title: A forward-design approach to increase the production of poly-3-hydroxybutyrate in genetically engineered Escherichia coli.
abstract: Biopolymers, such as poly-3-hydroxybutyrate (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 would be desirable. In this study, we describe the model-guided design and experimental validation of several engineered P(3HB) producing operons. In particular, we describe the characterization of a hybrid phaCAB operon that consists of a dual promoter (native and J23104) and RBS (native and B0034) design. P(3HB) production at 24 h 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. Additionally, we describe the utilization of non-recyclable waste as a low-cost carbon source for the production of P(3HB).
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Medicine, South Kensington Campus, London, United Kingdom.
authors.forename: Richard
authors.initials: R
authors.lastname: Kelwick
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Medicine, South Kensington Campus, London, United Kingdom.
authors.forename: Margarita
authors.initials: M
authors.lastname: Kopniczky
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Life Sciences, South Kensington Campus, London, United Kingdom.
authors.forename: Iain
authors.initials: I
authors.lastname: Bower
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Bioengineering, Imperial College London, South Kensington Campus, London, United Kingdom.
authors.forename: Wenqiang
authors.initials: W
authors.lastname: Chi
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Bioengineering, Imperial College London, South Kensington Campus, London, United Kingdom.
authors.forename: Matthew Ho Wai
authors.initials: MH
authors.lastname: Chin
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Life Sciences, South Kensington Campus, London, United Kingdom.
authors.forename: Sisi
authors.initials: S
authors.lastname: Fan
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Life Sciences, South Kensington Campus, London, United Kingdom.
authors.forename: Jemma
authors.initials: J
authors.lastname: Pilcher
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Life Sciences, South Kensington Campus, London, United Kingdom.
authors.forename: James
authors.initials: J
authors.lastname: Strutt
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Medicine, South Kensington Campus, London, United Kingdom.
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Webb
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Medicine, South Kensington Campus, London, United Kingdom.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Jensen
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Bioengineering, Imperial College London, South Kensington Campus, London, United Kingdom.
authors.forename: Guy-Bart
authors.initials: GB
authors.lastname: Stan
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Bioengineering, Imperial College London, South Kensington Campus, London, United Kingdom.
authors.forename: Richard
authors.initials: R
authors.lastname: Kitney
authors.affiliation: Centre for Synthetic Biology and Innovation, South Kensington Campus, London, United Kingdom; Department of Medicine, South Kensington Campus, London, United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: Freemont
mesh_terms: D019149:Bioreactors; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005798:Genes, Bacterial; D005818:Genetic Engineering; D006885:Hydroxybutyrates / Q000378:metabolism*; D009876:Operon; D011091:Polyesters / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D000094122:Polyhydroxybutyrates
chemical_list: D003470:Culture Media; D006885:Hydroxybutyrates; D011091:Polyesters; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates