A synthetic system for expression of components of a bacterial microcompartment

Frank Sargent*, Fordyce A. Davidson, Ciarán L. Kelly, Rachelle Binny, Natasha Christodoulides, David Gibson, Emelie Johansson, Katarzyna Kozyrska, Lucia Licandro Lado, Jane MacCallum, Rachel Montague, Brian Ortmann, Richard Owen, Sarah J. Coulthurst, Lionel Dupuy, Alan R. Prescott, Tracy Palmer

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)2427-2436
Number of pages10
JournalMicrobiology (United Kingdom)
Volume159
Issue numberPART11
Early online date1 Nov 2013
DOIs
Publication statusPublished - Nov 2013
Externally publishedYes

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