Dimethyl sulfoxide degrading bacterial agent, preparation and application thereof

Abstract

The application discloses a dimethyl sulfoxide high-efficiency degrading bacterial agent, preparation and application, and belongs to the technical field of biology.The microbial bacterial agent for treating dimethyl sulfoxide wastewater mainly comprises Brucella pseudogrignonensis (LBQ23-3), is identified as Brucella pseudogrignonensis through 16S rDNA sequence sequencing, has been preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC NO.28605 and the preservation date is October 10, 2023.The dimethyl sulfoxide degrading bacterial agent can grow by taking dimethyl sulfoxide as the only carbon source, and can be completely degraded in 48 hours when the dimethyl sulfoxide concentration is 2000 mg / L.

Description

Technical Field

[0001] This invention belongs to the field of biological wastewater treatment, specifically relating to the preparation and application of a composite bacterial agent for treating dimethyl sulfoxide wastewater. Background Technology

[0002] Dimethyl sulfoxide (DMSO) is a commonly used organosulfur compound. It is a colorless and transparent organic compound used as a solvent and stabilizer, and is widely applied in the petroleum, chemical, and metallurgical industries. DMSO-contaminated wastewater typically contains high concentrations of organosulfur compounds, making it difficult to treat.

[0003] For wastewater containing DMSO, common treatment methods include biological methods, adsorption methods, and chemical oxidation methods. Adsorption methods involve passing DMSO-containing wastewater through an adsorption material, where the DMSO is adsorbed. Studies show that low concentrations of DMSO are adsorbed more effectively, while high concentrations are not adsorbed completely, requiring more frequent replacement or regeneration of the adsorption material. Chemical oxidation uses oxidants to oxidize DMSO, but this produces significant amounts of dimethyl sulfide, dimethyl disulfide, and methanethiol, resulting in high reagent costs and secondary pollution requiring further treatment. Biological methods are currently one of the main treatment methods, introducing microorganisms adapted to DMSO to degrade it without secondary pollution. However, high concentrations of DMSO can have toxic effects on these microorganisms, reducing treatment effectiveness.

[0004] DMSO wastewater has poor biodegradability, with a BOD5 / COD ratio that can be as low as 0.02, making it unsuitable for direct treatment using conventional biochemical degradation methods. Biodegradation is widely used for the degradation of recalcitrant compounds due to its low cost, high efficiency, and the abundance of natural microbial resources. Lu Yu et al. screened a DMSO-degrading bacterium, identified as *Pseudomonas alkalophilus* (…). Pseudomonas alcaliphila The degradation characteristics of DMSO were studied. With an initial DMSO concentration of 200 mg / L, the degradation rate was 94.8% after 72 hours. Takako Murakami-Nitta et al. isolated a strain of *Cryptococcus terreus* (…). Cryptococcus humicolus ), and can completely degrade 50 mg / L DMSO in 48 hours. Zuo Xiaoyuan et al. screened a complex bacterial community from soil that could grow in DMSO inorganic salt medium and isolated a filamentous denitrifying bacterium ( Hyphom icrobium denitrificans This strain showed a 91.3% degradation rate of 100 mg / L DMSO in 24 hours, and a 39.8% degradation rate in 96 hours at a DMSO concentration of 18.8 mmol / L (1469 mg / L). Patent CN 116004474 B discloses a strain of *Aureobacterium intermedius* (…). Ochrobactrum intermediumIt can degrade 5000 mg / L of DMSO by 100% in 24 hours. However, there are still relatively few strains that can degrade DMSO, and most strains are not very effective at degrading DMSO concentrations above 1000 mg / L. Therefore, screening more types of high-concentration DMSO-degrading bacteria can help treat DMSO-containing wastewater and improve the adaptability of strains to different types of wastewater. Summary of the Invention

[0005] This invention addresses the problems existing in the prior art by disclosing a dimethyl sulfoxide degrading bacterial agent, its preparation, and its application, utilizing Brucella pseudogrigoniae (… Brucella pseudogrignonensis The microbial agent prepared by LBQ23-3 can completely degrade dimethyl sulfoxide concentration of 2000 mg / L in wastewater through aerobic biodegradation.

[0006] This invention is implemented as follows: A type of pseudogrinnioni (Brucella pseudogrinnioni) Brucella pseudogrignonensis LBQ23-3 is characterized in that it is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCCNO.28605 and deposit date of October 10, 2023.

