Biotechnology Letters

, Volume 35, Issue 8, pp 1223–1230 | Cite as

Oxygen supply in disposable shake-flasks: prediction of oxygen transfer rate, oxygen saturation and maximum cell concentration during aerobic growth

  • Sarah Schiefelbein
  • Alexander Fröhlich
  • Gernot T. John
  • Falco Beutler
  • Christoph Wittmann
  • Judith Becker
Original Research Paper

Abstract

Dissolved oxygen plays an essential role in aerobic cultivation especially due to its low solubility. Under unfavorable conditions of mixing and vessel geometry it can become limiting. This, however, is difficult to predict and thus the right choice for an optimal experimental set-up is challenging. To overcome this, we developed a method which allows a robust prediction of the dissolved oxygen concentration during aerobic growth. This integrates newly established mathematical correlations for the determination of the volumetric gas–liquid mass transfer coefficient (kLa) in disposable shake-flasks from the filling volume, the vessel size and the agitation speed. Tested for the industrial production organism Corynebacterium glutamicum, this enabled a reliable design of culture conditions and allowed to predict the maximum possible cell concentration without oxygen limitation.

Keywords

Corynebacterium glutamicum Disposable shake-flask Non-invasive optical sensor technology Oxygen transfer rate Volumetric gas–liquid mass transfer coefficient 

List of Symbols

Variables

µ

Specific growth rate (h−1)

c

Oxygen concentration (mmol l−1; %)

c*

Saturated oxygen concentration (c* = 0.232 mmol l−1 at 30 °C; c* = 0.212 mmol l−1 at 37 °C For relative expression, c* was set to 100 %) (mmol l−1; %)

kLa

Gas–liquid mass transfer coefficient (h−1)

n

Shaking frequency (h−1)

OTR

Oxygen transfer rate (mmol h−1)

OUR

Oxygen uptake rate (mmol h−1)

\( {\text{q}}_{{{\text{O}}_{2} }} \)

Specific oxygen uptake rate (mmol g−1CDW h−1)

Vmax

Maximum flask volume (ml)

X

Biomass concentration (g l−1)

Empirical parameters

Gaussian fitting

a, b, c, x0, y0

Parabolic fitting

a, b, c, d, y0

Supplementary material

10529_2013_1203_MOESM1_ESM.xlsx (25 kb)
Supplementary material 1 (XLSX 25 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Sarah Schiefelbein
    • 1
  • Alexander Fröhlich
    • 1
  • Gernot T. John
    • 2
  • Falco Beutler
    • 2
  • Christoph Wittmann
    • 1
  • Judith Becker
    • 1
  1. 1.Institute of Biochemical EngineeringTechnische Universität BraunschweigBraunschweigGermany
  2. 2.PreSens Precision Sensing GmbHRegensburgGermany

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