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Abstract
Bacterial microcompartments (BMCs) are organelles that host specific biochemical reactions for both anabolic and catabolic functions. Engineered morphologically diverse BMCs bearing heterologous enzymatic pathways have shown enhanced productivity for commodity chemicals, which makes BMCs an important focus for metabolic engineering. Gaining control of BMC assembly and incorporation of a heterologous enzymatic cargo has yet to be achieved in thermophiles. Herein, we address this by first conducting a detailed bioinformatic analysis of the propanediol utilization (pdu) operon in the thermophile Parageobacillus thermoglucosidasius. We then demonstrated, in vivo, the ability to assemble the native BMCs at an elevated temperature of 60 °C. Heterologous expression of Pdu shell proteins from P. thermoglucosidasius in Bacillus subtilis resulted in the assembly of a single tubular BMC with an average length of 1.4 μm; BMCs assembled after a 20 min induction of expression of the shell operons. Moreover, we show that it is possible to target the monomeric superfolder GFP (msfGFP) to the interior of the compartment by fusion of an N-terminal sequence of the propanediol utilization protein (PduP) of at least 24 amino acids. This study establishes the feasibility of constructing cell factories for small molecules in industrially important Bacillus and Geobacillus spp. by heterologous cargo-carrying BMC production and assembly. Additionally, the study provides experimental confirmation that BMCs are produced in thermophilic bacteria, which opens a path for future research on repurposing the native organelles to provide new functionality at elevated temperatures.
Original language | English |
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Pages (from-to) | 1642-1654 |
Number of pages | 13 |
Journal | ACS Synthetic Biology |
Volume | 8 |
Issue number | 7 |
Early online date | 26 Jun 2019 |
DOIs | |
Publication status | Published - 19 Jul 2019 |
Keywords
- Bacillus.
- Parageobacillus
- bacterial microcompartments
- biotechnology
- shell
- synthetic biology
- thermophiles
ASJC Scopus subject areas
- Biomedical Engineering
- Biochemistry, Genetics and Molecular Biology (miscellaneous)
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Dive into the research topics of 'Heterologous Microcompartment Assembly in Bacillaceae: Establishing the Components Necessary for Scaffold Formation'. Together they form a unique fingerprint.Projects
- 2 Finished
Profiles
-
David Leak
- Centre for Sustainable and Circular Technologies (CSCT)
- Water Innovation and Research Centre (WIRC)
- Department of Life Sciences - Professor Emeritus
- EPSRC Centre for Doctoral Training in Advanced Automotive Propulsion Systems (AAPS CDT)
Person: Research & Teaching, Affiliate staff, Honorary / Visiting Staff