Microcalorimetria: uma técnica aplicável ao estudo do diauxismo da Saccharomyces cerevisiae

In this work we first introduce the reader to the basic concepts of biology, bioenergetics and biochemistry, concerning the area of cell biology. Then we explain what diauxism is and an example of this phenomenon, applied to S. cerevisiae, is presented. Finally, thermograms obtained by microcalorimetry, from S. cerevisiae that undergo diauxism, are discussed from a biochemical point of view.

Estudos de proteção da célula de saccharomyces cerevisiae para utilização em reações de redução em meio orgânico

New methodologies for protection of Saccharomyces cerevisiae (FP) cells when supported in montmorillonite K10 (K10), recovered or not with gelatin (G) and in the presence or absence of sucrose (S) are presented. These systems were used for the enantioselective reduction of ethyl acetoacetate and a-chloroacetophenone in hexane, under FP/K10/G/S and FP/S at 20ºC during 24 hours, affording S-(+)-ethyl-3-hydroxybutanoate in 100% conversion and 99% ee, and R-(-)-2-chloro-1-phenylethanol 79% and 78% ee at 20 and 30 ºC, respectivelly.

O emprego de fermento de pão, Saccharomyces cerevisiae, na síntese de feromônios

The use of pheromones in integrated pest management has been increasing in the last years due to environmental concern. This development is accompanied by the search for simple, efficient and less aggressive synthetic methodologies for the preparation of pheromones. One of these methodologies includes microbiological reactions, more specifically biocatalytic reduction of carbonyl compounds using baker's yeast (Saccharomyces cerevisiae). This review presents the use of baker's yeast as an easy and cheap alternative to obtain enantiomerically enriched compounds employed in the synthesis of pheromones.

Pré-concentração de cádmio com Saccharomyces cerevisiae e determinação em águas fluviais usando espectrometria de emissão óptica com plasma indutivamente acoplado

A preconcentration method based on the use of Saccharomyces cerevisiae as sorbent material is proposed for the determination of Cd(II) in river water. The solid phase extraction was performed in batch mode and the determination of the analyte in the solid phase was easily carried out by introducing a slurry of the yeast (0.0625 g / 2.5 mL) directly into the ICP OES. A limit of detection of 0.11 µg L-1 and a sample throughput in the range of 4 - 54 sample h-1 were obtained. Determinations of cadmium in a certified sample and in real river water samples were in excellent agreement with the expected values.

Estudo do equilíbrio e cinética da biossorção do pb2+ por saccharomyces cerevisiae

The biosorption, based on the use of biomass for removal of ions is distinguished as an innovative and promising technology when compared with the traditional methods. In this context, the aim of the present work is to use Saccharomyces cerevisiae as biosorbent for the retention of Pb2+ metal ions. Factorial design was used for evaluation of the process. The observed equilibrium data were well described by Langmuir and Freundlich adsorption isotherms. The maximum adsorption capacity was 1486.88 mg/g. The results indicated that Saccharomyces cerevisiae is suitable for biosorption of Pb2+ metal ions.

Isolamento de polímeros da parede celular de Saccharomyces cerevisiae e avaliação da atividade antioxidante da manana-proteína isolada

Yeast cell wall contains polymers glucan and mannan-protein that have received much attention with respect to their biological activities. Conventional isolation process involving treatments with hot alkali and acids cause degradation of these polymers. The aim of this paper was to study a low-degrading process for the isolation of glucan and mannan-protein from S. cerevisiae cell wall comprising physic and enzymatic treatments. Yeast cell glucan was obtained in a purity of 87.4% and a yield of 33.7%. The isolated mannan-protein presented antioxidant activity that was increased after thirty minutes of protease treatment. Antioxidant activity was determined by β-carotene/linoleate model system.

comparação entre processos em SHF e em SSF de bagaço de cana-de-açúcar para a produção de etanol por Saccharomyces cerevisiae

In this work, four different process configurations, including three simultaneous saccharification and fermentation (SSF) schemes and one separate hydrolysis and fermentation (SHF) scheme, were compared, at 8% water-insoluble solids, regarding ethanol production from steam-pretreated and alkali-delignified sugar cane bagasse. Two configurations included a 16 h lasting enzymatic presaccharification prior to SSF, and the third one was a classical SSF without presaccharification. Cellulose conversion was higher for the delignified bagasse, and higher in SSF experiments than in SHF. The highest cellulose-to-ethanol conversion (around 60% in 24 h) and maximum ethanol volumetric productivities (0.29-0.30 g/L.h) were achieved in the presaccharification-assisted SSF.

