In this dissertation, two key approaches for the design of catalytic peptides are explored: (i) the incorporation of amino acids from the catalytic triad and (ii) an approach involving metal ions. To identify these main design approaches, a comprehensive dataset based on the collection of published data about purely peptidic catalysts for ester hydrolysis was gathered and analyzed including both active and inactive sequences tested towards the p-NPA and p-NPP substrates providing an insight into the currently developed catalytic mechanisms. Additionally, this open-access dataset also includes the SMILESbased representation of peptides and it can be used for training of machine learning-based models. Moreover, the standard p-NPA method was optimized. This extensively employed assay for determining catalytic activity, initially designed for enzymatic reactions, is not directly applicable to peptides due to variations in reaction parameters, such as the concentrations of catalysts and reaction times, when comparing peptides to enzymes. Consequently, standardizing conditions, including temperature, pH, buffer choice, and concentrations of both substrate and peptide, became essential to ensure accurate and reliable results in peptide reactions. Furthermore, the synthesis of a set of histidine-rich linear and cyclic peptides allowed us to understand the impact of amino acid arrangement, cyclization, and inclusion of D-amino acids on their self-assembly, coordination of Zn2+ ions and ability to hydrolyze p-NPA. Investigating structural changes crucial to the functionality of short peptides contributes to the development of a new generation of catalytic peptides and advances our understanding of the potential of self-assembly in improving catalytic efficiency. Additionally, we investigated the feasibility of isolating the active site of an enzyme to develop a minimalist catalyst, with a specific focus on recognizing cysteine as a crucial component within the catalytic triad, essential for the effectiveness of the peptide catalyst. The synthesis of a set of cysteine-rich peptides showing tunable activity resulting from the sensitivity of the thiol groups to the redox environment could open promising opportunities for tailored enzymatic catalysis. The overall goal of this research is to gain a deeper insight into the relationship between the sequence and function of catalytic peptides, leading to the development of innovative peptide catalysts with potential applications in various fields, including biocatalysis, pharmaceutical synthesis, and other industrial processes.