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Semicontinuous cultivation of photoautotrophic cell suspension cultures in a 20 l airlift-reactor

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Abstract

An airlift-bioreactor system was established for semicontinuous growth of photosynthetically active plant cell suspension cultures in a controlled environment. The bioreactor unit was constructed as a conventional, internal draught tube airlift-reactor, which is characterized by a H D-1 ratio of 2.9, a ratio of the cross-sectional area of the riser to the cross-sectional area of the downcomer of 0.25 and a surface area of 0.435 m2 for illumination. Cultivation experiments could be scaled up to working volumes of maximal 20 1. Sixteen fluorescent tubes were fixed around the outer glass cylinder to provide cells continuously with light. An external cooling device was used to keep the temperature constantly at 27°C. Agitation as well as supply with CO2 was performed by injecting air enriched with CO2 through a ring-shaped sparger at the bottom of the vessel. A first set of experiments was carried out with a photoautotrophic culture of Chenopodium rubrum L. Cell material adapted to large scale culture conditions was used to inoculate a modified MS medium (Murashige & Skoog 1962) without any organic constituents. Under these conditions a biomass increase of 1870% was achieved in 18 days. Several physiological parameters (e.g. pigmentation, photosynthetic O2 evolution, carbohydrate content) were measured routinely to elucidate the growth characteristics of large-scale grown Chenopodium cells. Electron microscopic photographs from different phases of culture growth clearly demonstrate the pattern of cellular development. Special emphasis was placed upon the differentiation of chloroplast ultrastructure. The presented data confirm the feasibility of large-scale culture techniques with photosynthetic active plant cell cultures.

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Abbreviations

D:

diameter

DW:

dry weight

FW:

fresh weight

H:

height

KLa:

volumetric oxygen transfer coefficient (h-1)

MES:

2-(N-morpholino)-ethanesulfonic acid

μ:

specific growth rate (d-1)

PAR:

photosynthetically active radiation (400–700 nm)

Pepcase:

phosphoenolpyruvate carboxylase

Rubisco:

ribulose-1,5-biphosphate carboxylase/oxygenase

td :

doubling time (d)

vvm:

(aeration volume) (medium volume)-1 min-1

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Authors and Affiliations

  1. Institut für Entwicklungs- und Molekularbiologie der Pflanzen, Heinrich-Heine-Universität Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf 1, Germany

    Uwe Fischer, Uwe J. Santore & A. Wilhelm Alfermann

  2. Institut für Biochemie und Biotechnologie der Pflanzen, Westfälische-Wilhelms-Universität Münster, Hindenburgplatz 55, 48143, Münster 1, Germany

    Wolfgang Hüsemann & Wolfgang Barz

Authors
  1. Uwe Fischer
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  2. Uwe J. Santore
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  3. Wolfgang Hüsemann
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  4. Wolfgang Barz
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  5. A. Wilhelm Alfermann
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Dedicated to Prof. F.-C. Czygan on the Occasion of his 60th Birthday

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Fischer, U., Santore, U.J., Hüsemann, W. et al. Semicontinuous cultivation of photoautotrophic cell suspension cultures in a 20 l airlift-reactor. Plant Cell Tiss Organ Cult 38, 123–134 (1994). https://doi.org/10.1007/BF00033869

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