title: Mutant strains of Pichia pastoris with enhanced secretion of recombinant proteins.
abstract: Although Pichia pastoris is a popular protein expression system, it exhibits limitations in its ability to secrete heterologous proteins. Therefore, a REMI (restriction enzyme mediated insertion) strategy was utilized to select mutant beta-g alactosidase s upersecretion (bgs) strains that secreted increased levels of a β-galactosidase reporter. Many of the twelve BGS genes may have functions in intracellular signaling or vesicle transport. Several of these strains also appeared to contain a more permeable cell wall. Preliminary characterization of four bgs mutants showed that they differed in the ability to enhance the export of other reporter proteins. bgs13, which has a disruption in a gene homologous to Saccharomyces cerevisiae protein kinase C (PKC1), gave enhanced secretion of most recombinant proteins that were tested, raising the possibility that it has the universal super-secreter phenotype needed in an industrial production strain of P. pastoris.
authors.affiliation: Department of Biological Sciences, University of the Pacific, 3601 Pacific Avenue, Stockton, CA, 95211, USA.
authors.forename: Sasha
authors.initials: S
authors.lastname: Larsen
authors.forename: Jun
authors.initials: J
authors.lastname: Weaver
authors.forename: Katherine
authors.initials: K
authors.lastname: de Sa Campos
authors.forename: Rhobe
authors.initials: R
authors.lastname: Bulahan
authors.forename: Jackson
authors.initials: J
authors.lastname: Nguyen
authors.forename: Heather
authors.initials: H
authors.lastname: Grove
authors.forename: Amy
authors.initials: A
authors.lastname: Huang
authors.forename: Lauren
authors.initials: L
authors.lastname: Low
authors.forename: Namphuong
authors.initials: N
authors.lastname: Tran
authors.forename: Seth
authors.initials: S
authors.lastname: Gomez
authors.forename: Jennifer
authors.initials: J
authors.lastname: Yau
authors.forename: Thomas
authors.initials: T
authors.lastname: Ilustrisimo
authors.forename: Jessica
authors.initials: J
authors.lastname: Kawilarang
authors.forename: Jonathan
authors.initials: J
authors.lastname: Lau
authors.forename: Maivi
authors.initials: M
authors.lastname: Tranphung
authors.forename: Irene
authors.initials: I
authors.lastname: Chen
authors.forename: Christina
authors.initials: C
authors.lastname: Tran
authors.forename: Marcia
authors.initials: M
authors.lastname: Fox
authors.forename: Joan
authors.initials: J
authors.lastname: Lin-Cereghino
authors.forename: Geoff P
authors.initials: GP
authors.lastname: Lin-Cereghino
mesh_terms: D017930:Genes, Reporter; D060847:Metabolic Engineering; D016296:Mutagenesis; D009154:Mutation; D010843:Pichia / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D001616:beta-Galactosidase