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Medium-chain-length (MCL) polyhydroxyalkanoates (PHAs) of near homopolymeric composition are unnatural polymers, having almost identical repeating units throughout the polymer chain. These homopolymeric PHAs can be produced by β-oxidation defective bacterial hosts.  Escherichia coli  is an attractive workhorse for the production of such genetically engineered PHAs; however, achieving efficient production of the near homopolymers by β-oxidation defective strains is a major challenge because of a lack of process development studies. In order to address this issue, we investigated the optimization of carbon feeding for efficient production of MCL-PHAs by an  E. coli  strain with defective β-oxidation, LSBJ. Engineered bacteria were cultured in shake-flasks with intermittent feeding of a fatty acid substrate [either decanoate (C10) or dodecanoate (C12)] at various concentrations together with a co-carbon source (glucose, glycerol, or xylose) in order to support cell growth. It was found that feeding low concentrations of both fatty acids and co-carbon sources led to an enhanced production of MCL-PHAs. Additionally, the supplementation of yeast extract improved cell growth, resulting in achieving higher titers of MCL-PHA. As a result, poly(3-hydroxydecanoate) [P(3HD)] and poly(3-hydroxydodecanoate) [P(3HDD)] were produced up to 5.44 g/L and 3.50 g/L, respectively, as near homopolymers by employing the developed feeding strategy. To the best of our knowledge, we record the highest titer of P(3HD) ever reported so far.

Low Carbon Concentration Feeding Improves Medium-Chain-Length Polyhydroxyalkanoate Production in Escherichia coli Strains With Defective β-Oxidation