen leaves form rotten fallen leaf layers on the ground surface, but mountain springs and creeks under the mountains are still clear and clean, and microorganisms with strong purification capacity exist in rotten plants.
In a preferred embodiment, in the step S8, the components of the plate medium are as follows: NH4Cl1.0g, K2HPO4, 0.5g and MgCl 2. 0.2g, NaCl2.0g, 0.1g of yeast extract, 18-30 g of agar and 1000ml of water, and then carrying out aseptic operation after sterilization.
In a preferred embodiment, in step S13, the total amount of inoculated seed solution is about 1-5% of the mother solution, fermentation culture is performed, and fermentation production can be stopped when the number of bacteria is over 1 × 107/ml.
In a preferred embodiment, in step S10, the method co-isolates 18 strains of anaerobic bacteria, photosynthetic anaerobic bacteria with strong deodorizing ability belonging to the class of water-ring microorganisms, followed by qualitative deodorizing tests.
In a preferred embodiment: in the step S9, 50mL of ethanol or pentanol or 4% amino acid subjected to filter sterilization can be added, the pH value is adjusted to 7.0 by using 0.1L of P0 subjected to filter sterilization, the flat plate is poured while the solution is hot, and after the flat plate is solidified, the surface of the flat plate is scribed.
In summary, due to the adoption of the technical scheme, the invention has the beneficial effects that:
1. in the invention, the microbial antibacterial deodorization shows attractive application prospects for air resistance/sterilization and organic matter purification. The microbial deodorization technology not only has strong broad-spectrum sterilization performance, but also can purify organic matters in the air, such as ethylene, and the like, and has the characteristics of high efficiency, low cost and no harm to the environment and human bodies, thereby showing important application prospects in sterilization and purification.
2. According to the invention, when the deodorant is prepared, the plant essential oil and the activated carbon adsorption particles are added in the deodorant, so that peculiar smell generated during reaction of microorganisms in a treated object can be adsorbed, and the aromatic essential oil can make the treated object emit aromatic smell, thereby increasing the product characteristics.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example (b):
a compound microorganism deodorant and a preparation method thereof are characterized in that: the compound microbial deodorant and the preparation method thereof comprise the following steps:
s1, collecting and culturing original strains, namely collecting the original strains from mountains of natural forests in mountainous areas of Qinling mountains in Shaanxi province or original forest soil; in step S1, the sampling should be performed to determine the specific time, environment, and target object according to the factors of the day of screening, the distribution of microorganisms, the main characteristics of the strains, their ecological relationships, and the like. It is observed that many original forests in the mountainous area of the Qin mountains in Shaanxi province and mountains with rich vegetation are full of fallen leaves, the thick fallen leaves form rotten fallen leaf layers on the ground surface, but mountain springs and creeks flowing out of the mountains are still clear and clean, and microorganisms with strong purification capacity possibly exist in rotten plants;
s2, strain separation is carried out from the strains collected in the step S1;
s3, weighing 10g of soil from the sample, and filling the soil into a triangular flask filled with 250mL of sterile water; vibrating for 20min to make the microorganisms in the soil fully and uniformly distributed in the liquid;
s4, bottling aerobic bacteria and anaerobic bacteria in an aerobic triangular bottle; taking out 0.5mL of original solution from the triangular flask, and performing a series of dilutions according to ten times of dilutions;
s5, inoculating 0.5mL of the diluted solution from each dilution in a test tube containing 4.5mL of the culture medium; respectively dripping 0.2mL of liquid from the first 4 dilutions into an improved Gauss culture medium plate and a PDA plate, and uniformly pushing by using a glass scraper; culturing and separating medium temperature bacteria, actinomycetes, fungi and yeast at 30 ℃, and culturing and separating high temperature resistant or thermophilic microorganisms at 50 ℃;
s6, adding the collected sample into fresh pig manure according to the inoculation amount of 30 percent, and repeatedly enriching until the pig manure is odorless; adding odorless pig manure serving as an initial strain into simulated organic garbage (the inoculation amount is 30%), performing acclimation culture, and sampling from the treated simulated garbage;
s7, separating medium temperature bacteria, actinomycetes, fungi and yeast at 25 deg.C by streaking or serial dilution method with beef extract peptone, Gao 'S I, Gao' S II, and martin as culture medium, and separating high temperature resistant or thermophilic microorganisms at 50 deg.C; the simulated garbage comprises 50 percent of vegetables, 15 to 25 percent of rice, 5 to 25 percent of meat and 10 percent of rice hull as a loosening agent;
