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Gene

Title:		The effect of alpha-mating factor secretion signal mutations on recombinant protein expression in Pichia pastoris
Author(s):		Lin-Cereghino GP; Stark CM; Kim D; Chang J; Shaheen N; Poerwanto H; Agari K; Moua P; Low LK; Tran N; Huang AD; Nattestad M; Oshiro KT; Chang JW; Chavan A; Tsai JW; Lin-Cereghino J;
Address:		"Department of Biological Sciences, University of the Pacific Stockton, CA 95211, USA"
Journal Title:		Gene
Year:		2013
Volume:		20130221
Issue:		2
Page Number:		311 - 317
DOI:		10.1016/j.gene.2013.01.062
ISSN/ISBN:		1879-0038 (Electronic) 0378-1119 (Print) 0378-1119 (Linking)
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 alpha-mating factor secretion signal leader. Extensive site-directed mutagenesis of the prepro-region of the alpha-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 alpha-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"
Keywords:		"Amino Acid Sequence Blotting, Western Gene Deletion *Gene Expression Regulation, Fungal Genes, Reporter Horseradish Peroxidase/genetics/metabolism Lipase/genetics/metabolism Mating Factor Molecular Sequence Data Mutagenesis, Site-Directed *Mutation Peptid;"
Notes:		"MedlineLin-Cereghino, Geoff P Stark, Carolyn M Kim, Daniel Chang, Jennifer Shaheen, Nadia Poerwanto, Hansel Agari, Kimiko Moua, Pachai Low, Lauren K Tran, Namphuong Huang, Amy D Nattestad, Maria Oshiro, Kristin T Chang, John William Chavan, Archana Tsai, Jerry W Lin-Cereghino, Joan eng R15 GM065882/GM/NIGMS NIH HHS/ GM65882-03/GM/NIGMS NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Netherlands 2013/03/05 Gene. 2013 May 1; 519(2):311-7. doi: 10.1016/j.gene.2013.01.062. Epub 2013 Feb 21"