The transcriptional regulator NtrC controls glucose-6-phosphate dehydrogenase expression and polyhydroxybutyrate synthesis through NADPH availability in Herbaspirillum seropedicae

Sci Rep. 2017 Oct 19;7(1):13546. doi: 10.1038/s41598-017-12649-0.

Abstract

The NTR system is the major regulator of nitrogen metabolism in Bacteria. Despite its broad and well-known role in the assimilation, biosynthesis and recycling of nitrogenous molecules, little is known about its role in carbon metabolism. In this work, we present a new facet of the NTR system in the control of NADPH concentration and the biosynthesis of molecules dependent on reduced coenzyme in Herbaspirillum seropedicae SmR1. We demonstrated that a ntrC mutant strain accumulated high levels of polyhydroxybutyrate (PHB), reaching levels up to 2-fold higher than the parental strain. In the absence of NtrC, the activity of glucose-6-phosphate dehydrogenase (encoded by zwf) increased by 2.8-fold, consequently leading to a 2.1-fold increase in the NADPH/NADP+ ratio. A GFP fusion showed that expression of zwf is likewise controlled by NtrC. The increase in NADPH availability stimulated the production of polyhydroxybutyrate regardless the C/N ratio in the medium. The mutant ntrC was more resistant to H2O2 exposure and controlled the propagation of ROS when facing the oxidative condition, a phenotype associated with the increase in PHB content.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Chromatography, Gas
  • Gene Expression Regulation, Bacterial
  • Glucosephosphate Dehydrogenase / genetics
  • Glucosephosphate Dehydrogenase / metabolism*
  • Herbaspirillum / drug effects
  • Herbaspirillum / enzymology
  • Herbaspirillum / metabolism*
  • Hydrogen Peroxide / toxicity
  • Hydroxybutyrates / analysis
  • Hydroxybutyrates / metabolism*
  • Monosaccharides / metabolism
  • Mutagenesis
  • NADP / metabolism*
  • Nitrogen / metabolism
  • Oxidative Stress / drug effects
  • Polyesters / analysis
  • Polyesters / metabolism*
  • Reactive Oxygen Species / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*

Substances

  • Bacterial Proteins
  • Hydroxybutyrates
  • Monosaccharides
  • Polyesters
  • Reactive Oxygen Species
  • Transcription Factors
  • poly-beta-hydroxybutyrate
  • NADP
  • Hydrogen Peroxide
  • Glucosephosphate Dehydrogenase
  • Nitrogen