title: Purification and characterisation of an Aspergillus niger invertase and its DNA sequence.
abstract: A secreted invertase was purified 23-fold by ultrafiltration, ion-exchange, and gel filtration chromatography from the culture supernatant of 18 h sucrose-grown cultures of Aspergillus niger. The purified enzyme hydrolysed sucrose and raffinose but there was no detectable hydrolysis of inulin, melezitose or PNPG. Invertase activity was optimal at pH 5.5 and 50 degrees C. The molecular mass of reduced invertase was 115 kDa, as determined by SDS gel electrophoresis. The native molecular weight of between 225 kDa and 250 kDa, estimated by electrophoresis under non-denaturing conditions, suggests that the protein is a dimer of identical subunits. The suc1 gene encoding this protein was completely-sequenced. The translated sequence yields a protein of 566 amino acids with a calculated molecular mass of 61 kDa, suggesting that carbohydrates represent about 50% of the mass of the protein.
authors.affiliation: Laboratoire de Génétique, Université de Bordeaux II UA CNRS 542, Talence, France.
authors.forename: L M
authors.initials: LM
authors.lastname: Boddy
authors.forename: T
authors.initials: T
authors.lastname: Bergès
authors.forename: C
authors.initials: C
authors.lastname: Barreau
authors.forename: M H
authors.initials: MH
authors.lastname: Vainstein
authors.forename: M J
authors.initials: MJ
authors.lastname: Dobson
authors.forename: D J
authors.initials: DJ
authors.lastname: Ballance
authors.forename: J F
authors.initials: JF
authors.lastname: Peberdy
mesh_terms: D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D015139:Blotting, Southern; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004271:DNA, Fungal; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D015183:Restriction Mapping; D013379:Substrate Specificity; D013696:Temperature; D043324:beta-Fructofuranosidase
chemical_list: D004271:DNA, Fungal; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase