title: A synthetic system for expression of components of a bacterial microcompartment.
abstract: In general, prokaryotes are considered to be single-celled organisms that lack internal membrane-bound organelles. However, many bacteria produce proteinaceous microcompartments that serve a similar purpose, i.e. to concentrate specific enzymic reactions together or to shield the wider cytoplasm from toxic metabolic intermediates. In this paper, a synthetic operon encoding the key structural components of a microcompartment was designed based on the genes for the Salmonella propanediol utilization (Pdu) microcompartment. The genes chosen included pduA, -B, -J, -K, -N, -T and -U, and each was shown to produce protein in an Escherichia coli chassis. In parallel, a set of compatible vectors designed to express non-native cargo proteins was also designed and tested. Engineered hexa-His tags allowed isolation of the components of the microcompartments together with co-expressed, untagged, cargo proteins. Finally, an in vivo protease accessibility assay suggested that a PduD-GFP fusion could be protected from proteolysis when co-expressed with the synthetic microcompartment operon. This work gives encouragement that it may be possible to harness the genes encoding a non-native microcompartment for future biotechnological applications.
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Frank
authors.initials: F
authors.lastname: Sargent
authors.affiliation: Division of Mathematics, College of Art, Science & Engineering, University of Dundee, Dundee DD1 4HN, Scotland, UK.
authors.forename: Fordyce A
authors.initials: FA
authors.lastname: Davidson
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Ciarán L
authors.initials: CL
authors.lastname: Kelly
authors.affiliation: Division of Mathematics, College of Art, Science & Engineering, University of Dundee, Dundee DD1 4HN, Scotland, UK.|College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Rachelle
authors.initials: R
authors.lastname: Binny
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Natasha
authors.initials: N
authors.lastname: Christodoulides
authors.affiliation: School of Computing, College of Art, Science & Engineering, University of Dundee, Dundee DD1 4HN, Scotland, UK.
authors.forename: David
authors.initials: D
authors.lastname: Gibson
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Emelie
authors.initials: E
authors.lastname: Johansson
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kozyrska
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Lucia Licandro
authors.initials: LL
authors.lastname: Lado
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Jane
authors.initials: J
authors.lastname: MacCallum
authors.affiliation: School of Computing, College of Art, Science & Engineering, University of Dundee, Dundee DD1 4HN, Scotland, UK.
authors.forename: Rachel
authors.initials: R
authors.lastname: Montague
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Brian
authors.initials: B
authors.lastname: Ortmann
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Richard
authors.initials: R
authors.lastname: Owen
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Coulthurst
authors.affiliation: Ecological Sciences, The James Hutton Institute, Invergowrie, Dundee DD2 5DA, Scotland, UK.
authors.forename: Lionel
authors.initials: L
authors.lastname: Dupuy
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Alan R
authors.initials: AR
authors.lastname: Prescott
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Tracy
authors.initials: T
authors.lastname: Palmer
mesh_terms: D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D053858:Metabolic Networks and Pathways; D011409:Propylene Glycols / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012475:Salmonella / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*
chemical_list: D011409:Propylene Glycols; D011994:Recombinant Proteins