Solution structure of a bacterial microcompartment targeting peptide and its application in the construction of an ethanol bioreactor

ACS Synth Biol. 2014 Jul 18;3(7):454-465. doi: 10.1021/sb4001118. Epub 2014 Feb 24.

Abstract

Targeting of proteins to bacterial microcompartments (BMCs) is mediated by an 18-amino-acid peptide sequence. Herein, we report the solution structure of the N-terminal targeting peptide (P18) of PduP, the aldehyde dehydrogenase associated with the 1,2-propanediol utilization metabolosome from Citrobacter freundii. The solution structure reveals the peptide to have a well-defined helical conformation along its whole length. Saturation transfer difference and transferred NOE NMR has highlighted the observed interaction surface on the peptide with its main interacting shell protein, PduK. By tagging both a pyruvate decarboxylase and an alcohol dehydrogenase with targeting peptides, it has been possible to direct these enzymes to empty BMCs in vivo and to generate an ethanol bioreactor. Not only are the purified, redesigned BMCs able to transform pyruvate into ethanol efficiently, but the strains containing the modified BMCs produce elevated levels of alcohol.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde Oxidoreductases / chemistry
  • Aldehyde Oxidoreductases / genetics
  • Aldehyde Oxidoreductases / metabolism
  • Amino Acid Sequence
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Bioreactors*
  • Citrobacter freundii / enzymology
  • Ethanol / metabolism*
  • Magnetic Resonance Spectroscopy
  • Metabolic Engineering
  • Molecular Sequence Data
  • Peptides / chemistry*
  • Peptides / genetics
  • Peptides / metabolism
  • Protein Structure, Tertiary
  • Pyruvate Decarboxylase / chemistry
  • Pyruvate Decarboxylase / genetics
  • Pyruvate Decarboxylase / metabolism
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification

Substances

  • Bacterial Proteins
  • Peptides
  • Recombinant Proteins
  • Ethanol
  • Aldehyde Oxidoreductases
  • Pyruvate Decarboxylase