Determinação seletiva de tributilestanho na presença de Sn(IV) em amostras ambientais usando HG-ICP OES e Saccharomyces cerevisiae como material sorvente

A method for selective determination of tributyltin in the presence of Sn(IV) by combining hydride generation inductively coupled plasma optical emission spectrometry (HG-ICP OES) and solid phase extraction (SPE) using baker's yeast is proposed. The procedure is based on selective retention of TBT by the yeast at pH = 6. Detection limit of 1.9 µg L-1 and quantification limit of 6.3 µg L-1 were obtained. TBT and San(IV) were determined in the range of 0-25 µg L-1 and the proposed approach was applied to analyze river water, sea water and biological extracts, with recoveries of 114, 101 and 86%, respectively.

Estudo da fermentação do hidrolisado de batata-doce utilizando diferentes linhagens de Saccharomyces cerevisiae

Ethanol is the most suitable substitute for oil-based fuels. The performance of the fermentation is affected by several factors, therefore the aim of this work was to evaluate the efficiency of the fermentation of a hydrolyzed must of sweet potato using three strains of the Saccharomyces cerevisiae. It was also evaluated the effect of three forms of the processes conduction in the fermentation yield, efficiency and viability of yeast at the end process. Among the parameters evaluated, only the cell viability showed significant difference. The strain PE-2 would be the most suitable for the fermentation of the hydrolysed sweet potato.

Composição química de aguardentes de cana-de-açúcar fermentadas por diferentes cepas de levedura Saccharomyces cerevisiae

The aim of this study was to evaluate the chemical composition of sugar cane spirits, fermented by different commercial Saccharomyces cerevisiae yeast strains and double distilled by pot still. Sugar cane juices were separately fermented by yeasts CA-11, Y-904, BG-1, PE-2, SA-1 and CAT-1 and distilled by pot still according to the methodology used for whisky production. The alcoholic liquids from first and second distillations were analyzed for concentrations of ethanol, volatile acidity, aldehydes, esters, furfural, higher alcohols and methanol. The sugar cane spirits derived from fermentation by the different yeast strains presented distinct chemical compositions.

Efeito de Saccharomyces cerevisiae na produtividade de sorgo e na severidade de doenças foliares no campo

Saccharomyces cerevisiae é uma levedura com potencial para o controle de doenças de plantas, pois apresenta a capacidade de sintetizar compostos antibióticos, habilidade de competição por espaço e nutrientes no filoplano de muitas espécies vegetais, além de possuir elicitores na parede celular. O presente trabalho teve como objetivos avaliar a produtividade de duas cultivares de sorgo (Sorghum bicolor) tratadas com a levedura, e verificar o controle da antracnose, causada por Colletotrichum sublineolum e da mancha foliar, provocada por Exserohilum turcicum. Os experimentos foram conduzidos em campo, em um delineamento de blocos ao acaso, com sete tratamentos e quatro repetições. Aplicações semanais de S. cerevisiae (Fermento Biológico Fleischmann - 25 mg/ml) reduziram significativamente a antracnose em sorgo cv. Tx-398B e melhoraram a produtividade da mesma. Na cv. 910753, uma única aplicação com a levedura foi suficiente para reduzir o progresso da mancha foliar, porém a produtividade não foi elevada por qualquer um dos tratamentos. Conclui-se que S. cerevisiae apresenta um bom desempenho no campo para o controle de doenças foliares em sorgo, melhorando ou não comprometendo a produtividade da cultura, na dependência da cultivar utilizada.

O efeito da complexidade estrutural da fonte de nitrogênio no transporte de amônio em Saccharomyces cerevisiae

O estudo do efeito da complexidade estrutural da fonte de nitrogênio no transporte de amônio em Saccharomyces cerevisiae foi realizado cultivando-se o microrganismo em um meio mínimo contendo glicose e fontes de nitrogênio, variando de um simples sal de amônio (sulfato de amônio) a aminoácidos livres (casaminoácidos) e peptídeos (peptona). O transporte de amônio foi avaliado acompanhando-se a entrada do análogo metilamônio, utilizando duas metodologias diferentes: transporte de metilamônio radioativo e efluxo de potássio acoplado ao transporte de metilamônio em células crescidas em diferentes condições de cultivo. A cinética de transporte de amônio é detectada nos meios contendo peptona e amônio e não no meio suplementado com casaminoácidos, e o transporte medido em diferentes fases de crescimento sugere que o processo é mais estável em células crescidas em peptona. Os resultados descritos neste trabalho indicam que a complexidade estrutural interfere com a expressão do transportador do íon amônio e que a complementação do meio de cultura com uma fonte de nitrogênio na forma de peptídeos é a mais eficiente não só para a expressão do transportador de amônio, mas também de conferir maior estabilidade ao processo.

