KR20080047664A - Concentrating culture methods of nirifying bacteria for improving of nitrogen removal - Google Patents

Concentrating culture methods of nirifying bacteria for improving of nitrogen removal Download PDF

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KR20080047664A
KR20080047664A KR1020060117388A KR20060117388A KR20080047664A KR 20080047664 A KR20080047664 A KR 20080047664A KR 1020060117388 A KR1020060117388 A KR 1020060117388A KR 20060117388 A KR20060117388 A KR 20060117388A KR 20080047664 A KR20080047664 A KR 20080047664A
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nitrogen
nitrifying
nitrogen removal
wastewater
bacteria
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KR1020060117388A
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Korean (ko)
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권오범
윤주천
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(주)누리바이오텍
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • 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
    • C02F3/302—Nitrification and denitrification treatment
    • C02F3/303—Nitrification and denitrification treatment characterised by the nitrification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Abstract

A method for concentration-culturing a nitrifying bacteria is provided to remove nitrogen from wastewater smoothly and efficiently by nitrifying nitrogen ingredients in the wastewater. A method for concentration-culturing a nitrifying bacteria comprises a step of culturing Nitrosomonas europaea selectively for nitrifying wastewater containing highly concentrated nitrogen in a culture medium composition including 0.5-1.0g/l of KH2PO4, 10.0-15.0g/l of Na2HPO4, 2.0-5.0g/l of (NH4)2SO4, 0.1-1.0g/l of NaHCO3, 0.001-0.01g/l of CaCl2.2H2O, 0.005-0.01g/l of MgSO4.7H2O, and 0.0005-0.01g/l of Fe-EDTA. Further, a culturing condition is 2 to 3 ppm of oxygen concentration and 25 to 40 °C of temperature.

Description

질소제거 효율향상을 위한 질산화미생물 농화배양 방법{Concentrating culture methods of nirifying bacteria for improving of nitrogen removal.}Concentrating culture methods of nirifying bacteria for improving of nitrogen removal.

발명은 오폐수내에 존재하여 부영화 및 수질오염의 주원인 질소성분을 효율적으로 제거하기 위한 질산화 공정의 효율성을 높이는 질소제거 효율 향상을 위한 질산화미생물의 농화배양에 관한 것으로, 더욱 상세하게는 오폐수내에 존재하는 질소성분을 제거하기 위하여 질산화공정-> 탈질공정 -> 질소로 전환하는 공정에서 가장 중요한 요소인 질산화공정을 이끄는 질산화 미생물인 니트로소모나스 유로파(Nitrosomonas europaea)를 고농도로 배양하는 농화배양 방법과 이를 이용한 질소제거 공정의 효율증진 방법으로서 고농도로 농화배양된 질산화미생물인 니트로소모나스 유로파(Nitrosomonas europaea)는 오폐수내에 존재하는 질소성분을 질산화하여 질소제거 공정을 원활하고 효율 높게 제공함으로 오폐수내의 주요한 오염물질인 질소제거를 통하여 안정적인 질소제거 오폐수처리를 할 수 있는 이점을 갖는다.The present invention relates to the enrichment culture of nitrifying microorganisms to improve the efficiency of nitrogen removal to increase the efficiency of the nitrification process to efficiently remove nitrogen components, which are present in waste water and the main source of sublimation and water pollution. Nitrosomonas europaea , a nitrifying microorganism leading the nitrification process, which is the most important element in the nitrification process-> denitrification process-> nitrogen conversion process to remove nitrogen components, and the enrichment culture method using the same Nitrosomonas europaea , a nitrifying nitrification microorganism concentrated at high concentration, is a major pollutant in wastewater by nitrifying nitrogen in wastewater and providing nitrogen removal process with high efficiency. Stable through nitrogen removal Phosphorus nitrogen removal has the advantage that can be treated with waste water.

