Engineering a lipoxygenase from cyclocybe aegerita towards long chain polyunsaturated fatty acids

The basidiomycetous lipoxygenase Lox1 from Cyclocybe aegerita catalyzes the oxygenation of polyunsaturated fatty acids (PUFAs) with a high preference towards the C18-PUFA linoleic acid (C18:2 (o-6)). In contrast, longer PUFAs, generally not present in the fungal cell such as eicosatrienoic acid (C20:3(o-3)) and docosatrienoic acid (C22:3 (o-3)), are converted with drastically lower activities. With site-directed mutagenesis, we were able to create two variants with enhanced activities towards longer chain PUFAs. The W330L variant showed a ~ 20 % increased specific activity towards C20:3(o-3), while a ~ 2.5-fold increased activity against C22:3 (o-3) was accomplished by the V581 variant.