title: Introduction of an N-glycosylation site increases secretion of heterologous proteins in yeasts.
abstract: Saccharomyces cerevisiae is often used to produce heterologous proteins that are preferentially secreted to increase economic feasibility. We used N-glycosylation as a tool to enhance protein secretion. Secretion of cutinase, a lipase, and llama V(HH) antibody fragments by S. cerevisiae or Pichia pastoris improved following the introduction of an N-glycosylation site. When we introduced an N-glycosylation consensus sequence in the N-terminal region of a hydrophobic cutinase, secretion increased fivefold. If an N-glycosylation site was introduced in the C-terminal region, however, secretion increased only 1.8-fold. These results indicate that the use of N glycosylation can significantly enhance heterologous protein secretion.
authors.affiliation: Department of Molecular Cell Biology and the Institute of Biomembranes, Utrecht University, 3584 CH Utrecht, The Netherlands.
authors.forename: C M
authors.initials: CM
authors.lastname: Sagt
authors.forename: B
authors.initials: B
authors.lastname: Kleizen
authors.forename: R
authors.initials: R
authors.lastname: Verwaal
authors.forename: M D
authors.initials: MD
authors.lastname: de Jong
authors.forename: W H
authors.initials: WH
authors.lastname: Müller
authors.forename: A
authors.initials: A
authors.lastname: Smits
authors.forename: C
authors.initials: C
authors.lastname: Visser
authors.forename: J
authors.initials: J
authors.lastname: Boonstra
authors.forename: A J
authors.initials: AJ
authors.lastname: Verkleij
authors.forename: C T
authors.initials: CT
authors.lastname: Verrips
mesh_terms: D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D014644:Genetic Variation; D006031:Glycosylation; D006596:Hexosaminidases / Q000378:metabolism; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000378:metabolism; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*
chemical_list: D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D011994:Recombinant Proteins; D002265:Carboxylic Ester Hydrolases; C019952:cutinase; D006596:Hexosaminidases