Browsing by Author "Rupp, Steffen"

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Open Access
Fermentation and recovery of cellobiose lipids using foam fractionation
(2023) Oraby, Amira; Hug, Daniel; Weickardt, Isabell; Maerz, Lea; Nebel, Sabrina; Kurmann, Jasper; Rupp, Steffen; Tovar, Günter E. M.; Zibek, Susanne
Cellobiose lipids (CL) are glycolipids secreted by many Ustilaginaceae species in aerobic fermentations characterised by excessive foaming. While increasing CL concentrations remains an aim for its industrial production, excessive foaming during fermentation presents a challenge even at laboratory scale. Foam fractionation (FF) provides a solution to the foaming problem and facilitates the proceeding purification of CL. Here, we present a first CL fermentation process applying FF. With our set-up, we manage to exploit the excessive foaming for continuous product separation. The set-up includes a foam collecting vessel (FCV) with inserts for CL accumulation and foamate recirculation to minimise biomass and nutrient loss. Integrating a foam column (FC) into the fermenter headspace enabled foam enrichment, resulting in the recovery of > 90% of the produced CL from the separated fractions consisting of foam depositions in the fermenter headspace and the FCV. We also increased the fermenter filling volume and thus achieved a higher fermentation capacity. The separated CL fraction was purified via ethanol extraction to obtain CL with purities > 90%. We further examined the effects of different culture media constituents, including biomass and CL, on foam generation and decay and assessed the effect of FC geometries on product enrichment and recovery. In this work, a FF set-up is presented that enables a stable CL fermentation without additional foam mitigation methods. At the same time, the application of FF separated a fraction that was highly enriched in CL during fermentation, resulting in highly pure CL after a simple ethanol extraction.
Open Access
Human-based immune responsive in vitro infection models for validation of novel TLR4 antagonists identified by computational discovery
(2022) Merk, Helena; Amran-Gealia, Tehila; Finkelmeier, Doris; Kohl, Christina; Pichota, Isabelle; Stern, Noa; Rupp, Steffen; Goldblum, Amiram; Burger-Kentischer, Anke
Infectious diseases are still a major problem worldwide. This includes microbial infections, with a constant increase in resistance to the current anti-infectives employed. Toll-like receptors (TLRs) perform a fundamental role in pathogen recognition and activation of the innate immune response. Promising new approaches to combat infections and inflammatory diseases involve modulation of the host immune system via TLR4. TLR4 and its co-receptors MD2 and CD14 are required for immune response to fungal and bacterial infection by recognition of microbial cell wall components, making it a prime target for drug development. To evaluate the efficacy of anti-infective compounds early on, we have developed a series of human-based immune responsive infection models, including immune responsive 3D-skin infection models for modeling fungal infections. By using computational methods: pharmacophore modeling and molecular docking, we identified a set of 46 potential modulators of TLR4, which were screened in several tests systems of increasing complexity, including immune responsive 3D-skin infection models. We could show a strong suppression of cytokine and chemokine response induced by lipopolysacharide (LPS) and Candida albicans for individual compounds. The development of human-based immune responsive assays provides a more accurate and reliable basis for development of new anti-inflammatory or immune-modulating drugs.
Open Access
Life cycle assessment as a driver for process optimisation of cellobiose lipids fermentation and purification
(2024) Oraby, Amira; Briem, Ann-Kathrin; Bippus, Lars; Rupp, Steffen; Zibek, Susanne
Purpose: Cellobiose lipids (CL) are biosurfactants produced by various Ustilaginaceae species in aerobic fermentations. They show high potential for application as alternatives to conventional oleochemical- or petrochemical surfactants. To ensure their environmentally friendly performance, we aimed to assess CL production from a life cycle perspective at an early developmental stage to identify process steps that have the highest impact on the environment. With this information, optimisation approaches can be derived.
Materials and methods: Following a cradle-to-gate approach, we modelled the CL fermentation and purification process based on experimental data from the lab scale and process simulation data at a 10 m 3 scale. For LCA, the impact categories (IC) abiotic depletion potential (ADP), eutrophication potential, photochemical ozone creation potential, global warming potential, acidification potential, and the primary energy demand were calculated for all process steps. Based on the obtained results, process bottlenecks were identified, and alternative process scenarios varying the related process parameters were simulated. These were used to assess the environmental impact reduction potential (EIRP) of an optimised process and draw recommendations for experimental process optimisation.
