CN114015586B - Deodorization degradation composite microbial agent and preparation method and application thereof - Google Patents
Deodorization degradation composite microbial agent and preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the field of microorganism application, and particularly relates to a deodorizing and degrading composite microbial agent, and a preparation method and application thereof. The technical proposal is as follows: pichia pastoris (Pichia fermentans), which is preserved in China general microbiological culture Collection center (China) for 9 and 14 days in 2021, with the preservation addresses: the preservation number of the Beijing city Chaoyang area North Chen Xili No.1 and 3 is: CGMCC No.23387. Pichia pastoris and lactobacillus plantarum are combined and compounded to prepare the novel deodorant. The composite microbial inoculum takes the microecological principles of symbiosis, coexistence and co-Rong as guidance, and constructs a deodorizing and degrading microbial system taking a high-efficiency bacterial strain as a core, so that the growth of saprophytic microorganisms and pathogenic bacteria can be competitively inhibited, malodor generation is controlled or malodorous substances are directly degraded by occupying dominant ecological niches of the fecal microecological environment, the stability and durability of deodorizing and degrading effects are ensured, and the aim of completing the recycling treatment of the fecal sewage on the premise of not generating malodorous substances is achieved.
Description
Technical Field
The invention belongs to the field of microorganism application, and particularly relates to a deodorizing and degrading composite microbial agent, and a preparation method and application thereof.
Background
Under the condition of no artificial regulation, the toilet excrement rich in organic matters is decomposed to generate irritant gases such as ammonia under the aerobic condition and to generate malodorous substances such as volatile low-molecular fatty alcohol, amine and acid under the anoxic or anaerobic condition under the microbial action, so that a large amount of malodorous gases are released, and the use environment of the toilet is seriously influenced. And the natural transformation process of the excrement is slower. Therefore, the artificial regulation and strengthening are needed in the process of controlling and biodegradation of the excrement and sewage.
In the biological treatment process of the toilet excrement, the key point is to develop functional microorganism strains with high-efficiency excrement degradation and malodor control, scientifically compound the functional strains, and release high protease with the function of degrading organic matters through the compound strains, so that macromolecular organic matters in the excrement are degraded into small molecular organic matters such as sugar, fatty acid, amino acid and the like under proper external conditions. The biodegradation and the malodor control of the toilet feces are enhanced and controlled, so that the harmless and recycling treatment of the feces are realized.
The microbial deodorizing and degrading composite microbial agent has the advantages of environmental friendliness, low cost and the like, and is an important research direction for treating malodorous environment at present. The effect depends on the efficient colonization effect of the core strain, so that it is critical to obtain excellent strains with good deodorizing and degrading effects and ensure that the composite microbial inoculum can compete with indigenous bacteria to become dominant bacteria after being applied.
Disclosure of Invention
The invention aims to provide a deodorizing degradation composite microbial inoculant, and a preparation method and application thereof.
In order to achieve the aim of the invention, the invention adopts the following technical scheme: pichia pastoris (Pichia fermentans), which is preserved in China general microbiological culture Collection center (China) for 9 and 14 days in 2021, with the preservation addresses: the preservation number of the Beijing city Chaoyang area North Chen Xili No.1 and 3 is: CGMCC No.23387.
Correspondingly, a strain of Pichia pastoris (Pichia fermentans) has an 18S rDNA sequence shown in SEQ ID NO. 2.
Correspondingly, the pichia pastoris is applied to deodorization.
Correspondingly, the deodorant prepared by using the pichia pastoris.
Preferably, the deodorant is a microbial composite microbial agent, including lactobacillus plantarum (Lactobacillus plantarum).
Preferably, the lactobacillus plantarum deposit number is: CGMCC No. 23188.
Preferably, the lactobacillus plantarum: pichia pastoris = 1-2:1-5.
Preferably, the lactobacillus plantarum: pichia pastoris = 2:3.
Preferably, the concentration of viable bacteria in the deodorant is not less than 4.25X10 8 CFU/mL。
Correspondingly, the deodorant is applied to deodorization, and the dosage of the deodorant is as follows: the feed of the material to be treated is 7X 4.25X 10 per hundred grams 8 Deodorant with CFU viable bacteria amount.
