Enhanced production of polyhydroxyalkanoate with manipulable and reproducible 3-hydroxyvalerate fraction by high alcohol tolerant Cupriavidus malaysiensis USMAA2-4 transformant

Abstract

The domination of high-cost organic acids over other 3-hydroxyvalerate (3HV) precursors due to the wide preference among polyhydroxyalkanoates (PHA)-producing bacteria has limited the development of diverse poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] production processes. 1-pentanol is a low-cost 3HV precursor but is rarely employed due to the relatively low tolerance among PHA-producing bacteria. This study demonstrated P(3HB-co-3HV) production with manipulable and reproducible 3HV composition and 3HV yield from palm olein and 1-pentanol. Cupriavidus malaysiensis USMAA2-4_ABH16 is the transformant strain with acquired lipase genes that retains the high tolerance towards 1-pentanol of its wild-type, with a preference for 1-pentanol over valeric acid indicated by the sixfold higher 3HV yield than that from valeric acid. C. malaysiensis USMAA2-4_ABH16 was able to tolerate up to 0.15 wt% C 1-pentanol. Upon optimization using response surface methodology, 0.41‒0.52 g/g P(3HB-co-3HV) yield and 72‒89 wt% PHA content was achieved for 7, 9, 12 and 16 mol% 3HV, with 3HV yields of 0.30 g/g, 0.26 g/g, 0.23 g/g and 0.23 g/g, respectively. Up-scaling batch production by adopting the optimized concentrations of substrates for 12 mol% 3HV resulted in reproducible 3HV composition and 3HV yield on a 120-fold larger scale. The P(3HB-co-12 mol% 3HV) produced displayed higher flexibility than polypropylene and P(3HB-co-3HV) produced from different carbon sources. C. malaysiensis USMAA2-4_ABH16 could be practically applicable for sustainable and economically feasible P(3HB-co-3HV) production on an industrial scale from used palm olein with relatively similar oleic acid content with palm olein and 1-pentanol, with higher 3HV compositions achievable through fed-batch strategies owing to its high 1-pentanol tolerance.

Keywords: 1-Pentanol; Cupriavidus malaysiensis USMAA2-4 transformant; Palm olein; Poly(3-hydroxybutyrate-co-3-hydroxyvalerate); Polyhydroxyalkanoates (PHA); Response surface methodology (RSM).