OPENPichia: licence-free Komagataella phaffii chassis strains and toolkit for protein expression.

Autor: Claes K; Center for Medical Biotechnology, VIB, Ghent, Belgium. Katrien.Claes@vib-ugent.be.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium. Katrien.Claes@vib-ugent.be., Van Herpe D; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium.; Inbiose NV, Ghent, Belgium., Vanluchene R; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Roels C; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Van Moer B; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Wyseure E; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Vandewalle K; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Eeckhaut H; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Yilmaz S; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Vanmarcke S; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Çıtak E; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Fijalkowska D; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Grootaert H; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Lonigro C; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Meuris L; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Michielsen G; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., Naessens J; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., van Schie L; Center for Medical Biotechnology, VIB, Ghent, Belgium.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium., De Rycke R; Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.; BioImaging Core, VIB, Ghent, Belgium., De Bruyne M; Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.; BioImaging Core, VIB, Ghent, Belgium., Borghgraef P; BioImaging Core, VIB, Ghent, Belgium., Callewaert N; Center for Medical Biotechnology, VIB, Ghent, Belgium. Nico.Callewaert@vib-ugent.be.; Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium. Nico.Callewaert@vib-ugent.be.
Jazyk: angličtina
Zdroj: Nature microbiology [Nat Microbiol] 2024 Mar; Vol. 9 (3), pp. 864-876. Date of Electronic Publication: 2024 Mar 04.
DOI: 10.1038/s41564-023-01574-w
Abstrakt: The industrial yeast Komagataella phaffii (formerly named Pichia pastoris) is commonly used to synthesize recombinant proteins, many of which are used as human therapeutics or in food. However, the basic strain, named NRRL Y-11430, from which all commercial hosts are derived, is not available without restrictions on its use. Comparative genome sequencing leaves little doubt that NRRL Y-11430 is derived from a K. phaffii type strain deposited in the UC Davis Phaff Yeast Strain Collection in 1954. We analysed four equivalent type strains in several culture collections and identified the NCYC 2543 strain, from which we started to develop an open-access Pichia chassis strain that anyone can use to produce recombinant proteins to industry standards. NRRL Y-11430 is readily transformable, which we found to be due to a HOC1 open-reading-frame truncation that alters cell-wall mannan. We introduced the HOC1 open-reading-frame truncation into NCYC 2543, which increased the transformability and improved secretion of some but not all of our tested proteins. We provide our genome-sequenced type strain, the hoc1 tr derivative that we named OPENPichia as well as a synthetic, modular expression vector toolkit under liberal end-user distribution licences as an unencumbered OPENPichia resource for the microbial biotechnology community.
(© 2024. The Author(s).)
Databáze: MEDLINE
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