The invention has the following beneficial effects: the invention screens out a new lactobacillus plantarum and a new pichia pastoris. The two strains of microorganisms are excellent in deodorizing performance and strong in environment adaptability: the application temperature range is wide, and the deodorant can well grow and maintain excellent deodorant performance in a low-temperature or even lower-temperature environment of 5 ℃; the application humidity range is wide, and the growth is good in a wet or dry environment. The microorganism and the prepared composite microorganism microbial inoculum provided by the invention can effectively inhibit the growth of odorogens, pathogenic bacteria and other miscellaneous bacteria, efficiently degrade odor sources, especially the odor gases such as ammonia gas, hydrogen sulfide, TVOC and the like generated by toilet excrement, have obvious odor control and excrement degradation effects, and can be used for deodorizing in special environments with severe cold and anoxic conditions, easy icing in winter and weaker ecological systems.
Specifically, the deodorant provided by the invention has definite pertinence and is mainly used for treating excrement. The ammonia gas, hydrogen sulfide, TVOC and other malodorous gases can be released in the biological degradation process of the feces, the degradation and decomposition period of the feces is long, and parasites or pathogenic bacteria, a large number of mosquito and termite, maggots and the like can exist in the feces. Aiming at the three key problems of the fecal sewage, the invention provides a functional strain and a bacterial system which can efficiently and quickly degrade the fecal sewage and treat the fecal sewage odor, and the functional strain and the bacterial system are further prepared into a deodorant. The deodorant contains two microorganisms which are not only antagonistic, but also can act synergistically on the excrement malodor source. Moreover, the deodorizing and degrading effects of the two strains of microorganisms are obviously superior to those of the single strain and the random proportion through effective compounding optimization.
The composite microbial inoculum takes the microecological principles of symbiosis, coexistence and co-Rong as guidance, and constructs a deodorizing and degrading microbial system taking a high-efficiency bacterial strain as a core, so that the growth of saprophytic microorganisms and pathogenic bacteria can be competitively inhibited, malodor generation is controlled or malodorous substances are directly degraded by occupying dominant ecological niches of the fecal microecological environment, the stability and durability of deodorizing and degrading effects are ensured, and the aim of completing the recycling treatment of the fecal sewage on the premise of not generating malodorous substances is achieved.
Drawings
FIG. 1 is a phylogenetic tree diagram of Lactobacillus plantarum;
FIG. 2 is a phylogenetic tree diagram of Pichia pastoris;
FIG. 3 is a photomicrograph of Lactobacillus plantarum;
FIG. 4 is a schematic representation of the colony of Lactobacillus plantarum;
FIG. 5 is a photomicrograph of Pichia pastoris;
FIG. 6 is a schematic representation of colonies of Pichia pastoris.
Detailed Description
The invention provides a composite microbial inoculum which can degrade malodorous gas, has excellent deodorizing effect and wide application range. The composite microbial inoculum comprises lactobacillus plantarum (Lactobacillus plantarum) and pichia pastoris (Pichia pastoris).
The preferable scheme is as follows: the lactobacillus plantarum is: CGMCC No. 23188; the pichia pastoris is as follows: CGMCC No.23387.
The preferable scheme is as follows: the lactobacillus plantarum: pichia pastoris = 2:3.
The preferable scheme is as follows: under the use state of the composite microbial inoculum, the living microbial inoculum amount in the microbial inoculum is more than or equal to 4.25X10 8 CFU/mL。
The invention also provides a preparation process of the composite microbial inoculum, which comprises the following steps:
1. activation of the strain. The strain is restored to room temperature from the preservation state, inoculated on a solid flat plate by aseptic streaking, and subjected to 3-generation rejuvenation process, so that the strain is gradually adapted to the culture environment. The lactobacillus plantarum is inoculated into MRS solid culture for activation standby, and the pichia pastoris is inoculated into YPED solid culture medium for activation standby.
2. And (5) fermenting and culturing the strain. Inoculating activated lactobacillus plantarum into MRS liquid culture medium, inoculating Pichia pastoris into YPED liquid culture medium, fermenting and culturing at 30deg.C and 150r/min for more than 24h, and fermenting for 48h.
