Sphingobacterium mizutani and its application in biological treatment of methylpyridine wastewater

Abstract

The invention discloses a Sphingobacterium mizugami and a biological treatment method for methylpyridine wastewater, and belongs to the field of biological treatment of methylpyridine wastewater. In the treatment method, the wastewater is activated and cultured with solid bacteria for methylpyridine degradation in sequence. The process is simple to operate and has high reaction efficiency, and methylpyridine can be degraded. The methylpyridine degradation bacterial agent in the invention is a microbial agent prepared by Sphingobacterium mizugami. Sphingobacterium mizugami ( Sphingobacterium mizutaii ) was deposited in the General Microbiological Center of China Microorganism Culture Collection Administration Committee on July 25, 2024, with the deposit number: CGMCC NO.31454. It can treat 2-methylpyridine, 3-methylpyridine or 4-methylpyridine wastewater with an influent methylpyridine concentration of 2000 mg / L. The method of the present invention is a bio-augmentation of the common biological method, which broadens the scope of application of biological treatment of methylpyridine wastewater.

Description

Technical Field

[0001] The invention belongs to the field of biological wastewater treatment, and in particular relates to a biological treatment method for methylpyridine wastewater. Background Art

[0002] Methylpyridine refers to pyridine compounds with methyl groups at different substitution positions. There are three main types of methylpyridine: 2-methylpyridine, 3-methylpyridine, and 4-methylpyridine. 2-methylpyridine is mainly used in the synthesis of chemicals such as medicines, dyes, and rubbers, and is also used as a solvent and experimental reagent; 3-methylpyridine is mainly used in the fields of pesticides, medicines, feed additives, and spices. In recent years, domestic pyridine pesticides have developed rapidly, and the demand for 3-methylpyridine is very promising; 4-methylpyridine is mainly used in the production of the drug isoniazid, the antidote diphosphonium and diphosphonium, and is also used in the production of pesticides, dyes, rubber additives, and synthetic resins, and can also be used as a solvent. Methylpyridine in the environment mainly comes from industrial production, pharmaceutical synthesis, dye and pesticide synthesis, and agricultural use. Methylpyridine is easily soluble in water. With the discharge of industrial wastewater, methylpyridine has become a common pollutant in water.

[0003] Most of the research on the biodegradation of methylpyridine at home and abroad focuses on synergistic degradation, co-metabolism and degradation pathways, and there are relatively few studies on efficient methylpyridine degrading bacteria. Current research has shown that 2-methylpyridine can be biodegraded in anoxic environments. KaiserJP discovered the degradation mechanism of 2-methylpyridine through research. During aerobic degradation, the methyl group will first be hydroxylated, then carboxylated, and 2-methylpyridine will eventually become a short-chain fatty acid. Yu Jianming et al. studied Gordonia ZJJ and its application in the degradation of 3-methylpyridine. The bacteria can degrade 190 mg / L of 3-methylpyridine; Fan Feng et al. screened a strain of Staphylococcus xylous B1, which had a degradation rate of 89.3% for 100 mg / L of 2-methylpyridine. Mana et al. studied the anoxic degradation effect of 2-methylpyridine and its co-degradation performance with methanol. When the initial 2-methylpyridine concentration was 100 mg / L, the anoxic degradation rate of 2-methylpyridine as a single substrate was 2.9 [mg / g (MLSS)·h], and the degradation rate constant of 2-methylpyridine with methanol was 3.9 [mg / g (MLSS)·h]. He Miao of Tsinghua University studied typical organic matter in coking wastewater through the Wahoo meter determination method and found that pyridine and methylpyridine had a synergistic effect, and the degradation of pyridine was conducive to the degradation of methylpyridine.

[0004] Methylpyridine has an inhibitory effect on microorganisms. Studies have shown that the oxidation products of chloromethylpyridine bind to membrane proteins indiscriminately, affecting the activity of AMO (ammonia monooxygenase), thereby inhibiting ammonia-oxidizing microorganisms. Conventional biological methods for treating methylpyridine wastewater have disadvantages such as low influent concentration limit, unstable operation, and low degradation efficiency. Although there are relatively few studies on biological methods for treating methylpyridine wastewater, existing studies have shown that it is possible to achieve effective treatment of this type of wastewater by screening and utilizing specific high-efficiency degradation bacteria. Summary of the invention

[0005] The present invention aims at the problems existing in the prior art and discloses a method for biological treatment of methylpyridine wastewater, which can degrade methylpyridine. The method is a biological enhancement of a common biological method and broadens the scope of application of biological treatment of methylpyridine wastewater. In addition, the process is simple to operate and has a high reaction efficiency. The methylpyridine degradation agent is a microbial agent prepared by Sphingobacillus aquatica, which can treat 2-methylpyridine, 3-methylpyridine or 4-methylpyridine wastewater with an influent methylpyridine concentration of 2000 mg / L.

