show Abstracthide AbstractOdontotaenius disjunctus is a wood-feeding beetle that possesses a digestive tract with four main compartments each of which contains well-differentiated microbial populations, suggesting that anatomical properties and separation of these compartments may enhance energy extraction from woody biomass. Here, we demonstrate how spatially segregated microbiome functionality facilitates lignocellulose deconstruction and transformation in the passalid digestive tract.Using metagenome and metaproteome analyses, along with spectroscopic, isotopic and analytical chemistry approaches, we reconstructed the pathways for sequential lignocellulose transformation, the subsequent fermentation of lignocellulose products, and the distribution of key microbial catalysts throughout the passalid digestive tract.