A synthetic system for expression of components of a bacterial microcompartment
Sargent, Frank and Davidson, Fordyce A. and Kelly, Ciarán L. and Binny, Rachelle and Christodoulides, Natasha and Gibson, David and Johansson, Emelie and Kozyrska, Katarzyna and Lado, Lucia Licandro and MacCallum, Jane and Montague, Rachel and Ortmann, Brian and Owen, Richard and Coulthurst, Sarah J. and Dupuy, Lionel and Prescott, Alan R. and Palmer, Tracy,, 159, 2427-2436 (2013),
doi = https://doi.org/10.1099/mic.0.069922-0,
publicationName = Microbiology Society,
issn = 1350-0872,
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.,
language=,
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