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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2016
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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. |
format | Online Article Text |
id | pubmed-4816682 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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