Efficient expression of haloarchaeal nucleoside diphosphate kinase via strong porin promoter in moderately halophilic bacteria.
Enzymes from extremely halophilic archaea require high concentration of salts for their proper folding and consequently are expressed as an unfolded and inactive form in Escherichia coli. Moderate halophile, which accumulates protein stabilizers, i.e., compatible solutes, is an attractive host cell for the recombinant production of heterologous proteins, since such protein stabilizers may help folding of expressed proteins. Here, we succeeded in efficient expression and purification to homogeneity of recombinant haloarchaeal nucleoside diphosphate kinase (HsNDK) in moderate halophile using newly isolated strong porin promoter.
Nagayoshi
Chizuru
C
Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.
Tokunaga
Hiroko
H
Hayashi
Aya
A
Harazono
Hiroaki
H
Hamasaki
Kyoko
K
Ando
Ayumi
A
Tokunaga
Masao
M
D015153:Blotting, Western; D004926:Escherichia coli / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D019617:Halobacterium salinarum / Q000201:enzymology / Q000235:genetics*; D009701:Nucleoside-Diphosphate Kinase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D018272:Porins / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D018272:Porins; D011994:Recombinant Proteins; D009701:Nucleoside-Diphosphate Kinase