Metallohydrolases are metal-dependent enzymes that catalyze the hydrolytic cleavage of a range of biologically relevant substrates, including esters, organophosphates, polynucleotides, peptides, and CO2. It was recently demonstrated that the de novo designed Due Ferri single chain (DFsc) scaffold is capable of hydrolyzing both small molecule substrates and double-stranded DNA. To further understand the structural and electronic factors that contribute to the hydrolase reactivity in the DFsc system, a series of protein variants were constructed and evaluated for their ability to bind metal ions and perform hydrolytic cleavage reactions. Metal ions commonly used by nature, like Zn2+, as well as metals underutilized by nature, like Ti4+ and V5+ were examined. Incorporation of an active site tyrosine residue similar to that found in purple acid phosphatase, which is known to bind and stabilize hydrolysis-prone metals, resulted in increased rates of hydrolytic activity. Results from these studies can be used to further optimize the DFsc scaffold for hydrolytic cleavage reactions.

Hydrolytic reactivity of de novo designed mini-metalloenzymes

ACS Spring 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.