• 제목/요약/키워드: denitrification enzyme activity

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Effect of Oxidation-Reduction Potential on Denitrification by Ochrobactrum anthropi SY509

  • Song, Seung-Hoon;Yeom, Sung-Ho;Choi, Suk-Soon;Yoo, Young-Je
    • Journal of Microbiology and Biotechnology
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    • 제13권3호
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    • pp.473-476
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    • 2003
  • The effect of oxidation-reduction potential (ORP) level on the denitrification by Ochrobactrum anthropi SY509 was investigated under nongrowing condition. The maximum ORP level of nitrate-containing buffer solution was -70∼-80 mV, under which the denitrification took place. By decreasing the initial ORP level, denitrifying enzyme activity was greatly enhanced, which led to higher denitrification efficiency.

Characterization of Membrane-bound Nitrate Reductase from Denitrifying Bacteria Ochrobactrum anthropi SY509

  • Kim Seung-Hwan;Song Seung-Hoon;Yoo Young-Je
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제11권1호
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    • pp.32-37
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    • 2006
  • In this study, we have purified and characterized the membrane bound nitrate reductase obtained from the denitrifying bacteria, Ochrobactrum anthropi SY509, which was isolated from soil samples. O. anthropi SY509 can grow in minimal medium using nitrate as a nitrogen source. We achieved an overall purification rate of 15-fold from the protein extracted from the membrane fraction, with a recovery of approximately 12% of activity. The enzyme exhibited its highest level of activity at pH 5.5, and the activity was increased up to $70^{\circ}C$. Periplasmic and cytochromic proteins, including nitrite and nitrous oxide reductase, were excluded during centrifugation and were verified using enzyme essay. Reduced methyl viologen was determined to be the most efficient electron donor among a variety of anionic and cationic dyestuffs, which could be also used as an electron donor with dimethyl dithionite. The effects of purification and storage conditions on the stability of enzyme were also investigated. The activity of the membranebound nitrate reductase was stably maintained for over 2 weeks in solution. To maintain the stability of enzyme, the cell was disrupted using sonication at low temperatures, and enzyme was extracted by hot water without any surfactant. The purified enzyme was stored in solution with no salt to prevent any significant losses in activity levels.

온대지역 댐 하류의 탈질화 능력과 탈질화균 분포 (Denitrification Potential and Denitrifier Abundance in Downstream of Dams in Temperate Streams)

  • ;이승훈;;정석희;강호정
    • 미생물학회지
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    • 제50권2호
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    • pp.137-151
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    • 2014
  • 댐의 존재가 하천 생태계에 미치는 영향을 연구하기 위해 다양한 연구가 지금까지 수행되어 왔지만, 댐이 하류의 탈질화에 미치는 영향은 잘 알려져 있지 않다. 대한민국 낙동강의 댐 원류에서 탈질화 효소 활성도(잠재 탈질율)와 탈질균 분포(nirS, nirK, nosZ 유전자를 표지유전자로 사용)를 조사하였다. 자갈 혹은 모래로 채워진 하천의, 갈대가 우거진 하변지역과 강바닥의 침전물을 채취하여 조사하였다. 이 실험의 가설은 다음과 같다. (i) 하천 침전물의 N과 C의 사용유효량이 높을수록 대조군에 비해 미생물 군집의 탈질화 작용이 더욱 증진한다, (ii) 하천생태계마다 상이하게 나타나는 잠재 탈질율 간의 차이는 탈질 미생물의 양에 비례한다. 30여 년간 댐에 의해 수문학적으로 큰 차이가 있었고 또한 댐 하류의 저서에 무기질소와 용존유기탄소 농도가 대조군에 비해 매우 높았음에도 불구하고, 탈질균 군집의 양과 잠재 탈질율은 하천 간에 큰 차이가 없었다. 하지만 nirS 유전자와 nosZ 유전자의 양과 잠재 탈질율은 댐 하류에 존재하는 자갈이 많은 하변과 모래가 많은 하천 바닥에서 홍수빈도와 계절별 온도변동에 관련하여 크게 증가함을 알 수 있었다. nirK 유전자는 모든 시료에서 발견되지 않았다. Canonical correspondence analysis (CCA) 분석결과는 탈질균 군집 양과 영양염류 가용도와 잠재 탈질율 사이에는 약한 상관관계가 있음을 보여주었다.

