Mixed Waste Streams for Bioproduction: Exploring Bacterial Wax Ester Production in Nitrogen‐Rich Acidogenic Fermentate

Microbial lipids offer a promising alternative to petrochemicals, but high associated costs and low conversion efficiencies pose barriers to their commercialisation. In particular, sugar-based feedstocks are too expensive for the production of commodity chemicals, and recently attention has turned to volatile fatty acids (VFAs) as a cheaper, more widely available carbon source. Acidogenic fermentation can be used to produce high concentrations of VFAs from municipal and agricultural waste. By harnessing metabolically engineered Acinetobacter baylyi ADP1, the suitability of VFAs as sole carbon sources for wax ester (WE) production was investigated. These studies resulted in the highest WE accumulation in ADP1 achieved to date, at 37% of cell dry weight, and the first reported production of bacterial WEs from a raw, mixed waste stream, utilising fermentate as the sole carbon source. WE titres of over 160 mg/L from VFAs were achieved, highlighting the unique benefits of mixed feedstocks typically considered problematic for bioproduction. Finally, the potential advantages of employing fermentates rich in longer chain VFAs are explored. In synthetic media, WE titres up to 190 mg/L were achieved, but translation to fermentate was challenging, emphasising the need for continued research in this area.