This study demonstrates that amyloid fibrils of a de-novo designed peptide, PFK, serve as effective catalysts for the hydrolysis of β-lactam antibiotics. The catalytic activity of PFK amyloid fibrils occurs via a unique allosteric mechanism, mediated through formation of a coiled double fiber organization embedding the β-lactam substrate molecules and facilitating nucleophilic attacks by the sidechains of the lysine residues. PFK amyloid fibrils successfully catalyzed hydrolysis of varied β-lactams, including nitrocefin, amoxicillin, and penicillin, underscoring the generic nature of the catalytic platform. Practical utilization of the catalytic amyloids was further manifested through the display of the PFK fibrils on silica beads and usage of the amyloid fibril/bead constructs for water decontamination in a conventional filtration column. In conclusion, PFK catalytic amyloids furnish an innovative strategy for degradation of antibiotics in water, offering means for addressing this major environmental and health hazard. were purchased from Holland-Moran (Yehud, Israel). Ammonium acetate (99% purity) Potassium chloride (99% purity) and 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP, 99% purity), amoxicillin (95% purity) and Penicillin G sodium salt (96% purity) were purchased from Sigma-Aldrich (Rehovot, Israel). Acetonitrile (LC-MS grade, 99.9% purity) was obtained from Beith Dekel (Raanana, Israel, Manufactured by JTBaker). The fluorescent dye Amytracker-680 was supplied by Ebba biotech (Stockholm, Sweden). Glass beads of 2 mm dimeter were purchased from Mercury (Darmstadt, Germany),and silica microparticles in diameter of 10 µm were purchased from microparticles GmbH (Volmerstraße, Berlin). Anti-β-lactamase (polyclonal anti-βlactamase, with affinity to β-lactamase of E.Coli, LOT AB3738-I, UniProt P62593) was purchased from Mercury (Darmstadt, Germany).