[0007] Deposit date: October 10, 2023 Name of depositary institution: China General Microbiological Culture Collection Center, China Microbiological Culture Collection Committee Accession number: CGMCC NO.28605 Address of the depository: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing Classification and nomenclature: Brucella pseudogrigione ( Brucella pseudogrignonensis )LBQ23-3

[0008] <110> Jiangsu Lanbisheng Chemical Environmental Protection Co., Ltd. <120> A DMSO-degrading bacterial agent, its preparation and application <160> 1 <170> SIPOSequenceListing 1.0 <210> 1 <211> 1417 <212> DNA <213> Brucella pseudogrignonensis LBQ23-3 (CGMCC 28605) <400> 1 ggctcagaac gaacgctggc ggcaggctta accatgcaa gtcgagcgcc ccgcagggg 60 agcggcagac gggtgagtaa cgcgtgggaa tctacctttt gctacggaat aactcaggga 120 aacttgtgct ataccgtat gtgcccttcg ggggaagat ttcggcaa aggatgagcc 180 cgcgttggat tagctagttg gtggggtaaa ggccaccaa ggcgacgatc catagctggt 240 ctgagaggat gatcagccac actgggactg agacacggcc cagactccta cgggaggcag 300 cagtggggaa tattggacaa tgggcgcaag cctgatccag ccatgccgcg tgagtgatga 360 aggccctagg gttgtaaagc tcttcaccg gtgaagataa tgacggtaac cggagaagaa 420 gcccggcta acttcgtgcc agcagccgcg gtaatacgaa gggggctagc gttgttcgga 480 tttactgggc gtaaagcgca cgtaggcgga ctttaagtc aggggtgaa tcccagagct 540 caactctgga actgcctttg atactggag tcttgagtat ggtagaggtg agtggaatttc 600 cgagtgtaga gtgaaatttc gtagatattc ggaggacac cagtggcgaa ggcggctcac 660 tggaccatta ctgacgctga ggtgcgaaag cgtggggagc aacaggatt agataccctg 720 gtagtccacg ccgtaaacga tgaatgttag ccgtcggggt gtttacactt cggtggcgca 780 gctaacgcat taaacattcc gcctggggag tacggtcgca agattaaaac tcaaaggaat 840 tgacgggggc ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac gcgcagaacc 900 ttaccagccc ttgacatacc ggtcgcggac acagagatgt gtctttcagt tcggctggac 960 cggatacagg tgctgcatgg ctgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc1020 gcaacgagcg caaccctcgc ctttagttgc catcatttag ttgggcactc taaagggact1080 gccagtgata agctggagga aggtggggat gacgtcaagt cctcatggcc cttacgggct1140 gggctacaca cgtgctacaa tggtggtgac agtgggcagc aagcacgcga gtgtgagcta1200 atctccaaaa gccatctcag ttcggattgc actctgcaac tcgagtgcat gaagttggaa1260 tcgctagtaa tcgcggatca gcatgccgcg gtgaatacgt tcccgggcct tgtacacacc1320 gcccgtcaca ccatgggagt tggttctgcc cgaaggcact gtgctaaccg taaggaggca1380 ggtgaccacg gtagggtcag cgactggggt gaagtcg 1417 A preparation method of a dimethyl sulfoxide degrading bacterial agent, which is characterized in that the method is specifically as follows: Step 1: Solid plate culture: Place *Brucella pseudogrigonia* (…) on a solid plate. Brucella pseudogrignonensis LBQ23-3 was transferred to solid culture medium and incubated at 30°C for 36 hours to activate the strain; Step 2, Seed culture: Inoculate the strain activated in Step 1 into liquid culture medium and culture at 30℃ and 150-200 rpm for 24-36 hours with shaking until the strain is in the logarithmic growth phase. Step 3: Fermentation and scale-up of the strain: Inoculate the culture medium obtained in Step 2 with 10% of the culture solution and carry out fermentation culture; obtain a concentration of 1×10 10 ~1×10 11 CFU / mL fermentation product; Step 4: Add 20% protectant to the fermentation product obtained in Step 4 to prepare a dry powder inoculum.

