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    Life cycle assessment for early-stage process optimization of microbial biosurfactant production using kinetic models : a case study on mannosylerythritol lipids (MEL)
    (2024) Bippus, Lars; Briem, Ann-Kathrin; Beck, Alexander; Zibek, Susanne; Albrecht, Stefan
    Introduction: This study assesses the environmental impacts of mannosylerythritol lipids (MELs) production for process optimization using life cycle assessment (LCA). MELs are glycolipid-type microbial biosurfactants with many possible applications based on their surface-active properties. They are generally produced by fungi from the family of Ustilaginaceae via fermentation in aerated bioreactors. The aim of our work is to accompany the development of biotechnological products at an early stage to enable environmentally sustainable process optimization. Methods: This is done by identifying hotspots and potentials for improvement based on a reliable quantification of the environmental impacts. The production processes of MELs are evaluated in a cradle-to-gate approach using the Environmental Footprint (EF) 3.1 impact assessment method. The LCA model is based on upscaled experimental data for the fermentation and purification, assuming the production at a 10 m³ scale. In the case analyzed, MELs are produced from rapeseed oil and glucose, and purified by separation, solvent extraction, and chromatography. Results: The results of the LCA show that the provision of substrates is a major source of environmental impacts and accounts for 20% of the impacts on Climate Change and more than 70% in the categories Acidification and Eutrophication. Moreover, 33% of the impacts on Climate Change is caused by the energy requirements for aeration of the bioreactor, while purification accounts for 42% of the impacts respectively. For the purification, solvents are identified as the main contributors in most impact categories. Discussion: The results illustrate the potentials for process optimization to reduce the environmental impacts of substrate requirements, enhanced bioreactor aeration, and efficient solvent use in downstream processing. By a scenario analysis, considering both experimental adaptations and prospective variations of the process, the laboratory development can be supported with further findings and hence efficiently optimized towards environmental sustainability. Moreover, the presentation of kinetic LCA results over the fermentation duration shows a novel way of calculating and visualizing results that corresponds to the way of thinking of process engineers using established environmental indicators and a detailed system analysis. Altogether, this LCA study supports and demonstrates the potential for further improvements towards more environmentally friendly produced surfactants.
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    Growth behavior of selected Ustilaginaceae fungi used for mannosylerythritol lipid (MEL) biosurfactant production - evaluation of a defined culture medium
    (2020) Beck, Alexander; Zibek, Susanne
    Fungi of the Ustilaginaceae family are a promising source for many biotechnologically relevant products. Among these, mannosylerythritol lipid (MEL) biosurfactants have drawn a special interested over the last decades due to their manifold application possibilities. Nevertheless, there is still a knowledge gap regarding process engineering of MEL production. As an example, no reports on the use of a chemically defined culture medium have been published yet, although such a defined medium might be beneficial for scaling-up the production process toward industrial scale. Our aim therefore was to find a mineral medium that allows fast biomass growth and does not negatively affect the successive MEL production from plant oils. The results showed comparable growth performance between the newly evaluated mineral medium and the established yeast extract medium for all seven investigated Ustilaginaceae species. Final biomass concentrations and specific growth rates of 0.16-0.25 h-1 were similar for the two media. Oxygen demand was generally higher in the mineral medium than in the yeast extract medium. It was shown that high concentrations of vitamins and trace elements were necessary to support the growth. Increasing starting concentrations of the media by a factor of 10 resulted in proportionally increasing final biomass concentrations and up to 2.3-times higher maximum growth rates for all species. However, it could also lead to oxygen limitation and stagnant growth rates when too high medium concentrations were used, which was observed for Ustilago siamensis and Moesziomyces aphidis. Successive MEL production from rapeseed oil was effectively shown for 4 out of 7 organisms when the mineral medium was used for cell growth, and it was even enhanced for two organisms, M. aphidis and Pseudozyma hubeiensis pro tem., as compared to the established yeast extract medium. Conversion of rapeseed oil into MEL was generally improved when higher biomass concentrations were achieved during the initial growth phase, indicating a positive relationship between biomass concentration and MEL production. Overall, this is the first report on the use of a chemically defined mineral medium for the cell growth of Ustilaginaceae fungi and successive MEL production from rapeseed oil, as an alternative to the commonly employed yeast extract medium.
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    Comprehensive characterization and evaluation of the process chain and products from Euphausia superba exocuticles to chitosan
    (2023) Hahn, Thomas; Egger, Jeannine; Krake, Simon; Dyballa, Michael; Stegbauer, Linus; Seggern, Nils von; Bruheim, Inge; Zibek, Susanne
    Antarctic krill (Euphausia superba) is a source for compounds of high nutritive value. Within that process of extraction, exocuticles (shells) accumulate which are currently disposed. A valorization of the compounds of the exocuticle such as chitosan would be beneficial to avoid waste and to obtain a versatile polymer at the same time. In contrast to previous investigations focusing on chitosan production from whole krill, we applied and optimized process stages of the chitosan production from the exocuticles, performing a comprehensive analytical evaluation of the whole process, the side streams and the products for the first time. Degreasing was the first step resulting in a krill oil yield of 6.2% using ethanol. The fatty acid profile exhibited high contents of phospholipids (21.2%). Citric acid offered a demineralization efficiency of 93%. Deproteinization investigation revealed 2 M NaOH and 90°C for 2.5 h to be the best parameters, resulting in a deproteinization efficiency of 99.9% and a chitin content of 92.8%. The spectroscopic investigation indicated that the chitin has a crystallinity index of 76% and an acetylation degree of 88%. The deacetylation degrees of the resulting chitosans is determined to be 74%-88%, the molecular weight ranges from 102 to 126 kDa.
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    Ökobilanz für die Bioprozessoptimierung : Herstellung des Biotensids MEL
    (2024) Bippus, Lars; Briem, Ann-Kathrin; Beck, Alexander; Zibek, Susanne; Albrecht, Stefan
    This study evaluates the environmental impacts of producing mannosylerythritol lipids (MELs) using life cycle assessment (LCA) and kinetic models. MELs are microbial biosurfactants with various applications produced from biobased sources. The LCA results indicate that substrate provision, bioreactor aeration and solvent use for purification are major environmental impact sources. The findings highlight areas for improving the environmental sustainability of the production processes.