High levels of expression of heterologous proteins (from 5 to 15% of total cell proteins) in the budding yeast Saccharomyces cerevisiae have been obtained previously by the use of the inducible strong hybrid promoter UASGAL/CYC1, in batch as well in continuous cultures. However, in order to maximize the yield of heterologous proteins, a computer controlled fed‐batch fermentation is essential. For this reason we have developed a fed‐batch system based on a semiconductor gas detector that measures ethanol in the outflow gases. The optimal conditions are described for very high production (up to 1550 mg/liter), with both high productivity (up to 100–120 mg/liter/h) and high yield (up to 15 mg of protein/g of dry biomass), of heterologous protein driven by the UASGAL/CYC1 promoter in a completely computer controlled fed‐batch fermentation of budding yeast. However, high production was dependent upon the addition of a large amount of galactose. The process was improved by developing a new, more easily inducible, vector system obtained by subcloning the GAL4 gene

Alberghina, L., Porro, D., Martegani, E., Ranzi, B. (1991). Efficient production of recombinant DNA proteins in Saccharomyces cerevisiae by controlled high‐cell‐density fermentation. BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, 14(1), 82-92 [10.1111/j.1470-8744.1991.tb00168.x].

Efficient production of recombinant DNA proteins in Saccharomyces cerevisiae by controlled high‐cell‐density fermentation

ALBERGHINA, LILIA;PORRO, DANILO;MARTEGANI, ENZO;
1991

Abstract

High levels of expression of heterologous proteins (from 5 to 15% of total cell proteins) in the budding yeast Saccharomyces cerevisiae have been obtained previously by the use of the inducible strong hybrid promoter UASGAL/CYC1, in batch as well in continuous cultures. However, in order to maximize the yield of heterologous proteins, a computer controlled fed‐batch fermentation is essential. For this reason we have developed a fed‐batch system based on a semiconductor gas detector that measures ethanol in the outflow gases. The optimal conditions are described for very high production (up to 1550 mg/liter), with both high productivity (up to 100–120 mg/liter/h) and high yield (up to 15 mg of protein/g of dry biomass), of heterologous protein driven by the UASGAL/CYC1 promoter in a completely computer controlled fed‐batch fermentation of budding yeast. However, high production was dependent upon the addition of a large amount of galactose. The process was improved by developing a new, more easily inducible, vector system obtained by subcloning the GAL4 gene
Articolo in rivista - Articolo scientifico
SACCHAROMYCES-CEREVISIAE, yeast
English
82
92
11
Alberghina, L., Porro, D., Martegani, E., Ranzi, B. (1991). Efficient production of recombinant DNA proteins in Saccharomyces cerevisiae by controlled high‐cell‐density fermentation. BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, 14(1), 82-92 [10.1111/j.1470-8744.1991.tb00168.x].
Alberghina, L; Porro, D; Martegani, E; Ranzi, B
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/33286
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