Quantitative Physiology of Saccharomyces cerevisiae at Near-Zero Specific Growth Rates

Growth at near-zero specific growth rates is a largely unexplored area of yeast physiology. To investigate the physiology of Saccharomyces cerevisiae under these conditions, the effluent removal pipe of anaerobic, glucose-limited chemostat culture (dilution rate, 0.025 h⁻¹) was fitted with a 0.22-μm...

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Published in:Applied and Environmental Microbiology 2009-09, Vol.75 (17), p.5607-5614
Main Authors: Boender, Léonie G.M, de Hulster, Erik A.F, van Maris, Antonius J.A, Daran-Lapujade, Pascale A.S, Pronk, Jack T
Format: Article
Language:English
Subjects:
Acetic acid
Acetic Acid - metabolism
Anaerobiosis
Biological and medical sciences
Biomass
Cell culture
Cells
Energy Metabolism
Ethanol - metabolism
Fermentation
Fundamental and applied biological sciences. Psychology
Glucose
Glucose - metabolism
Glycerol - metabolism
Lactic Acid - metabolism
Microbiology
Physiology
Saccharomyces cerevisiae
Saccharomyces cerevisiae - growth & development
Saccharomyces cerevisiae - physiology
Studies
Succinic Acid - metabolism
Yeast
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Summary:Growth at near-zero specific growth rates is a largely unexplored area of yeast physiology. To investigate the physiology of Saccharomyces cerevisiae under these conditions, the effluent removal pipe of anaerobic, glucose-limited chemostat culture (dilution rate, 0.025 h⁻¹) was fitted with a 0.22-μm-pore-size polypropylene filter unit. This setup enabled prolonged cultivation with complete cell retention. After 22 days of cultivation, specific growth rates had decreased below 0.001 h⁻¹ (doubling time of >700 h). Over this period, viability of the retentostat cultures decreased to ca. 80%. The viable biomass concentration in the retentostats could be accurately predicted by a maintenance coefficient of 0.50 mmol of glucose g⁻¹ of biomass h⁻¹ calculated from anaerobic, glucose-limited chemostat cultures grown at dilution rates of 0.025 to 0.20 h⁻¹. This indicated that, in contrast to the situation in several prokaryotes, maintenance energy requirements in S. cerevisiae do not substantially change at near-zero specific growth rates. After 22 days of retentostat cultivation, glucose metabolism was predominantly geared toward alcoholic fermentation to meet maintenance energy requirements. The strict correlation between glycerol production and biomass formation observed at higher specific growth rates was not maintained at the near-zero growth rates reached in the retentostat cultures. In addition to glycerol, the organic acids acetate, D-lactate, and succinate were produced at low rates during prolonged retentostat cultivation. This study identifies robustness and by-product formation as key issues in attempts to uncouple growth and product formation in S. cerevisiae.
ISSN:0099-2240
1098-5336
1098-6596
DOI:10.1128/AEM.00429-09