Caracterização de populações de leveduras associadas à produção de cachaça artesanal e estudos bioquímicos de metabolismo de sacarose por linhagens de Saccharomyces cerevisiae

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias. Programa de Pós-Graduação em Ciência dos AlimentosA sacarose é o principal açúcar utilizado em processos industriais fermentativos, sua adequada metabolização pela levedura Saccharomyces cerevisiae é fundamental para a otimização desses processos, seja visando o maior rendimento em etanol, a produção de biomassa ou a formação d

Diversity of Saccharomyces cerevisiae strains isolated of the spontaneous fermentation of cachaça from northeastern Brazil / Diversidade de linhagens de Saccharomyces cerevisiae isoladas de fermentações espontâneas de cachaça do nordeste Brasileiro

Cachaça is a beverage obtained by distilling fermented sugar cane juice. The state of Bahia in northeastern Brazil is the second-largest producer of traditional cachaça, and this region has the potential to improve the quality and quantity of its beverage production. The aim of this study was to analyze the genetic diversity of Saccharomyces cerevisiae populations isolated from must in six distilleries in Bahia using mitochondrial DNA restriction fragment length polymorphism (mtDNA-RFLP). Among the three hundred and thirty S. cerevisiae strains isolated, mtDNA-RFLP analysis identified a total of 30 molecular patterns. Analysis of molecular variance (AMOVA) revealed that the greatest genetic variation was found among, rather than within, the populations. Population structure analyses showed the presence of three distinct gene pools, thereby corroborating the AMOVA results. This study represents an important contribution to better understanding the molecular characterization and genetic variability of S. cerevisiae strains during the fermentation of cachaça. The dominant molecular patterns identified here may be used to select S. cerevisiae strains that could improve the quality and volume of traditional cachaça production in Bahia.

Lignocellulose-degrading enzymes production by solid-state fermentation through fungal consortium among Ascomycetes and Basidiomycetes.

In this study, five fungal strains (Aspergillus niger SCBM1 ? Ni, Aspergillus fumigatus SCBM6 ? Fu, Trametes versicolor 561 ? Tr, Ganoderma lucidum 601 ? Ga and Pleurotus ostreatus PL06 ? Pl) were cultivated individually and in consortium for biosynthesis of lignocellulose-degrading enzymes by solid-state fermentation (SSF). The enzyme production was investigated using a 25?1 fractional factorial design, with a total of 16 experiments (F1?F16) using raw sugarcane bagasse and raw wheat bran as substrates. Among the enzymatic extracts produced, Ni (F1) exhibited the highest production of endoglucanase (82.70 U/gds) (units per gram of dry substrate), exoglucanase (80.48 U/gds), ?-xylosidase (145.01 U/gds) and manganese peroxidase (3.38 U/gds). For filter paper cellulase, Tr cocktail (F5) was the one that stood out (9.45 U/gds). Among the extracts produced in consortium, Ni + Tr + Pl (F6) presented the highest production of ?-glucosidase (171.09 U/gds), ?-xylosidase (139.99 U/gds) and manganese peroxidase (3.29 U/gds). For FPase, Ni + Fu + Ga (F12) exhibited the best production (10.46 U/gds). The highest xylanase biosynthesis (2582.38 U/gds) was obtained in Ni + Fu + Pl extract (F4). For laccase, the maximum biosynthesis (25.27 U/gds) was obtained in Tr + Ga + Pl (F13). The cocktails that presented the best enzyme production were: Ni (F1), Ni + Fu + Pl (F9), Ni + Tr + Pl (F6) and Ni + Ga + Pl (F10), demonstrating that the use of microbial consortium can be a promising alternative to obtain enzymatic cocktails with high synergism

Artificially-aged cachaça samples characterised by direct infusion electrospray ionisation mass spectrometry

