Comparative sugar utilisation and metabolism of mannose as co-substrate indicate flexibility in carbon metabolism in anaerobic gut fungi

Anaerobic gut fungi (AGF) are key degraders of plant biomass in ruminants, yet there is limited knowledge of how AGF respond to mixtures of plant-derived sugars. Here, we assessed monosaccharide and disaccharide utilisation by Neocallimastix frontalis CoB3, Caecomyces communis SHB, and Piromyces edwardsiae SHC, which are abundant in the rumen microbiome. While all AGF isolates shared a core set of sugars that supported growth, they had different hierarchies of uptake. Co-substrate experiments using glucose and lignocellulose-derived sugars revealed species-specific responses, with N. frontalis displaying a novel concentration-dependent co-utilisation of glucose and mannose, whereas growth of P. edwardsiae was inhibited under the same conditions, and C. communis exhibited growth inhibition in glucose and xylose co-substrate cultures. Together, these findings demonstrate functional diversity in monosaccharide and disaccharide metabolism amongst the AGF investigated here. Understanding such sugar utilisation phenotypes provides a foundation for evaluating AGF isolate suitability for lignocellulosic biomass valorisation.