최근, 오, 폐수에 의한 하천 및 지하수의 오염이 점차 심각해지면서 수질 보존을 위한 오,폐수 처리장의 처리 효율 증대의 필요성이 점차 부각되고 있다. 또한 산업폐수와 함께 가정 생활 하수와 분뇨, 축산폐수가 또 다른 수질오염을 가속시키는 한 원인이 되고 있다. 또한 분뇨, 오, 폐수 등에서는 폐수내 함유된 오염물질로 인해 수질이 오염될 뿐만 아니라 처리효율이 저하됨으로서 비용 발생이 높아지고 이에 따른 폐기물의 발생 증가도 수반된다. 우리나라는 점차 오염되는 수자원을 확보하기위한 일환으로서 2000년 8월 수질환경보전법이 개정됨에 따라, 전 산업체는 2003년 1월부터 질소 성분의 배출 농도를 일정농도 이하로 처리한 후 배출하여야 한다. 이러한 질소의 배출허용 기준은 지역별 차이점이 있는데 청정지역의 경우는 30ppm이하이고, 기타지역은 60ppm으로 규정되어 있다. 이러한 질소를 효율적으로 제거하기 위한 생물학적 질소제거 공법은 1914년 Arden 과 Lockett 에 의해 개발된 Fill and draw 방식의 활성슬러지 공정이 시초로서 이후 여러가지의 질소제거 공정이 개발되어 국내외적으로는 약 30여개의 질소제거 공법이 상용화 되고 있고 대부분 공법이 질산화 및 탈질미생물을 이용한 생물학적공법이다. 이러한 공법을 바탕으로하여A/O, A2O, Bardenpho, MUCT 등의 부유성장 공법과 최근에는 접촉여재를 이용한 질산화미생물의 농도 및 체류시간을 개선한 접촉산화공법이 TETRA Tech(미) 등에서 상용화되었다. 이러한 생물학적 질소제거 공정의 핵심은 반응조내에서 질산화미생물과 탈질미생물의 효율을 극대화하는데 초점이 맞추어져 있다. 질산화미생물은 다른 종속영양 bacteria 와 달리 배양조건이 까다롭고, 증식속도가 느려 미생물학적 연구나 제품화 사례는 매우 적으며, 국내의 경우 질산화 공정개발은 활발한 편이나 질산화 미생물 및 이를 직접 이용한 효율개선방안은 미흡한 실정이다. 대부분의 질소제거 공법이 질산화 및 탈질 미생물을 이용하는 생물학적 공법이며, 생물학적 질소제거 공법에 있어서 가장 중요한 요소가 질산화 및 탈질에 관여하는 미생물임에도 불구하고, 미생물 자체보다는 공정중심의 기술개발이 주류를 이루고 있다. 이중에서 더욱 중요한 요소는 질산화미생물의 우점화에 의한 질산화 공정이 전체 질소제거공정의 효율을 좌우한 다는 점이다. 그러나 질산화미생물은 타미생물에 비해 증식속도가 현저히 느려 현탁배양 방식으로는 우점화에 의한 MLSS 의 유지가 곤란한 한계성을 갖는다. 이에 따라 질산화 공정의 효율을 극대화할 목적으로 질산화조에 질산화미생물을 고농도화, 고정화 또는 유동화 담체를 이용한 방법으로 한국특허 출원 10-2000-007870, 한국특허 10-1997-0068564, 한국특허 출원 10-1999-0057474 등에서 이러한 질산화미생물의 우점화를 통한 방법을 제시하고 있으나, 질산화미생물을 우점화시키는 메카니즘이 불분명하고 이에 따라 처리효율의 진폭도 큰 문제점을 가지고 있다. 따라서 이러한 니트로소모나스 유로파(Nitrosomonas europaea)와 같은 질산화미생물은 오폐수처리의 질산화공정에서 안정적으로 고농도로 유지시키는 것이 필요로 한다.In recent years, as pollution of rivers and groundwater by wastewater and wastewater becomes more serious, the necessity of increasing the treatment efficiency of wastewater treatment plants for water quality preservation is gradually increasing. In addition to industrial wastewater, household sewage, manure and livestock wastewater are another cause of accelerated water pollution. In addition, in manure, sewage, waste water, etc., not only the water quality is contaminated by the pollutants contained in the waste water, but also the treatment cost is lowered, resulting in higher cost and thus increased waste generation. As a part of Korea's efforts to secure polluted water resources, as the Water Quality Conservation Act was amended in August 2000, all industries must discharge nitrogen after treating it with a certain concentration below a certain concentration from January 2003. These allowances for nitrogen emissions vary from region to region: 30 ppm or less in clean areas and 60 ppm in other areas. The biological nitrogen removal method for efficient removal of nitrogen is based on the fill and draw activated sludge process developed by Arden and Lockett in 1914, and various nitrogen removal processes have been developed since then. Nitrogen removal methods are commercialized, and most of them are biological methods using nitrification and denitrification microorganisms. Based on this method, floating growth methods such as A / O, A2O, Bardenpho, and MUCT and contact oxidation methods have been commercialized at TETRA Tech (US) in recent years to improve the concentration and residence time of nitrifier microorganisms using contact media. The core of this biological nitrogen removal process is focused on maximizing the efficiency of nitrifying and denitrifying microorganisms in the reactor. Unlike other heterotrophic bacteria, nitrifying microorganisms have difficult culture conditions and slow growth rate, so there are very few cases of microbiological studies or commercialization.In Korea, nitrification process is active, but nitrifying microorganisms and efficiency improvement methods using them are It is insufficient. Although most nitrogen removal methods are biological methods using nitrification and denitrification microorganisms, the most important factor in biological nitrogen removal processes is microorganisms involved in nitrification and denitrification, but process-oriented technology development is mainstream rather than microorganisms themselves. . More importantly, the nitrification process by dominant nitrifying microorganisms determines the efficiency of the entire nitrogen removal process. However, nitrifying microorganisms have a limitation that it is difficult to maintain MLSS due to dominance in suspension culture because the growth rate is significantly slower than that of other microorganisms. Accordingly, in order to maximize the efficiency of the nitrification process, a method using high concentration, immobilization, or fluidization carrier of nitrification microorganisms in the nitrification tank is disclosed in Korean Patent Application No. 10-2000-007870, Korean Patent 10-1997-0068564, and Korean Patent Application No. 10-1999 -0057474 and the like have been proposed a method through the dominant nitrifying microorganisms, but the mechanism for dominant nitrifying microorganisms is unclear and accordingly has a big problem in the amplitude of the treatment efficiency. Therefore, the nitrifying microorganisms such as Nitrosomonas europaea need to be stably maintained at a high concentration in the nitrification process of wastewater treatment.

