Unveiling the chromate stress response in the marine bacterium Bacillus enclensis AGM_Cr8: a multifaceted investigation.

In this study, we introduce Bacillus enclensis AGM_Cr8, a gram-positive marine bacterium isolated from the chronically polluted Versova Creek in Mumbai, India. AGM_Cr8 exhibits robust tolerance to chromate stress, thriving in marine agar media containing up to 3200 mg/l of hexavalent chromium [Cr(VI)], with the Minimum Inhibitory Concentration (MIC) established at 5000 mg/l. Notably, AGM_Cr8 also displays tolerance to other heavy metals, including Lead [Pb (II)] (1200 mg/l), Arsenic [As (III)] (400 mg/l), Cadmium [Cd(II)] (100 mg/l), and Nickel [Ni(II)] (100 mg/l). Scanning Electron Microscopy (SEM) reveals the presence of Cr(VI) on the bacterial surface, accompanied by the secretion of extracellular polymeric substances (EPSs) facilitating Cr(VI) sequestration. This observation is validated through Energy Dispersive Spectroscopy (EDS). Transmission Electron Microscopy (TEM) and Scanning Transmission Electron Microscopy-Energy Dispersive Spectroscopy (STEM-EDS) confirm internal bioaccumulation of Cr(VI). X-ray photoelectron spectroscopy (XPS) identifies distinct peaks around 579 and 576 eV, indicating the coexistence of Cr(VI) and Cr(III), implying a bioreduction mechanism. De novo genome sequencing identifies twenty-two chromate-responsive genes, including putative chromate transporters (srpC1 and srpC2), suggesting an efflux mechanism. Other identified genes encode NAD(P)H-dependent FMN-containing oxidoreductase, NADH quinone reductase, ornithine aminotransferase, transporter genes (natA, natB, ytrB), and genes related to DNA replication and repair (recF), DNA mismatch repair (mutH), and superoxide dismutase. We therefore, propose a chromate detoxification pathway that involves an interplay of chromate transporters, enzymatic reduction of Cr(VI) to Cr(III), DNA repair and role of antioxidants in response to chromate stress. We have highlighted the potential of AGM_Cr8 for bioremediation in chromium-contaminated environments, given its robust tolerance and elucidated molecular mechanisms for detoxification.