Expression in periplasmic space of Shewanella oneidensis.
A Shewanella expression system has been used for an overproduction of c-type multiheme proteins. The proteins were exported to the periplasmic space for the maturation. Since the periplasmic expression system is attractive, especially for protease-sensitive proteins, an expression vector containing a signal peptide was constructed for expressions in the periplasmic space of Shewanella oneidensis. To evaluate the system, two eukaryotic proteins which originally do not have signal sequences and are difficult to express in Escherichia coli, were selected. The first is human cytochrome c. Properties of the recombinant cytochrome c were identical to those previously reported, indicating the protein is intact. The other was potato calcium-dependent protein kinase. The protein was expressed in periplasmic space. These results indicated that the system is generally applicable for any protein expression including c-type cytochromes, protease-sensitive proteins and those with multi-disulfide bonds because of transportation to the periplasmic space.
Takayama
Yuki
Y
Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita 565-0871, Japan.
Akutsu
Hideo
H
D045304:Cytochromes c / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular; D019897:Periplasm / Q000378:metabolism*; D011494:Protein Kinases / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis; D020592:Shewanella / Q000378:metabolism*; D011198:Solanum tuberosum / Q000201:enzymology; D017871:Calcium-Calmodulin-Dependent Protein Kinases
D045304:Cytochromes c; D011494:Protein Kinases; D011994:Recombinant Proteins; D017871:Calcium-Calmodulin-Dependent Protein Kinases