Strategic Use of Vegetable Oil for Mass Production of 5-Hydroxyvalerate-Containing Polyhydroxyalkanoate from δ-Valerolactone by Engineered Cupriavidus necator

Although efforts have been undertaken to produce polyhydroxyalkanoates (PHA) with various monomers, the low yield of PHAs because of complex metabolic pathways and inhibitory substrates remains a major hurdle in their analyses and applications. Therefore, we investigated the feasibility of mass production of PHAs containing 5-hydroxyvalerate (5HV) using δ-valerolactone (DVL) without any pretreatment along with the addition of plant oil to achieve enough biomass. We identified that PhaC_BP-M-CPF4, a PHA synthase, was capable of incorporating 5HV monomers and that <i>C. necator</i> PHB^-4 harboring <i>phaC</i>_BP-M-CPF4 synthesized poly(3HB-<i>co</i>-3HHx-<i>co</i>-5HV) in the presence of bean oil and DVL. In fed-batch fermentation, the supply of bean oil resulted in the synthesis of 49 g/L of poly(3HB-<i>co</i>-3.7 mol% 3HHx-<i>co</i>-5.3 mol%5HV) from 66 g/L of biomass. Thermophysical studies showed that 3HHx was effective in increasing the elongation, whereas 5HV was effective in decreasing the melting point. The contact angles of poly(3HB-<i>co</i>-3HHx-<i>co</i>-5HV) and poly(3HB-<i>co</i>-3HHx) were 109 and 98°, respectively. In addition, the analysis of microbial degradation confirmed that poly(3HB-<i>co</i>-3HHx-<i>co</i>-5HV) degraded more slowly (82% over 7 days) compared to poly(3HB-<i>co</i>-3HHx) (100% over 5 days). Overall, the oil-based fermentation strategy helped produce more PHA, and the mass production of novel PHAs could provide more opportunities to study polymer properties.