Chemistry Department Seminar: Tenure Track Candidate
"Discovery and Characterization of unique Alanine Mutation 2-Enoate Reductase (BurkOYE)"
The Old yellow enzyme (OYE) superfamily is a pharmaceutically and industrially relevant set of enzymes composed of 100,000+ members. Within that superfamily lies an underexplored family of 4Fe-4S cluster containing multidomain OYEs, The 2-Enoate Reductase (2-ER) family. The 2-ER family is the second-largest family within the superfamily; however, < 0.01% of that family has been explored to date. It has previously been discovered that this family can perform enzymatic activity that differs from the non-4Fe-4S cluster containing OYEs (traditional OYEs). Herein I describe the use of bioinformatics and traditional biochemistry to discover and characterize a novel member of the 2-ER family. The 2-ER family was visualized and analyzed for sequence divergence using sequence similarity networks and cluster analysis. It was discovered that within the 2-ER family lies a subfamily of unique alanine mutation in the 4Fe-4S cluster binding domain. We hypothesize that this mutation could lead to novel enzymatic activity. To explore such a representative member containing this unique mutation, BurkOYE, was heterologously transformed, co-expressed, and purified. BurkOYE was tested against a 10-panel substate screen. From this screen, we discovered that BurkOYE showed activity towards traditional OYE substrates while, surprisingly, maintaining 2-ER substrates and unique demethylase activity.
*This seminar counts towards the chemistry major seminar attendance requirement.