title: Isolation and characterization of glucoamylase from a wastewater treatment yeast Hansenula fabianii J640, and construction of expression vector.
abstract: Hansenula fabianii J640 highly expresses an extracellular glucoamylase (GA). Here, we purified the GA and showed that it has pH and temperature optima of 5.0 and 50 °C, respectively, stable at temperatures up to 50 °C, and is inhibited by Ag(2+), Hg(2+), and Cu(2+). The gene was found in an expression library with anti-GA antibodies. A cDNA was found to encode 491 amino acids, including a putative signal peptide of 21 amino acids. Because of the gene's high expression, we used its promoter and terminator regions to improve a previously developed H. fabianii J640 expression system.
authors.affiliation: Graduate School of Biosphere Science, Hiroshima University, 1-4-4, Kagamiyama, Higashihiroshima, Hiroshima 739-8527, Japan.
authors.forename: Miyoshi
authors.initials: M
authors.lastname: Kato
authors.forename: Teruhito
authors.initials: T
authors.lastname: Kitajima
authors.forename: Haruyuki
authors.initials: H
authors.lastname: Iefuji
mesh_terms: D003001:Cloning, Molecular; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D005087:Glucan 1,4-alpha-Glucosidase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D012722:Sewage / Q000382:microbiology*; D015003:Yeasts / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D005656:Fungal Proteins; D012722:Sewage; D005087:Glucan 1,4-alpha-Glucosidase