Core Facility Hohenheim

Permanent URI for this collectionhttps://hohpublica.uni-hohenheim.de/handle/123456789/16626

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Changes of microorganism composition in fresh and stored bee pollen from Southern Germany
(2021) Friedle, Carolin; D’Alvise, Paul; Schweikert, Karsten; Wallner, Klaus; Hasselmann, Martin
Analysis of plant pollen can provide valuable insights into the existing spectrum of microorganisms in the environment. When harvesting bee-collected pollen as a dietary supplement for human consumption, timely preservation of the freshly collected pollen is fundamental for product quality. Environmental microorganisms contained in freshly collected pollen can lead to spoilage by degradation of pollen components. In this study, freshly collected bee pollen was sampled at different locations and stored under various storage conditions to examine the hypothesis that storage conditions may have an effect on the composition of microorganisms in pollen samples. The samples were analyzed using 16S and 18S amplicon sequencing and characterized by palynological analysis. Interestingly, the bacterial communities between pollen samples from different locations varied only slightly, whereas for fungal community compositions, this effect was substantially increased. Further, we noticed that fungal communities in pollen are particularly sensitive to storage conditions. The fungal genera proportion Cladosporium and Mycosphaerella decreased, while Zygosaccharomyces and Aspergillus increased during storage. Aspergillus and Zygosaccharomyces fractions increased during storage at 30 °C, which could negatively impact the pollen quality if it is used as a dietary supplement.
Exploring Phaeodactylum tricornutum for nutraceuticals: cultivation techniques and neurotoxin risk assessment
(2025) Ebbing, Tobias; Kopp, Lena; Frick, Konstantin; Simon, Tabea; Würtz, Berit; Pfannstiel, Jens; Schmid-Staiger, Ulrike; Bischoff, Stephan C.; Tovar, Günter E. M.; Faraloni, Cecilia; Touloupakis, Eleftherios
This study investigates the potential of the diatom Phaeodactylum tricornutum (PT) as a sustainable and nutritionally valuable food source, focusing on its ability to produce bioactive compounds such as eicosapentaenoic acid, fucoxanthin, chrysolaminarin (CRY) and proteins. PT was cultivated in a flat-plate airlift photobioreactor (FPA-PBR) illuminated with LEDs from two sides. The study aimed to monitor and minimize β-methylamino-L-alanine (BMAA) levels to address safety concerns. The data showed that the selected FPA-PBR setup was superior in biomass and EPA productivity, and CRY production was reduced. No BMAA was detected in any biomass sample during cultivation. By adjusting the cultivation conditions, PT biomass with different compositional profiles could be produced, enabling various applications in the food and health industries. Biomass from nutrient-repleted conditions is rich in EPA and Fx, with nutritional and health benefits. Biomass from nutrient-depleted conditions accumulated CRY, which can be used as dietary fiber. These results highlight the potential of PT as a versatile ingredient for human consumption and the effectiveness of FPA-PBRs with artificial lighting in producing high-quality biomass. This study also provides the basis for future research to optimize photobioreactor conditions to increase production efficiency and to tailor the biomass profiles of PT for targeted health-promoting applications.
Glucoselipid biosurfactant biosynthesis operon of Rouxiella badensis DSM 100043T: screening, identification, and heterologous expression in Escherichia coli
(2025) Harahap, Andre Fahriz Perdana; Treinen, Chantal; Van Zyl, Leonardo Joaquim; Williams, Wesley Trevor; Conrad, Jürgen; Pfannstiel, Jens; Klaiber, Iris; Grether, Jakob; Hiller, Eric; Vahidinasab, Maliheh; Perino, Elvio Henrique Benatto; Lilge, Lars; Burger, Anita; Trindade, Marla; Hausmann, Rudolf; Seo, Myung-Ji
