Characterization of the AINV gene and the encoded invertase from the dimorphic yeast Arxula adeninivorans.
The invertase-encoding of AINV gene Arxula adeninivorans was isolated and characterized. The gene includes a coding sequence of 2700 bp encoding a putative 899 amino acid protein of 101.7 kDa. The identity of the gene was confirmed by a high degree of homology of the derived amino acid sequence to that of alpha-glucosidases from different sources. The gene activity is regulated by carbon source. In media supplemented with sucrose induction of the AINV gene and accumulation of the encoded invertase in the medium was observed. In addition the extracellular enzyme level is influenced by the morphological status of the organism, with mycelia secreting the enzyme in titres higher than those observed in budding yeasts. The enzyme characteristics were analysed from isolates of native strains as well as from those of recombinant strains expressing the AINV gene under control of the strong A. adeninivorans -derived TEF1 promoter. For both proteins a molecular mass of 600 kDa was determined, a pH optimum at pH 4.5 and a temperature optimum at 55 degrees C. The preferred substrates for the enzyme included the ss-D-fructofuranosides sucrose, inulin and raffinose. Only a weak enzyme activity was observed for the alpha-D-glucopyranosides maltotriose, maltose and isomaltose. Thus the invertase primarily is a ss-fructosidase and not an alpha-glucosidase as suggested by the homology to such enzymes.
Institut für Pflanzengenetik und Kulturpflanzenforschung, Corrensstr. 3, D-06466 Gatersleben, Germany.
Erik
E
Böer
Thomas
T
Wartmann
Bianka
B
Luther
Renate
R
Manteuffel
Rüdiger
R
Bode
Gerd
G
Gellissen
Gotthard
G
Kunze
D000595:Amino Acid Sequence; D000818:Animals; D000908:Antibodies, Fungal / Q000096:biosynthesis; D001483:Base Sequence; D004271:DNA, Fungal / Q000235:genetics; D015870:Gene Expression; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015183:Restriction Mapping; D004718:Saccharomycetales / Q000201:enzymology* / Q000235:genetics* / Q000276:immunology; D013379:Substrate Specificity; D043324:beta-Fructofuranosidase / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
D000908:Antibodies, Fungal; D004271:DNA, Fungal; D011994:Recombinant Proteins; D043324:beta-Fructofuranosidase