Stress response proteins within biofilm matrixome protect the cell membrane against heavy metals-induced oxidative damage in a marine bacterium Bacillus stercoris GST-03.

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). Thermodynamic analysis showed negative ∆G (-16.02 kJ/mol for Pb and -17.45 kJ/mol for Cd) and binding dissociation constant (KD) (1530 ± 157 μM for Pb and 875 ± 97.4 μM for Cd), indicating a stronger binding affinity of biofilm matrixome to heavy metals. Pb stress led to overproduction of detoxification proteins (YnaI, KhtS, Bacillopeptidase F), competence and sporulation proteins (RapF, CSSF, XkdP), while Cd exposure leads to overproduction of proteins involved in protein misfolding repair (YlxX, cysteine-tRNA ligase, YacP), DNA repair (YfkN), and redox balance (cysteine synthase, YdiK). The findings highlight the resilience of B. stercoris GST-03 to heavy metal stress in biofilm mode.