Fluoride (F-) contamination of groundwater is a silent killer of human health that has now become a global problem. In the present study, we sought to isolate fluoride-resistant, urease producing bacteria from F-contaminated groundwater to evaluate their potential F-biomineralization. Strain MLN9, isolated from the fluoride-contaminated village of Madhabpur in Birbhum, could resist up to 5600mgL-1F-concentration. Taxonomic classification of strain MLN9 based on whole genome sequence identified it as Proteus columbaeMLN9. Strain MLN9 showed 88.9% of maximum F- removal efficiency at pH 8.0, 1.0gL-1CaCl2, and 10.0gL-1urea concentration. Scanning electron micrographs showed dense and porous biological crystal precipitates surrounding the surface of the bacterial cells, and the adherence of F-to the cell surface was confirmed by the energy dispersion spectrum. Moreover, the X-ray diffraction pattern showed that the F-precipitates on the cell surface were biological crystals of Ca5(PO4)3F and CaF2. Genomic analysis of strain MLN9 revealed the presence of crcBgene homologs possibly encoding F-transporters and urease regulatory proteins such as UreA, UreB, UreC, UreD, and UreG suggesting a role in defluoridation. Our findings provide new insights into the urease based MICP technology using a noble bacterium Proteus columbaeMLN9 for fluoride removal.