[0006] The present invention is achieved in that:

[0007] A method for biological treatment of methylpyridine wastewater, characterized in that the biological treatment method comprises the following steps:

[0008] Step 1: Activation and cultivation of methylpyridine-degrading bacteria:

[0009] 1.1. Add 1-5% by mass of solid agent of methylpyridine-degrading bacteria, 0.5-1.5 g / L of glucose, and tap water. The amount of tap water is added according to the proportion of solid agent of methylpyridine-degrading bacteria, that is, 1 kg of solid agent corresponds to 1-5 t of tap water. Add dilute sulfuric acid or liquid alkali to adjust the pH to 7-8, and culture in a culture expansion tank at 25-35°C for 48 hours to activate the bacteria. DO2-4 mg / L.

[0010] 1.2. After the activation of the bacteria, the bacterial density was tested under a microscope and the bacterial density reached 1×10 9 CFU / mL is the completion of activation culture.

[0011] Step 2: Biological treatment of wastewater:

[0012] 2.1. The wastewater containing methylpyridine adjusted to pH 7-8 is passed into the biological treatment tank, the temperature is controlled at 25-35°C, the reaction time and dissolved oxygen are controlled, and 10% activated cultured methylpyridine-degrading bacterial liquid is added to the biological treatment tank.

[0013] 2.2. When the biological treatment system is started, the influent methylpyridine concentration is less than 500 mg / L, or the TOC is less than 400 mg / L. During the first 1 to 3 days after the system is started, 200 to 300 mg / LCOD equivalent carbon source is added every day. The carbon source is glucose or methanol. No more carbon source is added after 3 days.

[0014] 2.3. Load increase and full load: Increase the load in a gradient from 300 to 500 mg / L of picoline concentration or from 300 to 400 mg / L of TOC, with TOC given priority. Replace 80% of the water in each batch until the influent load reaches the target influent picoline concentration. The full load influent picoline concentration can reach 2000 mg / L or the TOC concentration can reach 2000 to 2500 mg / L.

[0015] Furthermore, the hydraulic retention time (HRT) of the wastewater biological treatment is: when the influent methylpyridine concentration is 300-500 mg / L, the HRT is 24 h; when the influent methylpyridine concentration is 500-1000 mg / L, the HRT is 48 h; when the influent methylpyridine concentration is 1500-2000 mg / L, the HRT is 72 h.

[0016] Furthermore, the biological treatment system adopts an aerobic biological treatment process and uses membrane aeration to supply oxygen, with a dissolved oxygen (DO) of 2 to 4 mg / L.

[0017] Furthermore, the methylpyridine-degrading bacteria solid agent is characterized in that it is a microbial agent prepared by Sphingobacterium mizutani, and the Sphingobacterium mizutani is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with a deposit number of CGMCC NO.31454 and a deposit date of July 25, 2024.

[0018] Furthermore, the dosage and method of adding the methylpyridine-degrading bacteria are as follows: the methylpyridine-degrading bacteria agent is added into the reaction tank in 2 to 3 batches at a dosage of 10%, and aerated culture is carried out at 25 to 35° C., and the residence time is controlled to be 48 to 72 hours.

[0019] Furthermore, when the TOC degradation rate is greater than 85% and the TOC degradation effect is stable for 5 to 10 days, the picoline concentration is increased by 300 to 500 mg / L or the TOC concentration is increased by 300 to 400 mg / L, with TOC given priority. 50 to 80% of the water is replaced in each batch until the influent load reaches the target influent picoline concentration; the full-load influent picoline concentration can reach 2000 mg / L or the TOC concentration is 2000 to 2500 mg / L.

