title: High-level expression and characterization of Galactomyces geotrichum (BT107) lipase I in Pichia pastoris.
abstract: The mature lipI gene, encoding the lipase I from Galactomyces geotrichum BT107, was obtained by PCR from genomic DNA, sequenced and cloned into a Pichia pastoris expression vector. Clones containing multiple copies of lipI integrated in their genome were analyzed to achieve high-level expression of the recombinant lipase I. One strain with four or more copies of the expression cassette was able to produce more than 200mg/L of extracellular heterologous protein. The lipase I was partially purified using anion exchange chromatography and its activity on monounsaturated (triolein) and polyunsaturated (triEPA) triglycerides was analyzed by a novel HPLC-MS assay.
authors.affiliation: Department of Biotechnology, Puleva Biotech, Camino de Purchil, 66, 18004-Granada, Spain.
authors.forename: Layla
authors.initials: L
authors.lastname: Fernández
authors.forename: Ignacio
authors.initials: I
authors.lastname: Pérez-Victoria
authors.forename: Alberto
authors.initials: A
authors.lastname: Zafra
authors.forename: Pedro L
authors.initials: PL
authors.lastname: Benítez
authors.forename: Juan C
authors.initials: JC
authors.lastname: Morales
authors.forename: Javier
authors.initials: J
authors.lastname: Velasco
authors.forename: José L
authors.initials: JL
authors.lastname: Adrio
mesh_terms: D002851:Chromatography, High Pressure Liquid; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005800:Genes, Fungal / Q000235:genetics; D008049:Lipase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012440:Saccharomyces / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D008049:Lipase