Project description:Differential expression was determined in Calu-3 cells between mock infected and infected with H1N1 influenza virus A/Netherlands/602/2009 at nine time points post-infection. As a comparison, cells were also infected with A/CA/04/2009 H1N1 influenza virus at 4 time points post-infection.
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:Differential expression was determined in Calu-3 cells between mock infected and infected with H1N1 influenza virus A/Netherlands/602/2009 at nine time points post-infection. As a comparison, cells were also infected with A/CA/04/2009 H1N1 influenza virus at 4 time points post-infection. Cells were infected at an MOI of 3.0. For the A/Netherlands/602/09-infected and mock-infected cells, samples were collected at 0, 3, 7, 12, 18, 24, 30, 36, and 48 hours post-infection (h.p.i.). For the A/California/04/2009-infected cells, samples were collected at 0, 12, 24, and 48 h.p.i. Samples were collected in triplicate.
