High-Level Secretion of Glycosylated Invertase in the Methylotrophic Yeast, Pichia Pastoris

Abstract

The SUC2 gene from S. cerevisiae was transformed into the methylotrophic yeast Pichia pastoris and used to study the synthesis, post-translational processing, and secretion of invertase. SUC2 gene expression under the control of the alcohol oxidase (AOX1) promoter resulted in efficient secretion of biologically active invertase into the periplasmic space and growth medium of P. pastoris, giving the methylotrophic yeast a Suc+ phenotype. Invertase produced and secreted from an alcohol oxidase-deficient (aox1) host reached levels of up to 2.5 grams per liter of growth medium. The N-terminus of invertase secreted from P. pastoris was homogeneous and found to be identical to invertase secreted from S. cerevisiae. Invertase secreted from P. pastoris was glycosylated but contained significantly less carbohydrate than protein secreted by S. cerevisiae.

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Tschopp, J., Sverlow, G., Kosson, R. et al. High-Level Secretion of Glycosylated Invertase in the Methylotrophic Yeast, Pichia Pastoris. Nat Biotechnol 5, 1305–1308 (1987). https://doi.org/10.1038/nbt1287-1305

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