[0009] Furthermore, the solid culture medium in step one is prepared as follows: 15 g / L tryptone, 5 g / L soybean peptone, 5 g / L sodium chloride, and 20 g / L agar; dissolved in deionized water, pH 7-8, and sterilized at 115-121°C for 15-30 min.

[0010] Furthermore, the preparation method of the liquid culture medium in step two is as follows: tryptone 5-10 g / L, soybean peptone 1-5 g / L, glucose 1-2 g / L, sodium chloride 1-5 g / L, dissolved in deionized water, pH=7.0-8.0, sterilized at 115-121℃ for 15-30 min.

[0011] Furthermore, the fermentation culture medium for step three is as follows: 5-10 g / L tryptone, 1-5 g / L soybean peptone, 1-5 g / L glucose, 1-2 g / L NH4Cl, 0.1-0.2 g / L K2HPO4, 0.1-0.5 g / L KH2PO4, 1-5 mL trace element solution, 0.2% antifoaming agent, dissolved in distilled water, pH = 7.0-8.0, sterilized at 115-121℃ for 15-30 min.

[0012] Furthermore, the fermentation conditions in step three are as follows: tank temperature 30℃, tank pressure 0.01~0.1MPa, aeration rate 1:0.6~1.2, stirring speed 150rpm, and fermentation time 36~48h.

[0013] Furthermore, the protective agent formula in step five is: 35% yeast powder; 35% fish protein; and 30% trace elements.

[0014] Furthermore, the formula of the trace element solution is as follows: MgSO4 0.5 g / L; MnSO4 0.1 g / L; H3BO3 0.1 g / L; ZnSO4·7H2O 0.1 g / L; FeSO4·7H2O 0.3 g / L; pH=7.

[0015] This invention also discloses the application of dimethyl sulfoxide degrading bacterial agents, which is achieved through the following methods: The dimethyl sulfoxide (DMSO) degrading bacterial agent produced in the above steps is added at a mass ratio of 0.5-5‰, with 0.5 g / L glucose and 0.5-1 g / L sucrose (or 0.5 g / L glucose, 0.5 g / L sucrose, and 0.2 g / L DMSO), tap water, pH 7-8, and cultured in a bacterial culture tank at 30-35℃ with aeration (DO 2-4 mg / L) for 48 hours to activate the bacterial strain. Further, after activation, the bacterial density is measured; microscopic examination shows a density of 1×10⁻⁶. 9 A concentration of CFU / mL or higher indicates successful activation culture, resulting in a suspension of activated bacteria prepared with dimethyl sulfoxide (DMSO) degrading agent.

[0016] Furthermore, the activated bacterial suspension of the dimethyl sulfoxide degrading agent was added to an inorganic salt culture medium containing 2000 mg / L dimethyl sulfoxide at a mass ratio of 5-10‰. After 48 hours of aerobic treatment (30-35℃, DO 2-4 mg / L), dimethyl sulfoxide was completely degraded.

[0017] Adding a suspension of activated dimethyl sulfoxide (DMSO) degrading bacteria at a concentration of 5-10‰ to a pesticide production wastewater containing 2049 mg / L DMSO, the DMSO can be completely degraded by aerobic degradation (25-35℃, DO 2-4 mg / L) over 48 hours.

[0018] The advantages of this invention compared to the prior art are as follows: 1. The dimethyl sulfoxide degrading bacterial agent provided by this invention can treat high-concentration dimethyl sulfoxide wastewater by a fully biological method, with good effect, low cost and no secondary pollution.

[0019] 2. The dimethyl sulfoxide degrading bacterial agent provided by the present invention has strong tolerance to dimethyl sulfoxide and can rapidly degrade high concentrations of dimethyl sulfoxide. When the concentration of dimethyl sulfoxide is ≤2000mg / L, it can be completely degraded by aerobic degradation in 48 hours. Attached Figure Description

[0020] Figure 1 Brucella pseudogrigonia in Example 1 of this invention ( Brucella pseudogrignonensis LBQ23-3 colony morphology.

[0021] Figure 2 This shows the degradation of dimethyl sulfoxide at different concentrations over time in Example 2 of the present invention. Detailed Implementation

[0022] To make the objectives, technical solutions, and effects of this invention clearer and more explicit, the following examples provide a more detailed description of the invention. It should be noted that the specific embodiments described herein are merely illustrative and not intended to limit the scope of the invention.

