Metabolic flux analysis and the NAD(P)H/NAD(P) + ratios in chemostat cultures of Azotobacter vinelandii
García, Andres (Universidad Nacional Autònoma de México. Instituto de Biotecnologia)
Ferrer, Pau (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Albiol i Sala, Joan (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Castillo, Tania (Universidad Nacional Autònoma de México. Instituto de Biotecnologia)
Segura, Daniel (Universidad Nacional Autònoma de México. Instituto de Biotecnologia)
Peña, Carlos (Universidad Nacional Autònoma de México. Instituto de Biotecnologia)

Fecha:	2018
Resumen:	Azotobacter vinelandii is a bacterium that produces alginate and polyhydroxybutyrate (P3HB); however, the role of NAD(P)H/NAD(P) + ratios on the metabolic fluxes through biosynthesis pathways of these biopolymers remains unknown. The aim of this study was to evaluate the NAD(P)H/NAD(P) + ratios and the metabolic fluxes involved in alginate and P3HB biosynthesis, under oxygen-limiting and non-limiting oxygen conditions. The results reveal that changes in the oxygen availability have an important effect on the metabolic fluxes and intracellular NADPH/NADP + ratio, showing that at the lowest OTR (2. 4 mmol L −1 h −1), the flux through the tricarboxylic acid (TCA) cycle decreased 27. 6-fold, but the flux through the P3HB biosynthesis increased 6. 6-fold in contrast to the cultures without oxygen limitation (OTR = 14. 6 mmol L −1 h −1). This was consistent with the increase in the level of transcription of phbB and the P3HB biosynthesis. In addition, under conditions without oxygen limitation, there was an increase in the carbon uptake rate (twofold), as well as in the flux through the pentose phosphate (PP) pathway (4. 8-fold), compared to the condition of 2. 4 mmol L −1 h −1. At the highest OTR condition, a decrease in the NADPH/NADP + ratio of threefold was observed, probably as a response to the high respiration rate induced by the respiratory protection of the nitrogenase under diazotrophic conditions, correlating with a high expression of the uncoupled respiratory chain genes (ndhII and cydA) and induction of the expression of the genes encoding the nitrogenase complex (nifH). We have demonstrated that changes in oxygen availability affect the internal redox state of the cell and carbon metabolic fluxes. This also has a strong impact on the TCA cycle and PP pathway as well as on alginate and P3HB biosynthetic fluxes.
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Materia:	NAD(P)H/NAD(P) + ratios ; Metabolic flux analysis ; Oxygen availability ; Azotobacter vinelandii
Publicado en:	Microbial cell factories, Vol. 17 (January 2018) , art. 10, ISSN 1475-2859

DOI: 10.1186/s12934-018-0860-8
PMID: 29357933

13 p, 1.7 MB

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