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Production and purification of a soluble hydrogenase from Ralstonia eutropha H16 for potential hydrogen fuel cell applications

The soluble hydrogenase (SH) from Ralstonia eutropha H16 is a promising candidate enzyme for H(2)-based biofuel application as it favours H(2) oxidation and is relatively oxygen-tolerant. In this report, bioprocess development studies undertaken to produce and purify an active SH are described, base...

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Autores principales: Jugder, Bat-Erdene, Lebhar, Helene, Aguey-Zinsou, Kondo-Francois, Marquis, Christopher P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4816682/
https://www.ncbi.nlm.nih.gov/pubmed/27077052
http://dx.doi.org/10.1016/j.mex.2016.03.005
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author Jugder, Bat-Erdene
Lebhar, Helene
Aguey-Zinsou, Kondo-Francois
Marquis, Christopher P.
author_facet Jugder, Bat-Erdene
Lebhar, Helene
Aguey-Zinsou, Kondo-Francois
Marquis, Christopher P.
author_sort Jugder, Bat-Erdene
collection PubMed
description The soluble hydrogenase (SH) from Ralstonia eutropha H16 is a promising candidate enzyme for H(2)-based biofuel application as it favours H(2) oxidation and is relatively oxygen-tolerant. In this report, bioprocess development studies undertaken to produce and purify an active SH are described, based on the methods previously reported [1], [2], [3], [4]. Our modifications are: • Upstream method optimizations were undertaken on heterotrophic growth media and cell lysis involving ultrasonication. • Two anion exchangers (Q Sepharose and RESOURCE Q) and size exclusion chromatographic (Superdex 200) matrices were successfully employed for purification of a hexameric SH from R. eutropha. • The H(2) oxidizing activity of the SH was demonstrated spectrophotometrically in solution and also immobilized on an EPG electrode using cyclic voltammetry.
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spelling pubmed-48166822016-04-13 Production and purification of a soluble hydrogenase from Ralstonia eutropha H16 for potential hydrogen fuel cell applications Jugder, Bat-Erdene Lebhar, Helene Aguey-Zinsou, Kondo-Francois Marquis, Christopher P. MethodsX Biochemistry, Genetics and Molecular Biology The soluble hydrogenase (SH) from Ralstonia eutropha H16 is a promising candidate enzyme for H(2)-based biofuel application as it favours H(2) oxidation and is relatively oxygen-tolerant. In this report, bioprocess development studies undertaken to produce and purify an active SH are described, based on the methods previously reported [1], [2], [3], [4]. Our modifications are: • Upstream method optimizations were undertaken on heterotrophic growth media and cell lysis involving ultrasonication. • Two anion exchangers (Q Sepharose and RESOURCE Q) and size exclusion chromatographic (Superdex 200) matrices were successfully employed for purification of a hexameric SH from R. eutropha. • The H(2) oxidizing activity of the SH was demonstrated spectrophotometrically in solution and also immobilized on an EPG electrode using cyclic voltammetry. Elsevier 2016-03-22 /pmc/articles/PMC4816682/ /pubmed/27077052 http://dx.doi.org/10.1016/j.mex.2016.03.005 Text en © 2016 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Biochemistry, Genetics and Molecular Biology
Jugder, Bat-Erdene
Lebhar, Helene
Aguey-Zinsou, Kondo-Francois
Marquis, Christopher P.
Production and purification of a soluble hydrogenase from Ralstonia eutropha H16 for potential hydrogen fuel cell applications
title Production and purification of a soluble hydrogenase from Ralstonia eutropha H16 for potential hydrogen fuel cell applications
title_full Production and purification of a soluble hydrogenase from Ralstonia eutropha H16 for potential hydrogen fuel cell applications
title_fullStr Production and purification of a soluble hydrogenase from Ralstonia eutropha H16 for potential hydrogen fuel cell applications
title_full_unstemmed Production and purification of a soluble hydrogenase from Ralstonia eutropha H16 for potential hydrogen fuel cell applications
title_short Production and purification of a soluble hydrogenase from Ralstonia eutropha H16 for potential hydrogen fuel cell applications
title_sort production and purification of a soluble hydrogenase from ralstonia eutropha h16 for potential hydrogen fuel cell applications
topic Biochemistry, Genetics and Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4816682/
https://www.ncbi.nlm.nih.gov/pubmed/27077052
http://dx.doi.org/10.1016/j.mex.2016.03.005
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