Reactive oxygen species (ROS) are unstable metabolites produced during cellular respiration that can cause extensive damage to the body. Here we report a unique structural metalloprotein called RSAPp for the first time, which exhibits robust ROS-scavenging activity, high thermostability, and stress resistance. RSAPp is a previously uncharacterized DUF2935 (domain of unknown function, accession number: cl12705) family protein from Paenibacillus, containing a highly conserved four-helix bundle with binding sites for variable-valence metal ions (Mn2+/Fe2+/Zn2+). Enzymatic characterization results indicated that RSAPp displays the functionality of three different antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD). In particular, RSAPp exhibits a significant SOD-like activity that is remarkably effective in eliminating superoxide radicals (up to kcat/KM = 2.27 × 1011 mol-1 s-1), and maintains the catalytical active in a wide range of temperatures (25-100 °C) and pH (pH 2.0-9.0), as well as resistant to high temperature, alkali and acidic pH, and 55 different concentrations of detergent agents, chemical solvents, and inhibitors. These properties make RSAPp an attractive candidate for various industrial applications, including cosmetics, food, and pharmaceuticals.
Keywords: Biomacromolecule; DUF2935; Multiple catalytic activities; ROS scavenging; Unknown function protein.
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