WO2021117981A1 - 메탄과 메탄올 의존 공생박테리아와 바이오가스를 이용한 하수처리공정의 반류수내 고농도 암모니아 제거 방법 - Google Patents
메탄과 메탄올 의존 공생박테리아와 바이오가스를 이용한 하수처리공정의 반류수내 고농도 암모니아 제거 방법 Download PDFInfo
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- WO2021117981A1 WO2021117981A1 PCT/KR2020/001603 KR2020001603W WO2021117981A1 WO 2021117981 A1 WO2021117981 A1 WO 2021117981A1 KR 2020001603 W KR2020001603 W KR 2020001603W WO 2021117981 A1 WO2021117981 A1 WO 2021117981A1
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- methane
- methanol
- biogas
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- ammonia
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/30—Aerobic and anaerobic processes
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
Definitions
- the present invention relates to a method for removing high-concentration ammonia in the countercurrent of a sewage treatment process using methane- and methanol-dependent symbiotic bacteria and biogas, and more particularly, methane for reducing the nitrogen concentration in the countercurrent essentially generated in the sewage treatment process and a methanol-dependent microorganism consortium and an invention related to ammonia removal in a methane oxidation reactor including biogas.
- Reject water is essentially generated from the sludge dewatering filtrate of the sewage treatment plant and the anaerobic digester supernatant and sludge dewatering (FIG. 1).
- the amount of countercurrent generated during the sludge dewatering process is as small as 1.5-3% of the influent sewage, it accounts for more than 30% of the ammonium nitrogen inflow load into the sewage treatment plant treatment process.
- the return water entering the sewage treatment plant influent greatly increases the treatment plant load, and ammonium overload above the design load level may occur irregularly. Accordingly, it may interfere with the efficient operation of the entire nitrogen removal process, and incur large costs for additional installation of nitrification/denitrification systems or improvement of the sewage treatment process.
- methane-oxidizing bacteria using methane as a substrate oxidize methane to form methanol, and have the ability to denitrify and dephosphorize themselves.
- methanotrophs have been discovered, there has been no full-scale study on the denitrification process using them.
- the present invention provides a method for removing high concentration of ammonia in the countercurrent of a sewage treatment process using a methane and methanol dependent microbial consortium and biogas.
- the methane- and methanol-dependent microorganism consortium may be composed of methanotrophs and methylophilus.
- the methanotrophs may be composed of one or more genera of methylobacter, methylomonas, methylomonas_f_uc, and methylosarcina.
- the Methylophilus may be composed of M. flavus and M. Rhizosphaerae species (Species).
- the biogas may have a volume ratio of methane and oxygen of 1:1 to 1:1.25.
- PQQ-MDH pyrroloquinoline quinone-methanol dehydrogenase
- the synergistic action of methane and methanol dependent microbial consortium and biogas generated from sewage treatment plant are used to directly treat high-concentration ammonium without diluting the wastewater from the sewage treatment process to lower nitrogen load and improve nitrification efficiency to maintain good sewage effluent quality. This has the effect of improving the water quality of the discharge river.
- Figure 2 shows a schematic diagram of metabolic pathways in which ammonium is removed by methane and methanol dependent microbial consortium (methanotrophs and methylophilus) in countercurrent.
- SBR sequencing batch reactor
- Figure 5 shows the species compositional phylogenetic diagram of Gammaproteobacteria methanotrophs and Betaproteobacteria methylophius.
- SBR 8 is a sequencing batch reactor (SBR) operation results (152 days); a) Change in ammonium inflow and outflow concentration according to operation time, and b) Change in removal rate according to ammonium inflow concentration are shown.
- SBR sequencing batch reactor
- a method for removing high concentration of ammonia in countercurrent of a sewage treatment process using a methane and methanol dependent microbial consortium and biogas is provided.
- Another embodiment according to the present invention is characterized in that the methane- and methanol-dependent microorganism consortium is composed of methanotrophs and methylophilus.
- methanotrophs are composed of one or more genera of methylobacter, methylomonas, methylomonas_f_uc and methylosarcina.
- Methylophilus is composed of M. flavus and M. Rhizosphaerae species (Species).
- biogas has a volume ratio of methane and oxygen of 1:1 to 1:1.25.
