A protein disulfide isomerase gene fusion expression system that suppresses the aggregation of heterologous proteins

2000 ◽  
Vol 28 (5) ◽  
pp. A215-A215
Author(s):  
T. Kajino ◽  
C. Ohto ◽  
M. Muramatsu ◽  
S. Obata ◽  
Y. Yamada ◽  
...  
Keyword(s):  
Protein Disulfide Isomerase ◽  
Gene Fusion ◽  
Expression System ◽  
Fusion Expression ◽  
Heterologous Proteins ◽  
Disulfide Isomerase ◽  
Protein Disulfide

scholarly journals A Protein Disulfide Isomerase Gene Fusion Expression System That Increases the Extracellular Productivity ofBacillus brevis

2000 ◽  
Vol 66 (2) ◽  
pp. 638-642 ◽  
Author(s):  
Tsutomu Kajino ◽  
Chikara Ohto ◽  
Masayoshi Muramatsu ◽  
Shusei Obata ◽  
Shigezo Udaka ◽  
...  
Keyword(s):  
Protein Disulfide Isomerase ◽  
Gene Fusion ◽  
Expression System ◽  
Biologically Active ◽  
Fusion Expression ◽  
Fusion Partner ◽  
Heterologous Proteins ◽  
Bacillus Brevis ◽  
Disulfide Isomerase ◽  
Protein Disulfide

ABSTRACT We have developed a versatile Bacillus brevisexpression and secretion system based on the use of fungal protein disulfide isomerase (PDI) as a gene fusion partner. Fusion with PDI increased the extracellular production of heterologous proteins (light chain of immunoglobulin G, 8-fold; geranylgeranyl pyrophosphate synthase, 12-fold). Linkage to PDI prevented the aggregation of the secreted proteins, resulting in high-level accumulation of fusion proteins in soluble and biologically active forms. We also show that the disulfide isomerase activity of PDI in a fusion protein is responsible for the suppression of the aggregation of the protein with intradisulfide, whereas aggregation of the protein without intradisulfide was prevented even when the protein was fused to a mutant PDI whose two active sites were disrupted, suggesting that another PDI function, such as chaperone-like activity, synergistically prevented the aggregation of heterologous proteins in the PDI fusion expression system.

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