Introduction of an N-glycosylation site increases secretion of heterologous proteins in yeasts.
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.
Sagt
C M
CM
Department of Molecular Cell Biology and the Institute of Biomembranes, Utrecht University, 3584 CH Utrecht, The Netherlands.
Kleizen
B
B
Verwaal
R
R
de Jong
M D
MD
Müller
W H
WH
Smits
A
A
Visser
C
C
Boonstra
J
J
Verkleij
A J
AJ
Verrips
C T
CT
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*
D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D011994:Recombinant Proteins; D002265:Carboxylic Ester Hydrolases; C019952:cutinase; D006596:Hexosaminidases