EPT

Trichothecenes are a structurally diverse family of toxic secondary metabolites produced by certain species of multiple fungal genera. All trichothecene analogs share a core 12,13-epoxytrichothec-9-ene (EPT) structure but differ in presence, absence and types of substituents attached to various positions of EPT. Formation of some of the structural diversity begins early in the biosynthetic pathway such that some producing species have few trichothecene biosynthetic intermediates in common. Cytochrome P450 monooxygenases (P450s) play critical roles in formation of trichothecene structural diversity. Within some species, relaxed substrate specificities of P450s allow individual orthologs of the enzymes to modify multiple trichothecene biosynthetic intermediates. It is not clear, however, whether the relaxed specificity extends to biosynthetic intermediates that are not produced by the species in which the orthologs originate. To address this knowledge gap, we used a mutant complementation-heterologous expression analysis to assess whether orthologs of three trichothecene biosynthetic P450s (TRI11, TRI13 and TRI22) from Fusarium sporotrichioides, Trichoderma arundinaceum, and Paramyrothecium roridum can modify trichothecene biosynthetic intermediates that they do not encounter in the organism in which they originated. The results indicate that TRI13 and TRI22 could not modify the intermediates that they do not normally encounter, whereas TRI11 could modify an intermediate that it does not normally encounter. These findings indicate that substrate promiscuity varies among trichothecene biosynthetic P450s. One structural feature that likely impacts the ability of the P450s to use biosynthetic intermediates as substrates is the presence and absence of an oxygen atom attached to carbon atom 3 of EPT.

Climate change can affect biological assemblages by shifting their species' geographic range and changing species richness. Aquatic insects represent more than half of the freshwater animal species but have been neglected mainly in climate change assessments, particularly in tropical ecosystems. Among the aquatic insect taxa, Ephemeroptera, Plecoptera, and Trichoptera (EPT) are well-known bioindicators of environmental changes and encompass an essential metric for rivers and streams' biomonitoring. Here, we use ecological niche models to project the impact of climate change on the distribution range and richness of EPT in the Atlantic Forest biodiversity hotspot. We found EPT to be at high risk from future climate change, with Plecoptera as the order of greatest concern. We projected range contraction of ca. 90 % of the analyzed EPT genera, resulting in a reduction in the richness of EPT genera under future climatic conditions. We projected >50 % contraction in the distribution of 50 % of Plecoptera, ≈14 % of Trichoptera, and ≈7 % of Ephemeroptera genera. The remaining climatically suitable regions in the Atlantic Forest are concentrated in the high-altitude areas, which may act as climate refuges for EPT biodiversity in the future. The projected changes in EPT's distribution range and richness may impact biomonitoring programs conducted in tropical ecosystems. Restricting EPT's geographic distribution may undermine its potential as a bioindicator and influence the composition of EPT assemblages at reference sites, which may lead to shifting baseline conditions. We reinforce the importance of considering future climatic conditions when planning long-term biomonitoring and priority areas for conservation.

Capturing the carbon in volatile fatty acids (VFA) produced from the anaerobic digestion (AD) of sewage sludge has the potential to not only provide economic benefits but also reduce greenhouse gas production. This study demonstrates a chemical-free method to collect VFA from an AD instead of methane that involves electrochemical pretreatment (EPT) of sludge. Experimental results show that applying 15 V EPT for 45 min enhances acidogenesis and selectively inhibits methanogenesis, leading to a substantial VFA accumulation (2563.1 ± 307.9 mg COD/L) and achieving 2.5 times more carbon fixation than via methane production. Interfacial thermodynamic analysis shows that EPT induces a decrease in both the repulsive electrostatic energy (from 152.9 kT to 12.2 kT) and the energy barrier (from 57.0 kT to 2.6 kT) in the sludge, leading to increased sludge aggregation and entrapment of microorganisms. Molecular docking sheds lights on how the methanogens interacts with the organic matter released from EPT (e.g., alanine-tRNA ligase), showing that these interactions potentially interfere with the proteins that are associated with the activities of the methanogens and the electron transfer pathways, thereby impeding methanogenesis. Integrating EPT into AD therefore facilitates the recovery of valuable VFA and the capture of carbon from freshwater sludge, providing notable economic and environmental benefits in sewage sludge treatment.