TY - JOUR
T1 - Cloning and shake flask expression of hrIDS-Like in Pichia pastoris
AU - Landázuri, Patricia
AU - Poutou-Piñales, Raúl A.
AU - Acero-Godoy, Jovanna
AU - Córdoba-Ruiz, Henry A.
AU - Echeverri-Peña, Olga Y.
AU - Sáenz, Homero
AU - Delgado, Julio M.
AU - Barrera-Avellaneda, Luis A.
PY - 2009/6/17
Y1 - 2009/6/17
N2 - 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.
AB - 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.
KW - Human recombinant protein
KW - Hunter syndrome
KW - Iduronate-2-sulfate sulfatase
KW - MPS II
KW - Pichia pastoris
UR - http://www.scopus.com/inward/record.url?scp=67650498704&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:67650498704
SN - 1684-5315
VL - 8
SP - 2871
EP - 2877
JO - African Journal of Biotechnology
JF - African Journal of Biotechnology
IS - 12
ER -