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Abstract
Manganese (III, IV) oxides are key environmental oxidants, yet the enzymatic mechanisms of Mn(II) oxidation in Pseudomonas putida GB-1 remain unclear. This study localized Mn(II)-oxidizing activity to the outer membrane and identified a multi-copper oxidase (Mco) via MS/MS, though deletion of mco did not abolish activity, indicating multiple enzymes are involved. Reactive oxygen species, including superoxide and hydrogen peroxide, were shown to play essential roles in catalysis. Two hypothetical outer membrane proteins (2552, 2553) were found critical for Mn(II) oxidation and associated with MnxG, suggesting a complex multi-protein system. These findings advance understanding of bacterial manganese oxidation pathways.