Two-Stage Acidic-Alkaline Hydrothermal Pretreatment of Lignocellulose for the High Recovery of Cellulose and Hemicellulose Sugars

The focus of this work was to develop a combined acid and alkaline hydrothermal pretreatment of lignocellulose that ensures high recovery of both hexose and pentose. Dilute sulfuric acid and lime pretreatments were employed sequentially. Process performance was optimized in terms of catalyst concentration, retention time, and temperature using response surface methodology. Medium operational conditions in the acid stage and harsh conditions in the alkaline stage were desirable with optimal performance at 0.73wt% H2SO4, 150°C, 6.1min in the first stage, and 0.024g lime/g biomass, 202°C, 30min in the second stage. In comparison to single-stage pretreatments with high recovery of either glucose or xylose, two-stage process showed great promises with >80% glucose and >70% xylose recovery. In addition, the method greatly improved ethanol fermentation with yields up to 0.145g/g Miscanthus, due to significantly reduced formation of inhibitory by-products such as weak acids, furans, and phenols. Supplementing biomimetic acids would further increase glucose yield by up to 15% and xylose yield by 25

Whole cell biosynthesis of a functional oligosaccharide, 2′-fucosyllactose, using engineered Escherichia coli

BACKGROUND: 2'-Fucosyllactose (2-FL) is a functional oligosaccharide present in human milk which protects against the infection of enteric pathogens. Because 2-FL can be synthesized through the enzymatic fucosylation of lactose with guanosine 5′-diphosphate (GDP)-l-fucose by α-1,2-fucosyltransferase (FucT2), an 2-FL producing Escherichia coli can be constructed through overexpressing genes coding for endogenous GDP- l-fucose biosynthetic enzymes and heterologous fucosyltransferase. RESULTS: The gene for FucT2 from Helicobacter pylori was introduced to the GDP- l-fucose producing recombinant E. coli BL21 star(DE3) strain. However, only small amount of 2-FL was produced in a batch fermentation because the E. coli BL21star(DE3) strain assimilated lactose instead of converting to 2-FL. As an alternative host, the E. coli JM109(DE3) strain which is incapable of assimilating lactose was chosen as a 2-FL producer. Whole cell biosynthesis of 2-FL from lactose was investigated in a series of batch fermentations using various concentrations of lactose. The results of batch fermentations showed that lactose was slowly assimilated by the engineered E. coli JM109(DE3) strain and 2-FL was synthesized without supplementation of another auxiliary sugar for cell growth. A maximum 2-FL concentration of 1.23 g/l was obtained from a batch fermentation with 14.5 g/l lactose. The experimentally obtained yield (g 2-FL/g lactose) corresponded to 20% of the theoretical maximum yield estimated by the elementary flux mode (EFM) analysis. CONCLUSIONS: The experimental 2-FL yield in this study corresponded to about 20% of the theoretical maximum yield, which suggests further modifications via metabolic engineering of a host strain or optimization of fermentation processes might be carried out for improving 2-FL yield. Improvement of microbial production of 2-FL from lactose by engineered E. coli would increase the feasibility of utilizing 2-FL as a prebiotic in various foods

Modulation of Antioxidant Activity Enhances Photoautotrophic Cell Growth of <i>Rhodobacter sphaeroides</i> in Microbial Electrosynthesis

Global warming is currently accelerating due to an increase in greenhouse gas emissions by industrialization. Microbial electrosynthesis (MES) using electroactive autotrophic microorganisms has recently been reported as a method to reduce carbon dioxide, the main culprit of greenhouse gas. However, there are still few cases of application of MES, and the molecular mechanisms are largely unknown. To investigate the growth characteristics in MES, we carried out growth tests according to reducing power sources in Rhodobacter sphaeroides. The growth rate was significantly lower when electrons were directly supplied to cells, compared to when hydrogen was supplied. Through a transcriptome analysis, we found that the expression of reactive oxygen species (ROS)-related genes was meaningfully higher in MES than in normal photoautotrophic conditions. Similarly, endogenous contents of H2O2 were higher and peroxidase activities were lower in MES. The exogenous application of ascorbic acid, a representative biological antioxidant, promotes cell growth by decreasing ROS levels, confirming the inhibitory effects of ROS on MES. Taken together, our observations suggest that reduction of ROS by increasing antioxidant activities is important for enhancing the cell growth and production of CO2-converting substances such as carotenoids in MES in R. sphaeroides</i