Results and discussion: The obtained results showed that the fermentation caused ~ 73% of ADP and more than 85% of all other ICs. The major contributor was the electricity consumption for continuous fermenter aeration. Thus, reducing the fermentation duration from the initial 14 to 5 days would result in a decrease in all investigated ICs of up to ~ 27-52%. An increase in CL concentration results in a decrease in all ICs of a similar magnitude due to the higher yield per batch at comparable energy and material consumption. Although the share of purification process steps to all ICs is overall relatively small, implementing foam fractionation for in situ product recovery showed an additional EIRP of 18-27% in all purification IC shares.
Conclusions: The conducted LCA showed that overall, more EIRP can be achieved by optimising fermentation process parameters compared to purification process steps. This is mainly due to the long fermentation duration and large energy consumption for fermenter aeration. This highlights the importance of using LCA as a driver for process optimisation to identify process steps with high EIRP. While some of the results are specific to CL, other obtained results can be transferred to other fermentations.
Open Access
Optimization and kinetic modeling of a fed-batch fermentation for mannosylerythritol lipids (MEL) production with moesziomyces aphidis
(2022) Beck, Alexander; Vogt, Franziska; Hägele, Lorena; Rupp, Steffen; Zibek, Susanne
Mannosylerythritol lipids are glycolipid biosurfactants with many interesting properties. Despite the general interest in those molecules and the need for a robust process, studies on their production in bioreactors are still scarce. In the current study, the fermentative production of MEL in a bioreactor with Moesziomyces aphidis was performed using a defined mineral salt medium. Several kinetic process parameters like substrate consumption rates and product formation rates were evaluated and subsequently enhanced by increasing the biomass concentration through an exponential fed-batch strategy. The fed-batch approaches resulted in two to three fold increased dry biomass concentrations of 10.9-15.5 g/L at the end of the growth phase, compared with 4.2 g/L in the batch process. Consequently, MEL formation rates were increased from 0.1 g/Lh up to around 0.4 g/Lh during the MEL production phase. Thus, a maximum concentration of up to 50.5 g/L MEL was obtained when oil was added in excess, but high concentrations of residual fatty acids were also present in the broth. By adjusting the oil feeding to biomass-specific hydrolysis and MEL production rates, a slightly lower MEL concentration of 34.3 g/L was obtained after 170 h, but at the same time a very pure crude lipid extract with more than 90% MEL and a much lower concentration of remaining fatty acids. With rapeseed oil as substrate, the ideal oil-to-biomass ratio for full substrate conversion was found to be around 10 goil/gbiomass. In addition, off-gas analysis and pH trends could be used to assess biomass growth and MEL production. Finally, kinetic models were developed and compared to the experimental data, allowing for a detailed prediction of the process behavior in future experiments.
Open Access
Techno-economic analysis as a driver for optimisation of cellobiose lipid fermentation and purification
(2022) Oraby, Amira; Rupp, Steffen; Zibek, Susanne
Cellobiose lipids (CL) are glycolipids synthesized by Ustilaginaceae species with potential application as detergents or in cosmetics. This study identified process optimisation potential for CL fermentation based on process modelling and techno-economic analysis. Using a stoichiometric equation based on laboratory data, we calculated the maximum possible CL yield YP/S of 0.45 gCL·gglucose -1 at the biomass yield of 0.10 gBiomass·gglucose -1 with an Ustilago maydis strain. Due to substrate inhibition that may occur at high glucose concentrations, a fed-batch process to increase biomass and CL concentrations was considered in our model. Simulation of different process scenarios showed that the choice of aeration units with high oxygen transfer rates and adaptation of power input to oxygen uptake can significantly decrease electricity consumption. We further assessed scenarios with different fermentation media and CL purification methods, suggesting additional process optimisation potential. Here the omission of vitamins from the fermentation medium proved to be a possible mean to enhance process economy, without compromising CL productivity.