Production of fungal alpha-amylase by Saccharomyces kluyveri in glucose-limited cultivations

Journal of Biotechnology
Kasper MøllerLisbeth Olsson

Abstract

Heterologous protein production by the yeast Saccharomyces kluyveri was investigated under aerobic glucose-limited conditions. Alpha-amylase from Aspergillus oryzae was used as model protein and the gene was expressed from a S. cerevisiae 2 micro plasmid. For comparison, strains of both S. kluyveri and S. cerevisiae were transformed with the same plasmid, which led to secretion of active alpha-amylase in both cases. The S. cerevisiae 2 micro plasmid was found to be stable in S. kluyveri as evaluated by a constant alpha-amylase productivity in a continuous cultivation for more than 40 generations. S. kluyveri and S. cerevisiae secreted alpha-amylase with similar yields during continuous cultivations at dilution rates of 0.1 and 0.2 h(-1) (4.8-5.7 mg (g dry weight)(-1)). At a dilution rate of 0.3 h(-1) the metabolism of S. kluyveri was fully respiratory, whereas S. cerevisiae produced significant amounts of ethanol. A fed-batch cultivation was carried out with S. kluyveri where the biomass concentration reached 85 g l(-1) and the alpha-amylase concentration reached 320 mg l(-1). Even though S. kluyveri could be grown to high cell density, it was also observed that it has a high maintenance coefficient, which resulted in low bioma...Continue Reading

References

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Apr 19, 2003·FEMS Yeast Research·Kasper MøllerLisbeth Olsson
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Citations

Jul 14, 2009·Genome Research·Célia PayenCécile Neuvéglise
Jun 16, 2009·Genome Research·UNKNOWN Génolevures ConsortiumEmmanuel Talla
Feb 28, 2019·Recent Patents on Biotechnology·Satya Eswari Jujjavarapu, Swasti Dhagat
Dec 3, 2011·Nucleosides, Nucleotides & Nucleic Acids·A RasmussenJ Piškur
Oct 12, 2018·Preparative Biochemistry & Biotechnology·Renpeng DuZhijiang Zhou
Feb 29, 2016·Food Science and Biotechnology·Belgacem NailiRadhouane Kammoun
May 15, 2021·Frontiers in Cellular and Infection Microbiology·Dibyajit LahiriRina Rani Ray

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