Trehalose accumulation, invertase activity and physiological characteristics of yeasts isolated from 24 h fermentative cycles during the production of artisanal Brazilian cachaça

Trehalose accumulation, invertase activity and physiological characteristics of 86 yeast isolates from short fermentative cycles during the production of cachaça in three artisanal distilleries of the State of Minas Gerais were studied. Among these isolates, 70 percent were able to grow at temperatures between 40 and 42ºC. Only Saccharomyces cerevisiae isolates were able to grow over 40ºC. Lower temperatures (<40ºC) favoured the growth of yeasts such as Candida parapsilosis-like, C. maltosa-like, Kloeckera japonica, S. exiguus and C. bombicola-like. The isolates from all three distilleries were ethanol tolerant, produced invertase, and accumulate trehalose in the presence of glucose. The strains isolated from distillery A presented more resistance to ethanol (around 84.2 percent of the strains were able to grow in the presence of 12 percent ethanol) when compared to the ones from distilleries C and B (9.5 percent and no strain, respectively). The strains of S. cerevisiae isolated from the three distilleries presented a higher capacity to produce invertase and accumulate trehalose in the presence of glucose. Based on the results of thermal and ethanol stress experiments, it was possible to identify strong relationship between intracellular trehalose accumulation and cell viability. The increase in cell viability was even more pronounced when the strains were subjected to a pre-treatment at sublethal temperatures

Physiological diversity and trehalose accumulation in Schizosaccharomyces pombe strains isolated from spontaneous fermentations during the production of the artisanal Brazilian cachaça.

Twenty-seven Schizosaccharomyces pombe isolates from seven cachaça distilleries were tested for maximum temperature of growth and fermentation, osmotolerance, ethanol resistance, invertase production, and trehalose accumulation. Two isolates were selected for studies of trehalose accumulation under heat shock and ethanol stress. The S. pombe isolates were also characterized by RAPD-PCR. The isolates were able to grow and ferment at 41 degrees C, resisted concentrations of 10% ethanol, and grew on 50% glucose medium. Four isolates yielded invertase activity of more than 100 micromol of reducing sugar x mg(-1) x min(-1). The S. pombe isolates were able to accumulate trehalose during stationary phase. Two isolates, strains UFMG-A533 and UFMG-A1000, submitted to a 15 min heat shock, were able to accumulate high trehalose levels. Strain UFMG-A533 had a marked reduction in viability during heat shock, but strain UFMG-A1000 preserved a viability rate of almost 20% after 15 min at 48 degrees C. No clear correlation was observed between trehalose accumulation and cell survival during ethanol stress. Strain UFMG-A1000 had higher trehalose accumulation levels than strain UFMG-A533 under conditions of combined heat treatment and ethanol stress. Molecular analysis showed that some strains are maintained during the whole cachaça production period; using the RAPD-PCR profiles, it was possible to group the isolates according to their isolation sites.

Trehalose accumulation, invertase activity and physiological characteristics of yeasts isolated from 24 h fermentative cycles during the production of artisanal Brazilian cachaça

Trehalose accumulation, invertase activity and physiological characteristics of 86 yeast isolates from short fermentative cycles during the production of cachaça in three artisanal distilleries of the State of Minas Gerais were studied. Among these isolates, 70% were able to grow at temperatures between 40 and 42ºC. Only Saccharomyces cerevisiae isolates were able to grow over 40ºC. Lower temperatures ( 40ºC) favoured the growth of yeasts such as Candida parapsilosis-like, C. maltosa-like, Kloeckera japonica, S. exiguus and C. bombicola-like. The isolates from all three distilleries were ethanol tolerant, produced invertase, and accumulate trehalose in the presence of glucose. The strains isolated from distillery A presented more resistance to ethanol (around 84.2% of the strains were able to grow in the presence of 12% ethanol) when compared to the ones from distilleries C and B (9.5% and no strain, respectively). The strains of S. cerevisiae isolated from the three distilleries presented a higher capacity to produce invertase and accumulate trehalose in the presence of glucose. Based on the results of thermal and ethanol stress experiments, it was possible to identify strong relationship between intracellular trehalose accumulation and cell viability. The increase in cell viability was even more pronounced when the strains were subjected to a pre-treatment at sublethal temperatures.

As características fisiológicas, acúmulo de trealose e produção de invertase de 86 leveduras isoladas do ciclo fermentativo durante a produção artesanal de cachaça em três destilarias no Estado de Minas Gerais foram estudadas. Destes isolados 70% foram capazes de crescer acima de 40 e 42ºC. Somente os isolados de Saccharomyces cerevisiae foram capazes de crescer acima de 40ºC. Baixas temperaturas ( 40ºC) favoreceram o crescimento de leveduras como Candida parapsilosis-like, C. maltosa-like, Kloeckera japonica, S. exiguus and C. bombicola-like. Os isolados das três destilarias foram tolerantes ao etanol, produziram invertase e acumularam trealose na presença de glicose. As linhagens isoladas da destilaria A apresentaram maior resistência ao etanol (cerca de 84.2% das linhagens foram capazes de crescer na presença de 12% de etanol) quando comparadas àquelas das destilarias B e C (nenhuma linhagem e 9,5%, respectivamente). As linhagens de S. cerevisiae isoladas das três destilarias apresentaram uma alta capacidade para produzir invertase e acumular trealose na presença de glicose. Com base nos resultados dos experimentos de estresse térmico e de etanol, foi possível observar uma forte relação entre acúmulo de trealose intracelular e viabilidade celular. O aumento da viabilidade celular foi mais pronunciado quando as linhagens foram submetidas a um pré-tratamento em temperaturas sub-letais.