[0020] Microbiological deposit information:

[0021] Deposit date: July 25, 2024

[0022] Name of depository: China National Microbiological Culture Collection Administration General Microbiology Center

[0023] Deposit number: CGMCC NO.31454

[0024] Address of the depository: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing

[0025] Classification name: Sphingobacterium mizutaii

[0026] Sequence Listing

[0027] <110> Jiangsu Lanbisheng Chemical Environmental Protection Co., Ltd.

[0028] <120> Sphingobacterium mizutani and its application in biological treatment of methylpyridine wastewater

[0029] <160> 1

[0030] <170> SIPOSequenceListing 1.0

[0031] <210> 1

[0032] <211> 1464

[0033] <212> DNA

[0034] <213> Sphingobacterium mizutaii.'CGMCC NO.31454' <400> 1

[0035] ggctcaggat gaacgctagc ggcaggccta atacatgcaa gtcggacggg atccatcggt60agcttgctac cgatggtgag agtggcgcac gggtgcgtaa cgcgtgagca acctgcccat120atcaggggga tagcccggag aaatccggat taacaccggt tgacccgct ttccggcatc180gggaggtagt caaatattca taggatatgg atgggctcgc gtgacattag ctagttggtg240gggtaacggc ccaccaaggc gacgatgtct aggggctctg agaggagaat cccccacact300ggtactgaga cacggaccag actcctacgg gaggcagtagcat taaggat tggtcaatgg360gggcaaccct gaaccagcca tgccgcgtgc aggacgactg ccctatgggt tgtaaactgc420ttttgttagg gaataaaccc cgctacgtgt agcgggctga atgtacctaa agaataagga480tcggctaact ccgtgcccgc atccggattt540attgggttta aagggtgcgt aggcggcact ttaagtcagg ggtgaaagac ggcagctcaa600ctgtcgcagt gcccttgata ctgaagtgct tgaatgcggt tgaagacggc ggaatgagac660aagtagcggt gaaatcggc agctgtctaa720gccgttattg acgctgatgc acgaaagcgt ggggatcgaa caggattaga taccctggta780gtccacgccc taaacgatga tgactcgatg tttgcgatat accgtaagcg tccaagcgaa840agcgttaagt catcacctg gggacgcgcgcgtgaaactcaaa ggaattgacg900ggggcccgca caagcggagg agcatgtggt ttaattcgat gatacgcgag gaaccttacc960cgggcttgaa agttatactgaa gggcgcagag acgcgcccgt ccttcgggac aggaaactag1020gtgctgcatg gctgtcgtca gctcgtgccg tgaggtgttg ggttaagtcc cgcaacgagc1080gcaaccccta tgtttagttg ccagcacgtc aaggtgggga ctctaaacag actgcctgcg1140caagcagaga ggaaggcggg gacgacgtca agtcatcatg gcccttacgt ccggggctac1200acacgtgcta caatggatgg tacagcgggc agctacacag caatgtggtg ccaatctcga1260aaagccattc acagttcgga tcggggtctg caactcgacc ccgtgaagtt ggattcgcta1320gtaatcgcgt atcagcaatg acgcggtgaa tacgttcccg ggccttgtac acaccgcccg1380tcaagccatg aaagctgggg gtacctaaag catgtaaccg caaggagcgt gttagggtaa1440aaccggtaat tggggctaag tcgt 1464

[0036] The beneficial effects of the present invention compared with the prior art are:

[0037] 1. The present invention can treat methylpyridine wastewater by a fully biological method, with good effect, low cost and no secondary pollution.

[0038] 2. The methylpyridine-degrading bacteria used in the present invention are microbial agents prepared by Sphingobacterium mizutaii, which was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration on July 25, 2024. It can treat methylpyridine wastewater, and when the methylpyridine concentration is 2000 mg / L, it can be efficiently biodegraded, and the organic matter degradation rate is higher than 95%.

[0039] 3. In the treatment method of the present invention, the wastewater is sequentially activated and cultured with a solid bacterial strain for degradation of picoline--the wastewater is biologically treated. The process is simple to operate and has a high reaction efficiency, and picoline can be degraded. The present invention can treat 2-picoline, 3-picoline or 4-picoline wastewater with an influent picoline concentration of 2000 mg / L. The method of the present invention is a biological enhancement of a common biological method, which broadens the scope of application of biological treatment of picoline wastewater. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 This is the colony morphology of Sphingobacterium mizutaii of the present invention. DETAILED DESCRIPTION

[0041] In order to make the purpose, technical solution and effect of the present invention clearer and more specific, the present invention is further described in detail by enumerating examples below. It should be noted that the specific implementation described here is only used to explain the present invention and is not used to limit the present invention.

