Heilongjiang Province is a major soybean production area in China. To improve soil structure and increase soybean yield, this study examined the effects of combined biochar and chemical fertilizer application on the biochemical properties of soil in a maize-soybean rotation system. The research were conducted from 2021 to 2022 at Heshan Farm Science Park in Heilongjiang Province, this field plot experiment utilized two soybean varieties, Heihe 43 (a high-protein variety) and Keshan 1 (a high-oil variety). In 2021, two plots with similar fertility levels were selected for planting soybeans and maize. In 2022, a maize-soybean rotation was implemented with five treatments: conventional fertilization (CK), increased biochar+reduced fertilizer 1 (F1+B), reduced fertilizer 1 (F1), increased biochar+reduced fertilizer 2 (F2+B), and reduced fertilizer 2 (F2). The study systematically analyzed the effects of combined biochar and chemical fertilizer application on soil chemical properties and microbial characteristics. Over 2 years, results showed that combined application effectively improved soil chemical traits. Compared to conventional fertilization (CK) and reduced fertilization (F1, F2), t he combined application of biochar and chemical fertilizer (F1+B, F2+B) increased soil pH, EC and the absolute value of zeta potential of soil surface, the CEC of soil significantly increased by 15.6-44.3%, the soil surface charge density and the soil surface charge quantity significantly increased by 16.4-73.5%. The combined application of biochar and chemical fertilizer also effectively enhanced the abundance and diversity of soil microbes. Dominant bacterial groups in soybean field soils under different treatments included Actinobacteria, Acidobacteria, Chloroflexi, and Proteobacteria; while dominant fungal groups were Ascomycota, Basidiomycota, and Mortierellomycota. Alpha and Beta diversity analyses revealed that the F1+B treatment significantly enhanced the species richness and diversity of bacteria and fungi in the soil, increasing the proportion and evenness of dominant and beneficial genera.