Document Type

Paper- Restricted to Campus Access

Publication Date

4-23-2020

Faculty Mentor

Amanda Reig

Abstract

A little less than half the proteins found in nature utilize metal ions to perform catalytic reactions. The metal of interest is manganese that is usually found in the enzyme arginase, a binuclear metallohydrolase protein that is a hydrolytic enzyme cleaving phosphate or ester bonds. This enzyme specifically cleaves L-arginine into L-ornithine and urea. A way to study how arginase performs its reactions is to use the synthetic model D e Novo Due Ferri single chain (DFsc) proteins. Preliminary trials of phosphate cleavage assays were performed to understand if any variation of the metal, protein, or buffer is performing the hydrolytic reaction with the substrate, bis(4-nitrophenyl) phosphate (BNPP). Michaelis-Menten kinetics were performed to predict the rate of the product formation from the enzyme at different concentrations of substrate. Mutagenesis experiments were also performed to mutate specific amino acid residues of the parent protein to closely mimic the natural arginase protein to allow stronger binding of the manganese ion. By understanding how these DFsc enzymes work and perform, new synthetic enzymes could be designed for drug discovery and biotechnology.

Comments

Presented as part of the Ursinus College Celebration of Student Achievement (CoSA) held April 23 – April 30, 2020.

The downloadable file is a poster with recorded audio commentary.

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