Effect of cultivation parameters on the production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) and poly(3-hydroxybutyrate-4-hydroxybutyrate-3-hydroxyvalerate) by Cupriavidus necator using waste glycerol

Short-chain polyhydroxyalkanoate co-polymers (poly(3-hydroxybutyrate-co-4-hydroxybutyrate)) (P(3HB-co-4HB)) and terpolymers (poly(3-hydroxybutyrate-4-hydroxybutyrate-3-hydroxyvalerate)) (P(3HB-4HB-3HV)) were produced using high-cell density fed-batch cultures of Cupriavidus necator DSM 545. C-source for growth and 3HB synthesis was waste glycerol (GRP) from a biodiesel plant. Incorporation of 4HB monomers was promoted by γ-butyrolactone (GBL). Propionic acid (PA), a stimulator of 4HB accumulation, increased the 4HB molar ratio 2-fold, but also acted as 3HV precursor, yielding P(3HB-4HB-3HV). Dissolved oxygen (DOC) was a key parameter for % PHA accumulation and volumetric productivity (Prod _vol). 4HB molar ratio increased in the presence of PA and with extended accumulation time. By manipulating DOC and cultivation time, P(3HB-4HB) with between 11.4 and 21.5molar% of 4HB were attained. Similarly, P(3HB-4HB-3HV) was obtained with 4HB molar% between 24.8% and 43.6% and 3HV% from 5.6% to 9.8%. Mw varied between 5.5×10 ⁵ and 1.37×10 ⁶Da. PHA production from GRP helps reducing production costs with concomitant GRP valorization.