Due to the increasing usage of silver nanoparticles in biomedical applications, it becomes important to understand their interaction with biomolecules such as protein. In the present work, we have investigated the interaction of bovine serum albumin (BSA) with silver nanorods (SNRs) using different spectroscopic techniques. The interaction between BSA and SNRs can happen through non-covalent interactions like electrostatic and hydrophobic forces. Upon the interaction of SNRs with various concentrations of BSA, a small red-shift was found in the localized surface plasmon resonance (LSPR) peak of SNRs from UV–visible spectroscopic studies, indicating the adsorption of BSA on the surface of SNRs. The UV–visible absorption spectral studies confirmed the formation of a ground-state BSA-conjugated SNR system. The number of binding sites (n = 1.86) and binding constant (K = 8.719 × 104 L·mol− 1) were obtained to check the strength of conjugation of BSA in presence of different concentrations of SNRs from fluorescence quenching analysis. Further, DLS and zeta potential analyses proved significant binding of BSA towards the SNRs. The secondary structural changes in BSA in the presence of SNRs were obtained using circular dichroism (CD) spectroscopy and Fourier transform infrared spectroscopy (FT-IR). The unfolding behavior of BSA and BSA-conjugated SNR complex were established using the surfactant, sodium dodecyl sulfate (SDS). Finally, the influence of SNRs in the hydrolysis of BSA was investigated by assaying the esterase activity. The esterase activity of BSA was determined by the hydrolysis of p-nitrophenyl acetate (p-NPA) to 4-nitrophenol. © 2017 Elsevier B.V.