Establishing Mixotrophic Growth of Cupriavidus necator H16 on CO2 and Volatile Fatty Acids

The facultative chemolithoautotroph Cupriavidus necator H16 is able to grow aerobically either with organic substrates or H2 and CO2 s and it can accumulate large amounts of (up to 90%) poly (3-hydroxybutyrate), a polyhydroxyalkanoate (PHA) biopolymer. The ability of this organism to co-utilize volatile fatty acids (VFAs) and CO2 as sources of carbon under mixotrophic growth conditions was investigated and PHA production was monitored. PHA accumulation was assessed under aerobic conditions, with either individual VFAs or in mixtures, under three different conditions—with CO2 as additional carbon source, without CO2 and with CO2 and H2 as additional sources of carbon and energy. VFAs utilisation rates were slower in the presence of CO2. PHA production was significantly higher when cultures were grown mixotrophically and with H2 as an additional energy source compared to heterotrophic or mixotrophic growth conditions, without H2. Furthermore, a two-step VFA feeding regime was found to be the most effective method for PHA accumulation. It was used for PHA production mixotrophically using CO2, H2 and VFA mixture derived from an anaerobic digestor (AD). The data obtained demonstrated that process parameters need to be carefully monitored to avoid VFA toxicity and low product accumulation.