The implementation of single-use technologies offers several major advantages, e.g. prevention of cross-contamination, especially when spore-forming microorganisms are present. This study investigated the application of a single-use bioreactor in batch fermentation of filamentous fungus Penicillium sp. (IBWF 040-09) from the Institute of Biotechnology and Drug Research (IBWF), which is capable of intracellular production of a protease inhibitor against parasitic proteases as a secondary metabolite. Several modifications to the SU bioreactor were suggested in this study to allow the fermentation in which the fungus forms pellets. Simultaneously, fermentations in conventional glass bioreactor were also conducted as reference. Although there are significant differences in the construction material and gassing system, the similarity of the two types of bioreactors in terms of fungal metabolic activity and the reproducibility of fermentations could be demonstrated using statistic methods. Under the selected cultivation conditions, growth rate, yield coefficient, substrate uptake rate, and formation of intracellular protease-inhibiting substance in the single-use bioreactor were similar to those in the glass bioreactor.

Purification of mRNA with oligo(dT)‐functionalized magnetic particles involves a series of magnetic separations for buffer exchange and washing. Magnetic particles interact and agglomerate with each other when a magnetic field is applied, which can result in a decreased total surface area and thus a decreased yield of mRNA. In addition, agglomeration may also be caused by mRNA loading on the magnetic particles. Therefore, it is of interest how the individual steps of magnetic separation and subsequent redispersion in the buffers used affect the particle size distribution. The lysis/binding buffer is the most important buffer for the separation of mRNA from the multicomponent suspension of cell lysate. Therefore, monodisperse magnetic particles loaded with mRNA were dispersed in the lysis/binding buffer and in the reference system deionized water, and the particle size distributions were measured. A concentration‐dependent agglomeration tendency was observed in deionized water. In contrast, no significant agglomeration was detected in the lysis/binding buffer. With regard to magnetic particle recycling, the influence of different storage and drying processes on particle size distribution was investigated. Agglomeration occurred in all process alternatives. For de‐agglomeration, ultrasonic treatment was examined. It represents a suitable method for reproducible restoration of the original particle size distribution.

Productive biofilms are gaining growing interest in research due to their potential of producing valuable compounds and bioactive substances such as antibiotics. This is supported by recent developments in biofilm photobioreactors that established the controlled phototrophic cultivation of algae and cyanobacteria. Cultivation of biofilms can be challenging due to the need of surfaces for biofilm adhesion. The total production of biomass, and thus production of e.g. bioactive substances, within the bioreactor volume highly depends on the available cultivation surface. To achieve an enlargement of surface area for biofilm photobioreactors, biocarriers can be implemented in the cultivation. Thereby, material properties and design of the biocarriers are important for initial biofilm formation and growth of cyanobacteria. In this study, special biocarriers were designed and additively manufactured to investigate different polymeric materials and surface designs regarding biofilm adhesion of the terrestrial cyanobacterium Nostoc flagelliforme (CCAP 1453/33). Properties of 3D-printed materials were characterized by determination of wettability, surface roughness, and density. To evaluate the influence of wettability on biofilm formation, material properties were specifically modified by gas-phase fluorination and biofilm formation was analyzed on biocarriers with basic and optimized geometry in shaking flask cultivation. We found that different polymeric materials revealed no significant differences in wettability and with identical surface design no significant effect on biomass adhesion was observed. However, materials treated with fluorination as well as optimized biocarrier design showed improved wettability and an increase in biomass adhesion per biocarrier surface.

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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.