Enzymes and biosurfactants of industrial interest produced by culturable fungi present in sediments of Boeckella Lake, Hope Bay, north-east Antarctic Peninsula.

This study characterized cultivable fungi present in sediments obtained from Boeckella Lake, Hope Bay, in the north-east of the Antarctic Peninsula, and evaluated their production of enzymes and biosurfactants of potential industrial interest. A total of 116 fungal isolates were obtained, which were classified into 16 genera within the phyla Ascomycota, Basidiomycota and Mortierellomycota, in rank. The most abundant genera of filamentous fungi included Pseudogymnoascus, Pseudeurotium and Antarctomyces; for yeasts, Thelebolales and Naganishia taxa were dominant. Overall, the lake sediments exhibited high fungal diversity and moderate richness and dominance. The enzymes esterase, cellulase and protease were the most abundantly produced by these fungi. Ramgea cf. ozimecii, Holtermanniella wattica, Leucosporidium creatinivorum, Leucosporidium sp., Mrakia blollopis, Naganishia sp. and Phenoliferia sp. displayed enzymatic index > 2. Fourteen isolates of filamentous fungi demonstrated an Emulsification Index 24% (EI24%) ≥ 50%; among them, three isolates of A. psychrotrophicus showed an EI24% > 80%. Boeckella Lake itself is in the process of drying out due to the impact of regional climate change, and may be lost completely in approaching decades, therefore hosts a threatened community of cultivable fungi that produce important biomolecules with potential application in biotechnological processes.

Diversity and enzymatic, biosurfactant and phytotoxic activities of culturable Ascomycota fungi present in marine sediments obtained near the South Shetland Islands, maritime Antarctica.

We studied the culturable fungal community recovered from deep marine sediments in the maritime Antarctic, and assessed their capabilities to produce exoenzymes, emulsifiers and metabolites with phytotoxic activity. Sixty-eight Ascomycota fungal isolates were recovered and identified. The most abundant taxon recovered was the yeast Meyerozyma guilliermondii, followed by the filamentous fungi Penicillium chrysogenum, P. cf. palitans, Pseudeurotium cf. bakeri, Thelebolus balaustiformis, Antarctomyces psychrotrophicus and Cladosporium sp. Diversity indices displayed low values overall, with the highest values obtained at shallow depth, decreasing to the deepest location sampled. Only M. guilliermondii and P. cf. palitans were detected in the sediments at all depths sampled, and were the most abundant taxa at all sample sites. The most abundant enzymes detected were proteases, followed by invertases, cellulases, lipases, carrageenases, agarases, pectinases and esterases. Four isolates showed good biosurfactant activity, particularly the endemic species A. psychrotrophicus. Twenty-four isolates of P. cf. palitans displayed strong phytotoxic activities against the models Lactuca sativa and Allium schoenoprasum. The cultivable fungi recovered demonstrated good biosynthetic activity in the production of hydrolytic exoenzymes, biosurfactant molecules and metabolites with phytotoxic activity, reinforcing the importance of documenting the taxonomic, ecological and biotechnological properties of fungi present in deep oceanic sediments of the Southern Ocean.

Fungal diversity in a sediment core from climate change impacted Boeckella Lake, Hope Bay, north-eastern Antarctic Peninsula assessed using metabarcoding.

We studied the fungal DNA present in a lake sediment core obtained from Trinity Peninsula, Hope Bay, north-eastern Antarctic Peninsula, using metabarcoding through high-throughput sequencing (HTS). Sequences obtained were assigned to 146 amplicon sequence variants (ASVs) primarily representing unknown fungi, followed by the phyla Ascomycota, Rozellomycota, Basidiomycota, Chytridiomycota and Mortierellomycota. The most abundant taxa were assigned to Fungal sp., Pseudeurotium hygrophilum, Rozellomycota sp. 1, Pseudeurotiaceae sp. 1 and Chytridiomycota sp. 1. The majority of the DNA reads, representing 40 ASVs, could only be assigned at higher taxonomic levels and may represent taxa not currently included in the sequence databases consulted and/or be previously undescribed fungi. Different sections of the core were characterized by high sequence diversity, richness and moderate ecological dominance indices. The assigned diversity was dominated by cosmopolitan cold-adapted fungi, including known saprotrophic, plant and animal pathogenic and symbiotic taxa. Despite the overall dominance of Ascomycota and Basidiomycota and psychrophilic Mortierellomycota, members of the cryptic phyla Rozellomycota and Chytridiomycota were also detected in abundance. As Boeckella Lake may cease to exist in approaching decades due the effects of local climatic changes, it also an important location for the study of the impacts of these changes on Antarctic microbial diversity.

