Laboratory scale production of the human recombinant iduronate 2-sulfate sulfatase-Like from Pichia pastoris

Clone IDS28 of the yeast Pichia pastoris expressing the human iduronate 2-sulfate sulfatase-Like (hIDSLike) was employed for low-scale production of the recombinant enzyme in a saline culture media without phosphate. The biological activity found was between 7.3 and 29.5 nmol h-1 mg-1 of total protein. It is about 1.73 to 7 times higher than the result obtained with the same clone in shake flask culture

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

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