Constitutive expression of Vitreoscilla haemoglobin in Sphingomonas elodea to improve gellan gum production

J Appl Microbiol. 2011 Feb;110(2):422-30. doi: 10.1111/j.1365-2672.2010.04896.x. Epub 2010 Nov 19.

Abstract

Aims: To improve a commercially used strain for gellan production by exogenous Vitreoscilla haemoglobin (VHb).

Methods and results: VHb gene was expressed in Sphingomonas elodea under the control of constitutive bla promoter. Biochemical activity of expressed VHb was confirmed by CO-difference spectra analysis that exhibited a characteristic absorption maximum at 419 nm. During cultivation, not only enhanced cell growth was detected, but also 20% improvement in gellan production was observed after 48 h of incubation, with a maximum yield of 16·82 g l(-1). Moreover, maximum sucrose conversion efficiency (g gellan per g sucrose) was 57·8, 20% higher than that of the parental strain. We further examined the polysaccharide production of VHb-expressing strain at different aeration levels in Erlenmeyer flasks. Again, in all cases, a significant enhancement of gellan production was observed, and the enhancement was more significant under oxygen-limiting conditions (up to 26·8%).

Conclusions: VHb exhibited positive effect on cell growth and gellan yield of S. elodea, especially under hypoxic conditions.

Significance and impact of the study: This is the first application of VHb as an effective metabolic engineering strategy in S. elodea to regulate cell growth and optimize gellan yield.

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Bioreactors
  • Fermentation
  • Polysaccharides, Bacterial / biosynthesis*
  • Sphingomonas / genetics
  • Sphingomonas / growth & development
  • Sphingomonas / metabolism*
  • Truncated Hemoglobins / genetics
  • Truncated Hemoglobins / metabolism*

Substances

  • Bacterial Proteins
  • Polysaccharides, Bacterial
  • Truncated Hemoglobins
  • hemoglobin protein, Vitreoscilla
  • gellan gum