BACKGROUNDHydroxyl radical‐mediated materials primarily liberate more reactive and acutely lethal hydroxyl radical (•OH) and act as potent bactericidal antibiotics, for example H2O2. Hydroxyl radical possess higher tendency than that of H2O2 to attack various biological molecules such as DNA, proteins and iron–sulfur clusters, and impair their proper functioning, actively leading to strongly potent bactericidal effect. To acquire the desired antimicrobial effect, high concentration of H2O2 is required that has found medically harmful to healthy tissues of humans.RESULTSWe herein report framework nucleic acid‐regulated DNAzyme cluster (FDC) – that is, peroxidase‐like hemin‐bound G‐quadruplex (G4/H) DNAzyme – to amplify the catalytic reduction potential of G4/H complex, leading to high conversion rate of H2O2 to more reactive hydroxyl radical that potentially shows the same antibacterial efficiency at lower and safer H2O2 concentration. Specifically, we have grafted multiple copies of DNAzymes outside framework nucleic acid (FNA) to successively achieve 3–9 orders of magnitude enhancement in catalytic activity and antibacterial efficiency of FDC.CONCLUSIONOur designed FDC has successfully alleviated H2O2 toxicity and increased its efficiency as antibacterial material, as FDC amplified the catalytic reduction potential of G4/H DNAzyme, leading to high conversion rate of H2O2 to more reactive •OH that potentially shows the same antibacterial efficiency at lower and safer H2O2 concentration. © 2024 Society of Chemical Industry (SCI).