Diversity of Fungi Present in Permafrost in the South Shetland Islands, Maritime Antarctic.

We assess the fungal diversity present in permafrost from different islands in the South Shetland Islands archipelago, maritime Antarctic, using next-generation sequencing (NGS). We detected 1,003,637 fungal DNA reads representing, in rank abundance order, the phyla Ascomycota, Mortierellomycota, Basidiomycota, Chytridiomycota, Rozellomycota, Mucoromycota, Calcarisporiellomycota and Zoopagomycota. Ten taxa were dominant these being, in order of abundance, Pseudogymnoascus appendiculatus, Penicillium sp., Pseudogymnoascus roseus, Penicillium herquei, Curvularia lunata, Leotiomycetes sp., Mortierella sp. 1, Mortierella fimbricystis, Fungal sp. 1 and Fungal sp. 2. A further 38 taxa had intermediate abundance and 345 were classified as rare. The total fungal community detected in the permafrost showed high indices of diversity, richness and dominance, although these varied between the sampling locations. The use of a metabarcoding approach revealed the presence of DNA of a complex fungal assemblage in the permafrost of the South Shetland Islands including taxa with a range of ecological functions among which were multiple animal, human and plant pathogenic fungi. Further studies are required to determine whether the taxa identified are present in the form of viable cells or propagules and which might be released from melting permafrost to other Antarctic habitats and potentially dispersed more widely.

Fungi in the Antarctic Cryosphere: Using DNA Metabarcoding to Reveal Fungal Diversity in Glacial Ice from the Antarctic Peninsula Region.

We assessed fungal diversity present in glacial from the Antarctic Peninsula using DNA metabarcoding through high-throughput sequencing (HTS). We detected a total of 353,879 fungal DNA reads, representing 94 genera and 184 taxa, in glacial ice fragments obtained from seven sites in the north-west Antarctic Peninsula and South Shetland Islands. The phylum Ascomycota dominated the sequence diversity, followed by Basidiomycota and Mortierellomycota. Penicillium sp., Cladosporium sp., Penicillium atrovenetum, Epicoccum nigrum, Pseudogymnoascus sp. 1, Pseudogymnoascus sp. 2, Phaeosphaeriaceae sp. and Xylaria grammica were the most dominant taxa, respectively. However, the majority of the fungal diversity comprised taxa of rare and intermediate relative abundance, predominately known mesophilic fungi. High indices of diversity and richness were calculated, along with moderate index of dominance, which varied among the different sampling sites. Only 26 (14%) of the total fungal taxa detected were present at all sampling sites. The identified diversity was dominated by saprophytic taxa, followed by known plant and animal pathogens and a low number of symbiotic fungi. Our data suggest that Antarctic glacial ice may represent a hotspot of previously unreported fungal diversity; however, further studies are required to integrate HTS and culture approaches to confirm viability of the taxa detected.

Diversity of fungal DNA in lake sediments on Vega Island, north-east Antarctic Peninsula assessed using DNA metabarcoding.

We assessed the diversity of fungal DNA present in sediments of three lakes on Vega Island, north-east Antarctic Peninsula using metabarcoding through high-throughput sequencing (HTS). A total of 640,902 fungal DNA reads were detected, which were assigned to 224 taxa of the phyla Ascomycota, Rozellomycota, Basidiomycota, Chytridiomycota and Mortierellomycota, in rank order of abundance. The most abundant genera were Pseudogymnoascus, Penicillium and Mortierella. However, a majority (423,508, 66%) of the reads, representing by 43 ASVs, could only be assigned at higher taxonomic levels and may represent taxa not currently included in the sequence databases used or be new or previously unreported taxa present in Antarctic lakes. The three lakes were characterized by high sequence diversity, richness, and moderate dominance indices. The ASVs were dominated by psychrotolerant and cosmopolitan cold-adapted Ascomycota, Basidiomycota and Mortierellomycota commonly reported in Antarctic environments. However, other taxa detected included unidentified members of Rozellomycota and Chytridiomycota species not previously reported in Antarctic lakes. The assigned diversity was composed mainly of taxa recognized as decomposers and pathogens of plants and invertebrates.

