Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Identification of amino acid residues involved in substrate specificity of plant acyl-ACP thioesterases using a bioinformatics-guided approach | BMC Plant Biology BioMed Central Research article Identification of amino acid residues involved in substrate specificity of plant acyl-ACP thioesterases using a bioinformatics-guided approach Kimberly M Mayer1 2 and John Shanklin 1 Open Access Address Brookhaven National Laboratory Department of Biology Upton NY 11973 USA and 2University of North Carolina at Wilmington Center for Marine Science Wilmington NC 28409 USA Email Kimberly M Mayer - mayerk@ John Shanklin - shanklin@ Corresponding author Published 03 January 2007 Received 14 September 2006 BMC Plant Biology 2007 7 1 doi 1471 -2229-7-1 Accepted 03 January 2007 This article is available from http 1471-2229 7 1 2007 Mayer and Shanklin licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License http licenses by which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. 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. Results Substitutions at four of the positions 74 86 141 and 174 changed substrate specificity to varying degrees while changes