본 발명은 상기와 같은 문제점을 해결하기위하여 오폐수내의 질소제거 공정에서 가장 중요한 요소인 질산화 공정을 이끄는 질산화 미생물인 니트로소모나스 유로파(Nitrosomonas europaea)를 고농도로 배양하는 농화배양 방법과 이를 이용한 질소제거 공정의 효율증진 방법으로서, 질산화미생물인 니트로소모나스 유로파 (Nitrosomonas europaea)를 선택적으로 고농도로 농화배양하여 오폐수내에 존재하는 질소성분을 질산화하여 질소제거 공정을 원활하고 효율 높게 제공함을 목적으로 한다.The present invention provides a thickening culture method and a nitrogen removal process using a high concentration of nitrosomonas europaea , a nitrifying microorganism leading the nitrification process, which is the most important element in the nitrogen removal process in waste water to solve the above problems. As a method for improving the efficiency, Nitrosomonas europaea , which is a nitrifying microorganism, is selectively concentrated in high concentration to nitrate nitrogen components present in waste water to provide a smooth and efficient nitrogen removal process.

본 발명은 오폐수내에 존재하여 부영화 및 수질오염의 주원인 질소성분을 효율적으로 제거하기 위한 질산화 공정의 효율성을 높이는 질산화미생물인 니트로소모나스 유로파(Nitrosomonas europaea)를 선택적으로 배양하는 방법과 이를 고농도로 배양하는 농화배양 방법과 농화배양된 질산화미생물인 니트로소모나스 유로파(Nitrosomonas europaea)를 이용하여 질소제거 공정의 효율성을 높이는데 있다. The present invention provides a method of selectively cultivating nitrosomonas europaea , a nitrifying microorganism, which enhances the efficiency of nitrification process to efficiently remove nitrogen, which is the main source of sublimation and water pollution. Nitrosomonas europaea , an enriched nitrifying microorganism, is used to increase the efficiency of the nitrogen removal process.

질산화는 2단계로 이루어 지며 첫번째 단계에서는 암모니아 산화 박테리아로 암모니아성 질소를 아질산성 질소로 산화시키며, 두 번째 단계에서는 질산염 산화 박테리아로 아질산성 질소를 질산성 질소로 산화 시킨다. 질산화 미생물들은 니트로소모나스(Nitrosomonas sp.)가 대표적 이며 질산염 산화박테리아는 니트로박터(Nitrobacter sp)가 대표적이다. 모든 생물학적 질소제거 공정은 질산화 반응의 성공적인 수행여부에 의해 좌우된다. 따라서 발명자들은 Nitrification consists of two stages: in the first stage, ammonia-oxidizing bacteria oxidize ammonia nitrogen to nitrite nitrogen, and in the second stage, nitrate-oxidizing bacteria oxidize nitrite nitrogen to nitrate nitrogen. Nitrogenated microorganisms are represented by Nitrosomonas sp. And nitrate bacteria are represented by Nitrobacter sp. All biological nitrogen removal processes depend on the successful performance of the nitrification reaction. So the inventors