- the methylophilus while assimilating carbon into formaldehyde through PQQ-MDH (pyrroloquinoline quinone-methanol dehydrogenase) from methanol produced by methanotrophs using the RuMP pathway. and ammonia as nitrogen sources.
- PQQ-MDH pyrroloquinoline quinone-methanol dehydrogenase
- the present invention is an invention for direct treatment without diluting countercurrent water containing a small amount (1.5-3% of influent water) high concentration ammonium, which is inevitably generated in the sewage treatment process.
- the present invention is to directly treat high concentration of ammonium without diluting the countercurrent using the synergistic action of methane and methanol dependent microbial consortium and biogas generated from a sewage treatment plant.
- Methylophilus assimilated formaldehyde through the same RumP (ribulose monophosphate) cycle as Methanotrophs with methanol produced by Methanotrophs, and applied a synergistic mechanism to remove ammonium through the glutamate cycle of Methanotrophs and Methylophilus. It is an invention.
- Example 1 Culture of microbial bacteria consortium and biogas
- AMS ammonium mineral salts
- the concentration of CuSO 4 in the medium was 20 ⁇ M, and each medium was titrated to pH6.8 by adding 1.5 mL of phosphate buffer (26 g/L KH 2 PO 4 , 33 g/L Na 2 HPO 4 ).
- the cultured methane-oxidizing bacteria were separated by a centrifuge (Centrifuge-416; Dongseo Science, Ltd., Dangjin, Korea) at 2,700 ⁇ g, and the pellet was freeze-dried at -55 ° C (OPERN FDS-12003; Seoul, Korea).
- the water quality of the sewage treatment plant countercurrent used in the experiment replacing the AMS medium was pH 7.3, TSS 10,540.0 mg L -1 , VSS 7,560.3 mg L -1 , TCODcr 7,636.0 mg L -1 , SCODcr 117.5 mg L -1 , TN 407.1 mg L -1 , NH 4 -N 398.6 mg L -1 , NO 2 -N 0.1 mg L -1 , NO 3 -N 0.9 mg L -1 , PO 4 -P 164.5 mg L - 1 , TP 254.4 mg L -1 , Alkalinity 2,500,0 mg L -1 , Cu 3.2 mg L -1 , Fe 238.3 mg L -1 .
- the water quality of the countercurrent used for the nitrogen removal characteristics analysis by the unit batch experiment was pH 7.8, CODcr 3021.2 mg L -1 , NH 4 -N 642.6 mg L -1 , NO 2 -N 0.07 mg L -1 , NO 3 -N 5.2 mg ⁇ L -1 , PO 4 -P 747.3 mg ⁇ L -1 , Cu 0.580 mg ⁇ L -1 It was stored at 4°C and used for unit experiments.
- the biogas used in the present invention is CH 4 : 66.9%, CO 2 : 31.3%, N 2 : 1.2% and O 2 : It is composed of 0.2%, and a number of 4.9-L high-pressure gas tanks. (GlobalGastec, Ltd., Buchun, Korea) was used for collection and storage and used for experiments.
- Type I methanotrophs directly assimilate ammonium and nitrate in return water into nitrogen sources necessary for growth, and at the same time, part of pMMO, an enzyme expressed to oxidize methane to methanol, has the same mechanism as AMO (Ammonia monooxygenase) of ammonia oxidizing bacteria (AOB) ammonium is oxidized and removed through the Like AMO, there are two metabolic pathways for hydroxylamine and nitrite, which are toxic intermediates of ammonium oxidation by MMO of methanotrophs.
- AMO Ammonia monooxygenase
- AOB ammonia oxidizing bacteria
- hydroxylamine produced as in ammonia-oxidizers is first oxidized to nitrite, then the produced nitrite is reduced to nitric oxide, and then nitric oxide is converted to nitrous oxide (Fig. 2).
- a, c The other is a pathway (b, c in FIG. 2) through which hydroxylamine is converted to nitrous oxide through nitric oxide.
- a portion of the hydroxylamine formed reduces hydroxylamine back to ammonia through hydroxylamine reductase enzymes, a unique detoxification mechanism by methanotrophs.
- M. flavus and M. Rhizosphaerae which were detected as major constituents of Methylophilus in the present invention, were oxidized to formaldehyde through PQQ-MDH (pyrroloquinoline quinone-methanol dehydrogenase) of methanol produced by methnaotrophs using the RuMP pathway like methanotrophs. (Oxidizes methanol by PQQ-MDH) Assimilate carbon.
