13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16

Introduction. Cupriavidus necator H16 is a gram-negative bacterium, capable of lithoautotrophic growth by utilizing hydrogen as an energy source and fixing carbon dioxide (CO2) through Calvin-Benson-Bassham (CBB) cycle. The potential to utilize synthesis gas (Syngas) and the prospects of rerouting c...

Full description

Bibliographic Details
Main Authors:	Alagesan, Swathi, Minton, Nigel P., Malys, Naglis
Format:	Article
Language:	English
Published:	Springer 2018
Online Access:	http://eprints.nottingham.ac.uk/48536/ http://eprints.nottingham.ac.uk/48536/ http://eprints.nottingham.ac.uk/48536/ http://eprints.nottingham.ac.uk/48536/8/s11306-017-1302-z.pdf

id	nottingham-48536
recordtype	eprints
spelling	nottingham-485362017-12-06T02:08:49Z http://eprints.nottingham.ac.uk/48536/ 13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16 Alagesan, Swathi Minton, Nigel P. Malys, Naglis Introduction. Cupriavidus necator H16 is a gram-negative bacterium, capable of lithoautotrophic growth by utilizing hydrogen as an energy source and fixing carbon dioxide (CO2) through Calvin-Benson-Bassham (CBB) cycle. The potential to utilize synthesis gas (Syngas) and the prospects of rerouting carbon from polyhydroxybutyrate synthesis to value-added compounds makes C. necator an excellent chassis for industrial application. Objectives. In the context of lack of sufficient quantitative information of the metabolic pathways and to advance in rational metabolic engineering for optimized product synthesis in C. necator H16, we carried out a metabolic flux analysis based on steady-state 13C-labelling. Methods. In this study, steady-state carbon labelling experiments, using either D-[1-13C]fructose or [1,2-13C]glycerol, were undertaken to investigate the carbon flux through the central carbon metabolism in C. necator H16 under heterotrophic and mixotrophic growth conditions, respectively. Results. We found that the CBB cycle is active even under heterotrophic condition, and growth is indeed mixotrophic. While Entner-Doudoroff (ED) pathway is shown to be the major route for sugar degradation, tricarboxylic acid (TCA) cycle is highly active in mixotrophic condition. Enhanced flux is observed in reductive pentose phosphate pathway (redPPP) under the mixotrophic condition to supplement the precursor requirement for CBB cycle. The flux distribution was compared to the mRNA abundance of genes encoding enzymes involved in key enzymatic reactions of the central carbon metabolism. Conclusion. This study leads the way to establishing 13C-based quantitative fluxomics for rational pathway engineering in C. necator H16. Springer 2018-01 Article PeerReviewed application/pdf en cc_by http://eprints.nottingham.ac.uk/48536/8/s11306-017-1302-z.pdf Alagesan, Swathi and Minton, Nigel P. and Malys, Naglis (2018) 13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16. Metabolomics, 14 (9). pp. 1-9. ISSN 1573-3890 https://link.springer.com/article/10.1007%2Fs11306-017-1302-z doi:10.1007/s11306-017-1302-z doi:10.1007/s11306-017-1302-z
repository_type	Digital Repository
institution_category	Local University
institution	University of Nottingham Malaysia Campus
building	Nottingham Research Data Repository
collection	Online Access
language	English
description	Introduction. Cupriavidus necator H16 is a gram-negative bacterium, capable of lithoautotrophic growth by utilizing hydrogen as an energy source and fixing carbon dioxide (CO2) through Calvin-Benson-Bassham (CBB) cycle. The potential to utilize synthesis gas (Syngas) and the prospects of rerouting carbon from polyhydroxybutyrate synthesis to value-added compounds makes C. necator an excellent chassis for industrial application. Objectives. In the context of lack of sufficient quantitative information of the metabolic pathways and to advance in rational metabolic engineering for optimized product synthesis in C. necator H16, we carried out a metabolic flux analysis based on steady-state 13C-labelling. Methods. In this study, steady-state carbon labelling experiments, using either D-[1-13C]fructose or [1,2-13C]glycerol, were undertaken to investigate the carbon flux through the central carbon metabolism in C. necator H16 under heterotrophic and mixotrophic growth conditions, respectively. Results. We found that the CBB cycle is active even under heterotrophic condition, and growth is indeed mixotrophic. While Entner-Doudoroff (ED) pathway is shown to be the major route for sugar degradation, tricarboxylic acid (TCA) cycle is highly active in mixotrophic condition. Enhanced flux is observed in reductive pentose phosphate pathway (redPPP) under the mixotrophic condition to supplement the precursor requirement for CBB cycle. The flux distribution was compared to the mRNA abundance of genes encoding enzymes involved in key enzymatic reactions of the central carbon metabolism. Conclusion. This study leads the way to establishing 13C-based quantitative fluxomics for rational pathway engineering in C. necator H16.
format	Article
author	Alagesan, Swathi Minton, Nigel P. Malys, Naglis
spellingShingle	Alagesan, Swathi Minton, Nigel P. Malys, Naglis 13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16
author_facet	Alagesan, Swathi Minton, Nigel P. Malys, Naglis
author_sort	Alagesan, Swathi
title	13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16
title_short	13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16
title_full	13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16
title_fullStr	13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16
title_full_unstemmed	13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16
title_sort	13c-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in cupriavidus necator h16
publisher	Springer
publishDate	2018
url	http://eprints.nottingham.ac.uk/48536/ http://eprints.nottingham.ac.uk/48536/ http://eprints.nottingham.ac.uk/48536/ http://eprints.nottingham.ac.uk/48536/8/s11306-017-1302-z.pdf
first_indexed	2018-09-06T14:00:18Z
last_indexed	2018-09-06T14:00:18Z
_version_	1610866930722275328

13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16

Similar Items