Avoiding acid crash: From apple pomace hydrolysate to butanol through acetone-butanol-ethanol fermentation in a zero-waste approach

Abstract

Apple pomace (AP) is a lignocellulosic residue from the juice and cider industries that can be valorized in a multi-product biorefinery to generate multiple value-added compounds, including biofuels such as butanol. Butanol is produced biologically by acetone-butanol-ethanol (ABE) fermentation using bacteria of the genus Clostridium from sugar-based feedstocks. In this study, AP hydrolysate was used as a substrate for producing butanol by ABE fermentation. Various environmental factors influence the amount of butanol produced, but only under certain conditions the so-called 'acid crash', an undesirable phenomenon characterized by a total arrest of cell growth and solvent production, can be avoided. Operational parameters that may influence the prevention of acid crash occurrence, such as pH, CaCO₃ concentration and culture temperature, were optimized in C. beijerinckii CECT 508 cultures applying a Box-Behnken experimental design. The mathematical model of the fermentation found the optimal conditions of pH 7, 6.8 g/L of CaCO₃ and 30 °C, and this was validated in an independent experiment carried out at the optimal conditions, reaching 10.75 g/L of butanol. Also, the comparison of butanol production between the supernatant of the AP hydrolysate (10.57 g/L) and the full hydrolysate with solids (11.69 g/L) indicated that it is possible to eliminate the centrifugation step after hydrolysis, which may allow to reduce process costs and the full utilization of apple pomace, aiming a zero-waste approach.

Keywords: ABE fermentation; Biorefinery; Calcium carbonate; Clostridium beijerinckii CECT 508; Temperature; pH.