Glutamine synthetase gene glnA plays a vital role in curdlan biosynthesis of Agrobacterium sp. CGMCC 11546

Int J Biol Macromol. 2020 Dec 15;165(Pt A):222-230. doi: 10.1016/j.ijbiomac.2020.09.152. Epub 2020 Sep 25.

Abstract

Curdlan is a neutral linear exopolysaccharide produced by Agrobacterium spp. under nitrogen-limiting conditions. In this study, we explored the role of glnA in curdlan biosynthesis in Agrobacterium sp. CGMCC 11546. The curdlan production of the ΔglnA strain was impaired, decreasing by 93% compared with that of the wild-type strain after 96 h fermentation. Analysis of fermentation profiles revealed that cell growth and utilization of carbon and nitrogen sources were impaired in the ΔglnA strain. Transcriptome analysis indicated that various of genes involved in curdlan biosynthesis were downregulated after 24 h fermentation in the ΔglnA strain, particularly genes involved in heme synthesis and the electron transport chain, which are essential for energy generation. Metabolomics analysis revealed flavin adenine dinucleotide (FAD) and adenosine diphosphate (ADP) accumulation in the ΔglnA strain, suggesting insufficient energy supply. Furthermore, glnA overexpression led to an 18% increase in the curdlan yield of the ΔglnA mutant compared with that of the wild-type strain after 96 h fermentation. Taken together, the findings demonstrate that glnA plays a vital role in curdlan biosynthesis by supplying ATP via regulating the expression of genes involved in heme synthesis and the electron transport chain.

Keywords: Agrobacterium; Curdlan; glnA.

MeSH terms

  • Agrobacterium / genetics
  • Agrobacterium / metabolism*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Glutamate-Ammonia Ligase / genetics
  • Glutamate-Ammonia Ligase / metabolism*
  • Mutation
  • beta-Glucans / metabolism*

Substances

  • Bacterial Proteins
  • beta-Glucans
  • curdlan
  • glutamine synthetase I
  • Glutamate-Ammonia Ligase