In Saccharomyces cerevisiae, protein secretion into the growth medium depends on environmental factors.
In the budding yeast Saccharomyces cerevisiae the cell wall, mainly composed of mannoproteins and glucans, constitutes a barrier to protein excretion in the growth medium. In this paper we have studied the effects of different environmental parameters on excretion of Escherichia coli beta-galactosidase obtained by exploiting the glucoamylase II signal sequence. Excretion of the unglycosylated beta-galactosidase was detectable only in cells grown in rich medium, was affected by temperature (36 degrees C > 30 degrees C >> 24 degrees C) and slightly stimulated by reducing agents. On the contrary, glycosylated proteins, such as alpha-galactosidase and glucoamylase II, were excreted to a good extent under all tested conditions of medium composition, growth temperature and pH. These data indicate that optimization of environmental parameters may help the excretion of heterologous proteins, offering advantages for protein purification.
Dipartimento di Fisiologia e Biochimica Generali, Università degli Studi di Milano, Italy.
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Rossini
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Porro
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Brambilla
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Venturini
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Ranzi
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Vanoni
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Alberghina
D003470:Culture Media; D004926:Escherichia coli / Q000201:enzymology; D005087:Glucan 1,4-alpha-Glucosidase / Q000378:metabolism*; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013696:Temperature; D014170:Transformation, Genetic; D000519:alpha-Galactosidase / Q000378:metabolism*; D001616:beta-Galactosidase / Q000378:metabolism*
D003470:Culture Media; D011994:Recombinant Proteins; D000519:alpha-Galactosidase; D001616:beta-Galactosidase; D005087:Glucan 1,4-alpha-Glucosidase