EcNikA, a versatile tool in the field of artificial metalloenzymes

• Published in: Journal of inorganic biochemistry 2025-01, Vol.262, p.112740, Article 112740
• Main Authors: Marchi-Delapierre, Caroline, Cavazza, Christine, Ménage, Stéphane
• Format: Article
• Language: English
• Subjects: Artificial metalloenzymes; Biochemistry, Molecular Biology; Catalysis; Catalytic Domain; Chemical Sciences; Cross-linked enzyme crystals; iron; Life Sciences; Metalloproteins - chemistry; Metalloproteins - metabolism; Nickel - chemistry; Oxidation; Oxidation-Reduction; Ruthenium
• ISSN: 0162-0134; 1873-3344; 1873-3344
• DOI: 10.1016/j.jinorgbio.2024.112740

This review describes the multiple advantages of using of EcNikA, a nickel transport protein, in the design of artificial metalloenzymes as alternative catalysts for synthetic biology. The rationale behind the strategy of artificial enzyme design is discussed, with particular emphasis on de novo active site reconstitution. The impact of the protein scaffold on the artificial active site and thus the final catalytic properties is detailed, highlighting the considerable aptitude of hybrid systems to catalyze selective reactions, from alkene to thioether transformations (epoxidation, hydroxychlorination, sulfoxidation). The different catalytic approaches – from in vitro to in cristallo – are compared, revealing the considerable advantages of protein crystals in terms of stabilization and acceleration of reaction kinetics. The versatility of proteins, based on metal and ligand diversity and medium/physical conditions, are thus illustrated for oxidation catalysis. [Display omitted] •Combining an artificial active site with NikA, an E. coli nickel transport protein.•The protein scaffold decouples the activity of the artificial active site.•Compared to in-solution conditions, the protein scaffold stabilizes ligand topology.•An in cristallo approach leads to greater stabilization than the in vitro approach.•Reaction mechanisms are accessible thanks to X-ray crystallography.