There has been alarming depletion of manganese (Mn) reserves owing to the ongoing extensive mining operations for catering the massive industrial demand of this element. Moreover, the mining operations have been leading to the generation of Mn-rich waste, thereby contaminating both terrestrial and aquatic bodies. The current scenario necessitates the development of alternative processes for bioremediation as well as economic recovery of Mn from mining wastes. The present investigation aims to report the bioleaching of Mn by Lysinibacillus sp. from mining waste residues in the context of mine waste remediation. Results confirmed that the native isolate had a high Mn biosolubilization potential with a solubilizing efficiency of 84% at the end of a 21-day study under optimized conditions of pulp density 2% (< 150-μm particle size), pH 6.5, and temperature 30 °C. Fourier transform infrared spectroscopy (FTIR) studies followed by liquid chromatography mass spectrometry (LC-MS) analysis were used to ascertain the change in microbial protein conformation, configuration, and protein identification. The results revealed the expression of heat shock proteins (HSP) from the family HSP which is predominantly expressed in bacteria during stress conditions. This study represents the application of native bacterial strain in Mn biosolubilization. We foresee the utility of proteomics-based studies to provide a methodological framework to the underlying mechanism of metal solubilization, thereby facilitating the two-tier benefit of recovery of Mn from alternative sources as well as bioremediation of waste having high manganese content.