Expression of cloned Saccharomyces diastaticus glucoamylase under natural and inducible promoters.
Any one of three homologous genes - STA1, STA2 and STA3 - encoding glucoamylase isozymes I, II and III respectively, allows the Saccharomyces species to utilize starch as a sole carbon source. We show in this paper that glucoamylase II production can be increased 4-fold over the level produced by STA2 strains, by using a two-step fermentation and a yeast strain transformed with a high-copy-number plasmid carrying the STA2 gene. The accumulation of anomalous STA2 mRNA species, mainly differing at their 5' ends, and saturation of step(s) in the secretory pathway appear to be among the major factors limiting glucoamylase expression in synthetic media.
Vanoni
M
M
Dipartimento di Fisiologia e Biochimica Generali, Università degli Studi di Milano, Italy.
Lotti
M
M
Alberghina
L
L
D003001:Cloning, Molecular; D003470:Culture Media / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005285:Fermentation; D005786:Gene Expression Regulation; D005800:Genes, Fungal; D005087:Glucan 1,4-alpha-Glucosidase / Q000096:biosynthesis / Q000235:genetics*; D015151:Immunoblotting; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics; D005810:Multigene Family; D009710:Nucleotide Mapping; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012331:RNA, Fungal / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012440:Saccharomyces / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D003470:Culture Media; D007527:Isoenzymes; D012331:RNA, Fungal; D012333:RNA, Messenger; D005087:Glucan 1,4-alpha-Glucosidase