3. The initial concentration of the strain broth was adjusted. Adjusting the initial fermentation liquor concentration of lactobacillus plantarum and pichia pastoris to OD 600 =0.5, viable bacteria concentration 4.25×10 8 CFU/mL。
4. Organic compounding of bacterial liquid. And uniformly mixing the regulated lactobacillus plantarum and pichia pastoris according to the viable bacteria ratio of 2:3 to obtain the liquid deodorizing and degrading composite microbial inoculum. Can be directly used or made into powder, granule or other forms of preparations or used as one of other components of physical or chemical deodorant which has no conflict with corresponding microorganisms according to actual needs.
The invention also provides a use method of the composite microbial inoculum. Will beThe composite microbial inoculum is directly put into the material to be treated and is evenly mixed with the material to be treated. And the fillers such as wood dust, rice husk, straw powder and the like can be simultaneously added according to actual conditions and uniformly mixed. It should be noted that: the amount of the composite microbial inoculum is related to the weight of the material to be treated, and each hundred grams of the material to be treated is preferably added with the composition containing 7 multiplied by 4.25 multiplied by 10 8 CFU viable count of composite microbial inoculum. Taking liquid composite microbial inoculum as an example, the adding amount is 7mL of composite microbial inoculum per 100g of material to be treated.
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. The technical means used in the examples are conventional means well known to those skilled in the art unless otherwise indicated.
The formula of the culture medium related by the invention is as follows:
(1) MRS liquid medium: 10g/L of peptone, 10g/L of beef extract, 5g/L of yeast extract, 5g/L of glucose, 5g/L of sodium acetate, 2g/L of diammonium citrate, 1g/L of tween 80, 0.58g/L of magnesium sulfate, 0.05g/L of manganese sulfate, 2g/L of dipotassium hydrogen phosphate and pH=6.2. MRS solid medium is added with 20g/L agar based on liquid medium.
(2) YPED liquid medium: yeast powder 10g/L, peptone 20g/L, glucose 20g/L, bengalne 0.03mL/L, pH=6.0. YPED solid medium was supplemented with 20g/L agar based on liquid medium.
Embodiment one: screening, identifying and preserving microorganisms in composite microbial agents
1. The screening method of the microorganisms comprises the following steps: the lactobacillus plantarum and the pichia pastoris provided by the invention are obtained by screening a fecal sample through a dilution coating method. The following microorganisms having deodorizing effect were obtained by repeated screening from a large number of fecal samples: 4 strains of bacillus, 2 strains of saccharomycetes and 5 strains of acidogenic bacteria. Further experiments found that: the lactobacillus plantarum and the pichia pastoris provided by the invention can grow at a low temperature of 10 ℃ or even 5 ℃ and keep good deodorizing performance. Meanwhile, the lactobacillus plantarum can also resist high temperature of more than 40 ℃ and keep good deodorizing and acid producing performances.
2. And (5) identifying and preserving the microbial molecules. The lactobacillus plantarum is subjected to 16S rRNA sequencing, the result is shown as SEQ ID NO:1, and according to a phylogenetic tree (figure 1), the lactobacillus plantarum has the closest relationship with Lactobacillus plantarum strain CIP 103151 (NR 104573.1) and has 99.93 percent of sequence similarity. The 18S rDNA molecular identification of the pichia pastoris is shown as SEQ ID NO:2, and according to a phylogenetic tree (figure 2), the closest relationship between the pichia pastoris and Pichia fermentans culture CBS:4807 (KY 104537.1) is found, and the sequence similarity is 99.31%.
The lactobacillus plantarum is preserved in China general microbiological culture collection center (CGMCC) for 14 days 9 and 2021, and the preservation address is: the preservation number of the Beijing city Chaoyang area North Chen Xili No.1 and 3 is: CGMCC No. 23188, classified and named as: lactobacillus plantarum (Lactobacillus plantarum). The Pichia pastoris is preserved in China general microbiological culture collection center (CGMCC) for 14 days 9 and 2021, and the preservation address is: the preservation number of the Beijing city Chaoyang area North Chen Xili No.1 and 3 is: CGMCC NO.23387, classified and named as: pichia pastoris (Pichia fermentans) is fermented.
