Novel strategy for protein production using a peptide tag derived from Bacillus thuringiensis Cry4Aa.
Numerous proteins cannot be sufficiently prepared by ordinary recombinant DNA techniques because they are unstable or have deleterious effects on the host cell. One idea to prepare such proteins is to produce them as protein inclusions. Here we developed a novel system to effectively prepare proteins by using peptide tags derived from the insecticidal Cry toxin of a soil bacterium, Bacillus thuringiensis. Fusion with this peptide tag, designated 4AaCter, facilitates the formation of protein inclusions of glutathione S-transferase in Escherichia coli without losing the enzyme activity. Application of 4AaCter to the production of syphilis antigens TpN15, TpN17 and TpN47 from Treponema pallidum yielded excellent results, including a dramatic increase in the production level, simplification of the product purification and high reactivity with syphilis antibody. The use of 4AaCter may provide an innovational strategy for the efficient production of proteins.
Hayakawa
Tohru
T
Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan.
Sato
Shinya
S
Iwamoto
Shigehisa
S
Sudo
Shigeo
S
Sakamoto
Yoshiki
Y
Yamashita
Takaaki
T
Uchida
Motoaki
M
Matsushima
Kenji
K
Kashino
Yohko
Y
Sakai
Hiroshi
H
D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D001413:Bacillus thuringiensis / Q000737:chemistry / Q000378:metabolism*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D004731:Endotoxins / Q000737:chemistry / Q000378:metabolism*; D005982:Glutathione Transferase / Q000096:biosynthesis*; D006460:Hemolysin Proteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data
D000942:Antigens, Bacterial; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D005982:Glutathione Transferase