A synthetic system for expression of components of a bacterial microcompartment

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

来源：Microbiology-SGM, 2013, 159(Pt_11): 2427-2436.

DOI：10.1099/mic.0.069922-0

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.

• 出版日期2013-11