Understanding interactions between proteins and nanoparticles (NPs) along with the underlying structural and dynamic information is of utmost importance to exploit nanotechnology for biomedical applications. Upon adsorption onto the NP surface, proteins form a well-organized layer that dictates the identity of the NP-protein complex named corona and governs its biological pathways. Given the high biological relevance, in-depth molecular investigations and applications of NPs-protein corona complexes are still scarce, especially since different proteins form unique patterns of corona and hence identification of biomolecular motifs at the interface is critical. In this work, we provide molecular insights and structural characterizations of the bionano interface of a popular food-based protein, bovine beta-lactoglobulin (β-LG), with gold nanoparticles (AuNPs) and the formation of corona complexes by combined molecular simulations and complementary experiments. Two major binding sites in β-LG were identified to be driven by citrate-mediated electrostatic interactions, while the associated binding kinetics and conformational changes in secondary structures were also characterized. More importantly, the superior stability of the corona led us to further explore its biomedical applications with examples of smart-phone based point-of-care biosensing of Escherichia coli (E. coli) and computed tomography (CT) of gastrointestinal (GI) tract through oral administration to probe GI tolerance and functions. Considering the biocompatibility, edible nature and efficient excretion through defecation, AuNPs-β-LG corona complexes have shown promising perspectives in future in vitro and in vivo clinical settings.