Pseudomonas rhizophila S211 as a microbial cell factory for direct bioconversion of waste cooking oil into medium-chain-length polyhydroxyalkanoates

The present study examines the use of waste cooking oil (WCO) as a substrate for medium-chain-length polyhydroxyalkanoates (mcl-PHA) production by Pseudomonas rhizophila S211. The genome analysis revealed that the S211 strain has a mcl-PHA cluster (phaC1ZC2DFI) encoding two class II PHA synthases (PhaC1 and PhaC2) separated by a PHA depolymerase (PhaZ), a transcriptional activator (PhaD) and two phasin-like proteins (PhaFI). Genomic annotation also identified a gene encoding family I.3 lipase that was able to hydrolyze plant oils and generate fatty acids as favorable carbon sources for cell growth and PHA synthesis via β-oxidation pathway. Using a three-variable Doehlert experimental design, the optimum conditions for mcl-PHA accumulation were achieved in 10% of WCO-based medium with an inoculum size of 10% and an incubation period of 48 h at 30 °C. The experimental yield of PHA from WCO was 1.8 g/L close to the predicted yield of 1.68 ± 0.14 g/L. Moreover, ¹H nuclear magnetic resonance spectroscopy analysis confirmed the extracted mcl-PHA. Overall, this study describes P. rhizophila as a cell factory for biosynthesis of biodegradable plastics and proposes green and efficient approach to cooking oil waste management by decreasing the cost of mcl-PHA production, which can help reduce the dependence on petroleum-based plastics.