Identification and characterization of glucoamylase from the fungus Thermomyces lanuginosus.
The glucoamylase from the thermophilic fungus Thermomyces lanuginosus has a molecular weight of 66 kDa and was characterized with isoelectric point, pH and temperature optimum of 3.8-4.0, 5.0 and 70 degrees C, respectively. In addition, the activation energy is 60.4 kJ/mol, Km is 3.5 mM and kcat is 25.3 s(-1). The glucoamylase was partially sequenced on the protein level, and the complete glucoamylase gene including its promoter (but excluding its terminator region) was cloned and sequenced. The glucoamylase protein comprises 617 amino acid residues and shows 60% identity with the glucoamylase from the thermophilic fungus Talaromyces emersonii. cDNA encoding Thermomyces lanuginosus glucoamylase was expression cloned into Pichia pastoris, producing approximately 7.4 U/ml. It was concluded that alternative mRNA splicing as it might occur in Aspergillus niger glucoamylase is not responsible for the occurrence of different glucoamylase isoforms in Thermomyces lanuginosus.
Thorsen
Thor S
TS
Department of Microbiology, University of Copenhagen, Sølvgade 83H, DK-1307 Copenhagen K, Denmark.
Johnsen
Anders H
AH
Josefsen
Knud
K
Jensen
Bo
B
D001203:Ascomycota / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004795:Enzyme Stability; D005087:Glucan 1,4-alpha-Glucosidase / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D003904:Mitosporic Fungi / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D013696:Temperature
D005087:Glucan 1,4-alpha-Glucosidase