Project description:Aspergillus oryzae is used in solid-state fermentation (SSF) to produce plant-based foods. To this end, the substrate is inoculated with spores of this fungus. So far, the effect of inoculum size on SSF with A. oryzae has primarily focused on the production of specific enzymes. Therefore, the aim of this study was to examine the impact of inoculum size on the full secretome, combined with enzyme activity assays, assessment of colonization, substrate degradation, and sporulation. To this end, A. oryzae was grown for 7 days on whole yellow pea (Pisum sativum). Fluorescence microscopy with a GFP-expressing A. oryzae strain showed that peas had been colonized externally and internally, irrespective of inoculum size. Yet, the highest inoculum size resulted in a stronger pea biomass reduction when compared to the lowest inoculum size. By contrast, sporulation decreased with increasing inoculum size. Notably, proteomics revealed no effect of inoculum size on the protein profiles of aqueous extracts of the colonized peas. Amylases and proteases were the most abundant secreted proteins, which was consistent with their high activity in the aqueous extracts. Proteomics also identified β-1,3-glucanases and chitinases, indicating hyphal lysis. Indeed, 10–19% of the fungal proteins detected in the aqueous extracts lacked signal peptides. These data contribute to our understanding of colonization of substrates by A. oryzae and may be used to optimize SSF with this food grade fungus.
Project description:In order to obtain the transcriptomic information about roof/dorsal and floor/ventral endothelial cells (ECs) of the dorsal aorta (DA) in zebrafish, we isolated single ECs from the DA roof and the floor respectively using the Tg(flk1:eGFP;flk1:NLS-Eos) zebrafish after photoconversion, and then performed scRNA sequencing. These single ECs were isolated at two different time points, 21 hpf and 28 hpf, when the lumen of the DA is established and the EHT begins , respectively.