Rational flux-tuning of Halomonas bluephagenesis for co-production of bioplastic PHB and ectoine

Nat Commun. 2020 Jul 3;11(1):3313. doi: 10.1038/s41467-020-17223-3.

Abstract

Ectoine, a compatible solute synthesized by many halophiles for hypersalinity resistance, has been successfully produced by metabolically engineered Halomonas bluephagenesis, which is a bioplastic poly(3-hydroxybutyrate) producer allowing open unsterile and continuous conditions. Here we report a de novo synthesis pathway for ectoine constructed into the chromosome of H. bluephagenesis utilizing two inducible systems, which serve to fine-tune the transcription levels of three clusters related to ectoine synthesis, including ectABC, lysC and asd based on a GFP-mediated transcriptional tuning approach. Combined with bypasses deletion, the resulting recombinant H. bluephagenesis TD-ADEL-58 is able to produce 28 g L-1 ectoine during a 28 h fed-batch growth process. Co-production of ectoine and PHB is achieved to 8 g L-1 ectoine and 32 g L-1 dry cell mass containing 75% PHB after a 44 h growth. H. bluephagenesis demonstrates to be a suitable co-production chassis for polyhydroxyalkanoates and non-polymer chemicals such as ectoine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids, Diamino / biosynthesis*
  • Biomass
  • Biosynthetic Pathways / genetics
  • Chromatography, Liquid / methods
  • Halomonas / genetics
  • Halomonas / growth & development
  • Halomonas / metabolism*
  • Hydroxybutyrates / metabolism*
  • Metabolic Engineering / methods
  • Polyesters / metabolism*
  • Polyhydroxyalkanoates / chemistry
  • Polyhydroxyalkanoates / metabolism
  • Tandem Mass Spectrometry / methods

Substances

  • Amino Acids, Diamino
  • Hydroxybutyrates
  • Polyesters
  • Polyhydroxyalkanoates
  • ectoine