In this study, we report for the first time, the fabrication of hybrid dual 2D-nanohybrid structure as an interface system for electrochemical biosensing, bioelectronics, and electrobiocatalysis devices through self-assembly combination of gold nanoparticles (AuNPs) with hybrid 2D materials consisting of boron nitride (BN) and tungsten disulphide (WS2). Horseradish peroxidase (HRP) was immobilized on the hybrid dual 2D-nanoparticle system to form a biointerface system. Structural characterization showed high crystallinity in the fabricated structure, while morphological characterization confirmed the high surface area of the hybrid material and the presence of well-dispersed gold nanoparticles (AuNPs). Electrochemical characterization also confirmed that the fabricated HRP/BN/WS2/AuNPs/GC bioelectrode exhibited excellent electron transfer properties at the interface. The electrobiocatalytic activity of the nanohybrid interface structure was studied using hydrogen peroxide (H2O2) as a model analyte. The fabricated bioelectrode exhibited a wide linear range from 0.15 mM to 15.01 mM towards hydrogen peroxide detection with a limit of detection of 3.0 mM (S/N = 3) and a sensitivity of 19.16 mu A/M/cm(2). A density functional theory (DFT) calculation was also used to validate the charge transport mobility and conductivity of BN/Au/WS2(001) nanohybrid structure as biosensing interface material. The DFT calculations combined with the experimental studies showed that the BN/Au/WS2(001) nanocomposite enhances the performance of the nanocomposite as sensor due to the introduction new electronic states emanating from the N 2p orbital. This work thus set the stage for the use BN/WS2/AuNPs nanohybrid system as interface material for bioelectronics and biosensing devices fabrications and applications.