Expression of spinach plastocyanin in E. coli

FEBS Letters

Olausson, Torbjörn¹  Nordling, Margareta¹  Lundberg, Lennart G.¹ 

[1] Department of Biochemistry and Biophysics, Chalmers University of Technology and University of Göteborg, S-4 J 2 96 Göteborg, Sweden

关键词: Plastocyanin;    Blue-copper;    Expression;    E. coli;    Photosynthesis;    IPTG;    isopropyl-β-D-thiogalactopyranoside;    RBS;    ribosome binding site;    SDS-PAGE;    sodium dodecyl sulphate-poly-acrylamide gel electrophoresis;    EPR;    electronic paramagnetic resonance;    CD;    circular dichroism;   

DOI  :  10.1016/0014-5793(90)80517-M

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

An expression vector designed for overexpression of plastocyanin in the periplasmic space of E. coli has been developed. The vector contains the signal peptide sequence of Pseudomonas aeruginosa azurin and the mature sequence of spinach plastocyanin. The precursor is efficiently translocated to the periplasmic space and correctly processed to mature plastocyanin. No detectable amount of plastocyanin was present in the cytoplasmic or in the membrane fraction. A large scale preparation of the recombinant plastocyanin in a 20 litre fennentor yielded approximately 30 mg of pure plastocyanin. The recombinant protein obtained from E. coli shows CD, EPR and optical properties identical to plastocyanin isolated from spinach.

【 授权许可】

Unknown