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A segment of DNA encoding Saccharomyces cerevisiae inorganic pyrophosphatase (ppa gene) was amplified by the polymerase chain reaction. The pSCH1 and pSCB6 plasmids containing the ppa gene were obtained. Transformation of the E. coli BL21 strain with the resulting recombinant plasmids and selection of clones having extremely high expression of inorganic pyrophosphatase (PPase) were carried out. Superproduction...
An E. coli inorganic pyrophosphatase overproducer and a method for a large-scale production of the homogeneous enzyme are described. The inorganic pyrophosphatase was crystallized in the form containing one subunit of a homohexameric molecule per asymmetric unit: space group R32, a = 110.4 Å, c = 76.8 Å. The electron density map to 2.5 Å resolution phased with Eu- and Hg-derivatives (figure of merit,...
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