Multi-Bioindicators to Assess Soil Microbial Activity in the Context of an Artificial Groundwater Recharge with Treated Wastewater: A Large-Scale Pilot Experiment

  • Michel, Caroline;Joulian, Catherine;Ollivier, Patrick;Nyteij, Audrey;Cote, Remi;Surdyk, Nicolas;Hellal, Jennifer;Casanova, Joel;Besnard, Katia;Rampnoux, Nicolas;Garrido, Francis
    • Journal of Microbiology and Biotechnology
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    • 제24권6호
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    • pp.843-853
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    • 2014
  • In the context of artificial groundwater recharge, a reactive soil column at pilot-scale (4.5 m depth and 3 m in diameter) fed by treated wastewater was designed to evaluate soil filtration ability. Here, as a part of this project, the impact of treated wastewater filtration on soil bacterial communities and the soil's biological ability for wastewater treatment as well as the relevance of the use of multi-bioindicators were studied as a function of depth and time. Biomass; bacterial 16S rRNA gene diversity fingerprints; potential nitrifying, denitrifying, and sulfate-reducing activities; and functional gene (amo, nir, nar, and dsr) detection were analyzed to highlight the real and potential microbial activity and diversity within the soil column. These bioindicators show that topsoil (0 to 20 cm depth) was the more active and the more impacted by treated wastewater filtration. Nitrification was the main activity in the pilot. No sulfate-reducing activity or dsr genes were detected during the first 6 months of wastewater application. Denitrification was also absent, but genes of denitrifying bacteria were detected, suggesting that the denitrifying process may occur rapidly if adequate chemical conditions are favored within the soil column. Results also underline that a dry period (20 days without any wastewater supply) significantly impacted soil bacterial diversity, leading to a decrease of enzyme activities and biomass. Finally, our work shows that treated wastewater filtration leads to a modification of the bacterial genetic and functional structures in topsoil.

탈수소효소(脫水素酵素), 탈질균(脫窒菌) 및 황산환원균(黃酸還元菌)의 정량(定量)을 통(通)한 온산공단(溫山工團)과 마니산(摩尼山) 산림토양(山林土壤)의 동태(動態) 조사(調査) (Investigation on Forest Soil Dynamics at Onsan Industrial Estate and Mt. Mani by the Assay of Dehydrogenase Activity, Denitrifying and Sulfur-Reducing Bacteria)

  • 박현
    • 한국산림과학회지
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    • 제87권1호
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    • pp.106-112
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    • 1998
  • 본 연구는 토양의 화학적 특성과 탈질소, 황 환원에 관여하는 세균류 및 탈수소효소 활성의 관계를 살펴보고, 궁극적으로는 산림 토양생태계의 동태를 평가하는 도구로 탈수소효소를 활용할 수 있는지에 대하여 고찰한 것이다. 오염지역으로 여겨지는 온산공단 주변과 해안의 청정지역인 강화도 마니산에서 각각 침엽수림과 활엽수림을 산정, 총 4개 임분을 대상으로 본 연구를 수행하였는데, 마니산 토양은 온산공단 주변에 비하여 상대적으로 유기물, 질소, 유효인산 등의 함량이 높아 미생물의 활동에 유리한 것으로 여겨졌다. 한편, 탈질균이나 황산환원균은 지역간 차이가 없었던 반면, 탈수소효소의 활성은 지역이나 시기에 따른 변화를 뚜렷이 나타내어 두 종류의 미생물 정량기법에 비하여 훨씬 민감하게 나타났다. 또한, 탈수소효소는 미생물의 활성과 깊은 관계가 있는 것으로 여겨지는 유기물함량(r=0.53. p=0.004), 전질소(r=0.41. p=0.008)나 C/Ava. P 비율(r=-0.52. p=0.001) 등과 비교적 높은 상관관계를 나타내었다. 따라서, 비슷한 토성을 지녀야 비교가 가능하다는 다른 연구자들의 지적을 감안하여 수행한다면, 토양생태계의 동태를 파악하는 지표로서 탈수소효소를 활용할 수 있을 것으로 여겨진다.

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