Cloning and shake flask expression of hrIDS-Like in Pichia pastoris

  • P Landázuri
  • RA Poutou-Piñales
  • J Acero-Godoy
  • HA Córdoba- Ruiz
  • OY Echeverri-Peña
  • H Sáenz
  • JM Delgado
  • LA Barrera- Avellaneda
Keywords: Iduronate-2-sulfate sulfatase, MPS II, Pichia pastoris, human recombinant protein, Hunter syndrome.


The human Iduronate-2-sulfate sulfatase (hIDS-Like) was cloned into the methylotrophic yeast Pichia pastoris under the control of alcohol oxidase promoter (AOX1) and the -mating factor signal peptide (a-factor). Six clones were identified by PCR. Using clone IDS28, the enzyme was secreted into the culture medium, yielding a protein with an activity of 4.213 of total protein-1 at 72 h, in 0.5% v/v methanol. Several bands were revealed by western-blot, indicating that a P. pastoris processing was slightly different than in mammalian cells.

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eISSN: 1684-5315