Two-Stage Acidic-Alkaline Hydrothermal Pretreatment of Lignocellulose for the High Recovery of Cellulose and Hemicellulose Sugars

ISSN:0273-2289ISSN:1559-029

Molecular Profiling and Optimization Studies for Growth and PHB Production Conditions in Rhodobacter sphaeroides

In the recent climate change regime, industrial demand for renewable materials to replace petroleum-derived polymers continues to rise. Of particular interest is polyhydroxybutyrate (PHB) as a substitute for polypropylene. Accumulating evidence indicates that PHB is highly produced as a carbon storage material in various microorganisms. The effects of growth conditions on PHB production have been widely studied in chemolithotrophs, particularly in Rhodobacter. However, the results on PHB production in Rhodobacter have been somewhat inconsistent due to different strains and experimental conditions, and it is currently unclear how diverse environmental factors are linked with PHB production. Here, we report optimized growth conditions for PHB production and show that the growth conditions are closely related to reactive oxygen species (ROS) regulation. PHB accumulates in cells up to approximately 50% at the highest level under dark-aerobic conditions as opposed to light aerobic/anaerobic conditions. According to the time-course, PHB contents increased at 48 h and then gradually decreased. When observing the effect of temperature and medium composition on PHB production, 30 °C and a carbon/nitrogen ratio of 9:1 or more were found to be most effective. Among PHB biosynthetic genes, PhaA and PhaB are highly correlated with PHB production, whereas PhaC and PhaZ showed little change in overall expression levels. We found that, while the amount of hydrogen peroxide in cells under dark conditions was relatively low compared to the light conditions, peroxidase activities and expression levels of antioxidant-related genes were high. These observations suggest optimal culture conditions for growth and PHB production and the importance of ROS-scavenging signaling with regard to PHB production

On-demand frequency tunability of fluidic antenna implemented with gallium-based liquid metal alloy

We investigated frequency tunability of a microfluidic-based antenna using on-demand manipulation of a gallium-based liquid metal alloy. The fluidic antenna was fabricated by polydimethylsiloxane (PDMS) and filled with the gallium-based liquid metal alloy (Galinstan®). It is composed of a digital number “7”-shaped feedline, and a square-shaped and a digital number “6”-shaped patterns, which are all implemented with the liquid metal. The gallium-based liquid metal was adhered to the channel surface due to its viscous oxide layer originating from the gallium oxide forming when it exposed to the air environment. We treated the liquid metal with hydrochloric acid solution to remove the oxide layer on the surface resulting in easy movement of the liquid metal in the channel, as the liquid metal surface has been transformed to be non-wettable. We controlled the physical length of the liquid metal slug filled in feedline with an applied air pressure, resulting in tuning the resonant frequency ranging from 2.2 GHz to 9.3 GHz. The fluidic antenna properties using the liquid metal’s electrical conductivity and mobility were characterized by measuring the return loss (S11), and also simulated with CST Microwave Studio

Programming characteristics of phase change random access memory using phase change simulations

We present a new simulation methodology for analyzing programming characteristics of a chalcogenide based phase-change device, phase change random access memory (PRAM), which is a next-generation non-volatile memory. Using the new simulation methodology, we analyze the initialization of chalcogenide material (ICM) of the mechanism and propose the next generation PRAM scheme. Front the results of the phase change simulation, the process conditions for ICM for stable operation are presented. Also. the self-heating confined structure to overcome the inherent limitation of high operation power is proposed that resolves the operating power limitation associated with PRAM development