title: Expression of active yeast pyruvate decarboxylase in Escherichia coli.
abstract: We have shown by appropriate modification of the translational signals and using the strong T7 RNA polymerase promoter phi 10, that a cloned Saccharomyces cerevisiae pyruvate decarboxylase gene (pdc1) can be expressed in Escherichia coli. This protein, which migrated as a single band on SDS-polyacrylamide gels, was found to have a subunit molecular mass of approximately 62 kDa, similar to that of the enzyme produced by yeast. Polyclonal antibodies raised against purified yeast pyruvate decarboxylase recognized this bacterially produced protein. We found that this recombinant enzyme is active, indicating that the homotetramer encoded by the pdc1 gene is functional.
authors.affiliation: Centre for Molecular Biology and Biotechnology, University of Queensland, Brisbane, Australia.
authors.forename: J M
authors.initials: JM
authors.lastname: Candy
authors.forename: R G
authors.initials: RG
authors.lastname: Duggleby
authors.forename: J S
authors.initials: JS
authors.lastname: Mattick
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004271:DNA, Fungal; D004926:Escherichia coli / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D008969:Molecular Sequence Data; D010957:Plasmids; D011767:Pyruvate Decarboxylase / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D012995:Solubility; D014170:Transformation, Genetic
chemical_list: D004271:DNA, Fungal; D011767:Pyruvate Decarboxylase