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

Patricia Landázuri, Raúl A. Poutou-Piñales, Jovanna Acero-Godoy, Henry A. Córdoba-Ruiz, Olga Y. Echeverri-Peña, Homero Sáenz, Julio M. Delgado, Luis A. Barrera-Avellaneda

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

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 α-mating factor signal peptide (α-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 nmol.h-1.mg 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.

Original languageEnglish
Pages (from-to)2871-2877
Number of pages7
JournalAfrican Journal of Biotechnology
Volume8
Issue number12
StatePublished - 17 Jun 2009

Keywords

  • Human recombinant protein
  • Hunter syndrome
  • Iduronate-2-sulfate sulfatase
  • MPS II
  • Pichia pastoris

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