NDLI: Axenic Cultures of Nitrosomonas europaea and Nitrobacter winogradskyi in Autotrophic Conditions: a New Protocol for Kinetic Studies

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Axenic Cultures of Nitrosomonas europaea and Nitrobacter winogradskyi in Autotrophic Conditions: a New Protocol for Kinetic Studies

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Farges, B. Poughon, L. Roriz, D. Creuly, C. Dussap, C-G Lasseur, C.
Description	Country affiliation: France Author Affiliation: Farges B ( Laboratoire de Génie Chimique et Biochimique, Polytech Clermont-Ferrand, Clermont Université, 24 Avenue des Landais, BP 206 Aubière Cedex, France.)
Abstract	As a part of a natural biological N-cycle, nitrification is one of the steps included in the conception of artificial ecosystems designed for extraterrestrial life support systems (LSS) such as Micro-Ecological Life Support System Alternative (MELiSSA) project, which is the LSS project of the European Space Agency. Nitrification in aerobic environments is carried out by two groups of bacteria in a two-step process. The ammonia-oxidizing bacteria (Nitrosomonas europaea) realize the oxidation of ammonia to nitrite, and the nitrite-oxidizing bacteria (Nitrobacter winogradskyi), the oxidation of nitrite to nitrate. In both cases, the bacteria achieve these oxidations to obtain an energy and reductant source for their growth and maintenance. Furthermore, both groups also use $CO_{2}$ predominantly as their carbon source. They are typically found together in ecosystems, and consequently, nitrite accumulation is rare. Due to the necessity of modeling accurately conversion yields and transformation rates to achieve a complete modeling of MELiSSA, the present study focuses on the experimental determination of nitrogen to biomass conversion yields. Kinetic and mass balance studies for axenic cultures of Nitrosomonas europaea and Nitrobacter winogradskyi in autotrophic conditions are performed. The follow-up of these cultures is done using flow cytometry for assessing biomass concentrations and ionic chromatography for ammonium, nitrite, and nitrate concentrations. A linear correlation is observed between cell count and optical density (OD) measurement (within a 10 % accuracy) validating OD measurements for an on-line estimation of biomass quantity even at very low biomass concentrations. The conversion between cell count and biomass concentration has been determined: $7.1 × 10^{12}$ cells g dry matter $(DM)^{−1}$ for Nitrobacter and $6.3 × 10^{12}$ cells g $DM^{−1}$ for Nitrosomonas. Nitrogen substrates and products are assessed redundantly showing excellent agreement for mass balance purposes and conversion yields determination. Although the dominant phenomena are the oxidation of $NH_{4}^{+}$ into nitrite (0.95 mol mol $N^{−1}$ for Nitrosomonas europaea within an accuracy of 3 %) and nitrite into nitrate (0.975 mol mol $N^{−1}$ for Nitrobacter winogradskyi within an accuracy of 2 %), the Nitrosomonas europaea conversion yield is estimated to be 0.42 g DM $mol N^{−1},$ and Nitrobacter winogradskyi conversion yield is estimated to be 0.27 g DM $mol N^{−1}.$ The growth rates of both strains appear to be dominated by the oxygen transfer into the experimental setups.
File Format	HTM / HTML
ISSN	02732289
Issue Number	5
Volume Number	167
e-ISSN	15590291
Journal	Applied Biochemistry and Biotechnology
Language	English
Publisher	Springer
Publisher Date	2012-07-01
Publisher Place	United States
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Biochemistry Discipline Biotechnology Autotrophic Processes Axenic Culture Methods Nitrobacter Growth & Development Nitrosomonas Europaea Ammonia Metabolism Batch Cell Culture Techniques Flow Cytometry Kinetics Nitrites Optical Phenomena Oxidation-reduction Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Medicine Molecular Biology Environmental Engineering Biochemistry Bioengineering Applied Microbiology and Biotechnology Biotechnology

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Axenic Cultures of Nitrosomonas europaea and Nitrobacter winogradskyi in Autotrophic Conditions: a New Protocol for Kinetic Studies

Metaproteomics, Heterotrophic Growth, and Distribution of Nitrosomonas europaea and Nitrobacter winogradskyi after Long-Term Operation of an Autotrophic Nitrifying Biofilm Reactor

Ammonia stimulates growth and nitrite-oxidizing activity of Nitrobacter winogradskyi

Limited impact of free ammonia on Nitrobacter spp. inhibition assessed by chemical and molecular techniques.

Maintenance Energy Demand and Starvation Recovery Dynamics of Nitrosomonas europaea and Nitrobacter winogradskyi Cultivated in a Retentostat with Complete Biomass Retention

Application of statistical techniques for elucidating flow cytometric data of batch and fed-batch cultures.

Influence of free ammonia on completely autotrophic nitrogen removal over nitrite (CANON) process.

Kinetics of nitrite oxidation by Nitrobacter winogradskyi

Kinetic and product distribution analysis of NO* reductase activity in Nitrosomonas europaea hydroxylamine oxidoreductase.

Axenic Cultures of Nitrosomonas europaea and Nitrobacter winogradskyi in Autotrophic Conditions: a New Protocol for Kinetic Studies

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Axenic Cultures of Nitrosomonas europaea and Nitrobacter winogradskyi in Autotrophic Conditions: a New Protocol for Kinetic Studies

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Axenic Cultures of Nitrosomonas europaea and Nitrobacter winogradskyi in Autotrophic Conditions: a New Protocol for Kinetic Studies