Produccion de beta-glucosidasas por cultivos de bacterias termofilas indigenas del altiplano boliviano / beta-Glucoside production by thermophilic bacteria indigenous the bolivian altiplano culture

Las beta-glucosidasas son enzimas que poseen actividad hidrolitica y transferasa o transglucosidasa. Tienen diversas aplicaciones; en la biosintesis de oligosacaridos, produccion de etanol utilizando residuos agricolas y en la industria de vinos. La aplicacion industrial, sin embargo, requiere estabilidad a temperaturas elevadas, por lo que los microorganismos termofilos tienen gran interes. El proposito de esta investigacion es el de optimizar el medio de cultivo anaerobio de bacterias termofilas, para aumentar la produccion de beta-glucosidasas. Esta enzima es producida por tres aislados bacterianos: FT3, 2B y P5 los cuales fueron aislados de la region andina de Bolivia. El aislado bacteriano FT3 mostro una actividad beta-glucosidasa de 0,35 [UI/mL]. Se tomaron como variables dentro de la optimizacion del medio de cultivo las fuentes de nitrogeno y de carbono, y el pH. Asi tambien se probaron dos sistemas de cultivo: celulas libres y encapsuladas. Empleando extracto de levadura como fuente de nitrogeno se obtuvo una actividad de 0,52 [UI/mL]. En la optimizacion del pH del medio de cultivo se obtuvo una actividad de 0,81 [UI/mL] a pH 5. Como fuente de carbono se eligieron los hidrolizados de paja de trigo y paja de quinoa lleg¨¢ndose a obtener actividades de 1,27 y 1,34 [UI/mL] respectivamente. Se establecio que la localizacion celular de la enzima beta-glucosidasa es extracelular y presenta estabilidad hasta una temperatura de 80 ºC y un pH de 7.

The beta-glucosidases possess hydrolytic and transferase activity or transglucosidase. They have various applications; such as biosynthesis of oligosaccharides, production of ethanol using agricultural residues and wine industry. However for industrial application, stability to high temperatures is needed. Therefore a great interesting in the thermophile microorganism study exist. The purpose of this research is to optimize the culture medium of thermophilic anaerobic bacteria to increase the production of beta-glucosidase. This enzyme is produced by three isolate bacterial FT3, 2B and P5 which were isolated from the Andean region of Bolivia. FT3 isolate showed beta-glucosidase activity of 0.35 [IU/mL]. In regards to the optimization of culture medium variables such as nitrogen source, carbon source and pH were taken into account and also the combination with free and encapsulated bacterial cells. Yeast extract was the selected source of nitrogen obtaining an activity of 0.52 [IU/ mL]. The optimal pH was 5 obtaining an activity of 0.81 [IU/mL]. The selected carbon source was the hydrolyzed wheat straw and quinoa straw obtaining activities of 1.27 and 1.34 [IU/mL], respectively. The cellular localization of beta-glucosidase enzyme is extracellular and provides stability to temperature of 80 ºC and stability at pH 7.

Ocorrência e biossíntese de frutooligossacarídeos em banana / Occurrence and biosynthesis of fructooligosaccharides in banana

A banana tem sido comumente indicada como uma boa fonte de frutooligossacarídeos (FOS), que são considerados componentes funcionais de alimentos. Contudo, diferenças significantes em suas quantidades têm sido referidas na literatura. Portanto, uma parte do trabalho foi destinada à identificação e quantificação de FOS durante o amadurecimento de cultivares de bananas pertencentes aos grupos genômicos mais comumente cultivados no Brasil. Considerando as diferencas de cultivar, estágio do amadurecimento e metodologia usada para análise de FOS, os conteúdos dos acúcares foram analisados por cromatografia líquida de alta performance (HPAEC-PAD) e cromatografia a gás (CG-MS). Uma pesquisa inicial entre oito cultivares no estágio maduro, mostrou acúmulo de 1-cestose, primeiro membro da série de FOS, em todas elas (quantidades entre 297 e 1600 ´MUg/g M. S´). A nistose, o segundo membro, foi detectado somente no cultivar Prata. Com bases nestes dados, foram escolhidas cinco cultivares, para que fossem analisadas durantes todo o amadurecimento. Os resultados mostraram uma forte correlação entre a síntese de 1-cestose e um nível específico de sacarose (~200mg/g M.S)...

The toxicity of a crude enzyme extract from Gliomastix murorum in Swiss Albino mice.

A crude enzyme extract from a fungus, Gliomastix murorum, could be used in the synthesis of oligosaccharides that are essential to the food and drug industries. This extract may contaminate such products and lead to serious health problems. An investigation on the possible toxicity and mutagenic effect of the extract from this fungal isolate was carried out in Swiss Albino mice. One hundred and 50% of the crude enzyme extract were injected intraperitoneally into the mice every 2 days for 30 days. Normal saline (0.9%), cultivation medium, and cyclophosphamide (80 mg/kg body weight) were given to the control groups. The results indicated that the white blood cell count, serum creatinine, and uric acid of the treated mice were significantly higher than those of the controls (p<0.05), whereas the serum urea-N was lower. For the micronucleus test, mice treated with the extract, especially the group received 100% crude enzyme extract, showed a higher number of micronuclei in polychromatic erythrocytes, as compared to controls. Nevertheless, the micronucleus values were not as high as those found in mice treated with cyclophosphamide, the mutagenic agent. It can be concluded from the results that crude enzyme extract had minor toxic effects on various organ systems tested; more extensive investigation on the safe use of this extract is therefore necessary.