Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) production from engineered Ralstonia eutropha using synthetic and anaerobically digested food waste derived volatile fatty acids

Abstract

Ralstonia eutropha Re2133/pCB81 is able to utilize various volatile fatty acids (VFAs) (acetate, butyrate, lactate, and propionate) for polyhydroxyalkanoates (PHAs) production. Acetate and lactate resulted in poly(3-hydroxybutyrate) P(3HB) production, butyrate in poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) P(3HB-co-3HHx), and propionate in poly(3-hydroxybutyrate-co-3-hydroxyvalerate) P(3-HB-co-3HV). Various biomass yields i.e. (Y_x/s, 0.131 ± 0.02 g/g acetate, 0.221 ± 0.02 g/g butyrate, 0.222 ± 0.05 g/g lactate, and 0.225 ± 0.04 g/g propionate) and PHA yields (Y_p/s, 0.01 ± 0.001 g/g acetate, 0.11 ± 0.004 g/g butyrate, 0.03 ± 0.001 g/g lactate, and 0.18 ± 0.005 g/g propionate) were observed with the different organic acids. When all the organic acids were mixed together R. eutropha Re2133/pCB81 had the following order of preference; lactate > butyrate > propionate > acetate. A response surface design study showed that in mixtures butyrate is the main organic acid involved in PHA production and acts as a precursor for HHx monomer units to produce copolymer P(3HB-co-3HHx). Food waste ferment (FWF) without any additional nitrogen source and precursors resulted in P(3HB-co-3HHx) accumulation (52 ± 4% w/w with 18.5 ± 3% HHx fraction).

Keywords: Butyrate; Copolymer; Polyhydroxyalkanoates; Ralstonia eutropha; Volatile fatty acids.