Mutant strains of Pichia pastoris with enhanced secretion of recombinant proteins

Authors

Sasha Larsen, University of the Pacific
Jun Weaver, University of the Pacific
Katherine de Sa Campos, University of the Pacific
Rhobe Bulahan, University of the Pacific
Jackson Nguyen, University of the Pacific
Heather Grove, University of the Pacific
Amy Huang, University of the Pacific
Lauren Low, University of the Pacific
Namphuong Tran, University of the Pacific
Seth Gomez, University of the Pacific
Jennifer Yau, University of the Pacific
Thomas Ilustrisimo, University of the Pacific
Jessica Kawilarang, University of the Pacific
Jonathan Lau, University of the Pacific
Maivi Tranphung, University of the Pacific
Irene Chen, University of the Pacific
Christina Tran, University of the Pacific
Marcia Fox, University of the Pacific
Joan Lin-Cereghino, University of the PacificFollow
Geoff P. Lin-Cereghino, University of the PacificFollow

Document Type

Article

Publication Title

Biotechnology Letters

Department

Biological Sciences

ISSN

0141-5492

Volume

35

Issue

11

DOI

10.1007/s10529-013-1290-7

First Page

1925

Last Page

1935

Publication Date

11-1-2013

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.

Recommended Citation

Larsen, S., Weaver, J., de Sa Campos, K., Bulahan, R., Nguyen, J., Grove, H., Huang, A., Low, L., Tran, N., Gomez, S., Yau, J., Ilustrisimo, T., Kawilarang, J., Lau, J., Tranphung, M., Chen, I., Tran, C., Fox, M., Lin-Cereghino, J., & Lin-Cereghino, G. P. (2013). Mutant strains of Pichia pastoris with enhanced secretion of recombinant proteins. Biotechnology Letters, 35(11), 1925–1935. DOI: 10.1007/s10529-013-1290-7
https://scholarlycommons.pacific.edu/cop-facarticles/719