The beneficial effects of biofuels as components of the worldwide energy supply are unquantifiable because they have versatile applications. However, an adequate understanding of the chemical properties of typical biomass is an integral aspect of maximizing the energy potentials because it is susceptible to biomass behavior during the conversion process, especially anaerobic digestion. Therefore, this study investigated the physicochemical characteristics of selected lignocellulose biomass, namely; cow dung, mango pulp, and Chromolaena odorata of Nigerian origin. The raw biomasses were characterized by proximate, calorific, ultimate, compositional, and microbial (for cow dung only) analyses using ASTM standards and equipment. Raw biomass characterization showed that cow dung, mango pulp, and Chromolaena odorata leaves recorded percentages; fixed carbon, volatile matter, and ash contents in addition to calorific values in the ranges of 6.22–7.25%, 5.02–7.79%, 1.14–1.91,% and 13.77–16.16 MJ/kg, respectively. The ultimate analysis of cow dung, mango pulp and Chromolaena odorata recorded carbon (43.08, 39.98, 41.69%); hydrogen (7.87, 6.74, 9.86%); nitrogen (1.53, 1.34, 1.51%); sulphur (0.46, 0.12, 0.25%) and oxygen (47.06, 51.82, 46.69%), respectively. Compositional analysis of the biomass gave percentages in the range of 7.47–11.37 for hemicellulose, 0.22–6.33 for lignin, and 3.71–12.03 for cellulose, while the microbial analysis of cow dung gave total bacteria counts of 5.78 × 108 and 3.93 × 105 cfu/g on wet and dry bases, respectively, which implied that it was rich in microbial colonies, evidently from the various species found, such as Escherichia coli, Staphylococcus aureus, Bacillus cereus, Pseudomonas aureginosa, Proteus morganii, and Micrococcus spp. In this regard, the physicochemical properties of selected biomass of Nigerian origin were established to conform with those of the literature and thus can be regarded as suitable feedstock for anaerobic digestion resulting in methane-rich biogas products.