Magnetische millifluidische Fraktionierung einer heterogenen Hefezellkultur zur statistisch relevanten Analyse der altersabhängigen Populationsentwicklung
Lebensmittelchemie
Zusammenfassung der Projektergebnisse
This project is focused on developing a millifluidic flow channel designed for the agedependent magnetophoretic fractionation of yeast cells, especially S. pastorianus ssp. carlsbergensis. This will allow a more detailed analysis of growth patterns and gene expression profiles, contributing to a more comprehensive understanding of yeast cellular aging and its role in industrial processes. To enable fractionation, yeast cells are selectively labelled by binding iron oxide nanoparticles (IONs) to chitin-enriched bud scars via a linker-protein. The IONs are synthesized and functionalized with a silica shell and ethylenediaminetetraacetic acid (EDTA) for stability and binding. The chitin-binding linker-protein is engineered and synthesized in E. coli, purified, and used to label the bud scars, allowing differentiation between mother and daughter cells. The binding capacity and agglomeration behavior of the particles are assessed in different buffers. Flow channel designs are tested for separation selectivity based on hydrodynamics and magnetic forces. A two-step approach is implemented to improve the overall process selectivity to > 90% by separating daughter cells first. Daughter cells and mother cells are compared in aerobic and anaerobic cultivations based on optical density and sugar consumption, showing higher growth rates and faster sugar consumption in daughter cells. In conclusion, this project offers a promising method for the age-based fractionation of cells due to a magnetophoretic sorting in a pinch-shaped millifluidic channel for a more profound knowledge of yeast cellular aging, allowing a first step towards improving the efficiency of yeast-based processes.
Projektbezogene Publikationen (Auswahl)
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Recombinant protein linker production as a basis for non-invasive determination of single-cell yeast age in heterogeneous yeast populations. RSC Advances, 11(51), 31923-31932.
Eigenfeld, Marco; Kerpes, Roland & Becker, Thomas
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The Effect of pH and Viscosity on Magnetophoretic Separation of Iron Oxide Nanoparticles. Magnetochemistry, 7(6), 80.
Wittmann, Leonie; Turrina, Chiara & Schwaminger, Sebastian P.
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Understanding the Impact of Industrial Stress Conditions on Replicative Aging in Saccharomyces cerevisiae. Frontiers in Fungal Biology, 2.
Eigenfeld, Marco; Kerpes, Roland & Becker, Thomas
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Autofluorescence prediction model for fluorescence unmixing and age determination. Biotechnology Journal, 17(12).
Eigenfeld, Marco; Kerpes, Roland; Whitehead, Iain & Becker, Thomas
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How to design a low-cost pilot scale magnetic bioseparation process for protein separation from complex mixtures using in-line process analytics. Separation and Purification Technology, 323, 124429.
Krolitzki, Eva; Steck, Sabrina; Nazifi, Andja; Abt, Michael; Schwaminger, Sebastian P. & Berensmeier, Sonja
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Quantification methods of determining brewer’s and pharmaceutical yeast cell viability: accuracy and impact of nanoparticles. Analytical and Bioanalytical Chemistry, 415(16), 3201-3213.
Eigenfeld, Marco; Wittmann, Leonie; Kerpes, Roland; Schwaminger, Sebastian & Becker, Thomas
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Studying the impact of cell age on the yeast growth behaviour ofSaccharomyces pastorianusvar.carlsbergensisby magnetic separation. Biotechnology Journal, 18(7).
Eigenfeld, Marco; Wittmann, Leonie; Kerpes, Roland; Schwaminger, Sebastian P. & Becker, Thomas
