Cloning, isolation and characterization of the Thermotoga maritima KDPG aldolase.

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Griffiths, JS; Wymer, NJ; Njolito, E; Niranjanakumari, S; Fierke, CA; Toone, EJ

Published Date

  • March 2002

Published In

Volume / Issue

  • 10 / 3

Start / End Page

  • 545 - 550

PubMed ID

  • 11814840

Pubmed Central ID

  • 11814840

Electronic International Standard Serial Number (EISSN)

  • 1464-3391

International Standard Serial Number (ISSN)

  • 0968-0896

Digital Object Identifier (DOI)

  • 10.1016/s0968-0896(01)00307-8

Language

  • eng