Bioprocess Development in Single‐Use Systems for Heterotrophic Marine Microalgae

Hillig, Friederike; Annemüller, Steffi; Chmielewska, Magdalena; Pilarek, Maciej; Junne, Stefan; Neubauer, Peter

doi:10.1002/cite.201200143

Cited by 21 publications

(15 citation statements)

References 43 publications

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“…From the bioprocess engineering point of view, PFCs are recognized as the most flexible liquid carriers/scavengers of gaseous compounds (O 2 and CO 2 , mainly) and were successfully applied in culture systems of various types of cells, i.e. microorganisms such as bacteria (Pilarek et al 2011;2013a), yeast (Pilarek et al 2006), fungi (Elibol and Ozer 2000) and microalgae (Hillig et al 2013(Hillig et al , 2014, animal (Shiba et al 1998;Rappaport 2003;Pilarek et al 2013bPilarek et al , 2014Douglas et al 2014) as well as plant cells (Wardrop et al 1996;Lowe et al 2003;Davey et al 2005;Pilarek and Szewczyk 2008;Sykłowska-Baranek et al 2014). …”

Section: Introductionmentioning

confidence: 99%

Perfluorodecalin-supported system enhances taxane production in hairy root cultures of Taxus x media var. Hicksii carrying a taxadiene synthase transgene

Sykłowska-Baranek

Pilarek

Bonfill

et al. 2014

Plant Cell Tiss Organ Cult

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Enhanced taxane production was observed in a hybrid, two liquid phases containing, cultures of Taxus x media var. Hicksii hairy root carrying a taxadiene synthase transgene, supported with liquid perfluorodecalin (PFD) in degassed or aerated form. The hairy root cultures were elicited with methyl jasmonate (MJ, 100 lM) or coronatine (COR, 1 lM), and fed with sucrose and L-phenylalanine. The root growth was not stimulated by PFD addition, irrespective of the day of its application (day 0 and 14). However, in the cultures elicited with MJ and performed in the presence of PFD the final root biomass accumulation was higher than in cultures performed without PFD while the opposite effect was observed in cultures supplemented with COR. The highest paclitaxel content in root biomass was determined at the end of the cultures elicited with MJ and supplemented with PFD-degassed at day 0 or 14, 1,440.8 and 1,432.5 lg g -1 DW, respectively. The highest total (i.e. intracellular ? extracellular: both in aqueous and PFD phases) paclitaxel yield in flasks (149.15 lg flask -1 ) was noted after the application of PFD-degassed at day 14. The other taxane detected was baccatin III, only in the root biomass, with the highest content (76.9 lg g -1 DW) observed under COR treatment. Although COR stimulated paclitaxel production with less efficiency than MJ, it resulted in higher paclitaxel excretion to the liquid phases of culture medium and PFD.

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Section: Introductionmentioning

confidence: 99%

Perfluorodecalin-supported system enhances taxane production in hairy root cultures of Taxus x media var. Hicksii carrying a taxadiene synthase transgene

Sykłowska-Baranek

Pilarek

Bonfill

et al. 2014

Plant Cell Tiss Organ Cult

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“…insoluble fraction of not-correctly folded protein molecules) achieved in milliliter-scale of dispersed PFD containing cultures of E. coli has been reported by Pilarek et al (2011). The same group has also informed of a 25 % increase in plasmid DNA yield biosynthesised in E. coli cells cultured in PFDsupported microliter-scale system (Grunzel et al, 2014;Pilarek et al, 2013). Such a huge range of culture volumes (from 10 ) which has been reviewed in Table 4 (as well as discussed above) clearly indicates that perfluorinated oxygen carriers are very flexible bioengineering tools for gas transfer intensification in scaling-up or scaling-down of bioprocesses.…”

Section: Pfc-mediated Oxygen Transfer Intensificationmentioning

confidence: 80%

“…microorganisms, e.g. bacteria (Pilarek et al, 2011;Pilarek et al, 2013a), yeast (Pilarek and Szewczyk, 2008;Pilarek et al, 2006), fungi (Elibol, 2001) or microalgae (Hillig et al, 2013;Hillig et al, 2014), or plant cells (Davey et al 2005;Pilarek and Szewczyk 2008;Sykłowska-Baranek et al 2014) as wells as animal (Douglas et al 2014;Pilarek et al, 2013b;Pilarek et al, 2014;Rappaport, 2003;Shiba et al, 1998) to successfully obtain higher cell densities or to identify positive influence on metabolite excretion due to enhanced the mass transfer effects in culture systems, mainly.…”