Ecological succession of fungal and bacterial communities in Antarctic mosses affected by a fairy ring disease.

We evaluated fungal and bacterial diversity in an established moss carpet on King George Island, Antarctica, affected by 'fairy ring' disease using metabarcoding. A total of 127 fungal and 706 bacterial taxa were assigned. Ascomycota dominated the fungal assemblages, followed by Basidiomycota, Rozellomycota, Chytridiomycota, Mortierellomycota and Monoblepharomycota. The fungal community displayed high indices of diversity, richness and dominance, which increased from healthy through infected to dead moss samples. A range of fungal taxa were more abundant in dead rather than healthy or fairy ring moss samples. Bacterial diversity and richness were greatest in healthy moss and least within the infected fairy ring. The dominant prokaryotic phyla were Actinobacteriota, Proteobacteria, Bacteroidota and Cyanobacteria. Cyanophyceae sp., whilst consistently dominant, were less abundant in fairy ring samples. Our data confirmed the presence and abundance of a range of plant pathogenic fungi, supporting the hypothesis that the disease is linked with multiple fungal taxa. Further studies are required to characterise the interactions between plant pathogenic fungi and their host Antarctic mosses. Monitoring the dynamics of mutualist, phytopathogenic and decomposer microorganisms associated with moss carpets may provide bioindicators of moss health.

Fungal diversity present on rocks from a polar desert in continental Antarctica assessed using DNA metabarcoding.

We evaluated the fungal diversity associated with carbonate veins and two types of salt encrustation in rocks in a polar desert region of continental Antarctica using DNA a metabarcoding approach. We detected 262,268 reads grouped into 517 amplicon sequence variants (ASVs) assigned to the phyla Ascomycota, Basidiomycota, Mortierellomycota and Mucoromycota. Fourteen ASVs belonging to the genera Trichosporon, Mortierella, Penicillium, Aspergillus, Cladosporium, Coprinellus, Pleurotus and Pseudogymnoascus were assessed to be dominant taxa. The fungal communities of the three habitats sampled displayed high diversity indices when compared with other habitats of Antarctica, although differing in detail, with the highest diversity indices in the gypsum crust type 2. Only 48 of the 517 ASVs (9.28%) were detected in all three habitats, including dominant, intermediate and minor components. In contrast with previous studies of fungal communities living in the ultra-extreme conditions of continental Antarctica, application of metabarcoding revealed the DNA of a rich and complex resident fungal community. The ASVs detected included fungi with different ecological roles, with xerophilic, human- and animal-associated, phytopathogenic, saprotrophic, mutualistic, psychrotolerant and cosmopolitan taxa. This sequence diversity may be the result of deposition of fungal propagules over time driven by air currents, precipitation or human activities in the region.

DNA Metabarcoding to Assess the Diversity of Airborne Fungi Present over Keller Peninsula, King George Island, Antarctica.

We assessed fungal diversity present in air samples obtained from King George Island, Antarctica, using DNA metabarcoding through high-throughput sequencing. We detected 186 fungal amplicon sequence variants (ASVs) dominated by the phyla Ascomycota, Basidiomycota, Mortierellomycota, Mucoromycota, and Chytridiomycota. Fungi sp. 1, Agaricomycetes sp. 1, Mortierella parvispora, Mortierella sp. 2, Penicillium sp., Pseudogymnoascus roseus, Microdochium lycopodinum, Mortierella gamsii, Arrhenia sp., Cladosporium sp., Mortierella fimbricystis, Moniliella pollinis, Omphalina sp., Mortierella antarctica, and Pseudogymnoascus appendiculatus were the most dominant ASVs. In addition, several ASVs could only be identified at higher taxonomic levels and may represent previously unknown fungi and/or new records for Antarctica. The fungi detected in the air displayed high indices of diversity, richness, and dominance. The airborne fungal diversity included saprophytic, mutualistic, and plant and animal opportunistic pathogenic taxa. The diversity of taxa detected reinforces the hypothesis that the Antarctic airspora includes fungal propagules of both intra- and inter-continental origin. If regional Antarctic environmental conditions ameliorate further in concert with climate warming, these fungi might be able to reactivate and colonize different Antarctic ecosystems, with as yet unknown consequences for ecosystem function in Antarctica. Further aeromycological studies are necessary to understand how and from where these fungi arrive and move within Antarctica and if environmental changes will encourage the development of non-native fungal species in Antarctica.

