Premium
Expression of spinach plastocyanin in E. coli
Author(s) -
Nordling Margareta,
Olausson Torbjörn,
Lundberg Lennart G.
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)80517-m
Subject(s) - plastocyanin , periplasmic space , spinach , signal peptide , escherichia coli , chemistry , biochemistry , cytoplasm , spheroplast , biology , recombinant dna , chloroplast , photosystem i , gene
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.