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Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases
Biological hydrogen production is based on activity of specific enzymes called hydrogenases. Hydrogenases are oxygen sensitive metalloenzymes containing Ni and/or Fe atoms at the active site, catalyzing reversible reduction of protons. Generally, [Fe-Fe] hydrogenases prefer proton reduction to molec...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5081567/ https://www.ncbi.nlm.nih.gov/pubmed/27786270 http://dx.doi.org/10.1038/srep36034 |
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author | Mangayil, Rahul Karp, Matti Lamminmäki, Urpo Santala, Ville |
author_facet | Mangayil, Rahul Karp, Matti Lamminmäki, Urpo Santala, Ville |
author_sort | Mangayil, Rahul |
collection | PubMed |
description | Biological hydrogen production is based on activity of specific enzymes called hydrogenases. Hydrogenases are oxygen sensitive metalloenzymes containing Ni and/or Fe atoms at the active site, catalyzing reversible reduction of protons. Generally, [Fe-Fe] hydrogenases prefer proton reduction to molecular hydrogen, a potential energy carrier molecule that can be produced by bioprocesses in sustainable manner. Thus, monitoring tools have been developed to study the relationship between [Fe-Fe] hydrogenases and biohydrogen production in bioreactors at DNA and RNA levels. In the present study, novel molecular tools are introduced for quantitative monitoring of clostridial [Fe-Fe] hydrogenases at the protein level. Aerobic and anaerobic biopanning (for inactive and active [Fe-Fe] hydrogenase, respectively) of phage displayed single-chain variable fragment (scFv) antibody libraries aided in isolating nine potential scFvs. The enriched antibodies demonstrated high specificity towards Clostridium spp. [Fe-Fe] hydrogenases allowing detection from pure and mixed cultures. Additionally, the antibodies showed different binding characteristics towards hydrogenase catalytic states, providing a possible means for functional detection of clostridial [Fe-Fe] hydrogenases. From hydrogenase-antibody interaction studies we observed that though antibody binding reduced the enzyme catalytic activity, it facilitated to retain hydrogen evolution from oxygen exposed hydrogenases. |
format | Online Article Text |
id | pubmed-5081567 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-50815672016-10-31 Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases Mangayil, Rahul Karp, Matti Lamminmäki, Urpo Santala, Ville Sci Rep Article Biological hydrogen production is based on activity of specific enzymes called hydrogenases. Hydrogenases are oxygen sensitive metalloenzymes containing Ni and/or Fe atoms at the active site, catalyzing reversible reduction of protons. Generally, [Fe-Fe] hydrogenases prefer proton reduction to molecular hydrogen, a potential energy carrier molecule that can be produced by bioprocesses in sustainable manner. Thus, monitoring tools have been developed to study the relationship between [Fe-Fe] hydrogenases and biohydrogen production in bioreactors at DNA and RNA levels. In the present study, novel molecular tools are introduced for quantitative monitoring of clostridial [Fe-Fe] hydrogenases at the protein level. Aerobic and anaerobic biopanning (for inactive and active [Fe-Fe] hydrogenase, respectively) of phage displayed single-chain variable fragment (scFv) antibody libraries aided in isolating nine potential scFvs. The enriched antibodies demonstrated high specificity towards Clostridium spp. [Fe-Fe] hydrogenases allowing detection from pure and mixed cultures. Additionally, the antibodies showed different binding characteristics towards hydrogenase catalytic states, providing a possible means for functional detection of clostridial [Fe-Fe] hydrogenases. From hydrogenase-antibody interaction studies we observed that though antibody binding reduced the enzyme catalytic activity, it facilitated to retain hydrogen evolution from oxygen exposed hydrogenases. Nature Publishing Group 2016-10-27 /pmc/articles/PMC5081567/ /pubmed/27786270 http://dx.doi.org/10.1038/srep36034 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Mangayil, Rahul Karp, Matti Lamminmäki, Urpo Santala, Ville Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases |
title | Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases |
title_full | Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases |
title_fullStr | Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases |
title_full_unstemmed | Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases |
title_short | Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases |
title_sort | recombinant antibodies for specific detection of clostridial [fe-fe] hydrogenases |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5081567/ https://www.ncbi.nlm.nih.gov/pubmed/27786270 http://dx.doi.org/10.1038/srep36034 |
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