Improved ethanol productivity and ethanol tolerance through genome shuffling of Saccharomyces cerevisiae and Pichia stipitis

Genome shuffling by recursive protoplast fusion between Saccharomyces cerevisiae and Pichia stipitis also known as Scheffersomyces stipitis resulted in a promising yeast hybrid strain with superior qualities than those of the parental strains in enhancing biofuel production. Our study focused on the substrate utilization, ethanol fermentation, and ethanol tolerance of the hybrids and the parental strains. The parental strain S. cerevisiae is limited to utilize only hexose sugars, and this leads to decrease in the ethanol yield when they are subjected to ethanol production from lignocellulosic biomass which is rich in pentose sugars. To overcome this limitation, we constructed a hybrid yeast strain through genome shuffling which can assimilate all the sugars present in the fermentation medium. After two rounds of recursive protoplast fusion, there was a higher increase in substrate utilization by hybrid SP2-18 compared to parental strain S. cerevisiae. SP2-18 was able to consume 34% of xylose sugar present in the fermentation medium, whereas S. cerevisiae was not able to utilize xylose. Further, the hybrid strain SP2-18 was able to reach an ethanol productivity of 1.03 g L−1 h−1, ethanol yield 0.447 g/g, and ethanol concentration 74.65 g L−1 which was relatively higher than that of the parental strain S. cerevisiae. Furthermore, the hybrid SP2-18 was found to be stable in the production of ethanol. The random amplified polymorphic DNA profile of the yeast hybrid SP2-18 shows the polymorphism between the parental strains indicating the migration of specific sugar metabolizing genes from P. stipitis, while the maximum similarity was with the parent S. cerevisiae

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Impact of formate on the growth and productivity of Clostridium ljungdahlii PETC and Clostridium carboxidivorans P7 grown on syngas

The current energy model based on fossil fuels is coming to an end due to the increase in global energy demand. Biofuels such as ethanol and butanol can be produced through the syngas fermentation by acetogenic bacteria. The present work hypothesizes that formate addition would positively impact kinetic parameters for growth and alcohol production in Clostridium ljungdahlii PETC and Clostridium carboxidivorans P7 by diminishing the need for reducing equivalents. Fermentation experiments were conducted using completely anaerobic batch cultures at different pH values and formate concentrations. PETC cultures were more tolerant to formate concentrations than P7, specially at pH 5.0 and 6.0. Complete growth inhibition of PETC occurred at sodium formate concentrations of 30.0 mM; however, no differences in growth rates were observed at pH 7.0 for the two strains. Incubation at formate concentrations lower than 2.0 mM resulted in increased growth rates for both strains. The most recognizable effects of formate addition on the fermentation products were the increase in the total carbon fixed into acids and alcohols at pH 5.0 and pH 6.0, as well as, a higher ethanol to total products ratio at pH 7.0. Taken all together, these results show the ability of acetogens to use formate diminishing the energy demand for growth, and enhancing strain productivity (AU)

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Saponin as regulator of biofuel: implication for ethnobotanical management of diabetes

There has been a sharp rise in the global prevalence of diabetes, obesity, and their comorbid conditions within the last decade prompting significant research into possible causes and cure via therapeutic intervention and lifestyle adjustments. Here, the molecular bases of antidiabetic plants used in the prehistorical treatment of diabetes and obesity are reviewed with particular focus on saponin as the phytotherapeutic principle. Until recently, the phytotherapeutic potentials of saponins have been masked in the heterogeneity of phytochemicals co-extractable during traditional preparations. With improved technique of purification and cutting edge biological assay methods, saponins have emerged as a regulator of primary biofuel availability through direct interaction with energy metabolism, cell signaling, and gene expression. Specific cases of lipoprotein lipase/peroxisome proliferator-activated receptor (PPAR)-gamma/phosphatidylinositide 3-kinase (PI-3-K)/protein kinase B (Akt) activation, adiponectin gene upregulation, fatty acid binding protein 4 repression (FABP4), and glucose transporter type 4 (Glut4) membrane exocytosis have been documented which provide molecular basis for hypocholesterolemic, hypoglycemic, and anti-obesity manifestations observed in experimental animals following saponin treatment. Although intensified research is required to characterize the pharmacophoric features in saponins exhibiting these interactions, however, this preliminary lead is valuable if the world will be free of diabetes, obesity, hypertension, hyperlipidemia, and atherosclerosis in no distant future

Influence of lipolysis and fatty acid availability on fuel selection during exercise

The aim of the present study was to investigate the influence of substrate availability on fuel selection during exercise. Eight endurance-trained male cyclists performed 90-min exercise at 70 % of their maximal oxygen uptake in a cross-over design, either in rested condition (CON) or the day after 2-h exercise practised at 70 % of maximal oxygen uptake (EX). Subjects were given a sucrose load (0.75 g kg−1 body weight) 45 min after the beginning of the 90-min exercise test. Lipolysis was measured in subcutaneous abdominal adipose tissue (SCAT) by microdialysis and substrate oxidation by indirect calorimetry. Lipid oxidation increased during exercise and tended to decrease during sucrose ingestion in both conditions. Lipid oxidation was higher during the whole experimental period in the EX group (p = 0.004). Interestingly, fuel selection, assessed by the change in respiratory exchange ratio (RER), was increased in the EX session (p = 0.002). This was paralleled by a higher rate of SCAT lipolysis reflected by dialysate glycerol, plasma glycerol, and fatty acids (FA) levels (p < 0.001). Of note, we observed a significant relationship between whole-body fat oxidation and dialysate glycerol in both sessions (r 2 = 0.33, p = 0.02). In conclusion, this study highlights the limiting role of lipolysis and plasma FA availability to whole-body fat oxidation during exercise in endurance-trained subjects. This study shows that adipose tissue lipolysis is a determinant of fuel selection during exercise in healthy subjects

Microbial populations associated with fixed- and floating-bed reactors during a two-stage anaerobic process

Microbial populations associated with methanogenic fixed- or floating-bed bioreactors used for anaerobic digestion of lignocellulosic waste were investigated. Fluorescent in situ hybridization (FISH) was used to characterize microorganisms in samples obtained from different heights in the reactors, which were operated in a semi-continuous manner (feeding and mixing once every 2 days). The FISH results showed that Methanosaeta concilii cells were most numerous at the bottom of both reactors. M. concilii cells were more abundant in the fixed-bed reactor (FXBR), which performed better than the floating-bed reactor (FLBR). Species of the Methanosarcina genera (mainly M. barkeri and M. mazei) were also observed in the FLBR but rarely in the FXBR. Methane production in each of the reactors ranged from 0.29 to 0.33 m3 CH(4)/kg COD(rem) (chemical oxygen demand removed). The removal of volatile fatty acids (VFA; 70-75 h) in the FXBR was more efficient than in the FLBR (AU)

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