Analysis of the alpha-amylase gene of Schwanniomyces occidentalis and the secretion of its gene product in transformants of different yeast genera.
We have cloned and characterized the alpha-amylase gene (AMY1) of the yeast Schwanniomyces occidentalis. A cosmid gene library of S. occidentalis DNA was screened in Saccharomyces cerevisiae for alpha-amylase secretion. The positive clone contained a DNA fragment harbouring an open reading frame of 1536 nucleotides coding for a 512-amino-acid polypeptide with a calculated Mr of 56,500. The deduced amino acid sequence reveals significant similarity to the sequence of the Saccharomycopsis fibuligera and Aspergillus oryzae alpha-amylases. The AMY l gene was found to be expressed from its original promoter in S. cerevisiae, Kluyveromyces lactis and Schizo-saccharomyces pombe leading to an active secreted gene product and thus enabling the different yeast transformants to grow on starch as a sole carbon source.
Strasser
A W
AW
Rhein Biotech GmbH, Düsseldorf, Federal Republic of Germany.
Selk
R
R
Dohmen
R J
RJ
Niermann
T
T
Bielefeld
M
M
Seeboth
P
P
Tu
G H
GH
Hollenberg
C P
CP
D000595:Amino Acid Sequence; D001203:Ascomycota / Q000201:enzymology*; D001483:Base Sequence; D003001:Cloning, Molecular; D015870:Gene Expression; D008969:Molecular Sequence Data; D014170:Transformation, Genetic; D015003:Yeasts / Q000201:enzymology*; D000516:alpha-Amylases / Q000032:analysis / Q000235:genetics* / Q000378:metabolism
D000516:alpha-Amylases