Construction and preliminary characterization of polyhistidine tagged phosphotriesterase

CHED 802

Briana Flaherty, briana.flaherty@gmail.com1, Angelo Romano1, Jasmine Furnish1, Andrew M. Slupe, Andrew.Slupe@boisetechnology.org2, and Henry A. Charlier Jr., hcharlier@chem.boisestate.edu1. (1) Department of Chemistry and Biochemistry, Boise State University, 1910 University Drive, Mail Stop 1520, Boise, ID 83725, (2) Boise Technology, 5465 East Terra Linda Way, Boise, ID 83687
Phosphotriesterase (PTE) catalyzes the breakdown of a variety of organophosphate pesticides and nerve agents. A HIS-tagged PTE expression system was implemented to facilitate a one step purification protocol using nickel affinity chromatography. This system yielded homogeneous and active PTE. Kinetic studies of the polyhistidine -modified PTE protein were done with parathion as a substrate to determine if the modification impacted enzyme function. The Km for parathion is comparable to that reported in the primary literature. The modified PTE is active in a carbontetrachloride/buffer biphasic system. Such a finding is a prelude to studies focused on understanding the decontamination kinetics of parathion hydrolysis in an organic/aqueous biphasic system. (Funded in part by Boise Technology, Inc., DTRA and NIH/P20RR016454)
 

Undergraduate Research Poster Session: Biochemistry
2:00 PM-4:00 PM, Monday, April 7, 2008 Morial Convention Center -- Hall A, Poster

Division of Chemical Education

The 235th ACS National Meeting, New Orleans, LA, April 6-10, 2008