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

The human lduronate-2-sulfate sulfatase (hlDS-Like) was cloned into the methylotrophic yeast Pichia pastoris under the control of alcohol oxidase promoter (AOX1) and the alpha -mating factor signal peptide ( alpha -factor). Six clones were identified by PCR. Using clone IDS28, the enzyme was secrete...

Full description

Saved in:
Bibliographic Details
Published in:African journal of biotechnology 2009-06, Vol.8 (12), p.2871-2877
Main Authors: Landazuri, P, Poutou-Pinales, R A, Acero-Godoy, J, Cordoba-Ruiz, HA, Echeverri-Pena, O Y, Saenz, H, Delgado, J M, Barrera-Avellaneda, LA
Format: Article
Language:English
Subjects:
Pichia pastoris
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The human lduronate-2-sulfate sulfatase (hlDS-Like) was cloned into the methylotrophic yeast Pichia pastoris under the control of alcohol oxidase promoter (AOX1) and the alpha -mating factor signal peptide ( alpha -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 super(-1).mg of total protein super(-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.
ISSN:1684-5315
1684-5315