Construction and characterization of a recombinant tripartite enzyme, galactose dehydrogenase/beta-galactosidase/galactokinase.
The in-frame gene fusion between 3 enzymes, galactose dehydrogenase, beta-galactosidase and galactokinase, is described. The purified artificial tripartite enzyme displayed all three enzymic activities. Two major forms of the hybrid protein were found, consisting of 4 and 8 subunits respectively, but other forms could also be identified. Each subunit was made up of one monomer each of galactose dehydrogenase, beta-galactosidase and galactokinase. Proximity effects exhibited by the hybrid enzyme could be demonstrated using [14C]galactose as a reporter molecule.
Ljungcrantz
P
P
Pure and Appied Biochemistry, Chemical Center, Lund, Sweden.
Bülow
L
L
Mosbach
K
K
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli; D005686:Galactokinase / Q000235:genetics* / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D005691:Galactose Dehydrogenases / Q000235:genetics* / Q000378:metabolism; D005798:Genes, Bacterial; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011489:Protein Denaturation; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D013329:Structure-Activity Relationship; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D005691:Galactose Dehydrogenases; C047959:galactose dehydrogenase; D005686:Galactokinase; D001616:beta-Galactosidase; D005690:Galactose