3. And (5) physiological and biochemical identification of microorganisms. The microscopic photograph of the lactobacillus plantarum is shown in fig. 3, and the photograph of the colony cultivated in the culture dish is shown in fig. 4; the micrograph of Pichia pastoris is shown in FIG. 5, and the photograph of the colonies cultured in the dish is shown in FIG. 6.
The physiological and biochemical identification results of lactobacillus plantarum are shown in table 1, and the physiological and biochemical identification results of pichia pastoris are shown in table 2. In the table "+" represents positive and "-" represents negative.
TABLE 1 physiological and biochemical identification results of Lactobacillus plantarum
TABLE 2 physiological and biochemical identification results of Pichia pastoris
Embodiment two: comparison of deodorizing effects of individual microorganisms and composite microbial agents
1. A simulator with a volume of 10L was produced, in which 200g of fresh manure was placed, and the device was closed. Setting 3 treatments, wherein 6mL of bacterial liquid is respectively added into a device filled with 200g of fresh feces, and the concentration of viable bacteria in the bacterial liquid is as follows: 4.25X10 8 CFU/mL. The added bacterial liquid in the treatment 1 is the lactobacillus plantarum bacterial liquid, the added bacterial liquid in the treatment 2 is the pichia pastoris bacterial liquid, and the added bacterial liquid in the treatment 3 is the lactobacillus plantarum and pichia pastoris mixed bacterial liquid mixed and combined in a volume ratio of 1:1. Meanwhile, the control group is not added with microbial inoculum. Three replicates were set for each set of experiments and incubated in a 30℃incubator with 60% ambient humidity.
2. After the treatments of 5d, 10d, 15d, 20d and 30d, each set of Volatile Solids (VS) ("method of water and wastewater monitoring analysis (fourth edition)), ammonia concentration (gas rapid detector), hydrogen sulfide concentration (gas rapid detector), total Volatile Organic Compound (TVOC) concentration (gas rapid detector) and comprehensive odor concentration (OU) (gas rapid detector) was measured, respectively. And evaluating degradation performance of each group of microbial agents according to the change condition of VS, and evaluating deodorizing effect of each group of microbial agents according to ammonia gas concentration, hydrogen sulfide concentration, TVOC concentration and comprehensive odor concentration. The measurement results are shown in Table 3. Each measurement index of the control group in table 3 shows the actual value of the corresponding index measured at the current time, and each measurement index of each treatment group shows the removal rate of the corresponding index measured at the current time; the method for calculating the removal rate comprises the following steps: (control measurement at current time-treatment measurement at current time)/control measurement at current time x 100%. In Table 3, the units of VS are mg/m for the ammonia concentration, hydrogen sulfide concentration and TVOC concentration in the control group 3 Each index unit of each treatment group was% (all were removal rates). The unit and removal rate calculation method of each index in the subsequent table are the same as those described above.
Table 3 shows deodorizing effect of each group of microbial agents
From the table, the VS removal rate is improved along with the time extension, and the content of residual organic solids in the fecal sewage tends to be reduced, so that the microorganisms provided by the invention have degradation effect on the solids in the fecal sewage. Fresh manure was odorless and tasteless after 10 days of treatment with treatment group 3. The effects of treatments 20d and 30d were not significantly different, so that the data from test 20d were the longest for all subsequent experiments.
Embodiment III: effect of preparation and use parameters on deodorant Effect
1. The proportion relation of the microbial dosage in the composite microbial agent.
Using the same device as the embodiment, each treatment device still contains 200g of fresh manure, and 9 treatments are arranged; meanwhile, the control group is prepared by taking the non-added bacteria liquid. All are placed in a constant temperature incubator at 30 ℃ for culture, and the environmental humidity is 60%. The Lactobacillus plantarum and Pichia pastoris compositions in each treatment are shown in Table 4. In Table 4, the values of the amounts of each microorganism refer to the volume to mass ratio of the amount of fecal sewage, for example, 1% means that 1% by 200, i.e., 2mL, of the corresponding microorganism fermentation broth was used in 200g fecal sewage. The viable bacteria concentration in each microbial fermentation broth was 4.25X10 8 CFU/mL. The same applies to the subsequent point where the volume to mass ratio of the microbial load is concerned.
