**ABSTRACT NOT FOR CITATION WITHOUT AUTHOR PERMISSION. The title, authors, and abstract for this completion report are provided below.  For a copy of the full completion report, please contact the author via e-mail at glasst@missouri.edu or via phone at (573)882-3813. Questions? Contact the GLFC via email at frp@glfc.org or via telephone at 734-662-3209.**



 Improving thiaminase measurements in Great Lakes forage fish


Timothy E, Glass1, James Zajicek2, Donald Tillitt2



1  Department of Chemistry, University of Missouri, 601 S. College Ave. Columbia, MO 65203

2  Biochemistry & Physiology Branch, Columbia Environmental Research Center, U.S. Geological Survey, Department of Interior, 4200 New Haven Rd., Columbia, MO 65201


January 2016




Two new fluorescent molecules have been developed for thiaminase: one used to measure thiaminase activity and the other one used to inactivate this interesting enzyme. The thiaminase sensor was designed based on a PET mechanism. The internal quenching of the fluorophore (6Hex) by the attachment of a pyrimidine group makes it non-emissive. Under the action of thiaminase (representative by bisulfite), the free pyridine is released, which results in an increase in fluorescence. In addition to the thiaminase sensor, a red fluorescent, selective, irreversible inhibitor has also been synthesized. The inactivation of this interesting molecule has been confirmed by both plate assays and fluorescent experiences. Inactivation was found to go to completion if given sufficiently long incubation, and to be irreversible.