66 Chemische Verfahrenstechnik
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The current work investigates the capability of a tailored multivariate curve resolution–alternating least squares (MCR-ALS) algorithm to analyse glucose, phosphate, ammonium and acetate dynamics simultaneously in an E. coli BL21 fed-batch fermentation. The high-cell-density (HCDC) process is monitored by ex situ online attenuated total reflection (ATR) Fourier transform infrared (FTIR) spectroscopy and several in situ online process sensors. This approach efficiently utilises automatically generated process data to reduce the time and cost consuming reference measurement effort for multivariate calibration. To determine metabolite concentrations with accuracies between ±0.19 and ±0.96·gL−l, the presented utilisation needs primarily — besides online sensor measurements — single FTIR measurements for each of the components of interest. The ambiguities in alternating least squares solutions for concentration estimation are reduced by the insertion of analytical process knowledge primarily in the form of elementary carbon mass balances. Thus, in this way, the established idea of mass balance constraints in MCR combines with the consistency check of measured data by carbon balances, as commonly applied in bioprocess engineering. The constraints are calculated based on online process data and theoretical assumptions. This increased calculation effort is able to replace, to a large extent, the need for manually conducted quantitative chemical analysis, leads to good estimations of concentration profiles and a better process understanding.
This study introduced an automated long-term fermentation process for fungals grown in pellet form. The goal was to reduce the overgrowth of bioreactor internals and sensors while better rheological properties in the fermentation broth, such as oxygen transfer and mixing time, can be achieved. Because this could not be accomplished with continuous culture and fed-batch fermentation, repeated-batch fermentation was implemented with the help of additional bioreactor internals (“sporulation supports”). This should capture some biomass during fermentation. After harvesting the suspended biomass, intermediate cleaning was performed using a cleaning device. The biomass retained on the sporulation support went through the sporulation phase. The spores were subsequently used as inocula for the next batch. The reason for this approach was that the retained pellets could otherwise cause problems (e.g., overgrowth on sensors) in subsequent batches because the fungus would then show undesirable hyphal growth. Various sporulation supports were tested for sufficient biomass fixation to start the next batch. A reproducible spore concentration within the range of the requirements could be achieved by adjusting the sporulation support (design and construction material), and an intermediate cleaning adapted to this.
The present work aimed at investigating an extraction protocol based on consecutive steps of isoelectric point (pH ~ 4.25) mediated gum swelling and deproteinisation as an alternative method to produce flaxseed gum extracts of enhanced techno-functional characteristics. The osidic and proximate composition, structure conformation, flow behaviour, dynamic rheological and thermal properties of gums isolated from brown and golden flaxseeds were assessed. Gum extraction under near-to-isoelectric point conditions did not impair the extraction yield, residual protein and ash content, whilst it resulted in minor changes in the sugar composition of the flaxseed gum extracts. The deconvolution of the GPC/SEC chromatographs revealed the presence of four major polysaccharidic populations corresponding to arabinoxylans, rhamnogalacturonan–I and two AX-RG-I composite fractions. The latter appeared to minimise the intra- and interchain polymer non-covalent interactions (hydrogen bonding) leading to a better solvation affinity in water and lyotropic solvents. Golden flaxseed gums exerted higher molecular weight (Mw = 1.34–1.15 × 106 Da) and intrinsic viscosities (6.63–5.13 dL g−1) as well as better thickening and viscoelastic performance than the brown flaxseed gum exemplars. Golden flaxseed gums exhibited a better thermal stability compared to the brown flaxseed counterparts and therefore, they are suitable for product applications involving severe heat treatments.