2015 IPFW Student Research and Creative Endeavor Symposium
 

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Faculty Sponsor

Dr. Eric Tippmann

Department/Program

Department of Chemistry

University Affiliation

Indiana University – Purdue University Fort Wayne

Abstract

An amino acyl t-RNA enzyme is one the most important and ancient of all enzymes. All organisms have them and their “job” is to make sure that the correct amino acid is inserted in the correct order of every protein. While our DNA determines the order of amino acids and proteins, it is this class of enzymes that makes sure the DNA’s code is followed with very few mistakes. Recently, these enzymes have been evolved away from their natural substrates so that they actually prefer non-natural amino acids. The larger goal of our project is to determine exactly how this occurred, but the more immediate goal is to validate a new assay for assessing these enzymes efficiency. To this end, we are using a wild type, or natural, enzyme from the bacteria M. jannascii to standardize the assay. This enzyme is known and its role in nature is to specifically insert the amino acid tyrosine into every growing protein as it is needed. The first goal is to obtain and purify the enzyme from bacterial expression cultures. The enzyme has a special affinity tag that facilitates its purification from the hundreds of other bacterial proteins. The second goal is to determine how efficiently the enzyme catalyzes the reaction with tyrosine. We will compare our results with the literature to validate the larger project’s goal. If successful, then we may use the new assay for other unknown enzymes made in our research laboratory.

Disciplines

Chemistry | Physical Sciences and Mathematics

Characterization of an amino-acyl tRNA enzyme from M. jannaschii for genetic applications

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Chemistry Commons

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