Fermentation performance of a Mexican native Clavispora lusitaniae strain for xylitol and ethanol production from xylose, glucose and cellobiose.

Lignocellulose hydrolysates are rich in fermentable sugars such as xylose, cellobiose and glucose, with high potential in the biotechnology industry to obtain bioproducts of higher economic value. Thus, it is important to search for and study new yeast strains that co-consume these sugars to achieve better yields and productivity in the processes. The yeast Clavispora lusitaniae CDBB-L-2031, a native strain isolated from mezcal must, was studied under various culture conditions to potentially produce ethanol and xylitol due to its ability to assimilate xylose, cellobiose and glucose. This yeast produced ethanol under microaerobic conditions with yields of 0.451 gethanol/gglucose and 0.344 gethanol/gcellobiose, when grown on 1% glucose or cellobiose, respectively. In mixtures (0.5% each) of glucose:xylose and glucose:xylose:cellobiose the yields were 0.367 gethanol/gGX and 0. 380 gethanol/gGXC, respectively. Likewise, in identical conditions, C. lusitaniae produced xylitol from xylose with a yield of 0.421 gxylitol/gxylose. In 5% glucose or xylose, this yeast had better ethanol and xylitol titers and yields, respectively. However, glucose negatively affected xylitol production in the mixture of both sugars (3% each), producing only ethanol. Xylose reductase (XR) and xylitol dehydrogenase (XDH) activities were evaluated in cultures growing on xylose or glucose, obtaining the highest values in cultures on xylose at 8 h (25.9 and 6.22 mU/mg, respectively). While in glucose cultures, XR and XDH activities were detected once this substrate was consumed (4.06 and 3.32 mU/mg, respectively). Finally, the XYL1 and XYL2 genes encoding xylose reductase and xylitol dehydrogenase, respectively, were up-regulated by xylose, whereas glucose down-regulated their expression.

A soft sensor based on online biomass measurements for the glucose estimation and control of fed-batch cultures of Bacillus thuringiensis.

On bioprocess engineering, experimental measurements are always a costly part of the modeling effort; therefore, there is a constant need to develop cheaper, simpler, and more efficient methodologies to exploit the information available. The aim of the present work was to develop a soft sensor with the capacity to perform reliable substrate predictions and control in microbial cultures of the fed-batch type, using mainly microbial growth data. This objective was achieved using dielectric spectroscopy technology for online monitoring of microbial growth and hybrid neural networks for online prediction of substrate concentration. The glucose estimator was integrated to a fuzzy logic controller to control the substrate concentration in a fed-batch experiment. Dielectric spectroscopy is a technology sensitive to the air volume fraction in the culture media and the turbulence generated by the agitation; however, the introduction of a polynomial function for the calibration of the permittivity signal allowed biomass estimations with an approximation error of 2%. The methodology presented in this work was successfully implemented for the glucose prediction and control of a fed-batch culture of Bacillus thuringiensis with an approximation error of 6%.

Expression of a codon-optimized ß-glucosidase from Cellulomonas flavigena PR-22 in Saccharomyces cerevisiae for bioethanol production from cellobiose.

Bioethanol is one of the main biofuels produced from the fermentation of saccharified agricultural waste; however, this technology needs to be optimized for profitability. Because the commonly used ethanologenic yeast strains are unable to assimilate cellobiose, several efforts have been made to express cellulose hydrolytic enzymes in these yeasts to produce ethanol from lignocellulose. The C. flavigenabglA gene encoding ß-glucosidase catalytic subunit was optimized for preferential codon usage in S. cerevisiae. The optimized gene, cloned into the episomal vector pRGP-1, was expressed, which led to the secretion of an active ß-glucosidase in transformants of the S. cerevisiae diploid strain 2-24D. The volumetric and specific extracellular enzymatic activities using pNPG as substrate were 155 IU L-1 and 222 IU g-1, respectively, as detected in the supernatant of the cultures of the S. cerevisiae RP2-BGL transformant strain growing in cellobiose (20 g L-1) as the sole carbon source for 48 h. Ethanol production was 5 g L-1 after 96 h of culture, which represented a yield of 0.41 g g-1 of substrate consumed (12 g L-1), equivalent to 76% of the theoretical yield. The S. cerevisiae RP2-BGL strain expressed the ß-glucosidase extracellularly and produced ethanol from cellobiose, which makes this microorganism suitable for application in ethanol production processes with saccharified lignocellulose.

