Utilization of nitrate abolishes the "Custers effect" in Dekkera bruxellensis and determines a different pattern of fermentation products

Nitrate is one of the most abundant nitrogen sources in nature. Several yeast species have been shown to be able to assimilate nitrate and nitrite, but the metabolic pathway has been studied in very few of them. Dekkera bruxellensis can use nitrate as sole nitrogen source and this metabolic characteristic can render D. bruxellensis able to overcome S. cerevisiae populations in industrial bioethanol fermentations. In order to better characterize how nitrate utilization affects carbon metabolism and the yields of the fermentation products, we investigated this trait in defined media under well-controlled aerobic and anaerobic conditions. Our experiments showed that in D. bruxellensis, utilization of nitrate determines a different pattern of fermentation products. Acetic acid, instead of ethanol, became in fact the main product of glucose metabolism under aerobic conditions. We have also demonstrated that under anaerobic conditions, nitrate assimilation abolishes the "Custers effect", in this way improving its fermentative metabolism. This can offer a new strategy, besides aeration, to sustain growth and ethanol production for the employment of this yeast in industrial processes

Cold exposure affects carbohydrates and lipid metabolism, and induces Hog1p phosphorylation in Dekkera bruxellensis strain CBS 2499

Dekkera bruxellensis is a yeast known to affect the quality of wine and beer. This species, due to its high ethanol and acid tolerance, has been reported also to compete with Saccharomyces cerevisiae in distilleries producing fuel ethanol. In order to understand how this species responds when exposed to low temperatures, some mechanisms like synthesis and accumulation of intracellular metabolites, changes in lipid composition and activation of the HOG-MAPK pathway were investigated in the genome sequenced strain CBS 2499. We show that cold stress caused intracellular accumulation of glycogen, but did not induce accumulation of trehalose and glycerol. The cellular fatty acid composition changed after the temperature downshift, and a significant increase of palmitoleic acid was observed. RT-PCR analysis revealed that OLE1 encoding for \u3949-fatty acid desaturase was up-regulated, whereas TPS1 and INO1 didn't show changes in their expression. In D. bruxellensis Hog1p was activated by phosphorylation, as described in S. cerevisiae, highlighting a conserved role of the HOG-MAP kinase signaling pathway in cold stress response

A new chemoenzymatic approach to the synthesis of Latanoprost and Bimatoprost

Bimatoprost (1) and Latanoprost (2) are prostaglandin analogues widely used for glaucoma treatment. We have developed a new chemoenzymatic synthesis for 1 and 2, which utilizes a highly stereoselective sequence of biotransformations catalyzed by enzymes belonging to a single microorganism (the yeast Pichia anomala). The original synthesis, starting from (-)-Corey lactone benzoate (3aR,4R,5R,6aS)-3, was modified by replacing three synthetic steps (Cdouble bond, longC reduction, stereoselective Cdouble bond, longO reduction and hydrolysis/deprotection of the benzoate ester) with a one-pot, three-enzymes reaction. The overall biotransformation gave good yields and it was highly stereoselective; noteworthy, by engineering the reaction medium, Cdouble bond, longC reduction could be modulated so that unsaturated (3aR,4R,5R,6aS,3\u2032S)-6 or saturated intermediate (3aR,4R,5R,6aS,3\u2032R)-7 could be preferentially obtained. \ua9 2014 Elsevier B.V. All rights reserved

Co-designing global target-seeking scenarios: A cross-scale participatory process for capturing multiple perspectives on pathways to sustainability

The United Nations 2030 Agenda catalysed the development of global target-seeking sustainability-oriented scenarios representing alternative pathways to reach the Sustainable Development Goals (SDGs). Implementing the SDGs requires connected actions across local, national, regional, and global levels; thus, target-seeking scenarios need to reflect alternative options and tensions across those scales. We argue that the design of global sustainability-oriented target-seeking scenarios requires a consistent process for capturing multiple and contrasting perspectives on how to reach the goals, including the perspectives from multiple scales (e.g. local, national, regional) and geographic regions (e.g. the Global South). Here we propose a novel approach to co-design global target-seeking scenarios, consisting of (a) capturing global perspectives on pathways to the SDGs through a review of existing global scenarios; (b) a multi-stakeholder process to obtain multiple sub-global perspectives on pathways to sustainability; (c) an analysis of convergences, and crucially, divergences between global and regional perspectives on pathways to reach the SDGs, feeding into the design of new target-seeking scenario narratives. As a case study, we use the results of the 2018 African Dialogue on The World in 2050, discussing the future of agriculture and food systems. The identified divergent themes emerging from our analysis included urbanization, population growth, agricultural practices, and the roles of different actors in the future of agriculture. The results challenge some of the existing underlying assumptions of the current sustainability-oriented global scenarios (e.g. population growth, urbanisation, agricultural practices), indicating the relevance and timeliness of the proposed approach. We suggest that similar approaches can be replicated in other contexts to better inform the process of sustainability-oriented scenario co-design across scales, regions and cultures. In addition, we highlight the implications of the approach for scenario quantification and the evolution of modeling tools