Fungi Present in Antarctic Deep-Sea Sediments Assessed Using DNA Metabarcoding.

We assessed fungal diversity in deep-sea sediments obtained from different depths in the Southern Ocean using the internal transcribed spacer 2 (ITS2) region of nuclear ribosomal DNA by metabarcoding through high-throughput sequencing (HTS). We detected 655,991 DNA reads representing 263 fungal amplicon sequence variants (ASVs), dominated by Ascomycota, Basidiomycota, Mortierellomycota, Mucoromycota, Chytridiomycota and Rozellomycota, confirming that deep-sea sediments can represent a hotspot of fungal diversity in Antarctica. The community diversity detected included 17 dominant fungal ASVs, 62 intermediate and 213 rare. The dominant fungi included taxa of Mortierella, Penicillium, Cladosporium, Pseudogymnoascus, Phaeosphaeria and Torula. Despite the extreme conditions of the Southern Ocean benthos, the total fungal community detected in these marine sediments displayed high indices of diversity and richness, and moderate dominance, which varied between the different depths sampled. The highest diversity indices were obtained in sediments from 550 m and 250 m depths. Only 49 ASVs (18.63%) were detected at all the depths sampled, while 16 ASVs were detected only in the deepest sediment sampled at 1463 m. Based on sequence identities, the fungal community included some globally distributed taxa, primarily recorded otherwise from terrestrial environments, suggesting transport from these to deep marine sediments. The assigned taxa included symbionts, decomposers and plant-, animal- and human-pathogenic fungi, suggesting that deep-sea sediments host a complex fungal diversity, although metabarcoding does not itself confirm that living or viable organisms are present.

Performance of xylose-fermenting yeasts in oat and soybean hulls hydrolysate and improvement of ethanol production using immobilized cell systems.

We investigated the fermentation of a mixture of oat and soybean hulls (1:1) subjected to acid (AH) or enzymatic (EH) hydrolyses, with both showing high osmotic pressures (> 1200 Osm kg-1) for the production of ethanol. Yeasts of genera Spathaspora, Scheffersomyces, Sugiymaella, and Candida, most of them biodiverse Brazilian isolates and previously untested in bioprocesses, were cultivated in these hydrolysates. Spathaspora passalidarum UFMG-CM-469 showed the best ethanol production kinetics in suspended cells cultures in acid hydrolysate, under microaerobic and anaerobic conditions. This strain was immobilized in LentiKats® (polyvinyl alcohol) and cultured in AH and EH. Supplementation of hydrolysates with crude yeast extract and peptone was also performed. The highest ethanol production was obtained using hydrolysates supplemented with crude yeast extract (AH-CYE and EH-CYE) showing yields of 0.40 and 0.44 g g-1, and productivities of 0.39 and 0.29 g (L h)-1, respectively. The reuse of the immobilized cells was tested in sequential fermentations of AH-CYE, EH-CYE, and a mixture of acid and enzymatic hydrolysates (AEH-CYE) operated under batch fluidized bed, with ethanol yields ranging from 0.31 to 0.40 g g-1 and productivities from 0.14 to 0.23 g (L h)-1. These results warrant further research using Spathaspora yeasts for second-generation ethanol production.

Whole-genome sequencing of the endemic Antarctic fungus Antarctomyces pellizariae reveals an ice-binding protein, a scarce set of secondary metabolites gene clusters and provides insights on Thelebolales phylogeny.