- PQQ-MDH pyrroloquinoline quinone-methanol dehydrogenase
- PQQ-MDH pyrroloquinoline quinone-methanol dehydrogenase
- Assimilate carbon Assimilate carbon.
- Figure 2 shows a schematic diagram of possible metabolic pathways in which ammonium is removed from countercurrent by methane and methanol dependent microbial consortium (methanotrophs and methylophilus) using the previously identified research results and the results of this study.
- the continuous SBR (sequencing batch reactor) reactor is equipped with a stirrer, a gas supply device and a pressure gauge, and the residence time is 6 - 12 hr depending on the experimental conditions while stirring at 20 to 25 ° C and operating at 150 rpm, and the precipitation time is It was set to 30 minutes, and MLSS was maintained at 3,700 - 4,400 mg ⁇ L -1 while SRT (sludge retention time) was set to 20 days to discharge microorganisms increasing by proliferation.
- the consumption of biogas and oxygen in the bioreactor was automatically injected from the gas tank by the pressure sensor (FIG. 3).
- the methane and methanol dependent mixed bacteria consortium separated and cultured from the sewage treatment plant sludge using AMS medium was replaced with a gas with a volume ratio of methane and oxygen in the air of 17.8 ml : 16.9 mL using biogas at a 24-hour cycle with AMS and Figure 4 shows the characteristics of the mixed bacteria consortium species after 6 days of culturing when re-cultured in countercurrent.
- Pseudomonas When pure air was injected without biogas injection (Fig. 4b), Pseudomonas was superior to 72.04% and Flavobacterium was dominant to 11.46%, and when cultured in biogas and AMS medium, Methylophilus, which was a dominant species with a high ratio, was significantly lowered to 1.33% ratio, and methanotrophs Also, only one type (oder) of Methylomonas was detected at a low rate of 0.44%.
- Pseudomonas which showed the dominant species, is a type of bacteria commonly present in activated sludge in sewage treatment plants and is mainly detected when separated and cultured from activated sludge. From this, it was observed that the mixed bacteria consortium was mainly reduced to activated sludge constituents without the supply of biogas.
- the methanotrophs that appeared were Gammaproteobacteria, ICM (intracellular multiplication) type, which expressed pMMO (particulate methane monooxygenase) type I methanotrophs, and were composed of methanotrophs that showed activity even when copper was present in high concentrations. It was observed that only Type I methanotroph existed because ammonium and copper with high countercurrent used as a culture medium were present.
- the species that appeared here are Gammaproteobacteria methanotrophs (Gamma-MOB), which belong to the methanotrophs taxa, which shows less growth inhibition or a tendency to promote growth, unlike Alphaproteobacteria methanotrophs (Alpha-MOB) when nitrogen is added to the growth environment. It can be seen that it consists only of species.
- Methylophilus As a dominant species, Methylophilus, which coexists clearly and is detected at a relatively high rate, is a denitrifying bacterium that uses methanol as a substrate and disappears from the microbial community when methanol is consumed. This is because this bacterium converts methanol to formaldehyde using PQQ-MDH and uses nitrate and ammonia as nitrogen sources while assimilating formaldehyde through the RuMP (ribulose monophosphate) pathway.
- RuMP ribulose monophosphate
- Methylophilus Rhizosphaerae Another dominant species, Methylophilus Rhizosphaerae, showed the same mechanism as a strictly aerobic gram-negative bacterium, and Methylophilus flavus was also an obligate methanol-utilizing gram-negative bacterium and strictly aerobic bacterium.
- Example 5 Bacteria Consortium species composition change according to methane and oxygen volume ratio
- the methane utilization rate of the bacteria consortium according to the methane per oxygen ratio was 0.098 mg-CH4 ⁇ mg-MLSS -1 when 1.25 was the highest (Table 1), and the ammonium removal rate was 52.1% when the ratio was 1.0 and when the ratio was 1.25. It was the highest at 58.2%. However, as the methane per oxygen ratio increased to 1.5, the removal efficiency was significantly lowered to 39.4%.
