Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2021950 | Protein Expression and Purification | 2006 | 9 Pages |
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 200 mg/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.
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Biochemistry
Authors
Layla Fernández, Ignacio Pérez-Victoria, Alberto Zafra, Pedro L. Benítez, Juan C. Morales, Javier Velasco, José L. Adrio,