Identification of amino acid residues involved in substrate specificity of plant acyl-ACP thioesterases using a bioinformatics-guided approach |
| |
Authors: | Kimberly M Mayer and John Shanklin |
| |
Institution: | (1) Brookhaven National Laboratory, Department of Biology, Upton, NY 11973, USA;(2) University of North Carolina at Wilmington, Center for Marine Science, Wilmington, NC 28409, USA |
| |
Abstract: | Background The large amount of available sequence information for the plant acyl-ACP thioesterases (TEs) made it possible to use a bioinformatics-guided
approach to identify amino acid residues involved in substrate specificity. The Conserved Property Difference Locator (CPDL)
program allowed the identification of putative specificity-determining residues that differ between the FatA and FatB TE classes.
Six of the FatA residue differences identified by CPDL were incorporated into the FatB-like parent via site-directed mutagenesis
and the effect of each on TE activity was determined. Variants were expressed in E. coli strain K27 that allows determination of enzyme activity by GCMS analysis of fatty acids released into the medium. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|