title: Efficient bacterial expression of fusion proteins and their selective processing by a recombinant Kex-1 protease.
abstract: A secreted, soluble variant of the Kex-1 endopeptidase from Kluyveromyces lactis has been produced and studied as a novel cleavage enzyme exhibiting high specificity for the Lys-Arg peptide. This highly selective, efficient enzyme is particularly adapted for use in manufacturing when a recombinant therapeutic protein, possessing its native N-terminus, has to be released in vitro from a bacterially-expressed fusion protein. In this paper, we describe the preparation of a Kex-1 variant using Saccharomyces cerevisiae and its application in the production of important therapeutic recombinant proteins such as human growth hormone, granulocyte colony-stimulating factor and interferon-alpha-2b.
authors.affiliation: Bio-Ker S.R.L., Sardinia Scientific and Technological Park, Building 3, 09010 Pula, Cagliari, Italy.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Pozzuolo
authors.forename: Umberto
authors.initials: U
authors.lastname: Breme
authors.forename: Barbara
authors.initials: B
authors.lastname: Salis
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Taylor
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Tonon
authors.forename: Gaetano
authors.initials: G
authors.lastname: Orsini
mesh_terms: D002268:Carboxypeptidases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics*; D005285:Fermentation; D017510:Protein Folding; D011683:Purine-Nucleoside Phosphorylase / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D011993:Recombinant Fusion Proteins; D011683:Purine-Nucleoside Phosphorylase; D001616:beta-Galactosidase; D002268:Carboxypeptidases; C078874:carboxypeptidase D