[0023] The main component of the dimethyl sulfoxide degrading bacterial agent of this invention is *Brucella pseudogrinnioni* (…). Brucella pseudogrignonensis LBQ23-3, microbial preparation with an effective viable count ≥10 10 The preparation method of CFU / g dimethyl sulfoxide degrading bacterial agent includes the following sequence and steps: Step 1: Solid plate culture: Place *Brucella pseudogrigonia* (…) on a solid plate. Brucella pseudogrignonensis LBQ23-3 was transferred to solid culture medium and incubated at 30℃ for 36 hours to activate the strain. The solid culture medium consisted of 15 g / L tryptone, 5 g / L soybean peptone, 5 g / L sodium chloride, and 20 g / L agar. The solution was dissolved in deionized water, pH 7–8, and sterilized at 121℃ for 20 minutes. Step 2, Seed Culture: Inoculate the strain activated in Step 1 into liquid culture medium and incubate at 30℃ with shaking at 150–200 rpm for 24–36 hours, until the strain reaches the logarithmic growth phase; Culture medium: 5-10 g / L tryptone, 1-5 g / L soybean peptone, 1-2 g / L glucose, 1-5 g / L sodium chloride, dissolved in deionized water, pH 7.0-8.0, sterilized at 121℃ for 20 min.

[0024] Step 3: Fermentation and scale-up of the strain: Inoculate the culture medium obtained in Step 2 with 10% of the culture solution and carry out fermentation culture; obtain a concentration of 1×10 10 ~1×10 11 CFU / mL fermentation product; fermentation expansion medium as follows: tryptone 5-10 g / L, soybean peptone 1-5 g / L, glucose 1-5 g / L, NH4Cl 1-2 g / L, K2HPO4 0.1-0.2 g / L, KH2PO4 0.1-0.5 g / L, trace element solution 1-5 mL, antifoaming agent 0.2%, dissolved in distilled water, pH=7.0-8.0, sterilized at 121℃ for 20 min.

[0025] Step 4: Add 20% protectant to the fermentation product obtained in Step 4 to prepare a dry powder inoculum. The protectant formula is: yeast powder 35%; fish protein 35%; trace elements 30%. The trace element solution formula is: MgSO4 0.5g / L; MnSO4 0.1g / L; H3BO3 0.1g / L; ZnSO4·7H2O 0.1g / L; FeSO4·7H2O 0.3g / L; pH=7.

[0026] The application of a dimethyl sulfoxide (DMSO) degrading bacterial agent is characterized in that the treatment method involves sequentially activating and cultivating the DMSO-degrading bacterial agent followed by wastewater treatment. The specific implementation scheme is as follows: Add 0.3-1% (w / w) of dimethyl sulfoxide (DMSO) degrading bacterial agent, 0.5 g / L glucose, 0.5 g / L sucrose, and tap water (pH 7-8) to a culture tank at 30-35℃ with aeration (DO 2-4 mg / L) for 48 hours to activate the bacterial strain. Further, after activation, measure the bacterial density; if it reaches 1×10⁻⁶, microscopic examination reveals a density of 1×10⁻⁶. 9 Once the concentration of CFU / mL reaches a certain level, the activation culture is complete, forming a suspension of activated bacteria with dimethyl sulfoxide degrading agent.

[0027] Adjust the wastewater to pH 7-8, and ensure the concentration of dimethyl sulfoxide in the system influent is less than 500 mg / L (or TOC less than 500 mg / L, with TOC taking priority). You can add 300 mg / L of sucrose to promote the growth of the microorganisms. The implementation method of using dimethyl sulfoxide (DMSO) degrading bacteria and polyurethane packing is as follows: The activated bacterial suspension of DMSO degrading bacteria is added to a reaction tank containing 70% polyurethane packing in 2-3 batches at a mass ratio of 5-10%. The residence time is controlled at 24-72 hours. Samples are taken to measure indicators such as COD, TOC, and DMSO. After the DMSO degradation rate is greater than 90%, the removal effect is stable for 5-10 days. The concentration of DMSO in the influent is gradually increased by a DMSO concentration gradient of 200-500 mg / L (or a TOC gradient of 300-500 mg / L, with TOC taking priority). The water volume is replaced by 50-80% in each batch until the influent load reaches the target influent DMSO concentration.