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Direct infusion electrospray ionisation mass spectrometry in the negative ion mode [ESI()-MS] was employed to evaluate the authenticity of aged cachacas, a traditional and valuable Brazilian alcoholic beverage prepared from the distillation of brewed sugarcane juice and aged in barrels made of common woods. Counterfeit samples were prepared by adding dyes, sawdust or essences to a freshly-distiled, much less valuable sample (white cachaca) to simulate the 1-2 years long natural ageing in wooden barrels. A simple visual inspection revealed remarkable differences between the ESI()-MS of the authentic samples (aged in oak or amburana casks) and the artificially-aged counterfeit samples. A set of diagnostic ions were detectable in the ESI()-MS of the authentic samples aged in oak (m/z 197, 241, 301 and 307) and amburana (m/z 271 and 377/379). This fast and direct methodology seems useful as a routine procedure to monitor this highly profitable and common counterfeit practice. (C) 2013 Elsevier Ltd. All rights reserved.Direct infusion electrospray ionisation mass spectrometry in the negative ion mode [ESI(−)-MS] was employed to evaluate the authenticity of aged cachaças, a traditional and valuable Brazilian alcoholic beverage prepared from the distillation of brewed sug1437781FAPEMIG - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE MINAS GERAISFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)sem informaçãosem informaçãosem informaçã

Hybrid Assembly Improves Genome Quality and Completeness of Trametes villosa CCMB561 and Reveals a Huge Potential for Lignocellulose Breakdown

Trametes villosa is a wood-decaying fungus with great potential to be used in the bioconversion of agro-industrial residues and to obtain high-value-added products, such as biofuels. Nonetheless, the lack of high-quality genomic data hampers studies investigating genetic mechanisms and metabolic pathways in T. villosa, hindering its application in industry. Herein, applying a hybrid assembly pipeline using short reads (Illumina HiSeq) and long reads (Oxford Nanopore MinION), we obtained a high-quality genome for the T. villosa CCMB561 and investigated its genetic potential for lignocellulose breakdown. The new genome possesses 143 contigs, N50 of 1,009,271 bp, a total length of 46,748,415 bp, 14,540 protein-coding genes, 22 secondary metabolite gene clusters, and 426 genes encoding Carbohydrate-Active enzymes. Our CAZome annotation and comparative genomic analyses of nine Trametes spp. genomes revealed T. villosa CCMB561 as the species with the highest number of genes encoding lignin-modifying enzymes and a wide array of genes encoding proteins for the breakdown of cellulose, hemicellulose, and pectin. These results bring to light the potential of this isolate to be applied in the bioconversion of lignocellulose and will support future studies on the expression, regulation, and evolution of genes, proteins, and metabolic pathways regarding the bioconversion of lignocellulosic residues

Additional file 3: of Effectiveness of ITS and sub-regions as DNA barcode markers for the identification of Basidiomycota (Fungi)

List of genera with sequences originated from type specimens and their PCI values (ITS, ITS1, ITS 2) and groups according to the barcode gap analysis. (DOCX 61 kb

Virome analyses of Hevea brasiliensis using small RNA deep sequencing and PCR techniques reveal the presence of a potential new virus

Abstract Background Hevea brasiliensis is an important commercial crop due to the high quality of the latex it produces; however, little is known about viral infections in this plant. The only virus described to infect H. brasiliensis until now is a Carlavirus, which was described more than 30 years ago. Virus-derived small interfering RNA (vsiRNAs) are the product of the plant’s antiviral defense triggered by dsRNA viral intermediates generated, during the replication cycle. These vsiRNAs are complementar to viral genomes and have been widely used to identify and characterize viruses in plants. Methods In the present study, we investigated the virome of leaf and sapwood samples from native H. brasiliensis trees collected in two geographic areas in the Brazilian Amazon. Small RNA (sRNA) deep sequencing and bioinformatic tools were used to assembly, identify and characterize viral contigs. Subsequently, PCR amplification techniques were performed to experimentally verify the presence of the viral sequences. Finally, the phylogenetic relationship of the putative new virus with related viral genomes was analyzed. Results Our strategy allowed the identification of 32 contigs with high similarity to viral reference genomes, from which 23 exhibited homology to viruses of the Tymoviridae family. The reads showed a predominant size distribution at 21 nt derived from both strands, which was consistent with the vsiRNAs profile. The presence and genome position of the viral contigs were experimentally confirmed using droplet digital PCR amplifications. A 1913 aa long fragment was obtained and used to infer the phylogenetic relationship of the putative new virus, which indicated that it is taxonomically related to the Grapevine fleck virus, genus Maculavirus. The putative new virus was named Hevea brasiliensis virus (HBrV) in reference to its host. Conclusion The methodological strategy applied here proved to be efficient in detecting and confirming the presence of new viral sequences on a ‘very difficult to manage’ sample. This is the second time that viral sequences, that could be ascribed as a putative novel virus, associated to the rubber tree has been identified