Search

Biomolecular condensation is a multipurpose cellular process that viruses use ubiquitously during their multiplication. Cauliflower mosaic virus replication complexes are condensates that differ from those of most viruses, as they are nonmembranous assemblies that consist of RNA and protein, mainly the viral protein P6. Although these viral factories (VFs) were described half a century ago, with many observations that followed since, functional details of the condensation process and the properties and relevance of VFs have remained enigmatic. Here, we studied these issues in Arabidopsis thaliana and Nicotiana benthamiana. We observed a large dynamic mobility range of host proteins within VFs, while the viral matrix protein P6 is immobile, as it represents the central node of these condensates. We identified the stress granule (SG) nucleating factors G3BP7 and UBP1 family members as components of VFs. Similarly, as SG components localize to VFs during infection, ectopic P6 localizes to SGs and reduces their assembly after stress. Intriguingly, it appears that soluble rather than condensed P6 suppresses SG formation and mediates other essential P6 functions, suggesting that the increased condensation over the infection time-course may accompany a progressive shift in selected P6 functions. Together, this study highlights VFs as dynamic condensates and P6 as a complex modulator of SG responses.

Volatile fatty acids (VFAs) appear to be an economical carbon feedstock for the cost-effective production of polyhydroxyalkanoates (PHAs). The use of VFAs, however, could impose a drawback of substrate inhibition at high concentrations, resulting in low microbial PHA productivity in batch cultivations. In this regard, retaining high cell density using immersed membrane bioreactor (iMBR) in a (semi-) continuous process could enhance production yields. In this study, an iMBR with a flat-sheet membrane was applied for semi-continuous cultivation and recovery of Cupriavidus necator in a bench-scale bioreactor using VFAs as the sole carbon source. The cultivation was prolonged up to 128 h under an interval feed of 5 g/L VFAs at a dilution rate of 0.15 (d−1), yielding a maximum biomass and PHA production of 6.6 and 2.8 g/L, respectively. Potato liquor and apple pomace-based VFAs with a total concentration of 8.8 g/L were also successfully used in the iMBR, rendering the highest PHA content of 1.3 g/L after 128 h of cultivation. The PHAs obtained from both synthetic and real VFA effluents were affirmed to be poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with a crystallinity degree of 23.8 and 9.6%, respectively. The application of iMBR could open an opportunity for semi-continuous production of PHA, increasing the feasibility of upscaling PHA production using waste-based VFAs.

Carrot pomace (CP) which is generated in a large volume in the juice production process, is rich in cellulose, hemicellulose, sugars, pectin, and minerals. However, in many previous investigations, only cellulose was purified and utilized while other components of CP were discarded as waste. Here, CP was valorized into fungal biomass and cellulose with the aim of utilizing all the CP components. Enzymatic pretreatments were applied to solubilize the digestible fraction of CP including hemicellulose, pectin, sucrose, and other sugars for fungal cultivation, while cellulose remained intact in the solid fraction. The dissolved fraction was utilized as a substrate for the cultivation of an edible fungus (Rhizopus delemar). Fungal cultivation was performed in shake flasks and bench-scale bioreactors. The highest fungal biomass concentration was obtained after pretreatment with invertase (5.01 g/L) after 72 h of cultivation (36 and 42% higher than the concentrations obtained after hemicellulase and pectinase treatments, respectively). Invertase pretreatment resulted in the hydrolysis of sucrose, which could then be taken up by the fungus. Carbohydrate analysis showed 28–33% glucan, 4.1–4.9% other polysaccharides, 0.01% lignin, and 2.7–7% ash in the CP residues after enzymatic pretreatment. Fourier transform infrared spectroscopy and thermogravimetric analysis also confirmed the presence of cellulose in this fraction. The obtained fungal biomass has a high potential for food or feed applications, or as a raw material for the development of biomaterials. Cellulose could be purified from the solid fraction and used for applications such as biobased-textiles or membranes for wastewater treatment, where pure cellulose is needed.

Supplementation of alternative carbon sources is a technological bottleneck, particularly in post-denitrification processes due to stringent effluent nitrogen levels. This study focuses on enhancing the sustainability of wastewater treatment practices by partially replacing conventionally used fossil-derived methanol with organic waste-derived volatile fatty acids (VFAs) in moving bed biofilm reactors (MBBRs). In this regards, results of denitrification batch assays with sequential or simultaneous addition of VFA effluent from acidogenic fermentation of potato starch residue (AD-VFAPPL) and chicken manure (AD-VFACKM), simulated synthetic VFAs solutions (sVFAs), and methanol as carbon source were presented and discussed. Although methanol has proven superior in the conversion of nitrate to nitrite, VFAs are more effective when it comes to reducing nitrite. Although solely added AD-VFAPPL had a slower denitrification capability (0.56 ± 0.13 mgNOx-N removed/m2/day) than methanol (1.04 ± 0.46 mgNOx-N removed/m2/day), up to 50% of the methanol can be replaced by waste-derived AD-VFAPPL and achieve comparable performance (1.08 ± 0.07 mgNOx-N removed/m2/day) with the pure methanol. This proves that the co-addition of VFAs together with methanol can fully compete with pure methanol in performance, providing a promising opportunity for wastewater treatment plants to potentially reduce their carbon footprint and become more sustainable in practice while benefiting from recovered nutrients from waste.