Hyperproduction of a bifunctional hybrid protein, metapyrocatechase-protein A, by gene fusion.
A hybrid protein between metapyrocatechase and Staphylococcal protein A was produced by recombinant DNA techniques. A plasmid carrying the fusion gene that encodes the hybrid protein was constructed and expressed in E. coli. Over 70% of soluble proteins of the cell extracts was estimated to be the hybrid protein. This fusion protein is about 65,000. Both the IgG-binding activity of protein A and the metapyrocatechase activity were found in the hybrid protein. The optimum pH of metapyrocatechase in the fusion protein was at around 6.5 and Km was 1.3 X 10(-5) M. A simple immuno-enzymometric assay was developed for anti-BSA antibody using the fusion protein.
Kobatake
E
E
Department of Bioengineering, Tokyo Institute of Technology, Japan.
Ikariyama
Y
Y
Aizawa
M
M
Miwa
K
K
Kato
S
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D050557:Catechol 2,3-Dioxygenase; D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D049308:Dioxygenases; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D008969:Molecular Sequence Data; D010105:Oxygenases / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D013205:Staphylococcal Protein A / Q000096:biosynthesis / Q000235:genetics*
D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D010105:Oxygenases; D049308:Dioxygenases; D050557:Catechol 2,3-Dioxygenase