Biochar is an effective soil amendment with capabilities of boosting carbon sequestration and enhancing soil fertility, thus enhancing plant growth and productivity. While numerous studies have documented the positive effects of biochar on improving soil properties, a number of studies have reported conflicting results. Therefore, the current study was conducted to evaluate the impact of Prosopis juliflora biochar (0, 2.5, 5.0, and 7.5 t ha−1) on soil biochemical properties in Coastal Kenya to ascertain biochar’s potential for soil fertility improvement. A randomized complete block design was used for setting up the experiment with three replicates, while Casuarina equisetifolia L. was planted as the test crop. Soil sampling for nutrient analysis was conducted quarterly for 12 months to assess nutrient dynamics under different biochar rates in the current study. Compared to soil untreated with Prosopis juliflora biochar, the results showed that there was a significant increase in soil pH by 21% following biochar utilization at the rate of 7.5 t ha−1. Total nitrogen was increased by 32% after the biochar application, whereas the total organic carbon was increased by four folds in comparison to biochar-untreated soil. Available phosphorus was increased by 264% following biochar application in comparison to the control treatment. In addition, the application of biochar resulted in an increment in the soil exchangeable cations (Ca2+, K+, Mg2+) across the assessment periods. Soil cation exchange capacity (CEC), bacteria and fungi were enhanced by 95, 33 and 48%, respectively, following biochar application at 7.5 t ha−1 in comparison to untreated soil. In conclusion, these results strongly suggest improvement of soil biochemical properties following Prosopis juliflora biochar application, thus providing potential for soil fertility improvement in regions such as the one in the study.