Section: Introductionmentioning

confidence: 99%

Liquid Perfluorochemicals as Flexible and Efficient Gas Carriers Applied in Bioprocess Engineering: An Updated Overview and Future Prospects

Pilarek¹

2014

Chemical and Process Engineering

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Fully synthetic, biochemically inert and water-immiscible liquid perfluorochemicals (PFCs) are recognised as flexible liquid carriers/scavengers of gaseous compounds (respiratory gases mainly, i.e. O 2 and CO 2 ) and increasingly applied in bioprocess engineering. A range of unmatched physicochemical properties of liquid PFCs, i.e. outstanding chemo-and thermostability, extremely low surface tension, simultaneous hydro-and lipophobicity, which result from carbon chain substitution with fluorine atoms (the most electronegative chemical element) and the presence of intramolecular C-F bonds (the strongest single bond known in organic chemistry) have been described in detail. Exceptional propensity to solubility of respiratory gases in liquid perfluorinated compounds has been widely discussed. Advantages and disadvantages of bioprocess applications of liquid PFCs in the form of a pure PFC as well as in an emulsified form have been pointed out. A liquid PFC-mediated mass transfer intensification in various types of microbial, plant cell and animal cell culture systems: from miniaturised microlitre-scale cultures, via biomaterial-based scaffolds containing culture systems, to litre-scale bioreactors, has been reviewed and elaborated on bearing in mind the benefits of bioprocesses.

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“…The CO 2 content was determined with an optical sensor (PreSens, Regensburg, Germany). Surprisingly, the positive influence of oxygenated PFD on DHA production, which was described in [88], was not visible when only a few samples were taken manually within a very short time (Fig. 5).…”

Section: Cultivation In Deep-well Platesmentioning

confidence: 80%

“…Nile Red staining intensity was highest at 180 rpm, and the distribution of the cells was narrow compared to results obtained at 315 rpm (Figure 8). This finding indicated that shear stress had a negative influence on the DHA content of the cells [88]. Hence, although a higher power input is achieved, which increases the gas transfer rate, the high shear forces act counter to the goal of increasing the product yield.…”

Section: Microalgae Cultivation In Ultra Yield Flasksmentioning

confidence: 94%

Cultivation of Marine Microorganisms in Single-Use Systems

Hillig

Pilarek

Junne

et al. 2013

Advances in Biochemical Engineering/Biotechnology

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: Marine cultures are an important source of novel substances and enzymes. As efforts to isolate strains from (deep) sea environments increase, the demand for methodology platforms to cultivate these organisms is also rising. Due to the high salt concentration and the shear sensitivity exhibited by some heterotrophic microalgae, single-use systems originally designed for the cultivation of mammalian cell lines can be a valuable alternative. Using the cultivation of the heterotrophic marine microalgae Crypthecodinium cohnii as an example, this chapter makes suggestions for experimental design, for improving process development by integrating parallel experiments, and for scaling-up and scaling-down methodologies. It describes how to identify suitable single-use systems and how to integrate a two-layer system with perfluordecalin to improve the gas transfer in deep-well plates. The process is also scaled up in several single-use systems. We also describe challenges in the process development to achieve sufficient oxygen transfer, monitoring, and control, and we discuss limitations such as corrosion, long-term stability, and leachables in single-use systems. Finally, we demonstrate a method for cheap, fast, and consistent process development for marine microorganisms.

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Bioprocess Development in Single‐Use Systems for Heterotrophic Marine Microalgae

Cited by 21 publications

References 43 publications

Perfluorodecalin-supported system enhances taxane production in hairy root cultures of Taxus x media var. Hicksii carrying a taxadiene synthase transgene

Perfluorodecalin-supported system enhances taxane production in hairy root cultures of Taxus x media var. Hicksii carrying a taxadiene synthase transgene

Liquid Perfluorochemicals as Flexible and Efficient Gas Carriers Applied in Bioprocess Engineering: An Updated Overview and Future Prospects

Cultivation of Marine Microorganisms in Single-Use Systems

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