Efeito de frações parcialmente purificadas de Saccharomyces cerevisiae na germinação de conídios e formação de apressórios por Colletotrichum sublineolum e Colletotrichum lagenarium

O trabalho teve por objetivo verificar o efeito de preparações ou de frações parcialmente purificadas obtidas de S. cerevisiae, autoclavada por 4 horas seqüencialmente, submetidas à cromatografia de troca iônica (CTI) utilizando tampão Tris-HCl ou bicarbonato de amônio, na germinação de esporos (GE) e formação de apressórios (FA) in vitro por C. lagenarium ou C. sublineolum. Para isto, 40 µL de cada preparação ou fração foram colocados em pocinhos de placa de ELISA, juntamente com 40 µL de uma suspensão de esporos (1 x 10(5) conídios/mL) de C. lagenarium ou de C. sublineolum. Após incubação, determinou-se GE e FA. Água destilada esterilizada foi utilizada como controle. Todas as preparações da levedura autoclavada promoveram estímulo da GE, sem a formação de apressórios por ambos os fitopatógenos. Frações provenientes da CTI, com tampão Tris-HCl, induziram a GE por C. sublineolum e C. lagenarium. Na FA de C. lagenarium houve estímulo pelas frações IV, V e VI, sem diferença, no entanto, na FA de C. sublineolum. Para as frações obtidas por CTI, utilizando tampão bicarbonato de amônio, houve estímulo da GE por C. lagenarium nas frações I e IV e efeito inibitório da germinação pelas frações V, VI e VII. Não houve FA na fração I e as demais frações apresentaram efeito inibitório da FA por C. lagenarium. As frações I e II estimularam a GE e a FA por C. sublineolum e demais frações apresentaram efeito inibitório. Assim, evidencia-se a importância da escolha de tampões no processo de purificação de frações de S. cerevisiae, o que pode resultar em frações que estimulem a germinação de esporos de fitopatógenos fúngicos ou em frações com atividade inibitória da germinação, podendo contribuir futuramente no controle de doenças causadas por esses fungos.

Photodynamic DNA damage induced by phycocyanin and its repair in Saccharomyces cerevisiae

In the present study, we analyzed DNA damage induced by phycocyanin (PHY) in the presence of visible light (VL) using a set of repair endonucleases purified from Escherichia coli. We demonstrated that the profile of DNA damage induced by PHY is clearly different from that induced by molecules that exert deleterious effects on DNA involving solely singlet oxygen as reactive species. Most of PHY-induced lesions are single strand breaks and, to a lesser extent, base oxidized sites, which are recognized by Nth, Nfo and Fpg enzymes. High pressure liquid chromatography coupled to electrochemical detection revealed that PHY photosensitization did not induce 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) at detectable levels. DNA repair after PHY photosensitization was also investigated. Plasmid DNA damaged by PHY photosensitization was used to transform a series of Saccharomyces cerevisiae DNA repair mutants. The results revealed that plasmid survival was greatly reduced in rad14 mutants, while the ogg1 mutation did not modify the plasmid survival when compared to that in the wild type. Furthermore, plasmid survival in the ogg1 rad14 double mutant was not different from that in the rad14 single mutant. The results reported here indicate that lethal lesions induced by PHY plus VL are repaired differently by prokaryotic and eukaryotic cells. Morever, nucleotide excision repair seems to play a major role in the recognition and repair of these lesions in Saccharomyces cerevisiae.

Evidence for a modulation of neutral trehalase activity by Ca2+ and cAMP signaling pathways in Saccharomyces cerevisiae

Saccharomyces cerevisiae neutral trehalase (encoded by NTH1) is regulated by cAMP-dependent protein kinase (PKA) and by an endogenous modulator protein. A yeast strain with knockouts of CMK1 and CMK2 genes (cmk1cmk2) and its isogenic control (CMK1CMK2) were used to investigate the role of CaM kinase II in the in vitro activation of neutral trehalase during growth on glucose. In the exponential growth phase, cmk1cmk2 cells exhibited basal trehalase activity and an activation ratio by PKA very similar to that found in CMK1CMK2 cells. At diauxie, even though both cells presented comparable basal trehalase activities, cmk1cmk2 cells showed reduced activation by PKA and lower total trehalase activity when compared to CMK1CMK2 cells. To determine if CaM kinase II regulates NTH1 expression or is involved in post-translational modulation of neutral trehalase activity, NTH1 promoter activity was evaluated using an NTH1-lacZ reporter gene. Similar ß-galactosidase activities were found for CMK1CMK2 and cmk1cmk2 cells, ruling out the role of CaM kinase II in NTH1 expression. Thus, CaM kinase II should act in concert with PKA on the activation of the cryptic form of neutral trehalase. A model for trehalase regulation by CaM kinase II is proposed whereby the target protein for Ca2+/CaM-dependent kinase II phosphorylation is not the neutral trehalase itself. The possible identity of this target protein with the recently identified trehalase-associated protein YLR270Wp is discussed.