TABLE 4 schematic Table of the composition of the microbial inoculum of each group
| Group of | Lactobacillus plantarum | |
| Group | ||
| 1 | 1％ | 1％ |
| Group 2 | 1％ | 2％ |
| Group 3 | 1％ | 3％ |
| Group 4 | 1％ | 4％ |
| Group 5 | 1％ | 5％ |
| Group 6 | 2％ | 1％ |
| Group 7 | 2％ | 3％ |
| Group 8 | 2％ | 5％ |
After 10d incubation (10 d incubation was performed according to example two, 10d was used for cost saving, 10 d) and each of the groups was assayed for VS, ammonia concentration, hydrogen sulfide concentration, TVOC concentration and OU concentration. The measurement results are shown in Table 5. Each measurement index of the control group shows the actual value of the corresponding index measured at the current time, and each measurement index of each treatment group shows the removal rate of the corresponding index measured at the current time.
Table 5 shows deodorizing effect of each group of microbial agents
The results show that: the deodorizing effect is best when lactobacillus plantarum and pichia pastoris are mixed according to the living bacteria amount ratio of 2:3. The compound bacteria in the subsequent experiments of this example all used a ratio of 2:3.
The stability of the composite microbial inoculum is tested at the same time in the embodiment: the prepared liquid compound bacteria agent is stored in room temperature environment, and the initial viable bacteria concentration is 4.25X10 8 CFU/mL, after 30d, viable bacteria concentration was 3.76X10 8 CFU/mL (2:3 viable bacteria ratio group is taken as an example; the viable bacteria concentration of the rest of the treatment groups is not significantly different from that of the treatment groups). The deodorizing effect of each group of composite microbial inoculum after being placed for 30d is not obvious from the deodorizing effect shown in table 5. The composite microbial inoculum provided by the invention has better stability.
2. The use amount of the composite microbial inoculum.
Uniformly compounding lactobacillus plantarum and pichia pastoris in a viable bacteria ratio of 2:3, wherein the viable bacteria concentration in each microbial fermentation broth is 4.25X10 8 CFU/mL; and then 6 treatments are designed according to different volume-mass ratios compared with the mass of the feces, and the adding gradients of the composite microbial inoculum in each treatment are respectively as follows: 1%, 3%, 5%, 7%, 10%, 15%. Culturing in a constant temperature incubator at 30deg.C with environmental humidity of 60%; sampling and detection are carried out at the time of culturing for 10d and 20d respectively. The results are shown in Table 6. Each measurement index of the control group shows the actual value of the corresponding index measured at the current time, and each measurement index of each treatment group shows the removal rate of the corresponding index measured at the current time. The results show that: the deodorizing effect is best when 7% of microorganism is added.
Table 6 shows deodorizing effect of each group of microbial agents
3. Influence of the method of use.
Uniformly compounding lactobacillus plantarum and pichia pastoris according to a viable cell mass ratio of 2:3, wherein the viable cell concentration in each microbial fermentation broth is 4.25X10 8 CFU/mL; the addition amount of the composite microbial inoculum is 7% or the addition amount equivalent to 7% of the addition amount of the liquid composite microbial inoculum. Except for the variables, the temperature conditions are set to be 30 ℃ and the humidity to be 65% under the other conditions, and the composite microbial inoculum is liquid. The test devices are placed under different environments, the temperature conditions are respectively designed to be 5 ℃, 10 ℃,20 ℃,30 ℃, 45 ℃ and the humidity conditions are respectively designed to be 35%, 50%, 65%, 80% and 90%, the use forms of the composite microbial inoculum are designed to be liquid, the embedding method is fixed (sodium alginate embedding method), and the spray drying is carried out. And simultaneously, setting a blank control group without adding the composite bacterial agent under different temperature and humidity conditions. Simultaneously, three lactobacillus plantarum sources of different factories and three pichia pastoris sources of different factories are purchased in the market, and the lactobacillus plantarum screened by the invention is replaced or the pichia pastoris screened by the invention is replaced respectively, or the lactobacillus plantarum screened by the invention and the pichia pastoris are replaced simultaneously, and the experiment is carried out simultaneously under the condition that the rest conditions are identical, so that the positive control group is obtained. The positive control group was set up as shown in Table 7. Commercially available lactobacillus plantarum include: CGMCC No.1.2437 (Lactobacillus plantarum), lactobacillus plantarum purchased from Jiangsu green family, lactobacillus plantarum purchased from Hebei Qiansheng organism; three commercially available pichia pastoris were all from Minghuata.