s8, weighing 1g of sample, adding 5ml of sterile water, fully oscillating and scattering to prepare a suspension liquid, then putting the suspension liquid and liquid culture into a large test tube of 40x180mm according to a ratio of 1: 5, adding a layer of sterilized liquid paraffin on the liquid surface to form anaerobism, placing the test tube at a temperature of 28-30 ℃ and under an illumination intensity of 2000Lux for anaerobic culture, wherein the culture solution is obviously dark red after three days; then, the culture solution is used for agar plate streaking separation and purification; in step S8, the plate medium was composed of NH4Cl1.0g, K2HPO4, 0.5g, and MgCl 2. 0.2g, NaCl2.0g, 0.1g of yeast extract, 18-30 g of agar and 1000ml of water, and performing aseptic operation after sterilization;
s9, taking the culture solution in the step S8, and adding filtered and sterilized NaHCO3 into the culture solution; 5.0/50mL water;
s10, using an epi-plane of sterilized liquid paraffin on the surface of the culture medium under the same culture conditions, wherein a red microcolony appears after three days, a single colony grows well after six days, and the single colony is picked and separated repeatedly until a pure single colony is obtained; in step S10, 18 strains of anaerobic bacteria are co-separated by the method, the anaerobic bacteria which have photosynthesis and strong deodorization capability belong to the class of water ring microorganisms, and then a deodorization qualitative test is carried out; the variety of media is numerous and the choice of which medium is the key to the success of the fermentation. Whether the composition and the proportion of the culture medium are proper or not has great influence on the growth of microorganisms, the selection of processes, the quality and the yield of products and the like. Different microorganisms are suitable for different culture media, the production application (or action) is different, and the selection standard of the culture media is different, for example, a spore culture medium is a common solid culture medium for breeding spores of strains, the requirement of the culture medium is that the strains can grow rapidly to produce more high-quality spores, and the culture medium is not easy to cause variation of the strains. Therefore, the basic preparation requirements of the spore culture medium are that the nutrition is not too rich (especially organic nitrogen source), otherwise, spores are not easy to generate; the concentration of the inorganic salt used is appropriate, otherwise the spore amount and color, etc. are affected. The seed culture medium is used for spore germination and strain growth and propagation, and has rich and complete nutrients easy to be absorbed and utilized by thallus. The contents of nitrogen source and vitamins are suitably high, but the total concentration is preferably slightly diluted so as to facilitate the reproduction of the bacteria. The fermentation medium is a medium for the growth, propagation and synthesis of the bacteria. It not only makes the seed grow quickly after inoculation to reach a certain thallus concentration, but also makes the growing bacteria synthesize the required product quickly. Therefore, the composition of the fermentation medium contains specific elements, preconditions, promoters and the like required for the product in addition to elements and compounds necessary for the growth of the microorganism;
s11: taking the strain with good deodorization performance, then carrying out propagation culture on the strain, and after culturing a mother solution, starting large-scale preparation;
s12: taking a fermentation tank, adding a 95% glucose solution into the fermentation tank, starting a high-temperature disinfection function inside the fermentation tank, and disinfecting and sterilizing the inside of the fermentation tank; then the strain seed liquid which is screened out in the step S11 and is subjected to propagation culture is added into a fermentation tank for culture; in the step S13, the total amount of the inoculated seed liquid is about 1-5% of the mother liquid, fermentation culture is carried out, and the fermentation production can be stopped when the number of bacteria is over 1 multiplied by 107/ml order of magnitude;
s13: and (4) adding plant aromatic essential oil and activated carbon adsorption particles into the fermentation liquor obtained in the step (S12), stirring and mixing uniformly, and then sealing and filling the solution in the fermentation tank to obtain the microbial deodorant.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (6)
1. A compound microorganism deodorant and a preparation method thereof are characterized in that: the compound microbial deodorant and the preparation method thereof comprise the following steps:
s1, collecting and culturing original strains, namely collecting the original strains from mountains of natural forests in mountainous areas of Qinling mountains in Shaanxi province or original forest soil;
s2, strain separation is carried out from the strains collected in the step S1;
s3, weighing 10g of soil from the sample, and filling the soil into a triangular flask filled with 250mL of sterile water; vibrating for 20min to make the microorganisms in the soil fully and uniformly distributed in the liquid;
s4, bottling aerobic bacteria and anaerobic bacteria in an aerobic triangular bottle; taking out 0.5mL of original solution from the triangular flask, and performing a series of dilutions according to ten times of dilutions;
