Microbiologia
URI permanente desta comunidadehttps://locus.ufv.br/handle/123456789/11840
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Item Optimizing and validating the production of ethanol from cheese whey permeate by Kluyveromyces marxianus UFV-3(Biocatalysis and Agricultural Biotechnology, 2014-04) Diniz, Raphael H. S.; Rodrigues, Marina Q. R. B.; Fietto, Luciano G.; Passos, Flávia M. L.; Silveira, Wendel B.The purpose of this study was to optimize the production of ethanol from cheese whey permeate using Kluyveromyces marxianus UFV-3. We used the response surface methodology (RSM) with a central composite rotational design (CCRD) to evaluate the effects of pH (4.5–6.5), temperature (30–45 °C), lactose concentration (50–250 g l^−1), and cell biomass concentration (A600 2–4). We performed 29 fermentations under hypoxia in cheese whey permeate and seven fermentations for the validation of the equation obtained via RSM. Temperature was the most significant factor in optimizing ethanol production, followed by pH, cell biomass concentration and lactose concentration. The conditions for producing ethanol at yields above 90% were as follows: temperature between 33.3 and 38.5 °C, pH between 4.7 and 5.7, cell biomass concentration between A600 2.4 and 3.3, and lactose concentration between 50 and 108 g l^−1. The equation generated from the optimization process was validated and exhibited excellent bias and accuracy values for the future use of this model in scaling up the fermentation process.Item Optimizing ethanol production by thermotolerant Kluyveromyces marxianus CCT 7735 in a mixture of sugarcane bagasse and ricotta whey(Food Science and Biotechnology, 2015-08) Ferreira, Priscila Gonçalves; Silveira, Fernando Augusto da; Santos, Raquel Cristina Vieira dos; Genier, Hugo Leonardo André; Diniz, Raphael Hermano Santos; Ribeiro Jr., José Ivo; Fietto, Luciano Gomes; Passos, Flávia Maria LopesThe simultaneous saccharification and fermentation (SSF) process is a promising strategy to obtain ethanol from cellulosic biomass. In this study, sugarcane bagasse was supplemented with ricotta whey to increase the sugar, vitamin, and trace metal concentrations in the fermentation medium. The optimum conditions for SSF ethanol production from a mixture of sugarcane bagasse and ricotta whey produced by Kluyveromyces marxianus CCT 7735 were evaluated considering five factors: cellulase concentration, cellulosic biomass concentration, pH, temperature, and agitation. The highest ethanol yield was 49.65 g/L with a cellulosic biomass of 80 g/L, pH value of 5.05, agitation at 65 rpm and temperature of 39.2°C. The results demonstrated that a mixture of the cellulosic residue of sugarcane bagasse and ricotta whey is promising for ethanol production because the ethanol yield in the mixture was higher than that in single substrate of sugarcane bagasse.Item Optimizing ethanol production by thermotolerant Kluyveromyces marxianus CCT 7735 in a mixture of sugarcane bagasse and ricotta wh(Food Science and Biotechnology, 2015-05-01) Ferreira, Priscila Gonçalves; Silveira, Fernando Augusto da; Santos, Raquel Cristina Vieira dos; Genier, Hugo Leonardo André; Diniz, Raphael Hermano Santos; Ribeiro Jr., José Ivo; Fietto, Luciano Gomes; Passos, Flávia Maria Lopes; Silveira, Wendel Batista daThe simultaneous saccharification and fermentation (SSF) process is a promising strategy to obtain ethanol from cellulosic biomass. In this study, sugarcane bagasse was supplemented with ricotta whey to increase the sugar, vitamin, and trace metal concentrations in the fermentation medium. The optimum conditions for SSF ethanol production from a mixture of sugarcane bagasse and ricotta whey produced by Kluyveromyces marxianus CCT 7735 were evaluated considering five factors: cellulase concentration, cellulosic biomass concentration, pH, temperature, and agitation. The highest ethanol yield was 49.65 g/L with a cellulosic biomass of 80 g/L, pH value of 5.05, agitation at 65 rpm and temperature of 39.2°C. The results demonstrated that a mixture of the cellulosic residue of sugarcane bagasse and ricotta whey is promising for ethanol production because the ethanol yield in the mixture was higher than that in single substrate of sugarcane bagasse.Item Construction of recombinant Kluyveromyces marxianus UFV-3 to express dengue virus type 1 nonstructural protein 1 (NS1)(Applied Microbiology and Biotechnology, 2010-04-02) Bragança, Caio Roberto Soares; Colombo, Lívia Tavares; Roberti, Alvaro Soares; Alvim, Mariana Caroline Tocantins; Cardoso, Silvia Almeida; Reis, Kledna Constancio Portes; Paula, Sérgio Oliveira de; Silveira, Wendel Batista da; Passos, Flavia Maria LopesThe yeast Kluyveromyces marxianus is a convenient host for industrial synthesis of biomolecules. However, despite its potential, there are few studies reporting the expression of heterologous proteins using this yeast. Here, we report expression of a dengue virus protein in K. marxianus for the first time. The dengue virus type 1 nonstructural protein 1 (NS1) was integrated into the K. marxianus UFV-3 genome at the LAC4 locus using an adapted integrative vector designed for high-level expression of recombinant protein in Kluyveromyces lactis. The NS1 gene sequence was codon-optimized to increase the level of protein expression in yeast. The synthetic gene was cloned in frame with K. lactis α-mating factor signal peptide, and the recombinant plasmid obtained was used to transform K. marxianus UFV-3 by electroporation. The transformed cells, selected in yeast extract peptone dextrose containing 200 μg mL−1 Geneticin, were mitotically stable. Analysis of recombinant strains by RT-PCR and protein detection using blot analysis confirmed both transcription and expression of extracellular NS1 polypeptide. After induction with galactose, the NS1 protein was analyzed by sodium dodecyl sulfate-PAGE and immunogenic detection. Protein production was investigated under two conditions: with galactose and biotin pulses at 24-h intervals during 96 h of induction and without galactose and biotin supplementation. Protease activity was not detected in post-growth medium. Our results indicate that recombinant K. marxianus is a good host for the production of dengue virus NS1 protein, which has potential for diagnostic applications.Item Genomic sequence of the yeast Kluyveromyces marxianus CCT 7735 (UFV-3), a highly lactose-fermenting yeast isolated from the Brazilian dairy industry(Genome Announcements, 2014-11-06) Silveira, Wendel B.; Diniz, Raphael H. S.; Cerdán, M. Esperanza; González-Siso, María I.; Souza, Robson de A; Vidigal, Pedro M. P.; Brustolini, Otávio J. B.; Prata, Emille R. B. de Almeida; Medeiros, Alexsandra C.; Paiva, Lílian C.; Nascimento, Moysés; Ferreira, Éder G.; Santos, Valdilene C. dos; Bragança, Caio R. S.; Fernandes, Tatiana A. R.; Colombo, Lívia T.; Passos, Flávia M. L.Here, we present the draft genome sequence of Kluyveromyces marxianus CCT 7735 (UFV-3), including the eight chromosomes and the mitochondrial genomic sequences.