Rethinking Cultural Relations and Exchange in the Critical Zone

Chapter for Managing Culture: Reflecting on Exchange in our Global Times, book edited by Raphaela Henze and Victoria Durrer, to be published by Palgrave: Sociology of the Arts

De-Novo Assembly and Analysis of the Heterozygous Triploid Genome of the Wine Spoilage Yeast Dekkera bruxellensis AWRI1499

Despite its industrial importance, the yeast species Dekkera (Brettanomyces) bruxellensis has remained poorly understood at the genetic level. In this study we describe whole genome sequencing and analysis for a prevalent wine spoilage strain, AWRI1499. The 12.7 Mb assembly, consisting of 324 contigs in 99 scaffolds (super-contigs) at 26-fold coverage, exhibits a relatively high density of single nucleotide polymorphisms (SNPs). Haplotype sampling for 1.2% of open reading frames suggested that the D. bruxellensis AWRI1499 genome is comprised of a moderately heterozygous diploid genome, in combination with a divergent haploid genome. Gene content analysis revealed enrichment in membrane proteins, particularly transporters, along with oxidoreductase enzymes. Availability of this assembly and annotation provides a resource for further investigation of genomic organization in this species, and functional characterization of genes that may confer important phenotypic traits

Slaughter weight rather than sex affects carcass cuts and tissue composition of Bisaro pigs

Carcass cuts and tissue composition were assessed in Bisaro pigs (n=64) from two sexes (31 gilts and 33 entire males) reared until three target slaughter body-weights (BW) means: 17 kg, 32 kg, and 79 kg. Dressing percentage and backfat thickness increased whereas carcass shrinkage decreased with increasing BW. Slaughter weight affected most of the carcass cut proportions, except shoulder and thoracic regions. Bone proportion decreased linearly with increasing slaughter BW, while intermuscular and subcutaneous adipose tissue depots increased concomitantly. Slaughter weight increased the subcutaneous adipose tissue proportion but this impaired intramuscular and intermuscular adipose tissues in the loin primal. The sex of the pigs minimally affected the carcass composition, as only the belly weight and the subcutaneous adipose tissue proportions were greater in gilts than in entire males. Light pigs regardless of sex are recommended to balance the trade-offs between carcass cuts and their non-edible compositional outcomes.Work included in the Portuguese PRODER research Project BISOPORC – Pork extensive production of Bísara breed, in two alternative systems: fattening on concentrate vs chesnut, Project PRODER SI I&DT Medida 4.1 “Cooperação para a Inovação”. The authors are grateful to Laboratory of Carcass and Meat Quality of Agriculture School of Polytechnic Institute of Bragança ‘Cantinho do Alfredo’. The authors are members of the MARCARNE network, funded by CYTED (ref. 116RT0503).info:eu-repo/semantics/publishedVersio

Insights into the Dekkera bruxellensis genomic landscape: comparative genomics reveals variations in ploidy and nutrient utilisation potential amongst wine isolates

The yeast Dekkera bruxellensis is a major contaminant of industrial fermentations, such as those used for the production of biofuel and wine, where it outlasts and, under some conditions, outcompetes the major industrial yeast Saccharomyces cerevisiae. In order to investigate the level of inter-strain variation that is present within this economically important species, the genomes of four diverse D. bruxellensis isolates were compared. While each of the four strains was shown to contain a core diploid genome, which is clearly sufficient for survival, two of the four isolates have a third haploid complement of chromosomes. The sequences of these additional haploid genomes were both highly divergent from those comprising the diploid core and divergent between the two triploid strains. Similar to examples in the Saccharomyces spp. clade, where some allotriploids have arisen on the basis of enhanced ability to survive a range of environmental conditions, it is likely these strains are products of two independent hybridisation events that may have involved multiple species or distinct sub-species of Dekkera. Interestingly these triploid strains represent the vast majority (92%) of isolates from across the Australian wine industry, suggesting that the additional set of chromosomes may confer a selective advantage in winery environments that has resulted in these hybrid strains all-but replacing their diploid counterparts in Australian winery settings. In addition to the apparent inter-specific hybridisation events, chromosomal aberrations such as strain-specific insertions and deletions and loss-of-heterozygosity by gene conversion were also commonplace. While these events are likely to have affected many phenotypes across these strains, we have been able to link a specific deletion to the inability to utilise nitrate by some strains of D. bruxellensis, a phenotype that may have direct impacts in the ability for these strains to compete with S. cerevisiae.Anthony R. Borneman, Ryan Zeppel, Paul J. Chambers, Chris D. Curti