Retainment of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) Properties from Oil-Fermented Cupriavidus necator Using Additional Ethanol-Based Defatting Process.

Engineering of Cupriavidus necator could enable the production of various polyhydroxyalkanoates (PHAs); particularly, poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(3HB-co-3HH)), a biopolymer with enhanced mechanical and thermal properties compared to poly(3-hydroxybutyrate) (PHB), can be efficiently produced from vegetable oils. However, challenges remain in the recovery process, particularly in removing residual oil and minimizing degradation of the polymer structure during extraction steps. This study investigated the effects of ethanol-based defatting on the recovery and polymeric properties of P(3HB-co-3HH). The proposed method involves the addition of ethanol to the cell broth to effectively remove residual oil. Ethanol improved the separation of microbial cells from the broth, thereby streamlining the downstream recovery process. Using ethanol in the washing step increased the recovery yield and purity to 95.7% and 83.4%, respectively (compared to 87.4% and 76.2% for distilled water washing), representing improvements of 8.3% and 7.2%. Ethanol washing also resulted in a 19% higher molecular weight compared to water washing, indicating reduced polymer degradation. In terms of physical properties, the elongation at break showed a significant difference: 241.9 ± 27.0% with ethanol washing compared to water (177.7 ± 10.3%), indicating ethanol washing retains flexibility. Overall, an ethanol washing step for defatting could simplify the recovery steps, increase yield and purity, and retain mechanical properties, especially for P(3HB-co-3HH) from oils.