TABLE 7 presentation of Positive control set settings
After the treatment groups and the control groups 10d and 20d were measured, the volatile solids VS, ammonia gas concentration, hydrogen sulfide concentration, TVOC concentration and OU concentration were measured. The specific results under the temperature change conditions are shown in Table 8. The results under humidity change conditions are shown in Table 9. The results of the formulation changes are shown in Table 10. Tables 8, 9, and 10 show the data for each treatment group and the blank group. Each measurement index of the blank control group shows the actual value of the corresponding index measured at the current time, and each measurement index of each treatment group shows the removal rate of the corresponding index measured at the current time.
Table 8 shows deodorizing effect of composite microbial inoculant at different temperatures
Table 9 shows deodorizing effect of composite microbial inoculant under different humidity
Table 10 shows deodorizing effect of composite microbial inoculant in different preparation forms
For the sake of brevity, specific data for each positive control group is not shown herein. The effect of the removal rate of each index of the positive control group 1-6 is 33% -42% of the corresponding effect of the treatment group in the range of 5-10 ℃, the effect of the removal rate of each index of the positive control group 7-15 is 0-12% of the corresponding effect of the treatment group in the invention, and the treatment effect of the positive control group 7-15 at 5 ℃ is 0. The removal rate effect of each index of each positive control group is 72-98% of the corresponding effect of the invention within the range of 20-30 ℃. The effect of the removal rate of each index of the positive control groups 1 to 6 is 55 to 71 percent of the corresponding effect of the treatment group of the invention at 45 ℃, and the effect of the removal rate of each index of the positive control groups 7 to 15 is 36 to 53 percent of the corresponding effect of the treatment group of the invention.
The results show that: the composite microbial inoculum provided by the invention not only survives under the extreme temperature environment of 5 ℃ and 45 ℃ and the extreme humidity environment of 35% and 90%, but also has excellent deodorizing performance. Meanwhile, the effect of the spray-dried composite microbial inoculum is only slightly weaker than that of a liquid form, and the application scene of the composite microbial inoculum is effectively expanded.
Embodiment four: deodorization effect display of composite microbial inoculum in practice
The composite microbial inoculum used in the embodiment is a liquid composite microbial inoculum obtained by compounding lactobacillus plantarum and pichia pastoris screened by the invention according to the ratio of 2:3, and the concentration of viable bacteria in the microbial inoculum is 4.25 multiplied by 10 8 CFU/mL。
1. And (5) selecting a factory in a double-flow area of the Chengdu city to sample human body excrement. A treatment group and a control group were set, each group being a fermentation tank storing 40kg of fecal sewage, and each group being repeated 3 times. 2.8mL of the compound bacteria agent is added into the treatment group, and the equivalent amount of sterile water is added into the control group. The samples were taken after 5d, 10d, 15d, 20d and 30d to determine VS, ammonia concentration, hydrogen sulfide concentration, TVOC concentration and comprehensive odor concentration, respectively, and the corresponding removal rates were calculated. The results are shown in Table 11 (the treatment group is the removal rate, and the control group is the actual measurement value).
Table 11 deodorizing effect display table
2. And (3) carrying out small-batch application effect test on 20 dry toilets in a dry land selected by Sichuan at first. The deodorization degradation composite microbial inoculum with the mass-volume ratio of 7% of the feces is respectively added into a toilet of 10 households, and the feces storage tank arranged under the dry toilet is not provided with an automatic stirring device, so that the composite microbial inoculum is respectively added in batches of 2d, 12d and 22d, the mass-volume ratio of 7% of the feces is added in total, and the mass of the feces is estimated by the volume of the feces storage tank. Taking another 10 users as a control group without adding microbial inoculum and with equal amount of sterile water, sampling after 5d, 10d, 15d, 20d and 30d respectively to measure VS, ammonia gas concentration, hydrogen sulfide concentration, TVOC concentration and comprehensive odor concentration, and calculating corresponding removal rate. The results are shown in Table 12.