The snow-covered surfaces of Antarctica comprise an extreme environment that favors the development of life forms with adaptations to adverse low-temperature habitats. The ability to survive and such temperatures might involve the production of antifreeze proteins and ice-binding proteins that attenuate the effects of intense cold temperatures. He, we sequenced and reconstructed the nuclear and mitochondrial genomes of the endemic Antarctic fungus Antarctomyces pellizariae UFMGCB 12416. We then have identified a putative ice-binding protein-coding gene, mapped the presence of secondary metabolite gene clusters, and reconstructed the phylogenetic relationships of a A. pellizariae with others Leotiomycetes from the alignment of hundreds of orthologous single-copy proteins. Our results will deepen the understanding of microbial ice-binding proteins and the genomic aspects of psychrophilic fungi. DATASET: The GenBank/EMBL/DDBJ accession number for the gene sequence of ice-binding protein from A. pellizariae determined in this study is MN867686. The Whole Genome Shotgun project of strain A. pellizariae UFMGCB 12416 has been deposited at DDBJ/ENA/GenBank under accession WCAA00000000. The version described in this paper is version WCAA01000000. The mitochondrial genome has been deposited under accession MT197497.

Diversity and distribution of cultivable fungi present in acid sulphate soils in chronosequence under para-periglacial conditions in King George Island, Antarctica.

We recovered 85 fungal isolates from the acid sulphate soils in chronosequence under para-periglacial conditions in King George Island, Antarctica. Thirty-two taxa belonging to the phylum Ascomycota, Basidiomycota and Mortierellomycota were identified. Mortierella amoeboidea, Mortierella sp. 2, Mortierella sp. 3, Penicillium sp. 2 and Penicillium sp. 3 dominated the sulphite soils. Despite the multi-extreme physic-chemical conditions of the sulphate soils (low pH, variable content of macro and micronutrients and organic matter), the fungal assemblages exhibited moderate diversity indices, which ranged according to the degree of soil development. Soils with more weathered and, consequently, with highest values of organic carbon shelter the most diverse fungal assemblages, which can be associated with the occurrence of sulphurisation and sulphide oxidation. Different taxa of Mortierella and Penicillium displayed broad pH (3-9) and temperature (5-35 °C) plasticity. The multi-extreme sulphite soils of Antarctica revealed the presence of moderate fungal diversity comprising cold cosmopolitan and psychrophilic endemic taxa. Among these, Mortierella and Penicillium, known to survive in extreme conditions such as low temperature and available organic matter, low pH and high concentrations of metals, might represent interesting techniques to be used in biotechnological processes such as bioleaching in metallurgy and phosphate solubilisation in agriculture.

Diversity, distribution, and ecology of viable fungi in permafrost and active layer of Maritime Antarctica.

We evaluated the diversity and distribution of viable fungi present in permafrost and active layers obtained from three islands of Maritime Antarctica. A total of 213 fungal isolates were recovered from the permafrost, and 351 from the active layer, which were identified in 58 taxa; 27 from permafrost and 31 from the active layer. Oidiodendron, Penicillium, and Pseudogymnoascus taxa were the most abundant in permafrost. Bionectriaceae, Helotiales, Mortierellaceae, and Pseudeurotium were the most abundant in the active layer. Only five shared both substrates. The yeast Mrakia blollopis represented is the first reported on Antarctic permafrost. The fungal diversity detected was moderate to high, and composed of cosmopolitan, cold-adapted, and endemic taxa, reported as saprobic, mutualistic, and parasitic species. Our results demonstrate that permafrost shelters viable fungi across the Maritime Antarctica, and that they are contrasting to the overlying active layer. We detected important fungal taxa represented by potential new species, particularly, those genetically close to Pseudogymnoascus destructans, which can cause extinction of bats in North America and Eurasia. The detection of viable fungi trapped in permafrost deserves further studies on the extension of its fungal diversity and its capability to expand from permafrost to other habitats in Antarctica, and elsewhere.

Fungi in glacial ice of Antarctica: diversity, distribution and bioprospecting of bioactive compounds.

