Diluted phosphoric acid pretreatment for production of fermentable sugars in a sugarcane-based biorefinery.

The influence of time (8-24 min), temperature (144-186 °C) and phosphoric acid concentration (0.05-0.20%, w/v) on the pretreatment of sugarcane bagasse in a 20 L batch rotary reactor was investigated. The efficiency of the pretreatment was verified by chemical characterization of the solid fraction of the pretreated bagasse and the conversion of cellulose to glucose by enzymatic hydrolysis. Models representing the percentage of cellulose, hemicelluloses, lignin, solubilized hemicellulose and the enzymatic conversion of cellulose to glucose were predictive and significant. Phosphoric acid concentration of 0.20% at temperature of 186 °C, during 8 and 24 min, was shown to be very effective in solubilizing hemicellulose from sugarcane bagasse, reaching solubilization of 96% and 98%, respectively. Relatively low amounts of inhibitors were produced, and the phosphoric acid remaining in the hemicellulosic hydrolysate is at adequate levels for supplying phosphorous requirement during subsequent fermentation.

Crabtree-negative characteristics of recombinant xylose-utilizing Saccharomyces cerevisiae.

For recombinant xylose-utilizing Saccharomyces cerevisiae, ethanol yield and productivity is substantially lower on xylose than on glucose. In contrast to glucose, xylose is a novel substrate for S. cerevisiae and it is not known how this substrate is recognized on a molecular level. Failure to activate appropriate genes during xylose-utilization has the potential to result in sub-optimal metabolism and decreased substrate uptake. Certain differences in fermentative performance between the two substrates have thus been ascribed to variations in regulatory response. In this study differences in substrate utilization of glucose and xylose was analyzed in the recombinant S. cerevisiae strain TMB3400. Continuous cultures were performed with glucose and xylose under carbon- and nitrogen-limited conditions. Whereas biomass yield and substrate uptake rate were similar during carbon-limited conditions, the metabolic profile was highly substrate dependent under nitrogen-limited conditions. While glycerol production occurred in both cases, ethanol production was only observed for glucose cultures. Addition of acetate and 2-deoxyglucose pulses to a xylose-limited culture was able to stimulate transient overflow metabolism and ethanol production. Application of glucose pulses enhanced xylose uptake rate under restricted co-substrate concentrations. Results are discussed in relation to regulation of sugar metabolism in Crabtree-positive and -negative yeast.

Growth, sporulation and production of bioactive compounds by Bacillus subtilis R14

The physiology of B. subtilis R14 was investigated in minimal medium under excess-oxygen and oxygen-limited conditions. Growth and efficient sporulation could be achieved in excess-oxygen culture on medium with readily metabolizable carbon and nitrogen sources, which allowed high growth rate and high biomass yield. A short transition phase between the exponential growth and sporulation could be attained by formulating a medium with a well-balanced C/N ratio. Under oxygen-limitation, but in the presence of essential nutrients (i.e. excess-nutrient cultivation), B. subtilis R14 produced bioactive compounds, which showed activity against several phytopathogenic bacteria. Under anaerobic condition, the organism did not grow neither through fermentation nor anaerobic respiration. However, addition of pyruvate to the medium allowed its growth through fermentation and anaerobic respiration. The knowledge acquired in this work could be relevant both for the design of a production process as well as for the formulation of an effective commercial biocontrol product.

A fisiologia de B. subtilis R14 foi investigada em meio mínimo em condições de excesso e limitação de oxigênio. Crescimento e esporulação eficientes foram obtidas em culturas com excesso de oxigênio em meio contendo fontes de carbono e nitrogênio de fácil assimilação, que permitiram alta taxa de crescimento e alto rendimento em biomassa. Uma fase de transição curta, entre o crescimento exponencial e a esporulação, pode ser obtida pela formulação de um meio com razão C/N balanceada. Sob limitação de oxigênio, mas em presença de nutrientes essenciais, B. subtilis R14 produziu compostos bioativos. Os compostos apresentaram atividade contra várias bactérias fitopatogênicas. Sob condição de anaerobiose, o microrganismo não cresceu, nem através de fermentação nem de respiração anaeróbica. A adição de piruvato ao meio permitiu o crescimento, tanto através de fermentação como de respiração anaeróbica. O conhecimento adquirido neste trabalho é relevante tanto para o projeto do processo de produção quanto para a formulação de um produto comercial eficiente para biocontrole.

Antagonism of Bacillus spp. against Xanthomonas campestris pv. campestris

Investigação sobre o antagonismo de oito isolados de Bacillus: B. subtilis R14, B. megaterium pv. cerealis RAB7, B. megaterium pv. cerealis C211, B. megaterium C116, Bacillus sp. RAB9, B. cereus C240, Bacillus sp. C11 e B. cereus C210, contra nove linhagens de X. campestris pv. campestris (bactéria responsável pela podridão negra das crucíferas) foi realizada para se verificar a participação de lipopeptídeos neste mecanismo. Testes de atividades antimicrobiana e hemolítica (surfactante) foram realizados, utilizando-se o método de difusão em ágar. Antibiose e hemólise foram positivas para quatro isolados de Bacillus: R14, RAB7, C116 e C210. A correlação observada entre as atividades antimicrobiana e a hemolítica indica que lipopeptídeos estão envolvidos no mecanismo de antibiose dos isolados investigados. As fermentações foram realizadas com os isolados que demonstraram melhores resultados nos testes de atividades antimicrobiana e hemolítica: R14, RAB7 e C116, para acompanhar o crescimento e a produção de compostos bioativos e surfactantes. As fermentações foram realizadas em mesa agitadora, usando frascos Fernbach. A produção de compostos bioativos e tensoativos foi observada durante a fase final de crescimento dos isolados Bacillus estudados.

Effect of the initial pH on the growth of a proteinase-producing alkalophilic Bacillus species

The subject matter of this work was the investigatioon of the growth of a proteinase-producing alkalophilic Bacillus species as a step towards the development of an industrial process for proteinase production. The basic medium consisted of molasses as the carbon/energy source and urea as the main nitrogen source. Growth in media with different initial pH values was investigated. It was shown that the microorganism was capable of changing the pH of the medium to a final value inside the range 8.5-10 and growth was observed for all initial pH values used. Growth at an initial pH near 10 showed, however, better growth and smoother growth curve. Maximum specific growth rate,umax, of 0.34 h was obtaneid under this condition