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

被引：128

作者：

Lin-Cereghino, Geoff P. ^{[1
]}

Stark, Carolyn M. ^{[1
]}

Kim, Daniel ^{[1
]}

Chang, Jennifer ^{[1
]}

Shaheen, Nadia ^{[1
]}

Poerwanto, Hansel ^{[1
]}

Agari, Kimiko ^{[1
]}

Moua, Pachai ^{[1
]}

Low, Lauren K. ^{[1
]}

Namphuong Tran ^{[1
]}

Huang, Amy D. ^{[1
]}

Nattestad, Maria ^{[1
]}

Oshiro, Kristin T. ^{[1
]}

Chang, John William ^{[1
]}

Chavan, Archana ^{[2
]}

Tsai, Jerry W. ^{[2
]}

Lin-Cereghino, Joan ^{[1
]}

机构：

[1] Univ Pacific, Dept Biol Sci, Stockton, CA 95211 USA

[2] Univ Pacific, Dept Chem, Stockton, CA 95211 USA

来源：

GENE | 2013年 / 519卷 / 02期

关键词：

Pichia pastoris; alpha-Mating factor secretion signal; Recombinant protein expression; METHANOL UTILIZATION PATHWAY; SACCHAROMYCES-CEREVISIAE; HETEROLOGOUS PROTEINS; DIRECTED EVOLUTION; YEAST; GENE; SEQUENCES; PRECURSOR; STRAINS; PEPTIDE;

D O I：

10.1016/j.gene.2013.01.062

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

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. (C) 2013 Elsevier B.V. All rights reserved.

引用

页码：311 / 317

页数：7

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