Publications [#234205] of Eric J. Toone

Journal Articles

  1. Griffiths, JS; Wymer, NJ; Njolito, E; Niranjanakumari, S; Fierke, CA; Toone, EJ, "Cloning, isolation and characterization of the Thermotoga maritima KDPG aldolase.", Bioorganic & medicinal chemistryMarch,, 2002, 10(3), 545-550 [11814840], [doi].
    (last updated on 2024/11/19)

    Abstract:
    The Thermotoga maritima aldolase gene has been cloned into a T7 expression vector and overexpressed in Escherichia coli. The preparation yields 470 UL(-1) of enzyme at a specific activity of 9.4 U mg(-1). During retroaldol cleavage of KDPG, the enzyme shows a k(cat) that decreases with decreasing temperature. A more than offsetting decrease in K(m) yields an enzyme that is more efficient at 40 degrees C than at 70 degrees C. The substrate specificity of the enzyme was evaluated in the synthetic direction with a range of aldehyde substrates. Although the protein shows considerable structural homology to KDPG aldolases from mesophilic sources, significant differences in substrate specificity exist. A preparative scale reaction between 2-pyridine carboxaldehyde and pyruvate provided product of the same absolute configuration as mesophilic enzymes, but with diminished stereoselectivity.