Production of 1-Kestose in Transgenic Yeast Expressing a Fructosyltransferase from Aspergillus foetidus

AUTOR(ES)

Rehm, Jochen

FONTE

American Society for Microbiology

RESUMO

Sucrose-inducible secretory sucrose:sucrose 1-fructosyltransferase (1-SST) from Aspergillus foetidus has been purified and subjected to N-terminal amino acid sequence determination. The enzyme is extensively glycosylated, and the active form is probably represented by a dimer of identical subunits with an apparent molecular mass of 180 kDa as judged from mobility in seminative acrylamide gels. The enzyme catalyzes fructosyl transfer from sucrose to sucrose producing glucose and 1-kestose. Oligosaccharides with a higher degree of polymerization are not obtained with sucrose as the substrate. The cDNA encoding the A. foetidus 1-SST has been cloned and sequenced. Sequence homology was found to be highest to levanases, but no hydrolytic activity was observed when levan was incubated with the enzyme. Expression of the cloned gene in an invertase-deficient mutant of Saccharomyces cerevisiae resulted in 1-kestose production, with 6-kestose and neokestose being side products of the reaction. Products were well distinguishable from those formed by yeast transformants expressing a cytosolic invertase.

Documentos Relacionados

• Purine Ribonucleosidase g from Aspergillus foetidus
• SYNTHESIS AND CHARACTERIZATION OF FRUCTOSYLTRANSFERASE FROM Aspergillus oryzae IPT-301 FOR HIGH FRUCTOOLIGOSACCHARIDES PRODUCTION
• Production of Se-methylselenocysteine in transgenic plants expressing selenocysteine methyltransferase
• Assimilation of Cellooligosaccharides by a Cell Surface-Engineered Yeast Expressing β-Glucosidase and Carboxymethylcellulase from Aspergillus aculeatus
• An Unstable Strain of Aspergillus foetidus Segregating Proline Auxotrophs