[0028] The dimethyl sulfoxide (DMSO) degrading bacterial agent provided by this invention can be used to treat DMSO wastewater. A suitable biochemical treatment process can be selected based on the actual site conditions: for DMSO concentrations ≤2000 mg / L, an aerobic biochemical process can be directly used, or other conventional biochemical combination processes can be applied; the aerobic reaction tank is inoculated with the DMSO degrading bacterial agent and polyurethane packing material at a volume ratio of 50-70%. The hydraulic retention time (HRT) is as follows: for influent DMSO concentrations less than 500 mg / L, HRT is 12 h; for influent DMSO concentrations of 1000 mg / L, HRT is 24 h; for influent DMSO concentrations of 1500 mg / L, HRT is 24-36 h; for influent DMSO concentrations of 2000 mg / L, HRT is 36-48 h.

[0029] The following are specific examples illustrating the application of the dimethyl sulfoxide degrading bacterial agent: Example

[0030] Strain acquisition and identification. The steps for obtaining and identifying strains are as follows: Step 1: Sample Collection and Strain Acclimation Activated sludge from the wastewater treatment biochemical tank of a pesticide production company in Zhejiang Province was added to an inorganic salt culture medium containing 500 mg / L dimethyl sulfoxide as the sole carbon and nitrogen source. The inoculum size of the activated sludge was 10% (V / V). The culture was acclimated and cultured for 5 days at 30℃ and 150 r / min in a constant temperature shaker to obtain acclimation culture solution 1. After the culture was completed, acclimation culture solution 1 was inoculated into an inorganic salt culture medium containing 500 mg / L dimethyl sulfoxide. The inoculum size of acclimation culture solution 1 was 10% (V / V). The culture was then cultured for 5 days to obtain acclimation culture solution 2. The above acclimation culture process was repeated continuously until the dimethyl sulfoxide degradation rate was greater than 90%, thus obtaining the dimethyl sulfoxide degrading bacteria acclimation culture solution.

[0031] The inorganic salt culture medium formula is as follows: KH2PO4 0.3 g / L, MgSO4·7H2O 0.2 g / L, FeSO4·7H2O 0.02 g / L, MnSO4·H2O 0.02 g / L, CaCl2·6H2O 0.01 g / L, pH 7.0.

[0032] Step 2: Strains Isolation and Purification Take 1 mL of the dimethyl sulfoxide degrading bacteria acclimatization culture and dilute it to 10 with autoclaved deionized water. -1 10 -2 10 -3 10 -4 10 -5 10 -6 10 -7 10 -8 10 -9The bacterial suspensions at a concentration of 1:1 were spread onto inorganic salt solid medium containing 500 mg / L dimethyl sulfoxide and incubated at 30°C for 2-3 days. After single colonies grew on the medium, the single colonies with the largest number and the same morphology were picked and purified by streak plating three times to obtain the target strain LBQ23-3, which was then stored at 4°C.

[0033] Step 3: Identification of strain LBQ23-3 Strain strain LBQ23-3 was inoculated onto beef extract peptone agar and incubated at 30℃ for 48 h. The colonies of strain LBQ23-3 were observed to be round, grayish-white, smooth, with regular edges, and opaque. (See attached image for colony morphology.) Figure 1 After Gram staining, it appears as a Gram-negative rod-shaped organism under a microscope. The 16S rDNA of strain LBQ23-3 was sequenced by PCR, and the 16S rDNA gene sequence of strain LBQ23-3 is shown in the sequence listing. BLAST comparison analysis using the NCBI gene bank was performed, and strain LBQ23-3 was compared with samples in the database. Brucella pseudogrignonensis The strain showed 100.00% homology. Combined with physiological and biochemical identification of strain LBQ23-3, it was determined that strain LBQ23-3 belongs to *Brucella pseudogrinnioni*, and was named... Brucella pseudogrignonensis .LBQ23-3.

[0034] The beef extract peptone solid medium is made by adding 20 g / L agar to the beef extract peptone medium; the beef extract peptone medium is made by adding 3 g of beef extract, 10 g of peptone, 5 g of NaCl, 1 L of distilled water, adjusting the pH to 7.5 ± 0.2, and autoclaving at 121 °C for 20 min.