Rouxiella badensis DSM 100043T had been previously proven to produce a novel glucoselipid biosurfactant which has a very low critical micelle concentration (CMC) as well as very good stability against a wide range of pH, temperature, and salinity. In this study, we performed a function-based library screening from a R. badensis DSM 100043T genome library to identify responsible genes for biosynthesis of this glucoselipid. The identified open reading frames (ORFs) were cloned into several constructs in Escherichia coli for gene permutation analysis and the individual products were analyzed using high-performance thin-layer chromatography (HPTLC). Products of interest from positive expression strains were purified and analyzed by liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) and nuclear magnetic resonance (NMR) for further structure elucidation. Function-based screening of 5400 clones led to the identification of an operon containing three ORFs encoding acetyltransferase GlcA (ORF1), acyltransferase GlcB (ORF2), and phosphatase/HAD GlcC (ORF3). E. coli pCAT2, with all three ORFs, resulted in the production of identical R. badensis DSM 100043T glucosedilipid with Glu-C10:0-C12:1 as the main congener. ORF2-deletion strain E. coli pAFP1 primarily produced glucosemonolipids, with Glu-C10:0,3OH and Glu-C12:0 as the major congeners, predominantly esterified at the C-2 position of the glucose moiety. Furthermore, fed-batch bioreactor cultivation of E. coli pCAT2 using glucose as the carbon source yielded a maximum glucosedilipid titer of 2.34 g/L after 25 h of fermentation, which is 55-fold higher than that produced by batch cultivation of R. badensis DSM 100043T in the previous study.
Multi-omics characterization of the monkeypox virus infection
(2024) Huang, Yiqi; Bergant, Valter; Grass, Vincent; Emslander, Quirin; Hamad, M. Sabri; Hubel, Philipp; Mergner, Julia; Piras, Antonio; Krey, Karsten; Henrici, Alexander; Öllinger, Rupert; Tesfamariam, Yonas M.; Dalla Rosa, Ilaria; Bunse, Till; Sutter, Gerd; Ebert, Gregor; Schmidt, Florian I.; Way, Michael; Rad, Roland; Bowie, Andrew G.; Protzer, Ulrike; Pichlmair, Andreas
Multiple omics analyzes of Vaccinia virus (VACV) infection have defined molecular characteristics of poxvirus biology. However, little is known about the monkeypox (mpox) virus (MPXV) in humans, which has a different disease manifestation despite its high sequence similarity to VACV. Here, we perform an in-depth multi-omics analysis of the transcriptome, proteome, and phosphoproteome signatures of MPXV-infected primary human fibroblasts to gain insights into the virus-host interplay. In addition to expected perturbations of immune-related pathways, we uncover regulation of the HIPPO and TGF-β pathways. We identify dynamic phosphorylation of both host and viral proteins, which suggests that MAPKs are key regulators of differential phosphorylation in MPXV-infected cells. Among the viral proteins, we find dynamic phosphorylation of H5 that influenced the binding of H5 to dsDNA. Our extensive dataset highlights signaling events and hotspots perturbed by MPXV, extending the current knowledge on poxviruses. We use integrated pathway analysis and drug-target prediction approaches to identify potential drug targets that affect virus growth. Functionally, we exemplify the utility of this approach by identifying inhibitors of MTOR, CHUK/IKBKB, and splicing factor kinases with potent antiviral efficacy against MPXV and VACV.
PKC regulates αKlotho gene expression in MDCK and NRK-52E cells
(2024) Wolf, Lisa; Vogt, Julia; Alber, Jana; Franjic, Domenic; Feger, Martina; Föller, Michael
Particularly expressed in the kidney, αKlotho is a transmembrane protein that acts together with bone hormone fibroblast growth factor 23 (FGF23) to regulate renal phosphate and vitamin D homeostasis. Soluble Klotho (sKL) is released from the transmembrane form and controls various cellular functions as a paracrine and endocrine factor. αKlotho deficiency accelerates aging, whereas its overexpression favors longevity. Higher αKlotho abundance confers a better prognosis in cardiovascular and renal disease owing to anti-inflammatory, antifibrotic, or antioxidant effects and tumor suppression. Serine/threonine protein kinase C (PKC) is ubiquitously expressed, affects several cellular responses, and is also implicated in heart or kidney disease as well as cancer. We explored whether PKC is a regulator of αKlotho. Experiments were performed in renal MDCK or NRK-52E cells and PKC isoform and αKlotho expression determined by qRT-PCR and Western Blotting. In both cell lines, PKC activation with phorbol ester phorbol-12-myristate-13-acetate (PMA) downregulated, while PKC inhibitor staurosporine enhanced αKlotho mRNA abundance. Further experiments with PKC inhibitor Gö6976 and RNA interference suggested that PKCγ is the major isoform for the regulation of αKlotho gene expression in the two cell lines. In conclusion, PKC is a negative regulator of αKlotho gene expression, an effect which may be relevant for the unfavorable effect of PKC on heart or kidney disease and tumorigenesis.

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