Characterization and physiological role of two types of chloroplastic fructose-1,6-bisphosphatases in Euglena gracilis

Arch Biochem Biophys. 2015 Jun 1;575:61-8. doi: 10.1016/j.abb.2015.04.002. Epub 2015 Apr 20.

Abstract

The chloroplastic fructose-1,6-bisphosphatase (FBPase) is a late-limiting enzyme in the Calvin cycle. In the present study, we isolated and characterized the cDNAs encoding two types of chloroplastic FBPase isoforms (EgFBPaseI and II) from Euglena gracilis. The Km values of recombinant EgFBPaseI and EgFBPaseII for fructose 1,6-bisphosphate (Fru 1,6-P2) were 165 ± 17 and 2200 ± 200 μM, respectively. The activity of EgFBPaseI was inhibited by 1mM H2O2 and recovered when incubated with DTT. The activity of EgFBPaseII was resistant to concentrations of H2O2 up to 1mM, which was distinct from those of EgFBPaseI and spinach chloroplastic FBPase. The suppression of EgFBPaseI gene expression by gene silencing markedly decreased photosynthetic activity and inhibited cell growth. The results of the present study clearly demonstrated that EgFBPaseI played a critical role in photosynthesis in Euglena chloroplasts.

Keywords: Euglena gracilis; Fructose-1,6-bisphosphatase; Photosynthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Chloroplasts / enzymology*
  • Euglena gracilis / enzymology*
  • Fructose-Bisphosphatase / chemistry
  • Fructose-Bisphosphatase / genetics
  • Fructose-Bisphosphatase / metabolism*
  • Molecular Sequence Data
  • Phylogeny
  • RNA Interference
  • Sequence Homology, Amino Acid

Substances

  • Fructose-Bisphosphatase