[0035] Step 4, bacterial strain Brucella pseudogrignonensis .LBQ23-3 Collection strain Brucella pseudogrignonensis LBQ23-3 was deposited on October 10, 2023, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC NO.28605. Example

[0036] The dimethyl sulfoxide (DMSO) degrading bacterial agent provided by this invention was inoculated at a mass ratio of 1‰ into an inorganic salt culture medium containing 200 mg / L DMSO, 0.2 g / L glucose, and 0.5 g / L sucrose to activate the bacterial strain. After inoculation, the culture was placed in a shaker at 30°C and 150 r / min for 48 h until the bacterial density reached 1 × 10⁻⁶. 9 CFU / mL, forming a dimethyl sulfoxide-degrading bacterial agent activation suspension.

[0037] The aforementioned dimethyl sulfoxide (DMSO) degrading bacterial agent activation suspension was inoculated at a 5% (v / v) inoculation rate into inorganic salt culture media containing DMSO concentrations of 500 mg / L, 1000 mg / L, 1500 mg / L, 2000 mg / L, 2500 mg / L, and 3000 mg / L, respectively. The media were incubated at 30°C in a shaker, and samples were taken at different time points to determine the residual DMSO content. The results are as follows: Figure 2 As shown, at a concentration of 500 mg / L dimethyl sulfoxide (DMSO), complete degradation occurred in 12 hours; at 1000 mg / L DMSO, complete degradation occurred in 24 hours; at 1500 mg / L DMSO, complete degradation occurred in 36 hours; and at 2000 mg / L DMSO, complete degradation occurred in 48 hours. Specific results are illustrated in the figure below. Figure 1 As shown.

[0038] In some embodiments, the inorganic salt culture medium may include any one nitrogen-containing inorganic substance selected from (NH4)2SO4, NH4Cl, K2HPO4, KH2PO4, NaCl, Na2SO4, MgSO4·7H2O, MnSO4·H2O, and FeSO4·7H2O, or a combination of one or more nitrogen-containing inorganic substances and any one or more other inorganic substances. In Example 2, the inorganic salt culture medium formulation is as follows: NH4Cl 0.3 g / L, KH2PO4 0.2 g / L, MgSO4·7H2O 0.1 g / L, FeSO4·7H2O 0.02 g / L, MnSO4·H2O 0.02 g / L, CaCl2·6H2O 0.01 g / L, pH 7.5. Example

[0039] The dimethyl sulfoxide (DMSO) degrading bacterial agent provided by this invention was added to wastewater containing DMSO. The wastewater was taken from a pesticide production company in Anhui Province. The raw wastewater had a COD of 5133 mg / L, TOC of 2328 mg / L, and DMSO of 2049 mg / L. The initial influent DMSO concentration was 500 mg / L. The pH was adjusted to 7.5, and 70% polyurethane packing was added. 0.2% of the DMSO degrading bacterial agent was added by mass. The wastewater was aerobically cultured at 30℃ (DO 2-4 mg / L). Samples were taken to measure COD, TOC, DMSO, and other indicators. The batch water exchange rate was 80%, and the HRT was 24-48 h. The DMSO degradation rate was greater than 90% and the removal effect was stable for 4-6 days. The influent DMSO concentration was gradually increased from 500 mg / L to 2049 mg / L of the raw wastewater. Each load of influent was maintained for 6-8 days, and the treated raw wastewater DMSO concentration was 2049 mg / L. After three batches of water were replaced (95% capacity), samples were taken at different time points after the water replacement to determine the residual dimethyl sulfoxide (DMSO) content. The average value of the data at the same residence time under three water replacement cycles (95% capacity) was calculated. The results showed that the raw water DMSO concentration was 2049 mg / L. After 36 hours, the DMSO concentration could be degraded to below 5 mg / L, and after 48 hours, the DMSO was completely degraded. After 48 hours, the effluent COD was reduced to 79-116 mg / L, with a COD removal rate of approximately 98%. After 48 hours, the effluent TOC was 29-38 mg / L, with a TOC removal rate of approximately 98%.

[0040] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements can be made without departing from the principle of the present invention, and these improvements should also be considered within the scope of protection of the present invention.

