Comparative properties of amplified external and internal invertase from the yeast SUC2 gene.
Saccharomyces cerevisiae external and internal invertases have been amplified by introducing the normal and modified SUC2 genes into yeast multicopy plasmids, which were then used to transform a yeast strain resistant to repression by glucose. Amino acid compositional analysis of these enzymes, in addition to end group sequencing, confirmed the DNA sequence data of Taussig and Carlson (Taussig, R., and Carlson, M. (1983) Nucleic Acids Res. 11, 1943-1954), indicating that both enzymes were encoded in the same gene. Comparison of the properties of carbohydrate-containing external invertase and its nonglycosylated internal form revealed that although the carbohydrate did not appear to influence the conformation of the peptide backbone, as determined by circular dichroism analyses, its presence considerably enhanced the ability of guanidine HCl-denatured external invertase to be renatured relative to internal invertase. The Mr of the internal enzymes was found to be greatly dependent on pH with the enzyme being a monomer at pH 9.4, a dimer at pH 8.3, and an apparent octamer at pH 4.9.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001483:Base Sequence; D050260:Carbohydrate Metabolism; D002845:Chromatography; D004274:DNA, Recombinant; D005784:Gene Amplification; D005800:Genes, Fungal; D005944:Glucosamine / Q000032:analysis; D006026:Glycoside Hydrolases / Q000235:genetics* / Q000302:isolation & purification; D019791:Guanidine; D006146:Guanidines; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances; D008970:Molecular Weight; D009154:Mutation; D010957:Plasmids; D011487:Protein Conformation; D011489:Protein Denaturation; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D043324:beta-Fructofuranosidase
D000596:Amino Acids; D004274:DNA, Recombinant; D006146:Guanidines; D046911:Macromolecular Substances; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D019791:Guanidine; D005944:Glucosamine