Yeasts with Fermentative Potential Associated with Fruits of Camu-Camu (Myrciaria dubia, Kunth) from North of Brazilian Amazon.

Considering the high biotechnological potential of yeasts associated to edible fruits, a screening for these microorganisms, capable of alcoholic fermentation, was performed in ripe fruits of camu-camu (Myrciaria dubia, Kunth). The fruits were collected from north of Brazilian Amazon, in the floodplain of the Cauamé River. Yeasts were isolated, and fermentation capability was evaluated using Durham tubes. Quantitative assays were performed to calculate ethanol yield (g g-1), specific growth rate (h-1), and ethanol productivity (g L-1·h-1). Taxonomic identification was performed by ribosomal gene nucleotide sequence analysis by alignment using BLASTN. A total of fifteen yeast colonies were isolated, and three of them presented promising ability to ferment glucose to ethanol. These isolates were identified as Candida orthopsilosis, Pichia kudriavzevii, and Meyerozyma caribbica. When cultured in broth containing 180 g·L-1 of glucose, M. caribbica CC003 reached 91.7 percent of the maximum theoretical ethanol concentration (84.4 g·L-1), presenting an ethanol yield and productivity of 0.4688 g·g-1 and 0.781 g·L-1·h-1, respectively. These results indicate a promising potential of this isolate for bioprocess applications. This paper is a rare report of C. orthopsilosis with endophytic habit because most of the references indicate it as a human pathogen. Besides this, M. caribbica is a promising fermenter for alcoholic beverages due to its osmotolerance and high ethanol yield. This is the first paper reporting endophytic yeasts associated with fruits of Myrciaria dubia.

Isolation and Characterization of Yeasts Able to Assimilate Sugarcane Bagasse Hemicellulosic Hydrolysate and Produce Xylitol Associated with Veturius transversus (Passalidae, Coleoptera, and Insecta).

Yeasts are an important component of insect gut microbial content, playing roles such as degradation of polymers and toxic compounds, biological control, and hormone, vitamin, and digestive enzyme production. The xylophagous beetle gut is a hyperdiverse habitat and a potential source of new species with industrial abilities such as enzyme production, pentose fermentation, and biodetoxification. In this work, samples of Veturius transversus (Passalidae, Coleoptera, and Insecta) were collected from the Central Amazon Rainforest. Their guts were dissected and a total of 20 microbial colonies were isolated using sugarcane bagasse hemicellulosic hydrolysate. They were identified as having 10 distinct biochemical profiles, and genetic analysis allowed identification as three clades in the genera Candida, Williopsis, and Geotrichum. All colonies were able to assimilate D-xylose and 18 were able to produce xylitol, especially a strain of Geotrichum, with a maximum yield of 0.502 g·g-1. These results agree with a previous prediction that the microbial community associated with xylophagous insects is a promising source of species of biotechnological interest.

Xylitol production and furfural consumption by a wild type Geotrichum sp

Background: Xylitol is a five carbons polyol with promising medical applications. It can be obtained from chemical D-xylose reduction or by microbial fermentation of Sugarcane Bagasse Hemicellulosic Hydrolysate. For this last process, some microbial inhibitors, as furfural, constitute severe bottleneck. In this case, the use of strains able to produce xylitol simultaneously to furfural neutralization is an interesting alternative. A wild-type strain of Geotrichum sp. was detected with this ability, and its performance in xylitol production and furfural consumption was evaluated. Furthermore, were analyzed its degradation products. Results: Geotrichum sp. produced xylitol from D-xylose fermentation with a yield of 0.44 g-g-1. Furfural was fully consumed in fermentation assay and when provided in the medium until concentration of 6 g-L-1. The furfural degradation product is not an identified molecule, presenting a molecular weight of 161 g-mol-1, an uncommon feature for the microbial metabolism of this product. Conclusion: This strain presents most remarkable potential in performing furfural consumption simultaneous to xylitol production. Subsequent efforts must be employed to establish bioprocess to simultaneous detoxification and xylitol production by Geotrichum sp.

