A synthetic system for expression of components of a bacterial microcompartment
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Author(s)
Type
Journal Article
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.
Date Issued
2013-11-01
Date Acceptance
2013-09-06
Citation
Microbiology, 2013, 159 (11), pp.2427-2436
ISSN
1350-0872
Publisher
Microbiology Society
Start Page
2427
End Page
2436
Journal / Book Title
Microbiology
Volume
159
Issue
11
Copyright Statement
This is an open access article published by the Microbiology Society under the Creative Commons Attribution License
License URL
Subjects
Escherichia coli
Genetic Vectors
Metabolic Networks and Pathways
Propylene Glycols
Recombinant Proteins
Salmonella
Microbiology
MD Multidisciplinary
Publication Status
Published