The goal was to elucidate the importance of the waste properties for effective exploitation in biogas production and for soil application, respectively, based on the physicochemical and microbial characterization of biowaste and the corresponding biogas residues. The following waste media were chosen: fruit waste, dairy sewage sludge, corn silage, grass silage, and grain brew to prepare three co-substrates for anaerobic bioconversion. The most satisfactory biogas yield was obtained from biowaste with the following composition: 25% fruit wastes, 25% dairy sewage sludge, 12% corn silage, and 38% grain brew. The study included functional and genetic diversity assessment through the characterization of the catabolic potential and structure of the microbial communities inhabiting the examined organic wastes and their relative biogas residues. The metabolome was based on the use of a Biolog ® plate. The elucidation of the metagenome employed the genetic structure of prokaryotes and involved denaturing gradient gel electrophoresis and next-generation sequencing analyses. The usefulness of metagenomics was emphasized by ecotoxicological evaluation of biowaste and in determining the accurate start-up community composition for biogas production, highlighting the pivotal role of anammox and hydrolytic bacteria as marker groups. The high importance of the great diversity of fungi was also revealed based on a functional approach.