Protein extraction method for the proteomic study of Zymomonas mobilis during production of ethanol and levans.

Zymomonas mobilis has aroused considerable interest owing to its rapid metabolism and efficiency in producing ethanol and by-products such as levans, sorbitol, and gluconic acid from simple sugars. We performed a proteomic analysis of Z. mobilis UFPEDA241 to provide a global profile of regulatory proteins. The choice of the methods of extraction and cell lysis are fundamental steps and of great importance for the detection and identification of intra- and extracellular proteins of a proteome. Strains were subjected to protein extraction methods using three different reagents: TRIzol, lysis buffer, and phenol. The optimum method was taken to be the one that produced the greatest quantity and quality of proteins in one dimension for further analysis in two dimensions during the production of ethanol and levans over 72 h. The results showed that the greatest amount of protein was obtained by the phenol method (1.44 ± 0.07 mg/mL), which was significantly different (P < 0.05) to the TRIzol (1.11 ± 0.01 mg/mL), and lysis buffer (0.93 ± 0.01 mg/mL) methods (both with P > 0.05). Fermentation at 20°C produced the highest level of levans, and using two-dimensional electrophoresis and mass spectrometry it was possible to identify 34 differentially expressed spots.

Application of fractional factorial design to levan production by Zymomonas mobilis

Levan is a non-toxic, biologically active, extra cellular polysaccharide composed solely by fructose units. Optimization of levan production by Zymomonas mobilis strain ZAG-12 employing a 2(4-1) fractional factorial design was performed to analyze the influence of the temperature (20, 25 e 30°C) agitation (50, 75 e 100 rpm), and the initial concentrations of both sucrose (150, 200 e 250 g.L-1) and yeast extract (2.0, 3.5 e 5.0g.L-1) on final levan concentration. Aerobic fermentation was performed batchwise in 500mL Pyrex flasks for 72 hours. Biomass, ethanol, levan and sucrose were determined at beginning and also at end of the fermentations. The experiments showed that the final levan concentration depended on initial sucrose concentration, temperature and agitation velocity and that the initial concentration of yeast extract did not influence levan production. However, when the production of ethanol and biomass were considered, it became evident that yeast extract was a significant variable. The best conditions for levan production occurred at 100 rpm agitation, 20°C and 250g.L-1 of initial sucrose resulting in 14.67g.L-1 of levan.

Levana é um polissacarídeo extracelular, biologicamente ativo, não tóxico, contendo em sua estrutura apenas frutose. A maximização da produção de levana, por via fermentativa, pela linhagem de Zymomonas mobilis ZAG-12, foi estudada utilizando-se um planejamento fatorial de dois níveis 2(4-1), variando-se as concentrações iniciais de sacarose (150, 200 e 250 g.L-1) , extrato de levedura (2.0, 3.5 e 5.0 g.L-1), temperatura (20, 25 e 30°C) e agitação (50, 75 e 100 rpm). As fermentações foram desenvolvidas por processos descontínuos em frascos Pyrex roscados, de 500 mL, contendo 300 mL de meio a base de sacarose, por 72 horas. No início e ao final do processo, foram dosados: biomassa, etanol, levana e sacarose como açúcares redutores totais. A análise dos dados mostra que o aumento da produção de levana depende tanto dos efeitos da concentração inicial de sacarose, temperatura e agitação, isoladamente, quanto da interação entre agitação e temperatura na faixa experimental estudada. O extrato de levedura não afeta a produção de levana, entretanto, quando a resposta é produção de etanol e biomassa, fica evidente que essa variável é significativa. Os resultados demonstraram que as melhores condições para a produção em batelada ocorreram com 250g/L de sacarose inicial, 100 rpm de agitação, a 20°C.

Saúde coletiva e direitos humanos: um diálogo possível e necessário / Public health and human rights: a possible and necessary dialogue

Aborda a história dos direitos humanos, na sua relaçäo com a saúde, em contraste, com a a história e a linguagem da saúde coletiva, mais desenvolvida e explorada no meio acadêmico. Visa oferecer informaçöes que venham otimizar o diálogo recém-inaugurado entre a saúde coletiva e os direitos humanos. (CACMS)