We identified cultivable fungi present in the glacial ice fragments collected in nine sites across Antarctica Peninsula and assessed their abilities to produce bioactive compounds. Three ice fragments with approximately 20 kg were collected, melted and 3 L filtered through of 0.45 µm sterilized membranes, which were placed on the media Sabouraud agar and minimal medium incubated at 10 °C. We collected 66 isolates classified into 27 taxa of 14 genera. Penicillium palitans, Penicillium sp. 1, Thelebolus balaustiformis, Glaciozyma antarctica, Penicillium sp. 7, Rhodotorula mucilaginosa, and Rhodotorula dairenensis had the highest frequencies. The diversity and richness of the fungal community were high with moderate dominance. Penicillium species were present in all samples, with Penicillium chrysogenum showing the broadest distribution. P. chrysogenum, P. palitans, and Penicillium spp. had trypanocidal, leishmanicidal, and herbicidal activities, with P. chrysogenum having the broadest and highest capability. 1H NMR signals revealed the presence of highly functionalized secondary metabolites in the bioactive extracts. Despite extreme environmental conditions, glacial ice harbours a diverse fungal community, including species never before recorded in the Arctic and Antarctica. Among them, Penicillium taxa may represent wild fungal strains with genetic and biochemical pathways that may produce new secondary bioactive metabolites.

Production of volatile compounds by yeasts using hydrolysed grape seed oil obtained by immobilized lipases in continuous packed-bed reactors.

Lipases CAL-B, TLL, and RML were used in the synthesis of free fatty acids of grape seed oil as heterogeneous substrate. The best enzyme was used to optimize the reaction variables temperature, enzyme content, and molar ratio of water:oil in batch reactions using experimental planning. The ideal conditions to produce free fatty acids using pure RML were 45 °C, 12:1 substrate molar ratio, and 15% enzyme, resulting in 66% of oil hydrolysis and a productivity of 0.54 mol L-1 min-1 in 4 h of reaction at 180 rpm. Repeated batches of reaction were performed testing the operational stability of RML, results showing that this enzyme could be used for at least 20 cycles keeping more than 80% of its initial activity, suggesting its potential use in industrial processes. The synthesis of free fatty acids was then evaluated in continuous reactions using packed-bed reactor (PBR). The highest productivity in the continuous process was 6.85 mol L-1 min-1, using only RML, showing an operational stability higher than 80% of its initial conversion capacity after 11 days of operation, at a flow rate of 0.13 mL min-1 at 45 °C. We evaluated the use of this hydrolyzed oil as substrate for lactone bioproduction using Galactomyces geotrichum UFMG-CM-Y3276, G. geotrichum UFMG-CM-Y3558, and Geotrichum klebahnii UFMG-CM-Y3014 screened for their oil-hydrolysis ability. Volatile compounds were qualitatively identified in GC-MS as γ-octalactone and γ-nonalactone.

Thiazole derivatives act on virulence factors of Cryptococcus spp.

Cryptococcosis is an opportunistic or primary fungal infection considered to be the most prevalent fatal fungal disease worldwide. Owing to the limited number of available drugs, it is necessary to search for novel antifungal compounds. In the present work, we assessed the antifungal efficacy of three thiazole derivatives (1, 2, and 3). We conducted in vitro and in vivo assays to investigate their effects on important virulence factors, such as capsule and biofilm formation. In addition, the phagocytosis index of murine macrophages exposed to compounds 1, 2, and 3 and the in vivo efficacy of 1, 2, and 3 in Galleria mellonella infected with Cryptococcus spp. were evaluated. All compounds exhibited antifungal activity against biofilms and demonstrated a reduction in biofilm metabolic activity by 43-50% for C. gattii and 26-42% for C. neoformans. Thiazole compounds promoted significant changes in the capsule thickness of C. gattii compared to that of C. neoformans. Further examination of these compounds suggests that they can improve the phagocytosis process of peritoneal murine macrophages in vitro, causing an increase in the phagocytosis rate. Survival percentage was examined in the invertebrate model Galleria mellonella larvae, and only compound 3 could increase the survival at doses of 5 mg/kg after infection with C. gattii (P = .0001) and C. neoformans (P = .0007), similar to fluconazole at 10 mg/kg. The results demonstrated that thiazole compounds, mainly compound 3, have potential to be used for future studies in the search for new therapeutics for cryptococcosis.

Thiosemicarbazone of lapachol acts on cell membrane in Paracoccidioides brasiliensis.

