On-line high performance liquid chromatography measurements of extracellular metabolites in an aerobic batch yeast (<Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>) culture |
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Authors: | Niina Tohmola Jouni Ahtinen Juha-Pekka Pitkänen Ville Parviainen Sakari Joenväärä Mika Hautamäki Peter Lindroos Jarno Mäkinen Risto Renkonen |
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Institution: | (1) Department of Chemical Engineering, Thammasat University, Pathumthani, 12120, Thailand;(2) Department of Electrical Engineering, Chiang Mai University, Chiang Mai, Thailand;(3) Department of Chemical Engineering, University of Waterloo, Waterloo, Canada |
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Abstract: | We constructed a bioprocess environment enabling automatic sampling from a bioreactor combined with a compact on-line high
performance liquid chromatography (HPLC) unit. This setup allowed us to measure extracellular glucose, ethanol, glycerol,
and acetate concentrations automatically at 5 min intervals during the cultivation. This environment also provides mechanical
measurement of the optical density (OD) of cells and enables us to collect and store (−35°C) samples for further off-line
analyses. Among the available devices, the performance of the sampling-analysis unit is by far the best with regard to speed
and number of analytes. Both the sampling and analysis phases are easily controlled by software; thus, providing a unique
environment to perform various bioprocess activity tasks, whether they would be cell line screening or optimisation of conditions
for growth and productivity. Complex research set-ups can be created and continuous automated measurements empower long-term
cultivations with a time series. We provide evidence for the applicability of this environment by performing three comparable
batch cultivations with Saccharomyces cerevisiae yeast and show that both the on-line sampling and analysis modes produce reliable data for further use in the monitoring
and controlling of bioprocesses. On-line data provided new insight into the dynamics of the diauxic shift during aerobic glucose
batch cultivation. When cell growth and carbon dioxide production ceased for the first time during the diauxic shift, acetate
accumulation and consumption of the remaining glucose below 0.15 g/L continued to occur for 1 h. At the same time, glycerol
and ethanol began to be consumed. Samples were also collected during cultivation for later analysis of intracellular metabolites
and to collect more valuable information about metabolism. |
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