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Verena Oehling
Integrated population and single-cell analyses for improving industrial lignocellulosic ethanol production with Saccharomyces cerevisiae
Cost-effective biotechnological processes based on renewable lignocellulosic biomass will be key in the transition from a fossil-based to a bio-based industry. Lignocellulosic biomass provides a mixture of hexose and pentose sugars for conversion via Saccharomyces cerevisiae to ethanol. Yet, potentially inhibiting chemical compounds like acetic acid (HAc) are released during the required biomass pretreatment for sugar release, which might decrease process efficiency. Knowledge about the physiological responses of S. cerevisiae to complex process conditions imposed by lignocellulosic feedstocks is thus essential for developing more efficient lignocellulose-based bioprocesses.
In this study, an integrated approach of population-based and single-cell analyses technologies was applied for revealing targets for process improvement of industrial lignocellulosic ethanol production. Single-cell analysis (SCA) delivers information about the physiology of single cells that is unattainable with traditional population-based methods and is thus valuable for accessing a mechanistic understanding of cellular processes. We introduce a novel electrical generator for single-cell cultivation in the microfluidic Envirostat system, called Envirostator. Moreover, a population-based cultivation setup was developed and evaluated for mimicking industrial production conditions at lab-scale. The developed setup was used for detailed investigations on HAc tolerance and pentose utilization of two recombinant industrial S. cerevisiae cell lineages expressing a conversion route for the two pentose sugars L-arabinose and D-xylose, respectively. Microfluidic single-cell technologies were finally applied for detailed investigations of robustness during HAc-imposed stress at the single-cell scale.
In this study, an integrated approach of population-based and single-cell analyses technologies was applied for revealing targets for process improvement of industrial lignocellulosic ethanol production. Single-cell analysis (SCA) delivers information about the physiology of single cells that is unattainable with traditional population-based methods and is thus valuable for accessing a mechanistic understanding of cellular processes. We introduce a novel electrical generator for single-cell cultivation in the microfluidic Envirostat system, called Envirostator. Moreover, a population-based cultivation setup was developed and evaluated for mimicking industrial production conditions at lab-scale. The developed setup was used for detailed investigations on HAc tolerance and pentose utilization of two recombinant industrial S. cerevisiae cell lineages expressing a conversion route for the two pentose sugars L-arabinose and D-xylose, respectively. Microfluidic single-cell technologies were finally applied for detailed investigations of robustness during HAc-imposed stress at the single-cell scale.
Schlagwörter: chemical biotechnology; bioethanol; yeast physiology; pentose utilization
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