On-line high performance liquid chromatography measurements of extracellular metabolites in an aerobic batch yeast (Saccharomyces cerevisiae) culture

Niina Tohmola (Corresponding Author), Jouni Ahtinen, Juha-Pekka Pitkänen, Ville Parviainen, Sakari Joenväärä, Mika Hautamäki, Peter Lindroos, Jarno Mäkinen, Risto Renkonen

Research output: Contribution to journalArticleScientificpeer-review

12 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)264-272
Number of pages9
JournalBiotechnology and Bioprocess Engineering
Volume16
Issue number2
DOIs
Publication statusPublished - 2011
MoE publication typeA1 Journal article-refereed

Fingerprint

High performance liquid chromatography
Metabolites
Yeast
Saccharomyces cerevisiae
Yeasts
High Pressure Liquid Chromatography
Sampling
Glucose
Glycerol
Acetates
Ethanol
Bioreactors
Growth
Carbon Dioxide
Density (optical)
Cell growth
Software
Metabolism
Cell Count
Time series

Keywords

  • diauxic shift
  • high performance liquid chromatography
  • HPLC
  • metabolite
  • on-line
  • Saccharomyces cerevisiae

Cite this

Tohmola, Niina ; Ahtinen, Jouni ; Pitkänen, Juha-Pekka ; Parviainen, Ville ; Joenväärä, Sakari ; Hautamäki, Mika ; Lindroos, Peter ; Mäkinen, Jarno ; Renkonen, Risto. / On-line high performance liquid chromatography measurements of extracellular metabolites in an aerobic batch yeast (Saccharomyces cerevisiae) culture. In: Biotechnology and Bioprocess Engineering. 2011 ; Vol. 16, No. 2. pp. 264-272.
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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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On-line high performance liquid chromatography measurements of extracellular metabolites in an aerobic batch yeast (Saccharomyces cerevisiae) culture. / Tohmola, Niina (Corresponding Author); Ahtinen, Jouni; Pitkänen, Juha-Pekka; Parviainen, Ville; Joenväärä, Sakari; Hautamäki, Mika; Lindroos, Peter; Mäkinen, Jarno; Renkonen, Risto.

In: Biotechnology and Bioprocess Engineering, Vol. 16, No. 2, 2011, p. 264-272.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - On-line high performance liquid chromatography measurements of extracellular metabolites in an aerobic batch yeast (Saccharomyces cerevisiae) culture

AU - Tohmola, Niina

AU - Ahtinen, Jouni

AU - Pitkänen, Juha-Pekka

AU - Parviainen, Ville

AU - Joenväärä, Sakari

AU - Hautamäki, Mika

AU - Lindroos, Peter

AU - Mäkinen, Jarno

AU - Renkonen, Risto

PY - 2011

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N2 - 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.

AB - 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.

KW - diauxic shift

KW - high performance liquid chromatography

KW - HPLC

KW - metabolite

KW - on-line

KW - Saccharomyces cerevisiae

U2 - 10.1007/s12257-010-0147-3

DO - 10.1007/s12257-010-0147-3

M3 - Article

VL - 16

SP - 264

EP - 272

JO - Biotechnology and Bioprocess Engineering

JF - Biotechnology and Bioprocess Engineering

SN - 1226-8372

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