Table 12 deodorizing effect display table
| Measurement index | VS | Ammonia concentration | Concentration of hydrogen sulfide | TVOC concentration | OU |
| 5d | 86.32 | 65.64 | 59.05 | 60.35 | 62.67 |
| Control group | 90.16 | 399.26 | 40.35 | 856 | 421 |
| 10d | 88.75 | 85.49 | 64.74 | 70.31 | 69.63 |
| Control group | 90.86 | 393.35 | 42.26 | 861 | 430 |
| 15d | 91.41 | 85.08 | 64.90 | 70.89 | 60.31 |
| Control group | 90.34 | 392.98 | 41.19 | 860 | 425 |
| 20d | 89.35 | 85.97 | 64.59 | 71.04 | 71.18 |
| Control group | 90.26 | 393.2 | 42.75 | 859 | 427 |
| 30d | 90.9 | 85.12 | 64.76 | 71.62 | 71.49 |
| Control group | 90.74 | 392.97 | 43.17 | 861 | 430 |
3. Six movable public toilets are selected in a tourist attraction of Hongyuan county in Sichuan A, and have the same-volume excrement storage tanks, three of which are added with composite microbial inoculum with mass and volume ratio of 7 percent of excrement, and the device is started and put into use. Three other mobile public toilets without microbial inoculum and with equal amount of sterile water are used as control. The samples were taken after 5d, 10d, 15d, 20d and 30d to determine VS, ammonia concentration, hydrogen sulfide concentration, TVOC concentration and comprehensive odor concentration, respectively, and the corresponding removal rates were calculated. The measurement results are shown in Table 13.
Table 13 deodorizing effect display table
The results show that: under the special environmental conditions of high cold and anoxic state, easy icing in winter and weaker ecological system, the composite microbial inoculum provided by the invention can still play roles in degrading feces and removing malodor, and has strong environmental adaptability.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications, variations, alterations, substitutions made by those skilled in the art to the technical solution of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the spirit of the design of the present invention.
Sequence listing
<110> institute of biological research for adult and capital of academy of sciences in China
Sichuan Emergency Convenience Technology Co., Ltd.
<120> a deodorizing degradation composite bacterial agent, and preparation method and application thereof
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1465
<212> DNA
<213> Lactobacillus plantarum (Lactobacillus plantarum)
<400> 1
ggcgatgtgc tatgatgcag tcgaacgaac tctggtattg attggtgctt gcatcatgat 60
ttacatttga gtgagtggcg aactggtgag taacacgtgg gaaacctgcc cagaagcggg 120
ggataacacc tggaaacaga tgctaatacc gcataacaac ttggaccgca tggtccgagt 180
ttgaaagatg gcttcggcta tcacttttgg atggtcccgc ggcgtattag ctagatggtg 240
gggtaacggc tcaccatggc aatgatacgt agccgacctg agagggtaat cggccacatt 300
gggactgaga cacggcccaa actcctacgg gaggcagcag tagggaatct tccacaatgg 360
acgaaagtct gatggagcaa cgccgcgtga gtgaagaagg gtttcggctc gtaaaactct 420
gttgttaaag aagaacatat ctgagagtaa ctgttcaggt attgacggta tttaaccaga 480
aagccacggc taactacgtg ccagcagccg cggtaatacg taggtggcaa gcgttgtccg 540
gatttattgg gcgtaaagcg agcgcaggcg gttttttaag tctgatgtga aagccttcgg 600
ctcaaccgaa gaagtgcatc ggaaactggg aaacttgagt gcagaagagg acagtggaac 660
tccatgtgta gcggtgaaat gcgtagatat atggaagaac accagtggcg aaggcggctg 720
tctggtctgt aactgacgct gaggctcgaa agtatgggta gcaaacagga ttagataccc 780
tggtagtcca taccgtaaac gatgaatgct aagtgttgga gggtttccgc ccttcagtgc 840
