Improved recombinant expression and purification of functional plant Rubisco

FEBS Letters

article

Robert H. Wilson¹  Gabriel Thieulin-Pardo¹  Franz-Ulrich Hartl²  Manajit Hayer-Hartl¹ 

[1] Chaperonin-assisted Protein Folding Group, Max Planck Institute of Biochemistry;Cellular Biochemistry Group, Max Planck Institute of Biochemistry

关键词: assembly;    folding;    molecular chaperones;    protein expression;    Rubisco;    Rubisco activase;    ;   

DOI  :  10.1002/1873-3468.13352

来源: John Wiley & Sons Ltd.

【 摘 要 】

Improving the performance of the key photosynthetic enzyme Ribulose-1,5- bisphosphate carboxylase/oxygenase (Rubisco) by protein engineering is a critical strategy for increasing crop yields. The extensive chaperone requirement of plant Rubisco for folding and assembly has long been an impediment to this goal. Production of plant Rubisco in Escherichia coli requires the coexpression of the chloroplast chaperonin and four assembly factors. Here, we demonstrate that simultaneous expression of Rubisco and chaperones from a T7 promotor produces high levels of functional enzyme. Expressing the small subunit of Rubisco with a C-terminal hexahistidine-tag further improved assembly, resulting in a ~ 12-fold higher yield than the previously published procedure. The expression system described here provides a platform for the efficient production and engineering of plant Rubisco.

【 授权许可】

Unknown