Abstract <p>The reduction of sulfurous species coupled with anaerobic ammonium oxidation (Sammox) processes plays a significant role in the geochemical cycling of sulfur, nitrogen, and carbon, as well as in the emergence of biochemical cycles. Anaerobic ammonium oxidation to nitrate (AAON) occurs within the microbiological Sulfammox process, which involves sulfate reduction coupled with anaerobic ammonium oxidation. We studied the AAON process in Sammox-driven chemical reaction networks (CRNs) by observing nitrate formation resulting from peptide function evolution during short-term hydrothermal and subsequent long-term experiments under ambient conditions. Small quantities of proteinogenic amino acids were produced during a 3-year reaction under ambient conditions, attributed to the autocatalysis of peptides formed in Sammox-driven CRNs after a 48-h reaction at 100^oC. After an additional 3 years of reaction, nitrate was detected in all treatment groups, suggesting that the “biological” Sulfammox process occurs within the systems. Peptides can function as proto-enzymes, while the formation of stable vesicle structures provides an optimal environment and conditions for the evolution of CRNs into enzymatic proto-metabolic systems. Prebiotic evolution may occur much more rapidly than previously believed.</p>