- FIG. 8 shows the results of SBR operation using methane and methanol dependent bacteria consortium and biogas for 152 days after receiving daily supply of countercurrent from a sewage treatment plant exhibiting countercurrent generation characteristics.
- the biogas supply was operated while automatically supplied by the sensor so that the methane per oxygen ratio in the biogas in air was maintained in the range of 1 - 1.25 according to the gas consumption rate of the SBR head space.
- the operation cycle of the SBR reactor was 6 hours of stirring operation and 30 minutes of precipitation, followed by discharging the treated supernatant, and the increasing mixed bacteria sludge was removed by periodically discharging to maintain 3,700 - 4,400 mg ⁇ L -1 based on the MLSS standard.
- the average concentration of influent is 877.3 mg ⁇ L -1 and the average effluent concentration is 359.9 mg ⁇ L -1, and as an average of 58.9% ( ⁇ 8.4) of ammonium is removed, the high concentration of ammonium in the countercurrent flowing back into the sewage treatment process It was confirmed that it can be lowered to the level of 59%.
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- Engineering & Computer Science (AREA)
- Environmental & Geological Engineering (AREA)
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Abstract
Description
CH4 : O2 ratio ( Biogas volume : mL) | 0.5(13.5) | 0.75(19.0) | 1.0(24.0) | 1.25(28.0) | 1.50(31.2) | 2.00(38.3) |
MLSS concentration (mg·L-1) | 3947.2± 23.4 | 4261.3± 32.7 | 4325.7± 35.9 | 4289.4± 26.1 | 4058.2± 43.6 | 3729.7± 38.7 |
Methane utilization rate (mg-CH4·mg-MLSS-1) | 0.021±0.006 | 0.067±0.009 | 0.073±0.005 | 0.098±0.007 | 0.068±0.011 | 0.032±0.014 |
NH4-N removal rate (%) | 38.3± 3.2 | 46.9± 5.7 | 52.1± 3.6 | 58.2± 4.4 | 39.4± 2.7 | 32.6± 4.1 |
Claims (6)
- 메탄 및 메탄올 의존미생물 컨소시엄(Methane and methanol dependent microbial consortium)과 바이오가스(biogas)를 이용한 하수처리공정의 반류수 내 고농도 암모니아 제거 방법.
- 제1항에 있어서, 상기 메탄 및 메탄올 의존미생물 컨소시엄은 methanotrophs 및 methylophilus로 구성되는 것을 특징으로 하는 하수처리공정의 반류수 내 고농도 암모니아 제거 방법.
- 제2항에 있어서, 상기 methanotrophs는 methylobacter, methylomonas, methylomonas_f_uc 및 methylosarcina 중 하나이상의 속(Genus)으로 구성되는 것을 특징으로 하는 하수처리공정의 반류수 내 고농도 암모니아 제거 방법.
- 제2항에 있어서, 상기 Methylophilus는 M. flavus와 M. Rhizosphaerae 종(Species)으로 구성되는 것을 특징으로 하는 하수처리공정의 반류수 내 고농도 암모니아 제거 방법.
- 제1항에 있어서, 상기 바이오가스는 메탄과 산소의 부피비가 1:1 내지 1:1.25 인 것을 특징으로 하는 하수처리공정의 반류수 내 고농도 암모니아 제거 방법.
- 제2항에 있어서, 상기 methylophilus는 RuMP pathway를 이용하여 methanotrophs에 의하여 생성되는 methanol을 PQQ-MDH(pyrroloquinoline quinone - methanol dehydrogenase)를 통하여 포름알데히드(formaldehyde)로 탄소를 동화(assimilation)하면서 nitrate와 ammonia를 질소원(nitrogen sources)으로 사용하는 것을 특징으로 하는 하수처리공정의 반류수 내 고농도 암모니아 제거 방법.
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CN114560596A (zh) * | 2022-03-01 | 2022-05-31 | 桂林理工大学 | 抗冲击碳捕捉农村污水一体化处理系统及方法 |
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CN113736717A (zh) * | 2021-11-03 | 2021-12-03 | 广东省科学院生态环境与土壤研究所 | 一株具有脱氮功能和缺氧抗逆性的甲烷氧化菌及其应用 |
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CN114560596A (zh) * | 2022-03-01 | 2022-05-31 | 桂林理工大学 | 抗冲击碳捕捉农村污水一体化处理系统及方法 |
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