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Selected Works of Geoffrey "Geoff" Lin-Cereghino
Article
The effect of α-mating factor secretion signal mutations on recombinant protein expression in Pichia pastoris.
Gene
  • Geoff P. Lin-Cereghino, University of the Pacific
  • 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 Pacific
Link
Document Type
Article
Department
Biological Sciences
DOI
10.1016/j.gene.2013.01.062
Publication Date
5-1-2013
Disciplines
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.

Citation Information
Geoff P. Lin-Cereghino, Carolyn Stark, Daniel Kim, Jennifer Chang, et al.. "The effect of α-mating factor secretion signal mutations on recombinant protein expression in Pichia pastoris." Gene Vol. 519 Iss. 2 (2013) p. 311 - 317 ISSN: 0378-1119
Available at: http://works.bepress.com/geoff-lin-cereghino/37/