Land use systems have a great impact on soil function and microbial diversity in tropical soils. Our study aimed to evaluate soil biochemical indicators and community composition and to assess the relationship between soil biochemical and microbial indicators and bacterial diversity of three agroecosystems (pine forest, soya and sugarcane) and native Cerrado forest in the Brazilian savanna. Soil biochemical indicators (soil organic matter and enzymes) and high-throughput sequencing of 16S rDNA were performed in two topsoil depths (0-5 cm and 5-10 cm). Soil microbial and enzyme activity showed that agricultural soil usage has a negative impact on soil function compared to native and pine forests. Results also revealed higher enzyme activities in 0-5 cm depth compared to 5-10 cm depth, but enzymatic activities depend on land use systems. Soil bacterial community was affected by land use systems and depth, revealing changes in structure and abundance of bacterial composition. Alpha-diversity indexes were higher in the agricultural systems than in the forests, however they showed a significant negative correlation with most of the studied soil microbial and biochemical indicators. Our research had brought new relevant information about the relationship between the soil biochemical indicators and the bacterial diversity in the Brazilian Cerrado.