Cloning and expression of a gene for an alpha-glucosidase from Saccharomycopsis fibuligera homologous to family GH31 of yeast glucoamylases.
Cloning of cDNA encoding an alpha-glucosidase from the dimorphous yeast Saccharomycopsis fibuligera and characterization of the gene product were performed. The cDNA of the putative alpha-glucosidase gene consists of 2,886 bp, which includes an open reading frame encoding a 19 amino acid signal peptide at the N-terminal end and a 944 amino acid mature protein with a predicted molecular mass of 105.4 kDa and pI value of 4.52. The deduced amino acid sequence shows a high degree of identity (70%) with two yeast glucoamylases, namely, the extracellular glucoamylase Gam from Schwanniomyces occidentalis and the cell surface glucoamylase Gca from Candida albicans. The recombinant product, synthesized in Saccharomyces cerevisiae, is localized on the cell surface and hydrolyses maltooligosaccharides exclusively without the ability to digest soluble starch, which is consistent with the specificity characteristic of alpha-glucosidase, EC. 3.2.1.20.
Hostinová
Eva
E
Institute of Molecular Biology, Slovak Academy of Sciences, Dúbravská cesta 21, 84551 Bratislava, Slovakia. eva.hostinova@savba.sk
Solovicová
Adriana
A
Gasperík
Juraj
J
D000595:Amino Acid Sequence; D002176:Candida albicans / Q000235:genetics; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics; D008565:Membrane Proteins / Q000032:analysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism; D016366:Open Reading Frames; D021382:Protein Sorting Signals / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D012442:Saccharomycopsis / Q000201:enzymology* / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013213:Starch / Q000378:metabolism; D013379:Substrate Specificity; D000520:alpha-Glucosidases / Q000235:genetics* / Q000378:metabolism*
D004271:DNA, Fungal; D005656:Fungal Proteins; D008565:Membrane Proteins; D009844:Oligosaccharides; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; C021705:maltooligosaccharides; D013213:Starch; D000520:alpha-Glucosidases; D005087:Glucan 1,4-alpha-Glucosidase