s5, inoculating 0.5mL of the diluted solution from each dilution in a test tube containing 4.5mL of the culture medium; respectively dripping 0.2mL of liquid from the first 4 dilutions into an improved Gauss culture medium plate and a PDA plate, and uniformly pushing by using a glass scraper; culturing and separating medium temperature bacteria, actinomycetes, fungi and yeast at 30 ℃, and culturing and separating high temperature resistant or thermophilic microorganisms at 50 ℃;
s6, adding the collected sample into fresh pig manure according to the inoculation amount of 30 percent, and repeatedly enriching until the pig manure is odorless; adding odorless pig manure serving as an initial strain into simulated organic garbage (the inoculation amount is 30%), performing acclimation culture, and sampling from the treated simulated garbage;
s7, separating medium temperature bacteria, actinomycetes, fungi and yeast at 25 deg.C by streaking or serial dilution method with beef extract peptone, Gao 'S I, Gao' S II, and martin as culture medium, and separating high temperature resistant or thermophilic microorganisms at 50 deg.C; the simulated garbage comprises 50 percent of vegetables, 15 to 25 percent of rice, 5 to 25 percent of meat and 10 percent of rice hull as a loosening agent;
s8, weighing 1g of sample, adding 5ml of sterile water, fully oscillating and scattering to prepare a suspension liquid, then putting the suspension liquid and liquid culture into a large test tube of 40x180mm according to a ratio of 1: 5, adding a layer of sterilized liquid paraffin on the liquid surface to form anaerobism, placing the test tube at a temperature of 28-30 ℃ and under an illumination intensity of 2000Lux for anaerobic culture, wherein the culture solution is obviously dark red after three days; then, the culture solution is used for agar plate streaking separation and purification;
s9 taking the culture solution from the step S8 and adding filter sterilized NaHCO into the culture solution3(ii) a 5.0/50mL water;
s10, using an epi-plane of sterilized liquid paraffin on the surface of the culture medium under the same culture conditions, wherein a red microcolony appears after three days, a single colony grows well after six days, and the single colony is picked and separated repeatedly until a pure single colony is obtained;
s11: taking the strain with good deodorization performance, then carrying out propagation culture on the strain, and after culturing a mother solution, starting large-scale preparation;
s12: taking a fermentation tank, adding a 95% glucose solution into the fermentation tank, starting a high-temperature disinfection function inside the fermentation tank, and disinfecting and sterilizing the inside of the fermentation tank; then the strain seed liquid which is screened out in the step S11 and is subjected to propagation culture is added into a fermentation tank for culture;
s13: and (4) adding plant aromatic essential oil and activated carbon adsorption particles into the fermentation liquor obtained in the step (S12), stirring and mixing uniformly, and then sealing and filling the solution in the fermentation tank to obtain the microbial deodorant.
2. The composite microbial deodorant and the method for producing the same according to claim 1, wherein: in the step S1, the sampling process determines the specific time, environment and target object according to the factors of the screened day, the distribution of microorganisms, the main characteristics of the strains and the ecological relationship thereof, and the like, and it is observed that many original forests in the mountainous area of the qin mountains in shanxi and mountains rich in vegetation thereof are full of fallen leaves, thick fallen leaves form rotten leaf layers on the ground surface, but mountain springs flowing out of the mountains and creeks under the mountains are still clear and clean, and microorganisms with strong purification capability exist in rotten plants.
3. The composite microbial deodorant and the method for producing the same according to claim 1, wherein: in the step S8, the components of the plate culture medium are NH4Cl1.0g, K2HPO4, 0.5g, MgCl2, 0.2g, NaCl2.0g, yeast extract 0.1g, agar 18-30 g and water 1000ml, and the plate culture medium is sterilized and then sterilized.
4. The composite microbial deodorant and the method for producing the same according to claim 1, wherein: in the step S13, the total amount of the inoculated seed liquid is about 1-5% of the mother liquid, fermentation culture is carried out, and the fermentation production can be stopped when the number of bacteria is over 1 multiplied by 107/ml order of magnitude.
5. The composite microbial deodorant and the method for producing the same according to claim 1, wherein: in the step S10, 18 strains of anaerobic bacteria are co-separated by the method, photosynthetic anaerobic bacteria with strong deodorization capability belong to the class of water ring microorganisms, and then a deodorization qualitative test is carried out.
6. The composite microbial deodorant and the method for producing the same according to claim 1, wherein: in the step S9, 50mL of ethanol or pentanol or 4% amino acid subjected to filter sterilization can be added, the pH value is adjusted to 7.0 by using 0.1L of P0 subjected to filter sterilization, the flat plate is poured while the solution is hot, and after the flat plate is solidified, the surface of the flat plate is scribed.
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Citations (5)
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