Developing biosorbents derived from agro-industrial biomass is considered as an economic and sustainable method for dealing with uranium-contaminated wastewater. The present study explores the feasibility of oxidizing a representative protein-rich biomass, brewer's spent grain (BSG), to an effective and reusable uranyl ion adsorbent to reduce the cost and waste generation during water treatment. The unique composition of BSG favors the oxidation process and yields in a high carboxyl group content (1.3 mmol/g) of the biosorbent. This makes BSG a cheap, sustainable, and suitable raw material independent from pre-treatment. The oxidized brewer's spent grain (OBSG) presents a high adsorption capacity of U(VI) of 297.3 mg/g (c0(U) = 900 mg/L, pH = 4.7) and fast adsorption kinetics (1 h) compared with other biosorbents reported in the literature. Infrared spectra (Fourier transform infrared), 13C solid-state nuclear magnetic resonance spectra, scanning electron microscopy, energy-dispersive X-ray spectroscopy, and thermogravimetric analysis were employed to characterize the biosorbents and reveal the adsorption mechanisms. The desorption and reusability of OBSG were tested for five cycles, resulting in a remaining adsorption of U(VI) of 100.3 mg/g and a desorption ratio of 89%. This study offers a viable and sustainable approach to convert agro-industrial waste into effective and reusable biosorbents for uranium removal from wastewater.