• Title/Summary/Keyword: 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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    • v.13 no.3
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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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    • v.11 no.1
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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.

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

  • Vo, Nguyen Xuan Que;Lee, Seung-Hoon;Doan, Tuan Van;Jung, Sokhee P.;Kang, Hojeong
    • Korean Journal of Microbiology
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    • v.50 no.2
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    • pp.137-151
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    • 2014
  • Various studies have been conducted to investigate effects of dams on river ecosystems, but less information is available regarding damming impacts on downstream denitrification. We measured denitrification enzyme activity (potential denitrification rate) and denitrifier abundances (using nirS, nirK, and nosZ as markers) in dammed headstreams of the Nakdong River in South Korea. Sediments in Phragmites-dominated riparian areas and in-stream areas across streams (dammed vs. reference) with different streambed materials (gravel and sand) were sampled occasionally. We hypothesized that (i) the higher available N and C contents in sediments downstream of dams foster larger denitrifier communities than in the reference system and (ii) differences in potential denitrification rates across the systems correspond with denitrifier abundances. Despite 30 years of different hydrological management with dams and greater inorganic N and DOC contents in sediments downstream of dams, compared to the references, abundances of denitrifier communities and potential denitrification rates within the whole sediment were not significantly different across the systems. However, nirS and nosZ denitrifier abundances and potential denitrification rates were considerably increased in specific sediments downstream of dams (gravelly riparian and sandy in-stream) with regard to flooding events and seasonal temperature variation. nirK was not amplified in all sediments. Canonical correspondence analyses (CCA) revealed that the relationship between abundances of denitrifier communities and nutrient availabilities and potential denitrification rates was a weak one.

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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    • v.24 no.6
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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.

Investigation on Forest Soil Dynamics at Onsan Industrial Estate and Mt. Mani by the Assay of Dehydrogenase Activity, Denitrifying and Sulfur-Reducing Bacteria (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))

  • Park, Hyun
    • Journal of Korean Society of Forest Science
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    • v.87 no.1
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    • pp.106-112
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    • 1998
  • This study was conducted to figure out the relationships among soil chemical properties and bacterial biomass related to denitrification and sulfur-reducing and the activity of dehydrogenase, and ultimately to consider the usefulness of dehydrogenase activity as a tool for evaluating the dynamics of forest soil ecosystem. Four sites were selected for the collection of soil samples within two regions(Onsan industrial estate as a polluted region and Mt. Mani at Kanghwa island as a clean area) with two forest types (coniferous and deciduous stands). The soils of Mt. Mani showed higher amount of organic matter, total nitrogen and available phosphorus than those collected from Onsan industrial estate, which indicated that the soils were more beneficial for microbial growth than those of Onsan. The dehydrogenase activity was more sensitive than the denitrifying bacteria or sulfur-reducing bacteria since the activity was significantly different between the regions and season while the two bacterial biomass were not significantly different between the two regions. In addition, the dehydrogenase activity showed relatively high correlation coefficients with organic matter(r=0.53, p=0.004), total nitrogen(r=0.41, p=0.008) and C/Ava. P-ratio(r=-0.52, p=0.001), which was thought to be closely related with microbial activity. Thus, the dehydrogenase activity was thought to be a useful index of soil ecosystem dynamics with considering that the technique need to be applied with the same soil texture for the comparison of the activity as other researchers indicated.

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