EnzyMime - Biocompatible and sustainable enzyme mimics based on metal complexes with peptide derivatives: synthesis, characterization and potential biological applications

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Abstract:

Despite the many advantages of the use of enzymes as catalysts, including high selectivity and stereocontrol, their large-scale application is limited by several drawbacks: low thermal stability and tolerance to different experimental conditions, poor substrate versatility, unsuitability to abiotic reactions and high cost of purification. As an attempt to overcome such vulnerabilities, enzyme mimics (EMs) has been investigating since long time.

The development of efficient EMs represents a formidable opportunity on the way of a green transition and a high grade of competitive innovation, both needful for our prosperity and wellbeing. Notably, in recent years, the research progress on EMs has led to significant developments of sustainable chemical synthesis protocols, at both laboratory and industrial scale level. These compounds have the potential to compete with natural enzymes and “classical” catalysts, finding application in a wide range of fields, from chemical and food industries to diagnostics and medicine or production of fuels and gaseous hydrogen.

EMs are usually designed to reproduce the catalytic sites of natural enzymes, following two possible approaches: imitating the enzyme functionality using metal complexes with similar activity, or mimicking the structure of the enzyme active site by introducing appropriate functional groups (e.g.: oligopeptides).

The present research project (EnzyMime) aims at designing a new class of de novo EMs, based on synthetic branched peptides. A biocompatible central scaffold represents the core of the EM structure, to which four identical oligopeptides are linked through the so-called “peptide welding technology” (PWT). This will allow the easy development of different EMs, where the potential catalytic site can be produced introducing appropriate amino acid sequences, able to bind the active metal ions. An example may be represented by the synthesis of different surrogates of Cu/Zn-SOD, Mn-SOD and Ni-SOD to catalyse dismutation of superoxide radicals.

The experimental approach will consist of: synthesis of several EMs and their catalytic characterization; complete characterization of the most promising metal/peptide systems, through potentiometry, voltammetry, spectrophotometry, spectrofluorimetry, mass spectrometry, nuclear magnetic resonance, electron paramagnetic resonance, theoretical calculations and structural investigation by X-ray crystallography. To help the crystallization of peptides and their metal complexes, a spacer containing a short alpha-helix folding sequence will also been added to the chelating sequences. Finally, in order to explore some possible practical applications in the field of human health and wellbeing, biological tests will be performed, including enzymatic stability in human plasma, interaction with nucleic acids, cytotoxicity against cancer cells, mobility through different types of membranes and antimicrobial activity.

Dettagli progetto:

Referente scientifico: Bellotti Denise

Fonte di finanziamento: Bando PRIN 2022 PNRR

Data di avvio: 30/11/2023

Data di fine: 30/11/2025

Contributo MUR: 60.200 €

Web site: https://sites.google.com/unife.it/enzymime/

Partner:

  • Università degli Studi di FERRARA (capofila)
  • Università degli Studi di PALERMO
  • Università degli Studi di SASSARI
  • Università degli Studi di Napoli Federico II
  • Università degli Studi di TRIESTE
  • Università degli Studi di TORINO