The effect of α-mating factor secretion signal mutations on recombinant protein expression in Pichia pastoris.

Authors

Geoff P. Lin-Cereghino, University of the PacificFollow
Carolyn Stark, University of the Pacific
Daniel Kim, University of the Pacific
Jennifer Chang, University of the Pacific
Nadia Shaheen, University of the Pacific
Hansel Poerwanto, University of the Pacific
Kimiko Agari, University of the Pacific
Pachai Moua, University of the Pacific
Lauren Low, University of the Pacific
Namphuong Tran, University of the Pacific
Amy Huang, University of the Pacific
Maria Nattestad, University of the Pacific
Kristin Oshiro, University of the Pacific
John Chang, University of the Pacific
Archana Chavan, University of the Pacific
Jerry Tsai, University of the Pacific
Joan Lin-Cereghino, University of the PacificFollow

Document Type

Article

Publication Title

Gene

Department

Biological Sciences

ISSN

0378-1119

Volume

519

Issue

2

DOI

10.1016/j.gene.2013.01.062

First Page

311

Last Page

317

Publication Date

5-1-2013

Abstract

The methylotrophic yeast, Pichia pastoris, has been genetically engineered to produce many heterologous proteins for industrial and research purposes. In order to secrete proteins for easier purification from the extracellular medium, the coding sequence of recombinant proteins is initially fused to the Saccharomyces cerevisiae α-mating factor secretion signal leader. Extensive site-directed mutagenesis of the prepro-region of the α-mating factor secretion signal sequence was performed in order to determine the effects of various deletions and substitutions on expression. Though some mutations clearly dampened protein expression, deletion of amino acids 57–70, corresponding to the predicted 3rd alpha helix of α-mating factor secretion signal, increased secretion of reporter proteins horseradish peroxidase and lipase at least 50% in small-scale cultures. These findings raise the possibility that the secretory efficiency of the leader can be further enhanced in the future.

Recommended Citation

Lin-Cereghino, G. P., Stark, C., Kim, D., Chang, J., Shaheen, N., Poerwanto, H., Agari, K., Moua, P., Low, L., Tran, N., Huang, A., Nattestad, M., Oshiro, K., Chang, J., Chavan, A., Tsai, J., & Lin-Cereghino, J. (2013). The effect of α-mating factor secretion signal mutations on recombinant protein expression in Pichia pastoris.. Gene, 519(2), 311–317. DOI: 10.1016/j.gene.2013.01.062
https://scholarlycommons.pacific.edu/cop-facarticles/720