Impact of long-term partial aeration on the removal of 2,4,6-trichlorophenol in an initially methanogenic fluidized bed bioreactor.

A fluidized bed bioreactor (FBBR) was operated for more than 1000 days under two regimes, Methanogenic (M) and Methanogenic-Aerobic (M-A), to remove 2,4,6-trichlorophenol (TCP) and phenol (Phe) from a synthetic wastewater, containing different amounts of TCP and Phe, using different aeration flow-rates (0, 2.13, and 1.06 NL O(2)/L.day). M conditions (80:20 mg/L of TCP:Phe, 0 NL O(2)/L.day) showed similar TCP and Phe removal (>95%). Nevertheless accumulation of 4-chlorophenol (4CP) up to 16 mg/L and Phe up to 4 mg/L was observed, while in M-A conditions (80:20 mg/L of TCP:Phe, 2.13 NL O(2)/L.day) TCP and Phe removal achieved 99.9(+)% and after 70 days no accumulation of intermediates were detected. The increase of TCP and Phe in the influent under M-A conditions from 80:20 to 120:30 mg/L of TCP:Phe did not negatively affect the removal of TCP, intermediates and Phe; in fact, they were similar to those in previous M-A conditions. The decrease in the oxygen flow rate from 2.13 to 1.06 NL O(2)/L.day had no negative effect on pollutant removals, which were as high as in previous two M-A conditions. The specific methanogenic activity of bioparticles of the fluidized bed decreased with long-term partial aeration, starting from 1.097 mmol CH(4)/h.g(TKN) in the M regime (day 60) to <0.02 mmolCH(4)/h.g(TKN) at day 1050, suggesting aerobic regime in the bioreactor rather than an M-A regime. In conclusion, complete removal of TCP and less chlorinated intermediates could be achieved in an initially methanogenic FBBR under conditions of partial aeration, although long-term operation seemed to negatively affect the methanogenic activity of biomass. It is also likely that after extended aeration the microbial community was finally enriched with strains with the ability to attack 2,4,6-TCP under aerobic conditions. This report represents the first evidence of a long exposure to oxygen of an anaerobic microbial consortium that efficiently remove TCP.

Resistencia a la exposición al oxígeno de lodos anaerobios suspendidos / Oxygen exposure resistance in suspended anaerobic sludge

La tolerancia al oxígeno de una biomasa anaerobia suspendida en presencia o ausencia de un sustrato primario (sacarosa) se evaluó en términos de la recuperación de la actividad metanogénica aceticlástica especial (AME) de la biomasa anaerobia y un índice de inhibición 50 por ciento (II50) asociado a la AME. Incubada en presencia de sacarosa. La biomasa anaerobia suspendida mostró resistencia a la exposición al oxígeno; la recuperación de la AME fue >45 por ciento para [O²] iniciales <20 por ciento en el espacio gaseoso, y de 10 a 12 por ciento para [O²] iniciales >20 por ciento en el espacio gaseoso. Cuando fue incubada son fuente, la biomasa suspendida fue mucho más inhibida después de la exposición al oxígeno para [O²] iniciales >20 por ciento. El efecto inhibitorio fue descrito por un II50 elevado /28,6) en contraste con un bajo II50 (5,9) cuando se incubó en presencia de sacarosa. La tolerancia de la biomasa suspendida en este trabajo parece ser del mismo orden de la biomasa anaerobia inmovilizada (gránulos anaerobios) en condiciones de incubación en presencia de sustrato: los II50 fueron 5,9 para todos anaerobios suspendidos (sacarosa) y 5,3 y 2,4 para todos granulares incubados con acetato etanol, respectivamente. La respiración aerobia heterótrofa de los lodos anaerobio floculentos incubados con sacarosa fue cerca de 4 veces mayor que la respiración basal, y la inhibición de la AME descrita por el II50 parece seguir una relación inversa con la respiración aerobia heterótrofa. La relación inversa entre II50 y respiración aerobia heterótrofa se ajustó para datos de todos granulares en la literatura y todos floculentos de este trabajo, y sigue un modelo semi-empírico general, con un coeficiente de correlación de 0,82. Esta relación parece reforzar que uno de los mecanismos principales de protección de los consorcios anaerobios contra la inhibición por oxígeno es la respiración aerobia heterótrofa