Claims

1. A Brucella inopinata bacterium (Brucella inopinata LBQ23-3) characterized in that, Brucella pseudogrignonensis It is deposited at the China General Microbiological Culture Collection Center, accession number: CGMCC NO.28605, and the deposit date is October 10, 2023. ​ 2. A method for preparing a dimethyl sulfoxide-degrading microbial agent, characterized by, The method is as follows: the dimethyl sulfoxide degrading agent contains Brucella pseudogrigonia ( Brucella pseudogrignonensis LBQ23-3, the preparation method is as follows: Step one, solid plate culture: the Brucella inopinata (LBQ23-3) was transferred to solid culture medium and cultured in a 30°C incubator for 36h to activate the strain; Brucella pseudogrignonensis Step two, liquid culture: the activated strain was inoculated into liquid culture medium and cultured at 30°C for 24h. Step 2, Seed culture: Inoculate the strain activated in Step 1 into liquid culture medium and culture at 30℃ and 150-200 rpm for 24-36 hours with shaking until the strain is in the logarithmic growth phase. Step 3: Fermentation and scale-up culture of the strain: Inoculate the seed liquid described in Step 2 into the fermentation medium at a ratio of 10% for fermentation culture; Step 4: Add 20% protectant to the fermentation product obtained in step 3 to prepare a dry powder inoculum.

3. The method for preparing the dimethyl sulfoxide degrading bacterial agent according to claim 2, characterized in that, The solid culture medium in step one is prepared as follows: 15 g / L tryptone, 5 g / L soybean peptone, 5 g / L sodium chloride, and 20 g / L agar; dissolved in deionized water, pH 7-8, and sterilized at 115-121℃ for 15-30 min.

4. The method for preparing the dimethyl sulfoxide degrading bacterial agent according to claim 2, characterized in that, The preparation method of the liquid culture medium in step two is as follows: tryptone 5-10 g / L, soybean peptone 1-5 g / L, glucose 1-2 g / L, sodium chloride 1-5 g / L, dissolved in deionized water, pH=7.0-8.0, sterilized at 115-121℃ for 15-30 min.

5. The method for preparing the dimethyl sulfoxide degrading bacterial agent according to claim 2, characterized in that, The fermentation culture medium for step three is as follows: tryptone 5-10 g / L, soybean peptone 1-5 g / L, glucose 1-5 g / L, NH4Cl 1-2 g / L, K2HPO4 0.1-0.2 g / L, KH2PO4 0.1-0.5 g / L, trace element solution 1-5 mL, antifoaming agent 0.2%, dissolved in distilled water, pH=7.0-8.0, sterilized at 115-121℃ for 15-30 min; the formula of the trace element solution is: MgSO4 0.5 g / L; MnSO4 0.1 g / L; H3BO3 0.1 g / L; ZnSO4·7H2O 0.1 g / L; FeSO4·7H2O 0.3 g / L; pH=7; The fermentation conditions in step three are as follows: tank temperature 30℃, tank pressure 0.01~0.1MPa, aeration rate 1:0.6~1.2, stirring speed 150rpm, and fermentation time 36~48h.

6. The method for preparing the dimethyl sulfoxide degrading bacterial agent according to claim 2, characterized in that, The protective agent formulation in step four is as follows: yeast powder 35%; fish peptone 35%; trace element solution 30%; the trace element solution formulation is as follows: MgSO4 0.5g / L; MnSO4 0.1g / L; H3BO3 0.1g / L; ZnSO4·7H2O 0.1g / L; FeSO4·7H2O 0.3g / L; pH=7.

7. A dimethyl sulfoxide degrading bacterial agent prepared by the preparation method according to any one of claims 2 to 6.

8. The dimethyl sulfoxide-degrading bacterial agent according to claim 7, characterized by, The effective viable bacteria quantity in the dimethyl sulfoxide degrading bacteria agent is ≥10 10 CFU / g.

9. Use of the dimethyl sulfoxide-degrading bacterial agent according to claim 7 in chemical industrial wastewater, characterized by, Adding dimethyl sulfoxide (DMSO) degrading bacteria to an inorganic salt culture medium containing 2000 mg / L DMSO at a ratio of 0.5-5% resulted in complete degradation of DMSO within 48 hours. Adding dimethyl sulfoxide (DMSO) degrading bacteria to a pesticide production wastewater containing 2049 mg / L DMSO at a ratio of 0.5-5‰ also resulted in complete degradation of DMSO within 48 hours.

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