Isolation, taxonomic identification and investigation of the biotechnological potential of wild-type Meyerozyma guilliermondii associated with amazonian termites able to ferment D-xylose / Isolamento, identificação taxonômica e investigação do potencial biotecnológico de linhagem selvagem de Meyerozyma guilliermondii fermentadora de d-xilose associada a cupins da Amazônia

A wild-type yeast that could ferment D-xylose was isolated from the abdominal content of Nasutitermes sp. collected in the Central Amazon rainforest using sugarcane bagasse hemicellulosic hydrolyzate (SBHH) as selective medium. The yeast was identified as Meyerozyma guilliermondii. Its ability to ferment D-xylose was assessed using liquid medium containing Durham tubes. A fermentometer assay showed a low ethanol yield using D-xylose as the carbon source. Cell viability after heat shock and ethanol shock was 39.8% and 56.0%, respectively. Cultivation in SBHH (pH = 5.0) showed its capability to perform saccharification of this substrate, increasing total reducing sugar concentration to 42.6%. The log phase was observed between 36 and 108 hours of cultivation with a highest specific growth rate (µMAX) of 0.10 h-1. After 120 hours, 79.5% of total reducing sugar was consumed giving a biomass yield of 0.52 g/g. The final pH of SBHH (7.6) showed that M. guilliermondii was able to neutralize the acids of this substrate. These results agree with some predictions in the early eighties, which stated that investigations about microbial content of termite guts would provide new tools for bioconversion of lignocellulosic biomass to fuels and other added-value chemicals. This work is the first report for this species associated with termites in the Amazonian habitat.

Uma levedura selvagem fermentadora de D-xilose foi isolada do conteúdo abdominal de Nasutitermes sp., coletado na Amazônia Central usando Hidrolisado Hemicelulósico de Bagaço de Cana-de-açúcar (HHCA) como meio seletivo. A levedura foi identificada como Meyerozyma guilliermondii. Sua capacidade de fermentar D-xilose foi avaliada usando meio líquido contendo tubos de Durham. O isolado demonstrou moderada tolerância ao calor e ao etanol, com viabilidade celular de 39,8% e 56,0%, respectivamente, após submetida a estes fatores limitantes. O ensaio em fermentômetro demonstrou baixo rendimento de etanol usando D-xilose como fonte de carbono. O cultivo em HHCA (pH = 5,0) demonstrou sua capacidade de executar sacarificação e neutralização deste substrato, com aumento da concentração de açúcar redutor total em 42,6% e elevação do pH para 7,6. A fase log foi observada entre 36 e 108 horas de cultivo, com máxima taxa de crescimento específico (µMAX) de 0,10 h-1. Depois de 120 horas, 79,5% do açúcar redutor total foi consumido, com rendimento de biomassa de 0,52 g/g. Estes resultados endossam as predições de alguns autores, os quais propuseram, no início dos anos 80, que a investigação da microbiota intestinal de cupins proveria novas ferramentas para utilização de biomassa lignocelulósica e seus derivados. Este trabalho é o primeiro a reportar a ocorrência de Meyerozyma guilliermondii associada a cupins da Amazônia Central.

Cultivation of Trichosporon mycotoxinivorans in sugarcane bagasse hemicellulosic hydrolyzate

Background: The yeast strain IB09 was isolated from the gut of Calosoma sp. (Carabidae, Coleoptera, Insecta) that were collected in the central Amazon rainforest. First, tolerance of the strain to ethanol and heat was tested. Then, IB09 was cultivated in a medium using sugarcane bagasse hemicellulosic hydrolyzate as a carbon source, and cell growth (OD600), specific growth rate (uMAX, h-1), biomass yield (Y B, g.g-1) and relative sugar consumption (RSC, percent) were evaluated. Taxonomic identification was determined by sequencing the ITS1 region of IB09 and comparing it to sequences obtained from the GenBank database (NCBI). Results: IB09 showed both ethanol tolerance and thermotolerance. Relative sugar consumption indicated that IB09 was able to perform saccharification of sugarcane bagasse hemicellulosic hydrolyzate, increasing the total reducing sugar concentration by approximately 50 percent. The μMAX value obtained was 0,20, indicating that cell growth was slow under the assessed conditions. Biomass yield was 0,701 g per g of consumed sugar, which is relatively high when compared with other findings in the literature. After 120 hrs of cultivation, 80,1 percent of total reducing sugar had been consumed. Sequencing of the ITS1 region identified IB09 as Trichosporon mycotoxinivorans. Conclusion: This is the first report to document this species in the central Amazon rainforest at this host. Trichosporon mycotoxinivorans has great biotechnological potential for use in the saccharification of sugarcane bagasse hemicellulosic hydrolyzate and for biomass production with this substrate as carbon source.