Cloning and expression of Hormoconis resinae glucoamylase P cDNA in Saccharomyces cerevisiae.
A cDNA coding for glucoamylase P of Hormoconis resinae was cloned using a synthetic oligonucleotide probe coding for a peptide fragment of the purified enzyme and polyclonal anti-glucoamylase antibodies. Nucleotide-sequence analysis revealed an open reading frame of 1848 base pairs coding for a protein of 616 amino-acid residues. Comparison with other fungal glucoamylase amino-acid sequences showed homologies of 37-48%. The glucoamylase cDNA, when introduced into Saccharomyces cerevisiae under the control of the yeast ADC1 promoter, directed the secretion of active glucoamylase P into the growth medium.
Vainio
A E
AE
Research Laboratories, Alko Ltd, Helsinki, Finland.
Torkkeli
H T
HT
Tuusa
T
T
Aho
S A
SA
Fagerström
B R
BR
Korhola
M P
MP
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D002963:Cladosporium / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D004271:DNA, Fungal; D015723:Gene Library; D005087:Glucan 1,4-alpha-Glucosidase / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D004271:DNA, Fungal; D005087:Glucan 1,4-alpha-Glucosidase