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Characterization of a rationally engineered phaCAB operon with a hybrid promoter design

Iain Bower, Bobby Wenqiang Chi, Matthew Ho Wai Chin, Sisi Fan, Margarita Kopniczky, Jemma Pilcher, James Strutt, Richard Kelwick, Alexander Webb, Kirsten Jensen, Guy-Bart Stan, Richard Kitney, Paul Freemont
doi: https://doi.org/10.1101/006106
Iain Bower
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
3Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK
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Bobby Wenqiang Chi
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
2Department of Bioengineering, Imperial College London, Kensington Campus, London SW7 2AZ, UK
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Matthew Ho Wai Chin
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
2Department of Bioengineering, Imperial College London, Kensington Campus, London SW7 2AZ, UK
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Sisi Fan
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
3Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK
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Margarita Kopniczky
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
3Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK
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Jemma Pilcher
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
3Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK
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James Strutt
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
3Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK
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Richard Kelwick
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
3Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK
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  • For correspondence: p.freemont@imperial.ac.uk r.kitney@imperial.ac.uk r.kelwick@imperial.ac.uk
Alexander Webb
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
3Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK
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Kirsten Jensen
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
3Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK
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Guy-Bart Stan
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
2Department of Bioengineering, Imperial College London, Kensington Campus, London SW7 2AZ, UK
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Richard Kitney
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
2Department of Bioengineering, Imperial College London, Kensington Campus, London SW7 2AZ, UK
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  • For correspondence: p.freemont@imperial.ac.uk r.kitney@imperial.ac.uk r.kelwick@imperial.ac.uk
Paul Freemont
1Centre for Synthetic Biology and Innovation, Imperial College London, Kensington Campus, London SW7 2AZ, UK
3Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK
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  • For correspondence: p.freemont@imperial.ac.uk r.kitney@imperial.ac.uk r.kelwick@imperial.ac.uk
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Abstract

Biopolymers, such as poly-3-hydroxy-butyrate (P(3HB)) are produced as a carbon store in an array of organisms and exhibit characteristics which are similar to oil-derived plastics, yet have the added advantages of biodegradability and biocompatibility. Despite these advantages, P(3HB) production is currently more expensive than the production of oil-derived plastics, and therefore more efficient P(3HB) production processes are required. In this study, we describe the model-guided design and experimental characterization of several engineered P(3HB) producing operons. In particular, we describe the characterization of a novel hybrid phaCAB operon that consists of a dual promoter (native and J23104) and RBS (native and B0034) design. P(3HB) production was around six-fold higher in hybrid phaCAB engineered Escherichia coli in comparison to E. coli engineered with the native phaCAB operon from Ralstonia eutropha H16. The hybrid phaCAB operon represents a step towards the more efficient production of P(3HB), which has an array of applications from 3D printing to tissue engineering.

Footnotes

  • The authors declare no conflict of interest and no competing financial interests.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted June 09, 2014.
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Characterization of a rationally engineered phaCAB operon with a hybrid promoter design
Iain Bower, Bobby Wenqiang Chi, Matthew Ho Wai Chin, Sisi Fan, Margarita Kopniczky, Jemma Pilcher, James Strutt, Richard Kelwick, Alexander Webb, Kirsten Jensen, Guy-Bart Stan, Richard Kitney, Paul Freemont
bioRxiv 006106; doi: https://doi.org/10.1101/006106
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Characterization of a rationally engineered phaCAB operon with a hybrid promoter design
Iain Bower, Bobby Wenqiang Chi, Matthew Ho Wai Chin, Sisi Fan, Margarita Kopniczky, Jemma Pilcher, James Strutt, Richard Kelwick, Alexander Webb, Kirsten Jensen, Guy-Bart Stan, Richard Kitney, Paul Freemont
bioRxiv 006106; doi: https://doi.org/10.1101/006106

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