Expression and regulation of glucoamylase from the yeast Schwanniomyces castellii

Expression of the 146-kilodalton (kDa) extracellular glucoamylase by the budding yeast Schwanniomyces castellii is induced by maltose and starch. By use of antiglucoamylase antisera, we found that this expression was regulated at the level of the mRNA, taking place within 30 min after exposure of yeast cells to the respective sugars. Polyacrylamide gel electrophoresis analysis of the in vitro-translated products of total RNA from maltose-treated cells established that the glucoamylase precursor was approximately 120 kDa in size. Stable glucoamylase transcript was not produced in cells exposed to glucose, 2-deoxyglucose, and heat shock. Cells exposed to these two sugars also degraded intracellular and extracellular glucoamylase. In the presence of sugars such as cellobiose, galactose, lactose, and xylose or in the absence of any carbohydrate, a low-level, constitutive-like expression of this preglucoamylase occurred. The nascent glucoamylase underwent at least two posttranslational modifications, resulting in a 138-kDa cell-associated form and the 146-kDa active form that was found free in the medium. These results suggest that glucoamylase expression is tightly regulated similarly to expression of the enzymes responsible for maltose metabolism in Saccharomyces yeasts.

[1]  V. Seligy,et al.  A temporally expressed gene from Schwanniomyces alluvius and detection of homologous sequences in other yeasts. , 1988, Gene.

[2]  S. Scherer,et al.  Differential glucoamylase expression in Schwanniomyces castellii induced by maltose , 1988 .

[3]  I. S. Pretorius,et al.  Transcriptional control of glucoamylase synthesis in vegetatively growing and sporulating Saccharomyces species , 1986, Molecular and cellular biology.

[4]  J. A. Barnett,et al.  The utilization of starch by yeasts , 1986, Yeast.

[5]  A. Nasim,et al.  Cloning and expression of a Saccharomyces diastaticus glucoamylase gene in Saccharomyces cerevisiae and Schizosaccharomyces pombe , 1986, Journal of bacteriology.

[6]  Kunihiro Matsumoto,et al.  Genetic analysis of the role of cAMP in yeast , 1985, Yeast.

[7]  G. Calleja,et al.  Secretion and export of amylolytic activities in Schwanniomyces alluvius , 1985 .

[8]  G. Stewart,et al.  ISOLATION AND CHARACTERISATION OF THE AMYLOTIC SYSTEM OF SCHWANNIOMYCES CASTELLII , 1984 .

[9]  B. Simões‐Mendes.  Purification and characterization of the extracellular amylases of the yeast Schwanniomyces alluvius , 1984 .

[10]  V. Seligy,et al.  The induction of alpha-amylase by starch in Aspergillus oryzae: evidence for controlled mRNA expression. , 1984, Canadian journal of biochemistry and cell biology = Revue canadienne de biochimie et biologie cellulaire.

[11]  A. Nasim,et al.  Thermosensitive export of amylases in the yeast Schwanniomyces alluvius , 1984 .

[12]  M. Desrochers,et al.  Extracellular proteins secreted by the basidiomycete Schizophyllum commune in response to carbon source , 1984, Journal of bacteriology.

[13]  I. Yamashita,et al.  Molecular Cloning of a Glucoamylase-producing Gene in the Yeast Saccharomyces , 1983 .

[14]  W. M. Ingledew,et al.  Isolation and characterization of Schwanniomyces alluvius amylolytic enzymes , 1982, Applied and environmental microbiology.

[15]  N. Uden,et al.  Selective isolation of depressed mutants of an α‐amylase yeast by the use of 2‐deoxyglucose , 1980 .

[16]  W. Rutter,et al.  Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. , 1979, Biochemistry.

[17]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[18]  H. Dellweg,et al.  Cyclic AMP and Catabolite Repression in Yeasts , 1974 .

[19]  C. Byus,et al.  Ribonucleic acid isolated by cesium chloride centrifugation. , 1974, Biochemistry.

[20]  R. van Wijk,et al.  Cyclic 3′, 5′‐amp in Saccharomyces carlsbergensis under various conditions of catabolite repression , 1971, FEBS letters.

[21]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[22]  S. Baird,et al.  Glucanase gene diversity in prokaryotic and eukaryotic organisms. , 1985, Bio Systems.

[23]  B. Svensson,et al.  Characterization of two forms of glucoamylase from aspergillus niger , 1982 .

[24]  G. I. Bourd,et al.  [Glucose catabolite repression of glucoamylase biosynthesis by the yeast Endomycopsis fibuligera]. , 1978, Prikladnaia biokhimiia i mikrobiologiia.