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Keywords:

  • Saccharomycopsis fibuligera;
  • Glucoamylase;
  • Gene cloning;
  • Nucleotide sequence

Abstract: The nucleotide sequence of the 2544-bp PstI fragment carrying the glucoamylase gene of Saccharomycopsis fibuligera KZ, designated as GLA1, has been determined. When compared with the nucleotide sequence of the GLU1 gene [1] one nucleotide substitution was found in the 321- bp of the 5′-flanking region; 24 nucleotides were altered within the 1557 bp of the structural gene causing the deduced protein products of both genes to differ in three amino acids in the signal-peptide region and in eight amino acids of the mature protein. Six nucleotide insertions and 27 substitutions were in the 663 bp of the 3′-flanking region. The gene product expressed and secreted in Saccharomyces cerevisiae into the functional enzyme was not homogeneous. In situ detection of the enzyme in a polyacrylamide gel revealed two dominant and three minor bands.