As part of our investigation of fatty acid metabolism in Trypanosoma brucei, we have expressed four acyl-CoA synthetase (TbACS) genes in Esherichia coli. The recombinant proteins, with His-tags on their C-termini, were purified to near homogeneity using nickel-chelate affinity chromatography. Although these enzymes are highly homologous, they have distinct specificities for fatty acid chain length. TbACS1 prefers saturated fatty acids in the range C11:0 to C14:0 and TbACS2 prefers shorter fatty acids, mainly C10:0. TbACS3 and 4, which have 95% sequence identity, have similar specificities, favouring fatty acids between C14:0 and C17:0. In addition, TbACS1, 3 and 4 function well with a variety of unsaturated fatty acids.
This content is only available as a PDF.
The Biochemical Society, London ©2001
2001
You do not currently have access to this content.