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 h...

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Veröffentlicht in:Journal of biotechnology 1990-10, Vol.16 (1), p.87-96
Hauptverfasser: Kobatake, Eiry, Ikariyama, Yoshihito, Aizawa, Masuo, Miwa, Kyoko, Kato, Seishi
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Sprache:eng
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Zusammenfassung: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 65000. 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 K m was 1.3 × 10 −5 M. A simple immuno-enzymometric assay was developed for anti-BSA antibody using the fusion protein.
ISSN:0168-1656
1873-4863
DOI:10.1016/0168-1656(90)90067-L