Extracellular production of human hepatitis B virus preS2 antigen as hybrid proteins with Bacillus subtilis alpha-amylases in high-salt-concentration media.
To produce PreS2 antigen of human hepatitis B virus extra-cellularly in Bacillus subtilis, it was fused with the COOH-termini of B. subtilis alpha-amylases of 522 (Amy+), 467 (Amy+) and 443 (Amy-) amino acids. Among them, alpha-amylase-A467, which has 467 amino acids, was a relatively stable carrier when the cells were cultured in Luria-Bertani (LB) medium. The alpha-amylase-A443-PreS2 hybrid protein (Amy-) was quickly degraded. The alpha-amylase-A522-PreS2 hybrid was most efficiently produced when a B. subtilis transformant of a protease-super-deficient mutant was cultured in the presence of 0.5 M sodium sulphate. The production of A522-PreS2 hybrid protein under such conditions reached 5-10 mg/l and was eight, and 200-500 times higher than those obtained by the transformants of an alkaline/neutral protease-deficient mutant of B. subtilis, and a wild-type strain, in LB medium, respectively.
Honda
K
K
Institute of Biological Sciences, University of Tsukuba, Ibaraki, Japan.
Fujieda
H
H
Ogawa
K
K
Imai
M
M
Yamamoto
H
H
Ikeda
T
T
Yamane
K
K
D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D003470:Culture Media; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D000516:alpha-Amylases / Q000096:biosynthesis* / Q000235:genetics
D003470:Culture Media; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C050807:presurface protein 2, hepatitis B surface antigen; D000516:alpha-Amylases