Introduction of an N-glycosylation site increases secretion of heterologous proteins in yeasts

Saccharomyces cerevisiae is often used to produce heterologous proteins tha t are preferentially secreted to increase economic feasibility. We used N-g lycosylation as a tool to enhance protein secretion. Secretion of cutinase, a lipase, and Ilama V-HH antibody fragments by S. cerevisiae or Pichin pas toris improved following the introduction of an N-glycosylation site. When we introduced an N-glycosylation consensus sequence in the N-terminal regio n of a hydrophobic cutinase, secretion increased fivefold. If an N-glycosyl ation site was introduced in the C-terminal region, however, secretion incr eased only 1.8-fold. These results indicate that the use of N glycosylation can significantly enhance heterologous protein secretion.