Biofilm formation is a key adaptive response of marine bacteria towards stress conditions. The protective mechanisms of biofilm matrixome proteins against heavy metals (Pb and Cd) induced oxidative damage in the marine bacterium Bacillus stercoris GST-03 was investigated. Exposure to heavy metals resulted in significant changes in cell morphology, biofilm formation, and matrixome composition. Biofilm-encased cells showed lower oxidative damage. Biofilm matrixome protein exhibited major conformational changes, with 100 % α-helix turned to 62.33 % and 69.64 % of random coil under Pb and Cd stress, respectively. Fluorescence quenching kinetics revealed slow interactions between biofilm matrixome proteins and heavy metals (Kq values <
2.0 × 10