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African Journal of Biotechnology

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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

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 -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 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.



AJOL African Journals Online