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

2013; Elsevier BV; Volume: 519; Issue: 2 Linguagem: Inglês

10.1016/j.gene.2013.01.062

1879-0038

Geoff P. Lin‐Cereghino, Carolyn M. Stark, Daniel H. Kim, Jennifer Chang, Nadia Shaheen, Hansel Poerwanto, Kimiko Agari, Pachai Moua, Lauren K. Low, NamPhuong Tran, Amy Huang, Maria Nattestad, Kristin T. Oshiro, John William Chang, Archana G. Chavan, Jerry Tsai, Joan Lin‐Cereghino,

Endoplasmic Reticulum Stress and Disease

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.