Paracoccidioidomycosis (PCM) is the most prevalent systemic mycosis in Latin American countries. Amphotericin B, sulfonamides, and azoles may be used in the treatment of PCM. However, the high toxicity, prolonged course of treatment, and significant frequency of disease relapse compromise their use. Therefore, there is a need to seek new therapeutic options. We conducted tests with thiosemicarbazone of lapachol (TSC-lap) to determine the antifungal activity and phenotypic effects against several isolates of Paracoccidioides spp. In addition, we evaluated the toxicity against murine macrophages and the ability to enhance phagocytosis. Further, we verified that TSC-lap was active against yeasts but did not show any interaction with the drugs tested. The TSC-lap showed no toxicity at the concentration of 40 µg/ml in macrophages, and at 15.6 µg/ml it could increase the phagocytic index. We observed that this compound induced in vitro ultrastructural changes manifested as withered and broken cells beyond a disorganized cytoplasm with accumulation of granules. We did not observe indications of activity in the cell wall, although membrane damages were noted. We observed alterations in the membrane permeability, culminating in a significant increase in K+ efflux and a gradual loss of the cellular content with increase in the concentration of TSC-lap. In addition, we showed a significant reduction of ergosterol amount in the Pb18 membrane. These data reinforce the possible mechanism of action of this compound to be closely associated with ergosterol biosynthesis and reaffirms the antifungal potential of TSC-lap against Paracoccidioides spp.

Antagonistic lactic acid bacteria in association with Saccharomyces cerevisiae as starter cultures for standardization of sour cassava starch production.

In order to improve cassava's palatability and reduce its toxicity, this root is fermented and applied in foods, such as sour cassava starch used to prepare cheese bread and biscuits. This fermentation occurs spontaneously with lactic acid bacteria (LAB) and yeasts. However, it remains an empirical process, with long duration and lack of product quality homogeneity. This work aims to use starter cultures in a pilot-scale fermentation process for the production of sour cassava starch. After differentiation of strains, Lactobacillus plantarum Lp3, which exhibited great total titratable acidity (TTA) (5.01 ± 0.05%) and antagonistic activity against Bacillus cereus, Escherichia coli and Salmonella Typhimurium, together with Lactobacillus brevis Lb9 (with lesser TTA values: 2.71 ± 0.10%, but amylolytic activity: 2.75 ± 0.61 mm) were tested as single and co-cultures with Saccharomyces cerevisiae UFMG-A1007. LAB and yeasts were inoculated at counts of 8 and 7 log10 CFU/g, respectively, and they remained until the 28th day only in co-culture, highlighting the importance of the yeast for the LAB viability. Although single cultures lead to higher acidity during fermentation, the final product acidity obtained with single cultures did not differ from the acidity obtained with L. plantarum Lp3 in association with S. cerevisiae UFMG-A1007. Therefore, this co-culture exhibited higher potential to be tested as a starter culture in industrial-scale fermentation studies because both microorganisms were in high counts until the end of fermentation and contributed to a final product safe for human consumption, with satisfactory acidity, expansion capacity, and physicochemical properties.

Influence of genetic background of engineered xylose-fermenting industrial Saccharomyces cerevisiae strains for ethanol production from lignocellulosic hydrolysates.

An industrial ethanol-producing Saccharomyces cerevisiae strain with genes of fungal oxido-reductive pathway needed for xylose fermentation integrated into its genome (YRH1415) was used to obtain haploids and diploid isogenic strains. The isogenic strains were more effective in metabolizing xylose than YRH1415 strain and able to co-ferment glucose and xylose in the presence of high concentrations of inhibitors resulting from the hydrolysis of lignocellulosic biomass (switchgrass). The rate of xylose consumption did not appear to be affected by the ploidy of strains or the presence of two copies of the xylose fermentation genes but by heterozygosity of alleles for xylose metabolism in YRH1415. Furthermore, inhibitor tolerance was influenced by the heterozygous genome of the industrial strain, which also showed a marked influenced on tolerance to increasing concentrations of toxic compounds, such as furfural. In this work, selection of haploid derivatives was found to be a useful strategy to develop efficient xylose-fermenting industrial yeast strains.