[0042] A method for biological treatment of methylpyridine wastewater, characterized in that in the treatment method, the wastewater is sequentially activated and cultured with methylpyridine-degrading solid bacteria → biological treatment of wastewater, and the specific implementation scheme is as follows:

[0043] The method comprises adding 1-5% methylpyridine-degrading bacteria and 1-1.5 g / L glucose in a mass ratio, adding tap water in a proportion of solid bacterial agent of methylpyridine-degrading bacteria, i.e., 1 kg solid bacterial agent corresponds to 1-5 t tap water, adding dilute sulfuric acid or liquid alkali to adjust the pH to 7-8, and culturing the bacteria in a bacterial expansion tank at 25-35° C. with aeration (DO 2-4 mg / L) for 48 hours to activate the bacteria; the methylpyridine-degrading bacteria is a microbial agent prepared from Sphingobacillus mizunatum.

[0044] After the activation of the strain, the bacterial density was tested and the microscopic examination showed that it reached 1×10 9 CFU / mL is the completion of activation culture.

[0045] Adjust the pH of the wastewater to 7-8, the initial system influent methylpyridine concentration is less than 500 mg / L, or TOC is less than 400 mg / L, TOC is prioritized; add 200-300 mg / LCOD equivalent carbon source every day on the 1st to 3rd day after the system is started, the carbon source is glucose or methanol, and no carbon source is added after 3 days; when the TOC degradation rate is greater than 85%, the water is influent at a stable load for 5-10 days, and then the load is increased by a gradient of methylpyridine concentration of 300-500 mg / L or TOC by a gradient of 300-400 mg / L, TOC is prioritized, and 80% of the water is replaced each batch until the influent load reaches the target influent methylpyridine concentration; the full load influent methylpyridine concentration can reach 2000 mg / L or the TOC concentration is 2000-2500 mg / L.

[0046] The dosage and method of methylpyridine-degrading bacteria are as follows: the activated bacterial solution of methylpyridine-degrading bacteria is added into the reaction tank in 2-3 batches at a dosage of 10%, and aerated culture is carried out at 25-35° C., and the residence time is controlled to be 48-72 hours.

[0047] The biological treatment process of methylpyridine wastewater can adopt aerobic biochemical process, using membrane aeration to supply oxygen, and the dissolved oxygen is 2-4 mg / L. The hydraulic retention time is: when the influent methylpyridine concentration is 300-500 mg / L, HRT is 24h; when the influent methylpyridine concentration is 500-1000 mg / L, HRT is 48h; when the influent methylpyridine concentration is 1500-2000 mg / L, HRT is 72h.

[0048] The following specific examples are given to describe the application of the biological treatment method for methylpyridine wastewater: the experimental methods in the following embodiments, unless otherwise specified, are carried out in accordance with the technical conditions provided by the present invention.

[0049] The activation method of the solid agent of methylpyridine-degrading bacteria in the following embodiment is as follows: adding methylpyridine-degrading bacteria at a mass ratio of 1‰, glucose 0.5g / L, tap water, adjusting the pH to 7-8, aerating culture at 25-35°C for 48h to activate the bacteria, DO2-4mg / L, to form an activated bacterial solution.

[0050] Example 1, 2-methylpyridine biological treatment

[0051] The TOC of wastewater from a certain project was 1941 mg / L, of which the concentration of 2-methylpyridine was 2144 mg / L. The aerobic biological treatment pool was inoculated with 10% methylpyridine-degrading bacteria to activate the bacterial solution, with an HRT of 72h. After 15 days of operation, the effluent was stable, with an effluent TOC of 28-37 mg / L and a TOC degradation rate of 98%. No 2-methylpyridine was detected.

[0052] Example 2, 3-methylpyridine biological treatment

[0053] The methylpyridine production wastewater of a certain project was mixed with low-concentration wastewater. The TOC of the mixed water was 1833 mg / L, of which the concentration of 3-methylpyridine was 2016 mg / L. The aerobic biological treatment pool was inoculated with 10% methylpyridine-degrading bacteria to activate the bacterial solution. The HRT was 72h. After 18 days of operation, the effluent was stable, with an effluent TOC of 33-49 mg / L and a TOC degradation rate of 97-98%. No 3-methylpyridine was detected.

