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Expression and characterization of triosephosphate isomerase from Epulopiscium sp. Type B
Author(s) -
DealLaroda Nikolas Antonio,
Piefer Andrew John
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.853.12
Subject(s) - triosephosphate isomerase , escherichia coli , biochemistry , isomerase , biology , dihydroxyacetone phosphate , amino acid , cloning (programming) , clostridium botulinum , gene , microbiology and biotechnology , enzyme , computer science , toxin , programming language
The main goals of this project are to express and characterize the putative Triosephosphate isomerase gene (TPI) from the bacterium Epulopiscium sp. Type B. This large bacterium (~250 µm in length) has never been cultured, grows in the gut of Naso tonganus (a surgeonfish), and may assist in the fish's algae digestion. TPI is responsible for the interconversion of dihydroxyacetone phosphate and glyceraldehyde 3‐phosphate. This catalytic step bridges the initial ATP priming stage and the energy production stage of glycolysis. PCR was performed to amplify TPI for subsequent cloning and expression in Escherichia coli . The Epulopiscium TPI protein contains approximately 247 amino acids and its theoretical molecular weight is 26,841 Daltons. Most characterized TPIs from other organisms are homodimeric in structure, but this remains to be determined for Epulopiscium TPI . Epulopiscium TPI was aligned to TPI from Clostridium botulinum, E. coli, and Homo sapiens , showing 91.5% amino acid conservation among these diverse organisms. Furthermore, the alignment shows that Epulopiscium TPI has 63.7 % identity with Clostridium botulinum , a 41.4 % identity with E. coli , and a 42.9 % identity with Homo sapiens . These studies may provide a glimpse into this unique organism's physiology through examination of a highly conserved enzymatic process. This project was funded by the Hartwick College Department of Chemistry.