title: Solution structure of a bacterial microcompartment targeting peptide and its application in the construction of an ethanol bioreactor.
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.
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent, CT2 7NJ, UK.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Lawrence
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent, CT2 7NJ, UK.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Frank
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent, CT2 7NJ, UK.
authors.forename: Sarah
authors.initials: S
authors.lastname: Newnham
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent, CT2 7NJ, UK.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Lee
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent, CT2 7NJ, UK.
authors.forename: Ian R
authors.initials: IR
authors.lastname: Brown
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent, CT2 7NJ, UK.
authors.forename: Wei-Feng
authors.initials: WF
authors.lastname: Xue
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent, CT2 7NJ, UK.
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Rowe
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent, CT2 7NJ, UK.
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Mulvihill
authors.affiliation: Department of Microbiology, University College Cork, Cork, Ireland.|Department of Pathology, University College Cork, Cork, Ireland.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Prentice
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent, CT2 7NJ, UK.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Howard
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent, CT2 7NJ, UK.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Warren
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019149:Bioreactors; D016971:Citrobacter freundii / Q000201:enzymology; D000431:Ethanol / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D060847:Metabolic Engineering; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011767:Pyruvate Decarboxylase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D010455:Peptides; D011994:Recombinant Proteins; D000431:Ethanol; D000445:Aldehyde Oxidoreductases; D011767:Pyruvate Decarboxylase