cDNA cloning and characterization of an alpha-glucosidase gene from potato (Solanum tuberosum L.)

• Published in: The Plant journal : for cell and molecular biology Vol. 13; no. 3; pp. 419 - 425
• Main Authors: Taylor, M.A, George, L.A, Ross, H.A, Davies, H.V
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science, Ltd 01.02.1998; Blackwell Science
• Subjects: alpha-glucosidase; alpha-Glucosidases - chemistry; alpha-Glucosidases - genetics; alpha-Glucosidases - metabolism; Amino Acid Sequence; amino acid sequences; Arabidopsis - enzymology; Arabidopsis - genetics; Arabidopsis thaliana; Biological and medical sciences; carbohydrate metabolism; Cloning, Molecular; complementary DNA; DNA, Complementary - genetics; DNA, Plant - genetics; enzyme activity; Enzymes; Fundamental and applied biological sciences. Psychology; genbank/aj0001374; Gene Expression; genes; Genes. Genome; Genetic Complementation Test; hexosyltransferases; histochemistry; Humans; leaves; maltose; Maltose - metabolism; Metabolism; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Molecular Weight; Mutation; nucleotide sequences; plant extracts; Plant physiology and development; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - growth & development; Saccharomyces cerevisiae - metabolism; Solanum tuberosum; Solanum tuberosum - enzymology; Solanum tuberosum - genetics; tubers; Enzyme; Solanum tuberosum; Maltose; Gene expression; Metabolism; Fungi; Enzymatic activity; Complementary DNA; α-Glucosidase; Dicotyledones; Glycosidases; Maltodextrin; Angiospermae; Hydrolases; Isolation; Ascomycetes; Spermatophyta; Complementation; Solanaceae; Mutation; O-Glycosidases; Saccharomyces cerevisiae; Aminoacid sequence; Thallophyta
• ISSN: 0960-7412; 1365-313X
• DOI: 10.1046/j.1365-313X.1998.00051.x

Using an Arabidopsis thaliana expressed sequence tag with sequence similarity to human lysosomal alpha-glucosidase as a probe, a potato cDNA was isolated. The cDNA encodes a polypeptide with an Mr value of 105,400 and the most significant matches of the deduced amino acid sequence are with members of family 31 of glucosyl transferase. The potato cDNA was expressed in a strain of Saccharomyces cerevisiae that is deficient in maltase activity and unable to grow using maltose as a carbon source (ABYSMAL81). Expression of the potato cDNA in the mutant yeast strain restores its ability to use maltose as a carbon source for growth. Additionally, alpha-glucosidase activity could be measured in extracts of the yeast cells following complementation. A range of maltodextrins were substrates for this activity. The steady-state expression level of the potato alpha-glucosidase gene was low in most tissues examined, the highest levels occurring in sprouting tubers and source leaves.