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Noble metal nanoparticles have attracted much interest in the heterogeneous catalysis. Particularly, efficient manipulation of the responsive catalytic properties of the metal nanoparticles is an interesting topic. In this work, a simple and efficient strategy is developed to regulate the pH‐responsive catalytic activities of glucose oxidase (GOx)‐mimicking gold nanoparticles (AuNPs). Four DNA strands (regulating strands) that differ slightly in sequences are used to interact non‐covalently with citrate‐capped AuNPs, resulting in markedly distinct pH‐dependent catalytic behavior of AuNPs. This is ascribed to the characteristic pH‐induced conformational change of the DNA strands that leads to the different adsorption capability to the NPs surface, as demonstrated by pH‐CD profiles of the respective DNA molecules. The pH‐dependent catalysis of AuNPs is also encoded with structural information of the double‐stranded …