질산화미생물이 선택적으로 배양 농화하는 조건을 선정하여 그에 적합한 배양배지의 조성물과 배양조건을 발견하여 본 발명의 기반조건을 설정하였고, 이것을 이용하여 확립된 농화배양 방법을 확립하여 질소제거공정의 기본이 되는 질산화 공정의 효율을 극대화하게 되었다.By selecting the conditions for selective culture enrichment of nitrifying microorganisms, the composition and culture conditions of the culture medium were found, and the basic conditions of the present invention were established. The efficiency of the nitrification process is maximized.

질산화미생물인 니트로소모나스(Nitrosomonas sp.)는 배양 및 농화를 통하여 니트로소모나스 유로파(Nitrosomonas europaea)가 질산화 공정의 효율증진에 가장 좋음을 발견하였다. Nitrosomonas sp., A nitrifying microorganism, found that Nitrosomonas europaea was the best method for enhancing the nitrification process through culture and concentration.

질산화 미생물인 니트로소모나스 유로파(Nitrosomonas europaea)는 독립영양세균으로서 배양조건이 까다롭고 증식속도가 늦은 단점을 가지고 있다. 따라서 이 질산화미생물을 배양하는데는 인산칼륨(KH2PO4), 일인산나트륨(Na2HPO4), 황산암모늄((NH4)2SO4), 탄산나트륨(NaHCO3)염화칼슘(CaCl2??2H2O), 황산마그네슘(MgSO4??7H2O),이디티에이철(Fe-EDTA)이 선택성과 농화성에 좋음을 발견하게 되었다. 또한 선택배양과 농화배양의 주요한 요인으로는 산소농도와 온도조건이 중요한데 산소농도는 2~3ppm, 온도조건은 25℃에서 40℃ 사이가 적당하다. Nitrosomonas europaea , a nitrifying microorganism, is an independent trophic bacterium that has difficult culture conditions and slow growth rate. Therefore, the culture of the nitrifying microorganisms included potassium phosphate (KH 2 PO 4 ), sodium monophosphate (Na 2 HPO 4 ), ammonium sulfate ((NH 4 ) 2 SO 4 ), sodium carbonate (NaHCO 3 ) calcium chloride (CaCl 2 ?? 2H 2 O), magnesium sulfate (MgSO 4 ˜7H 2 O), and editied iron (Fe-EDTA) were found to be good for selectivity and thickening. In addition, oxygen concentration and temperature conditions are important as selective factors and enrichment culture. Oxygen concentration is 2 ~ 3ppm, and temperature condition is suitable between 25 ℃ and 40 ℃.

이하 본 발명을 실시 예를 통해 더욱 상세히 기술하고자 한다. Hereinafter, the present invention will be described in more detail with reference to Examples.

실시 예1 : 니트로소모나스 유로파(Nitrosomonas europaea)의 선택배양 조건 비교.Example 1: Comparison of selective culture conditions of Nitrosomonas europaea .

Figure 112006509498993-PAT00001
Figure 112006509498993-PAT00001

<표-1>TABLE 1

상기 <표-1>에서 보는바와 같이 선택배지 1의 조성이 질산화 미생물인 니트로소모나스 유로파(Nitrosomonas europaea)의 성장과 암모니아 산화능에 우수한 결과를 보임을 나타내었다.As shown in Table 1, the composition of the selective medium 1 showed excellent results on the growth and ammonia oxidation ability of nitrosomonas europaea , which is a nitrifying microorganism.

실시예2. 본 발명의 질산화미생물 니트로소모나스 유로파(Nitrosomonas europaea)농화배양Example 2. Nitrogenated microorganism nitrosomonas europaea enrichment culture of the present invention

조성물과 기존 특허와의 비교         Comparison of composition with existing patent

피혁공장의 폐수(NH4-N 320ppm)를 이용하여 실험실에서 타사 제품과 비교하여 처리한 결과는 아래와 같다.The wastewater from the leather factory (NH 4 -N 320ppm) was treated in the laboratory compared to other companies' products.

