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NADPH‐related processes studied with a SoxR‐based biosensor in Escherichia coli

NADPH plays a crucial role in cellular metabolism for biosynthesis and oxidative stress responses. We previously developed the genetically encoded NADPH biosensor pSenSox based on the transcriptional regulator SoxR of Escherichia coli, its target promoter P(soxS) and eYFP as fluorescent reporter. He...

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Detalles Bibliográficos
Autores principales: Spielmann, Alina, Baumgart, Meike, Bott, Michael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6612552/
https://www.ncbi.nlm.nih.gov/pubmed/30585443
http://dx.doi.org/10.1002/mbo3.785
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author Spielmann, Alina
Baumgart, Meike
Bott, Michael
author_facet Spielmann, Alina
Baumgart, Meike
Bott, Michael
author_sort Spielmann, Alina
collection PubMed
description NADPH plays a crucial role in cellular metabolism for biosynthesis and oxidative stress responses. We previously developed the genetically encoded NADPH biosensor pSenSox based on the transcriptional regulator SoxR of Escherichia coli, its target promoter P(soxS) and eYFP as fluorescent reporter. Here, we used pSenSox to study the influence of various parameters on the sensor output in E. coliduring reductive biotransformation of methyl acetoacetate (MAA) to (R)‐methyl 3‐hydroxybutyrate (MHB) by the strictly NADPH‐dependent alcohol dehydrogenase of Lactobacillus brevis (LbAdh). Redox‐cycling drugs such as paraquat and menadione strongly activated the NADPH biosensor and mechanisms responsible for this effect are discussed. Absence of the RsxABCDGE complex and/or RseC caused an enhanced biosensor response, supporting a function as SoxR‐reducing system. Absence of the membrane‐bound transhydrogenase PntAB caused an increased biosensor response, whereas the lack of the soluble transhydrogenase SthA or of SthA and PntAB was associated with a strongly decreased response. These data support the opposing functions of PntAB in NADP(+) reduction and of SthA in NADPH oxidation. In summary, the NADPH biosensor pSenSox proved to be a useful tool to study NADPH‐related processes in E. coli.
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spelling pubmed-66125522019-07-16 NADPH‐related processes studied with a SoxR‐based biosensor in Escherichia coli Spielmann, Alina Baumgart, Meike Bott, Michael Microbiologyopen Original Articles NADPH plays a crucial role in cellular metabolism for biosynthesis and oxidative stress responses. We previously developed the genetically encoded NADPH biosensor pSenSox based on the transcriptional regulator SoxR of Escherichia coli, its target promoter P(soxS) and eYFP as fluorescent reporter. Here, we used pSenSox to study the influence of various parameters on the sensor output in E. coliduring reductive biotransformation of methyl acetoacetate (MAA) to (R)‐methyl 3‐hydroxybutyrate (MHB) by the strictly NADPH‐dependent alcohol dehydrogenase of Lactobacillus brevis (LbAdh). Redox‐cycling drugs such as paraquat and menadione strongly activated the NADPH biosensor and mechanisms responsible for this effect are discussed. Absence of the RsxABCDGE complex and/or RseC caused an enhanced biosensor response, supporting a function as SoxR‐reducing system. Absence of the membrane‐bound transhydrogenase PntAB caused an increased biosensor response, whereas the lack of the soluble transhydrogenase SthA or of SthA and PntAB was associated with a strongly decreased response. These data support the opposing functions of PntAB in NADP(+) reduction and of SthA in NADPH oxidation. In summary, the NADPH biosensor pSenSox proved to be a useful tool to study NADPH‐related processes in E. coli. John Wiley and Sons Inc. 2018-12-25 /pmc/articles/PMC6612552/ /pubmed/30585443 http://dx.doi.org/10.1002/mbo3.785 Text en © 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Spielmann, Alina
Baumgart, Meike
Bott, Michael
NADPH‐related processes studied with a SoxR‐based biosensor in Escherichia coli
title NADPH‐related processes studied with a SoxR‐based biosensor in Escherichia coli
title_full NADPH‐related processes studied with a SoxR‐based biosensor in Escherichia coli
title_fullStr NADPH‐related processes studied with a SoxR‐based biosensor in Escherichia coli
title_full_unstemmed NADPH‐related processes studied with a SoxR‐based biosensor in Escherichia coli
title_short NADPH‐related processes studied with a SoxR‐based biosensor in Escherichia coli
title_sort nadph‐related processes studied with a soxr‐based biosensor in escherichia coli
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6612552/
https://www.ncbi.nlm.nih.gov/pubmed/30585443
http://dx.doi.org/10.1002/mbo3.785
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