Proteomics of industrial fungi: trends and insights for biotechnology

Filamentous fungi are widely known for their industrial applications, namely, the production of food-processing enzymes and metabolites such as antibiotics and organic acids. In the past decade, the full genome sequencing of filamentous fungi increased the potential to predict encoded proteins enormously, namely, hydrolytic enzymes or proteins involved in the biosynthesis of metabolites of interest. The integration of genome sequence information with possible phenotypes requires, however, the knowledge of all the proteins in the cell in a system-wise manner, given by proteomics. This review summarises the progress of proteomics and its importance for the study of biotechnological processes in filamentous fungi. A major step forward in proteomics was to couple protein separation with high-resolution mass spectrometry, allowing accurate protein quantification. Despite the fact that most fungal proteomic studies have been focused on proteins from mycelial extracts, many proteins are related to processes which are compartmentalised in the fungal cell, e.g. β-lactam antibiotic production in the microbody. For the study of such processes, a targeted approach is required, e.g. by organelle proteomics. Typical workflows for sample preparation in fungal organelle proteomics are discussed, including homogenisation and sub-cellular fractionation. Finally, examples are presented of fungal organelle proteomic studies, which have enlarged the knowledge on areas of interest to biotechnology, such as protein secretion, energy production or antibiotic biosynthesis

Zooxanthellar Symbionts Shape Host Sponge Trophic Status through Translocation of Carbon

Sponges belonging to the genus Cliona are common inhabitants of many coral reefs, and as bioeroders, they play an important role in the carbonate cycle of the reef. Several Cliona species maintain intracellular populations of dinoflagellate zooxanthellae (i.e., Symbiodinium spp.), which also form symbioses with a variety of other invertebrates and protists (e.g., corals, molluscs, foraminifera). Unlike the case of coral symbioses, however, almost nothing is known of the metabolic interaction between sponges and their zooxanthella symbionts. To assess this interaction, we performed a tracer experiment to follow C and N in the system, performed a reciprocal transplant experiment, and measured the stable carbon isotope ratio of Cliona spp. with and without zooxanthellae to study the influence of environment on the interaction. We found strong evidence of a transfer of C from zooxanthellae to their sponge hosts but no evidence of a transfer of N from sponge to zooxanthellae. We also saw significant influences of the environment on the metabolism of the sponges. Finally, we observed significant differences in carbon metabolism of sponge species with and without symbionts. These data strongly support hypotheses of metabolic integration between zooxanthellae and their sponge host and extend our understanding of basic aspects of benthic-pelagic coupling in shallow-water marine environments

Variability in Fitness Trade-Offs Amongst Coral Juveniles With Mixed Genetic Backgrounds Held in the Wild

Novel restoration methods are currently under consideration worldwide to help coral reefs recover or become more resilient to higher temperature stress. Critical field-based information concerning the paradigm of “local is best” is lacking for many methods; information which is essential to determine the risk and feasibility associated with restoration. One method involves breeding corals from different reef regions with expected variation in heat tolerance and moving those offspring to new locations to enhance offspring survival; thereby augmenting local stock to enhance survival for anticipated warming. In this study, surviving colonies from the 2016 to 2017 mass bleaching events on the Great Barrier Reef (GBR) were reproductively crossed and they included colonies sourced from northern (three) and central (two) reefs. The gravid colonies of Acropora tenuis were collected across 6° of latitude, and they were spawned to produce a total of 17 purebred and hybrid crosses. Juvenile corals (3,748 individual colonies settled on 1,474 terracotta tiles) were deployed to Davies reef in the central GBR after 4 months of aquarium rearing. Survival, growth, and coral colour (as a proxy for bleaching) were assessed after 0, 91, and 217 days of field deployment. Overall, a high percentage of juveniles (17% ± 2.5 SE) survived relative to expected survival at the final census. Survival was significantly higher for central purebred crosses, hybrid crosses had intermediate survival while northern purebreds had the lowest survival. Colour and growth rates (0.001−0.006 mm2 day–1) were not significantly different amongst central, northern, or hybrid crosses but were of a reverse pattern compared to survival. On average, northern purebred crosses grew the fastest, followed by hybrid crosses, and then central purebred crosses. Modelled growth trajectories suggest that northern purebreds would take 8 years to grow to reproductive size, hybrids would take nine, and central purebreds would require 12. All deployed juvenile corals paled over time in the field although the colour of A. tenuis juveniles did not differ significantly amongst central, northern, or hybrid crosses. Growth and survival trade-off analysis showed that although most crosses did not outperform the native central juveniles, two of the eight hybrid crosses (SBxLS, DRxCU) demonstrated faster time to reproductive age and increased survival. Overall, reduced time to reach reproductive size and minimal trade-offs in at least two of the eight hybrids suggest that these crosses may accelerate and supplement recovery through natural re-seeding of genes sourced from northern reefs

IS VO2 SUPRESSED DURING NON-APNOEIC FACIAL SUBMERSION?

The mammalian dive response (DR) is described as oxygen-conserving based on measures of bradycardia, peripheral vasoconstriction and decreased ventilation (VE). Using a model of simulated diving, this study examined the effect of non-apnoeic facial submersions (NAFS) on oxygen consumption (VO2). 19 participants performed four, 2-minute NAFS with 8 minutes of rest between each. Two submersions were performed in 5°C water, two in 25°C water. HR was collected using chest strap monitors. A tube connected to the inspired port of a non-rebreathing valve allowed participants to breathe during facial submersion. Expired air was directed to a metabolic cart to determine VO2 and VE. Baseline (BL) HR, VO2 and VE values were determined by the average during the two minutes prior to facial submersion, cold shock response (CSR) values were the maximum during the first 30 seconds of facial submersion, and NAFS values were the minimum during the last 90 seconds of facial submersion. A two-way repeated measures ANOVA indicated: Both HR and VE were greater during the CSR (92.5±3.6 bpm, 16.3±0.8 L/min) compared to BL (78.9±3.2 bpm, 8.7±0.4 L/min), while both were decreased from BL during the NAFS (60.0±4.0 bpm, 6.0±0.4 L/min) (all, pThe accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

The sesquiterpene lactone parthenolide induces selective apoptosis of B-chronic lymphocytic leukemia cells in vitro.

We have studied the in vitro actions of the sesquiterpene lactone parthenolide (PTL) on cells isolated from patients with chronic lymphocytic leukemia (CLL). Dye reduction viability assays showed that the median LD(50) for PTL was 6.2 muM (n=78). Fifteen of these isolates were relatively resistant to the conventional agent chlorambucil but retained sensitivity to PTL. Brief exposures to PTL (1-3 h) were sufficient to induce caspase activation and commitment to cell death. Chronic lymphocytic leukemia cells were more sensitive towards PTL than were normal T lymphocytes or CD34(+) haematopoietic progenitor cells. The mechanism of cell killing was via PTL-induced generation of reactive oxygen species, resulting in turn in a proapoptotic Bax conformational change, release of mitochondrial cytochrome c and caspase activation. Parthenolide also decreased nuclear levels of the antiapoptotic transcription factor nuclear factor-kappa B and diminished phosphorylation of its negative regulator IkappaB. Killing of CLL cells by PTL was apparently independent of p53 induction. This is the first report showing the relative selectivity of PTL towards CLL cells. The data here warrant further investigation of this class of natural product as potential therapeutic agents for CLL