Secretion of active urokinase-type plasminogen activator from the yeast Yarrowia lipolytica

Abstract

In order to study the secretion of the human urokinase-type plasminogen activator, u-PA, from the yeast Yarrowia lipolytica, three kinds of integrative expression vector were constructed. These vectors differed only in their secretion control regions, pre-, pre-dip- (dipeptide stretch) or pre-dip-pro sequences of the alkaline extracellular protease, which were joined inframe to the human u-PA cDNA. The recombinant Y. lipolytica strains, transformed with the expression vectors, secreted the hyperglycosylated u-PA. A fibrin plate assay of the culture supernatants showed that the hyperglycosylated u-PA proteins could catalyze fibrinolysis, and that the pre-dip sequence was the most efficient secretory signal for the secretion of the u-PA from Y. lipolytica. This result suggests that Y. lipolytica can be developed as a potential host for the production of recombinant human u-PA.