Enhanced production of recombinant secretory proteins in Pichia pastoris by optimizing Kex2 P1' site.
Pichiapastoris is one of the most widely used expression systems for the production of recombinant secretory proteins. Its universal application is, however, somewhat hampered by its unpredictable yields for different heterologous proteins, which is now believed to be caused in part by their varied efficiencies to traffic through the host secretion machinery. The yeast endoprotease Kex2 removes the signal peptides from pre-proteins and releases the mature form of secreted proteins, thus, plays a pivotal role in the yeast secretory pathways. In this study, we found that the yields of many recombinant proteins were greatly influenced by Kex2 P1' site residues and the optimized P1's amino acid residue could largely determine the final amount of secretory proteins synthesized and secreted. A further improvement of secretory yield was achieved by genomic integration of additional Kex2 copies, which again highlighted the importance of Kex2 cleavage to the production of recombinant secretory proteins in Pichia yeast.
The Key Laboratory of Regenerative Biology and The Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
Song
S
Yang
Ye
Y
Kuang
Hongbo
H
Li
Yuehong
Y
Liu
Xiaoyan
X
Hui
Peng
P
Li
Zhiwu
Z
Jiang
Yulai
Y
Zhou
Yu
Y
Wang
Aimin
A
Xu
Shiwu
S
Li
Pentao
P
Liu
Donghai
D
Wu
D005656:Fungal Proteins / Q000378:metabolism*; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D008163:Luminescent Measurements; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D043484:Proprotein Convertases / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism*; D014170:Transformation, Genetic
D005656:Fungal Proteins; D011994:Recombinant Proteins; D043484:Proprotein Convertases