[0054] Example 3, 4-methylpyridine biological treatment

[0055] The methylpyridine production wastewater of a certain project was mixed with low-concentration wastewater. The TOC of the mixed water was 2519 mg / L, of which the 4-methylpyridine concentration was 2033 mg / L. The aerobic biological treatment pool was inoculated with 10% methylpyridine-degrading bacteria to activate the bacterial solution. The HRT was 72h. After 21 days of operation, the effluent was stable, with an effluent TOC of 37-60 mg / L and a TOC degradation rate of 97-98%. No 4-methylpyridine was detected.

[0056] The above description is only a preferred embodiment of the present invention. It should be pointed out that a person skilled in the art can make several improvements without departing from the principle of the present invention, and these improvements should also be regarded as within the protection scope of the present invention.

Claims

1. A Sphingobacterium mizutaii, characterized in that It is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number: CGMCC NO.31454 and the deposit date of July 25, 2024.

2. A method for biological treatment of picoline wastewater according to claim 1, characterized in that: A microbial agent for the degradation of methylpyridine solids prepared by Sphingobacillus mizugami.

3. A method for biological treatment of picoline wastewater, characterized in that: In combination with the bacterial agent described in claim 2, a biological treatment method is used to treat methylpyridine wastewater, and the specific steps are as follows: Step 1: Activation and cultivation of microbial agents for methylpyridine-degrading solid bacteria: Add 1-5% of methylpyridine-degrading bacteria solid agent, 0.5-1.5 g / L of glucose, and tap water. The amount of tap water is added according to the ratio of methylpyridine-degrading bacteria solid agent, that is, 1 kg of solid agent corresponds to 1-5 t of tap water. Add dilute sulfuric acid or liquid alkali to adjust the pH to 7-8. Aeration culture is carried out in a strain expansion tank at 25-35° C. for 48 hours to activate the strain. DO is 2-4 mg / L. After the strain is activated, the bacterial density is detected under a microscope. The bacterial density reaches 1×10 9 CFU / mL is the completion of activation culture, and the activated bacterial solution of methylpyridine-degrading bacteria after activation culture is obtained; Step 2: Biological treatment of wastewater: The pH of the wastewater containing picoline is adjusted to 7-8, and the wastewater is passed into a biological treatment tank for biological treatment. The activated bacterial solution of the picoline-degrading bacteria after 10% activation culture in the above step 1 is added to the biological treatment tank. After the bacteria are domesticated and cultured to form biofilm, the picoline concentration can reach 2000 mg / L or the TOC concentration is 2000-2500 mg / L.

4. A method for biological treatment of picoline wastewater according to claim 3, characterized in that: The methylpyridine-degrading bacteria solid agent is added into the reaction tank in 2 to 3 batches according to the dosage of 10% activated bacteria solution and aerated for 48 to 72 hours to fix the bacteria.

5. A method for biological treatment of picoline wastewater according to claim 3, characterized in that: The hydraulic retention time (HRT) of wastewater biological treatment is 24h for influent methylpyridine concentration of 300-500mg / L; 48h for influent methylpyridine concentration of 500-1000mg / L; and 72h for influent methylpyridine concentration of 1500-2000mg / L.

6. A method for biological treatment of picoline wastewater according to claim 3, characterized in that: The biological treatment in step 2 adopts an aerobic process, oxygen is supplied by membrane aeration, the dissolved oxygen (DO) is 2-4 mg / L, and the reaction temperature is 25-35°C.

7. A method for biological treatment of picoline wastewater according to claim 3, characterized in that: When the system is started, the influent methylpyridine concentration is less than 500 mg / L, or TOC is less than 400 mg / L, with TOC being given priority. On the first to third days after the system is started, 200 to 300 mg / LCOD equivalent carbon source is added every day, and the carbon source is glucose or methanol. After three days, no carbon source is added. After that, the methylpyridine concentration is increased by a gradient of 300 to 500 mg / L or TOC by a gradient of 300 to 400 mg / L, with TOC being given priority. 80% of the water is replaced in each batch until the influent load reaches the target influent methylpyridine concentration. The full load inlet methylpyridine concentration can reach 2000 mg / L or the TOC concentration is 2000-2500 mg / L.

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