The taxonomic and functional diversity of bacteria in seven different experimental variants applied to soil under a maize plantation was determined by means of next-generation sequencing and biochemical methods. The aim of the study was to discover differences in the structure of bacteria and the level of soil enzymatic activity (BIF—biochemical index of fertility) after the application of a biofertiliser made of lignocellulosic substrate and biochar containing various microorganisms (algae, mycorrhizal fungi of the Glomus genus or the consortium of Bacillus sp. bacteria). The chemical composition and yield of crops was a measurable indicator of the effectiveness of the fertilisers. The biofertilisers influenced both the structure and the percentage share of individual bacterial operational taxonomic units (OTU). The cultivation of maize also modified qualitative and quantitative changes in the soil bacterial microbiome. A canonical variate analysis (CVA) showed that the soil pH exhibited a minimal positive correlation with the soil enzymatic activity and selected plant parameters, with the exception of the biofertiliser variant with arbuscular mycorrhiza (AM). Moreover, the AM biofertiliser significantly increased the BIF value, the yield of maize seeds and the starch content in the plants. The comprehensive nature of the research allowed for a deepening and systematization of the existing knowledge on the influence of biochar with the addition of selected microorganisms on the biochemical parameters of the soil and the bacterial biodiversity of the soil environment. Additionally, the inclusion of the chemical, sanitary composition and yield of maize in the research brought a measurable view of the changes taking place in the soil and plant environment under the influence of the discussed factor. Apart from the agronomic aspect (integrated crop cultivation—Directive 2009/128/EC) of our study, it was also closely related to environmental protection, as it proved that biochar-based biofertilisers could be an alternative to mineral fertilisation.