Fig. 2.

The D6d activity of D6d mutants. The D6d mutants were generated by site-directed mutagenesis using one or more oligonucleotide primers as indicated by the numbers, which correspond with the primer numbers (d6d5-1–d6d5-35; Table 1). The mutants carrying D5d amino acid(s) were expressed in yeast S. cerevisiae in the presence of LA, and the fatty acid composition was measured as described in the Materials and Methods. D6d activity was determined as the conversion rate from LA to GLA and is shown as the average value relative to that of intact D6d, together with the standard deviation (n = 3). FLAG-tagged D6d proteins were detected by Western blotting of total yeast protein using anti-FLAG antibody.