Archaeal fructose-1,6-bisphosphate aldolases constitute a new family of archaeal type class I aldolase

J Biol Chem. 2001 Aug 3;276(31):28710-8. doi: 10.1074/jbc.M103447200. Epub 2001 May 31.

Abstract

Fructose-1,6-bisphosphate (FBP) aldolase activity has been detected previously in several Archaea. However, no obvious orthologs of the bacterial and eucaryal Class I and II FBP aldolases have yet been identified in sequenced archaeal genomes. Based on a recently described novel type of bacterial aldolase, we report on the identification and molecular characterization of the first archaeal FBP aldolases. We have analyzed the FBP aldolases of two hyperthermophilic Archaea, the facultatively heterotrophic Crenarchaeon Thermoproteus tenax and the obligately heterotrophic Euryarchaeon Pyrococcus furiosus. For enzymatic studies the fba genes of T. tenax and P. furiosus were expressed in Escherichia coli. The recombinant FBP aldolases show preferred substrate specificity for FBP in the catabolic direction and exhibit metal-independent Class I FBP aldolase activity via a Schiff-base mechanism. Transcript analyses reveal that the expression of both archaeal genes is induced during sugar fermentation. Remarkably, the fbp gene of T. tenax is co-transcribed with the pfp gene that codes for the reversible PP(i)-dependent phosphofructokinase. As revealed by phylogenetic analyses, orthologs of the T. tenax and P. furiosus enzyme appear to be present in almost all sequenced archaeal genomes, as well as in some bacterial genomes, strongly suggesting that this new enzyme family represents the typical archaeal FBP aldolase. Because this new family shows no significant sequence similarity to classical Class I and II enzymes, a new name is proposed, archaeal type Class I FBP aldolases (FBP aldolase Class IA).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Bacteria / enzymology
  • Bacteria / genetics
  • Base Sequence
  • Binding Sites
  • Fructose-Bisphosphate Aldolase / chemistry
  • Fructose-Bisphosphate Aldolase / classification
  • Fructose-Bisphosphate Aldolase / genetics*
  • Fructose-Bisphosphate Aldolase / metabolism
  • Kinetics
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Operon*
  • Phylogeny
  • Promoter Regions, Genetic
  • Protein Subunits
  • Pyrococcus / classification
  • Pyrococcus / enzymology*
  • Pyrococcus / genetics*
  • Pyrococcus furiosus / classification
  • Pyrococcus furiosus / enzymology
  • Pyrococcus furiosus / genetics
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Sequence Homology, Nucleic Acid
  • TATA Box
  • Thermoproteaceae / classification
  • Thermoproteaceae / enzymology*
  • Thermoproteaceae / genetics*
  • Transcription, Genetic

Substances

  • Protein Subunits
  • Recombinant Proteins
  • Fructose-Bisphosphate Aldolase