tgcagctaac gcattaagca ttccgcctgg ggagtacggc cgcaaggctg aaactcaaag 900
gaattgacgg gggcccgcac aagcggtgga gcatgtggtt taattcgaag ctacgcgaag 960
aaccttacca ggtcttgaca tactatgcaa atctaagaga ttagacgttc ccttcgggga 1020
catggataca ggtggtgcat ggttgtcgtc agctcgtgtc gtgagatgtt gggttaagtc 1080
ccgcaacgag cgcaaccctt attatcagtt gccagcatta agttgggcac tctggtgaga 1140
ctgccggtga caaaccggag gaaggtgggg atgacgtcaa atcatcatgc cccttatgac 1200
ctgggctaca cacgtgctac aatggatggt acaacgagtt gcgaactcgc gagagtaagc 1260
taatctctta aagccattct cagttcggat tgtaggctgc aactcgccta catgaagtcg 1320
gaatcgctag taatcgcgga tcagcatgcc gcggtgaata cgttcccggg ccttgtacac 1380
accgcccgtc acaccatgag agtttgtaac acccaaagtc ggtggggtaa ccttttagga 1440
accagccgcc taaggtcaca gcgtg 1465
<210> 2
<211> 436
<212> DNA
<213> Pichia pastoris (Pichia fermentans)
<400> 2
ccgtaggtga acctgcggaa ggatcattac tgtgatttat acaccacact gcgtgggcga 60
cacgaaacac cgaaaccgaa cgcacgccgt caagcaagaa atccacaaaa ctttcaacaa 120
cggatctctt ggttctcgca tcgatgaaga gcgcagcgaa atgcgatacc tagtgtgaat 180
tgcagccatc gtgaatcatc gagttcttga acgcacattg cgcccgctgg tattccggcg 240
ggcatgcctg tctgagcgtc gtttccttct tggagcggag cttcagacct ggcgggctgt 300
ctttgtggac ggcgcgccca aagcgagggg ccttctgcgc gaactagact gtgcgcgcgg 360
ggcggccggc gaacttatta ccaagctcga cctcaaatca ggcaggagta cccgctgaac 420
ttaagcatat caataa 436
Claims (9)
1. A strain of pichia pastoris (Pichia fermentans) which is characterized in that: the microbial strain is preserved in China general microbiological culture Collection center (China) for 9 and 14 days in 2021, and the preservation addresses are as follows: the preservation number of the Beijing city Chaoyang area North Chen Xili No.1 and 3 is: CGMCC No.23387.
2. The use of the fermented pichia pastoris according to claim 1 for deodorization.
3. A deodorant prepared using the fermented pichia pastoris of claim 1.
4. A deodorant according to claim 3, characterized in that: the deodorant is a microbial composite microbial agent and comprises lactobacillus plantarum (Lactobacillus plantarum).
5. The deodorant according to claim 4, wherein: the lactobacillus plantarum deposit number is: CGMCC No. 23188.
6. The deodorant according to claim 4 or 5, characterized in that: the lactobacillus plantarum: pichia pastoris = 1-2:1-5.
7. The deodorant according to claim 6, wherein: the lactobacillus plantarum: pichia pastoris = 2:3.
8. A deodorant according to any one of claims 3 to 5, characterized in that: the concentration of viable bacteria in the deodorant is more than or equal to 4.25X10 8 CFU/mL。
9. Use of a deodorant according to any one of claims 4 to 8 for deodorizing, characterized in that: the deodorant is used in the following amount: the feed of the material to be treated is 7X 4.25X 10 per hundred grams 8 Deodorant with CFU viable bacteria amount. .
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| CN114774296B (en) * | 2022-05-18 | 2024-03-26 | 黄冈师范学院 | Pichia pastoris strain HGC34 resistant to high Wen Jiye and application thereof in deodorization and degradation of livestock manure |
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| CN102861350A (en) * | 2012-06-11 | 2013-01-09 | 中国科学院成都生物研究所 | Compound biological agent for deodorizing public toilets, and preparation method and application of compound biological agent |
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