Production of Saccharomyces cerevisiae metacaspase (YCA1) containing site-directed mutations: importance of the calcium interaction site for activation and control of enzyme activity

• Grant number: 20/02244-8
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: May 01, 2020
• End date: December 31, 2020
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Mauricio Ferreira Marcondes Machado
• Grantee: Mariana Nascimento Romero Trujilho
• Host Institution: Pró-Reitoria Acadêmica. Universidade de Mogi das Cruzes (UMC). Campus da Sede Mogi das Cruzes. Mogi das Cruzes , SP, Brazil

Abstract

The present research project aims at the production of Saccharomyces cerevisiae (YCA1) metacaspase, including mutations in the site directed at the aspartate residues responsible for the interaction of alcohol with this enzyme. A YCA1 metacaspase undergoes automatic processing to become catalytically active, and uses calcium as an enzymatic co-factor both for its automatic processing and for carrying out its enzymatic activity, for this reason it intended to carry out a production of this enzyme using site mutations. directed at the four waste sections responsible for the interaction of YCA1 with calcium, they are Asp236, Asp252, Asp253 and Asp283 and thus try to elucidate the importance of the self-processing test and activity of this protease activity. Performs double mutants and mutants of all residues of interaction with calcium or in order to verify which are the crucial residues for an enzyme of YCA1, such an investment study for the rational design of molecules that inhibit and/or activate a YCA1 being of great importance for the development of possible new products.