PLoS ONE (Jan 2015)

A forward-design approach to increase the production of poly-3-hydroxybutyrate in genetically engineered Escherichia coli.

  • Richard Kelwick,
  • Margarita Kopniczky,
  • Iain Bower,
  • Wenqiang Chi,
  • Matthew Ho Wai Chin,
  • Sisi Fan,
  • Jemma Pilcher,
  • James Strutt,
  • Alexander J Webb,
  • Kirsten Jensen,
  • Guy-Bart Stan,
  • Richard Kitney,
  • Paul Freemont

DOI
https://doi.org/10.1371/journal.pone.0117202
Journal volume & issue
Vol. 10, no. 2
p. e0117202

Abstract

Read online

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).