5리터 용량의 비이커에 상기의 도축폐수 2리터를 채운 후 폭기를 시키며, 아래의 처리조건으로 2일, 4일간 시간 처리를 하고 상등액을 취하여 암모니성질소(NH4-N)를 측정하여 처리효율을 비교한 결과는 아래의 표 2.과 같다.Fill a 5 liter beaker with 2 liters of slaughtered wastewater and aeration. Treat it for 2 days and 4 days under the following treatment conditions and take the supernatant to measure ammonia nitrogen (NH 4 -N). The results of the comparison are shown in Table 2 below.

<표2><Table 2>

Figure 112006509498993-PAT00002
Figure 112006509498993-PAT00002

상기 <표-2>에서 보는바와 같이 본 발명의 질산화 농화배양물을 이용한 처리방법이 기존 특허방법에 비하여 14~15% 정도의 처리효율이 우수함을 알 수 있었다.As shown in Table 2, the treatment method using the nitrification enrichment culture of the present invention was found to have an excellent treatment efficiency of about 14 to 15% compared to the existing patent method.

본 발명에 따라 오폐수내에 존재하여 부영화 및 수질오염의 주원인 질소성분을 효율적으로 제거하기 위한 질산화 공정의 효율성을 높이는 질산화미생물인 니트로소모나스 유로파(Nitrosomonas europaea)를 선택적으로 배양하는 방법과 이를 고농도로 배양하는 농화배양 방법과 농화배양된 질산화미생물인 니트로소모나스 유로파(Nitrosomonas europaea)를 이용하여 질소제거 공정에 투입하여 효과를 비교한 결과 처리효율이 우수하고 안정적이었다.According to the present invention, a method of selectively cultivating nitrosomonas europaea , a nitrifying microorganism, which enhances the efficiency of nitrification process for efficiently removing nitrogen components, which are present in waste water, are the main sources of sublimation and water pollution, and high concentration thereof. Nitrosomonas europaea , a cultured thickening method and a cultured nitrifying microorganism, was used in the nitrogen removal process to compare the effects, and the treatment efficiency was excellent and stable.

본 발명을 적용함으로써 고함유 질소를 함유한 폐수처리가 높은 효율성을 가지게 되어, 오폐수내에 존재하는 질소성분을 질산화하여 질소제거 공정을 원활하고 효율 높게 제공함으로서 오폐수내의 주요한 오염물질인 질소제거를 통하여 안정적인 질소제거 오폐수처리를 할 수 있는 효과를 갖는다. By applying the present invention, wastewater treatment containing high nitrogen content has high efficiency, and it is stable through nitrogen removal, which is a major pollutant in wastewater, by nitrifying nitrogen components present in wastewater and providing a smooth and efficient nitrogen removal process. Nitrogen removal waste water treatment can be effected.

Claims (3)

고농도 질소함유 폐수를 질산화하기 위한 질산화 미생물 니트로소모나스 유로파(Nitrosomonas europaea)을 선택적으로 배양하여 농화배양하는 방법.A method for the enrichment culture by selectively culturing nitrifying microorganism Nitrosomonas europaea to nitrate high concentration nitrogen containing wastewater. 제1항에 있어서 선택적으로 배양하는 배양배지의 조성물로서 KH2PO4 0.5~1.0 g/l0.5 to 1.0 g / l KH 2 PO 4 as a composition of the culture medium to be selectively cultured according to claim 1 Na2HPO4 10.0~15.0 g/l, (NH4)2SO4 2.0~ 5.0 g/l, NaHCO3 0.1~1.0 g/l, CaCl2 Na 2 HPO 4 10.0-15.0 g / l, (NH 4 ) 2 SO 4 2.0-5.0 g / l, NaHCO 3 0.1-1.0 g / l, CaCl 2 2H2O 0.001~0.01 g/l, MgSO47H2O 0.005~0.01 g/l, Fe-EDTA 0.0005~0.01 g/l2H 2 O 0.001 ~ 0.01 g / l, MgSO 4 7H 2 O 0.005 ~ 0.01 g / l, Fe-EDTA 0.0005 ~ 0.01 g / l 을 사용하는 방법.How to use it. 제2항에 있어서 배양조건은 산소농도 2~3ppm, 온도조건은 25℃에서 40℃로 하는 방법.The method of claim 2, wherein the culture conditions are oxygen concentration 2-3 ppm, the temperature conditions are 25 ℃ to 40 ℃.
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