Characterization of Blue Agave Bagasse (BAB) as Raw Material for Bioethanol Production Processes by Gravimetric,Thermal, Chromatographic,X-ray Diffraction,Microscopy, and Laser Light Scattering Techniques

A detailed characterization of the main types of blue agave bagasse (BAB) obtained from the four largest tequila factories in the State of Jalisco (Mexico) is presented here. After milling/sieving the agave bagasses, two particle size fractions were identified, one rich in fibers and the other consisting of dust/fine particles. Both fractions were analyzed to determine the content of cellulose, hemicellulose, lignin, organic-soluble compounds, absorbed remaining sugars, minerals, and organic matter. After detailed analyses of both fractions by wet, thermal (thermo-gravimetric analysis (TGA)/differential thermo-gravimetric analysis (DTA)), and other methods (high-performance liquid chromatography (HPLC), microscopy, particle size by laser diffraction light scattering, and crystallinity by X-ray diffraction), a moderate-to-intensive method was devised for further processing the fibrous fraction, which had a high crystalline cellulose content, as well as for its subsequent enzymatic saccharification under well-defined moderate conditions. Alternative processing options were also devised for the dust/fine particle fraction, which has a moderate crystalline cellulose that is rich in adsorbed sugars and that has a high mineral matter content.     

Structural insights into the dynamics and function of the C-terminus of the E. coli RNA chaperone Hfq

The hexameric Escherichia coli RNA chaperone Hfq (Hfq(Ec)) is involved in riboregulation of target mRNAs by small trans-encoded RNAs. Hfq proteins of different bacteria comprise an evolutionarily conserved core, whereas the C-terminus is variable in length. Although the structure of the conserved core has been elucidated for several Hfq proteins, no structural information has yet been obtained for the C-terminus. Using bioinformatics, nuclear magnetic resonance spectroscopy, synchrotron radiation circular dichroism (SRCD) spectroscopy and small angle X-ray scattering we provide for the first time insights into the conformation and dynamic properties of the C-terminal extension of Hfq(Ec). These studies indicate that the C-termini are flexible and extend laterally away from the hexameric core, displaying in this way features typical of intrinsically disordered proteins that facilitate intermolecular interactions. We identified a minimal, intrinsically disordered region of the C-terminus supporting the interactions with longer RNA fragments. This minimal region together with rest of the C-terminal extension provides a flexible moiety capable of tethering long and structurally diverse RNA molecules. Furthermore, SRCD spectroscopy supported the hypothesis that RNA fragments exceeding a certain length interact with the C-termini of Hfq(Ec).     

Isolation and characterization of a new alkali-thermostable lipase cloned from a metagenomic library

The construction of a cosmid library from the biomass produced in an enriched Sequencing Fed-Batch Reactor allowed the isolation of a new lipase by functional screening. The open reading frame of 928 bp encoded a polypeptide of 308 amino acids with a molecular mass of 32.6 kDa. The amino acid sequence analysis revealed the presence of the conserved pentapeptide GXSXG essential for lipase activity. Alignment with known sequences of proteins showed no more than 52% identity with different lipases, confirming the discovery of a novel gene sequence. The lipase was cloned and expressed in Streptomyces lividans and further purified by a combination of hydrophobic interaction and size-exclusion chromatography. Spectrophotometric assays with different p-nitrophenyl esters demonstrated a preference for long-length acyl chains, especially p-nitrophenylmyristate (C14). Moreover, the enzyme presented an optimal activity at 60°C and at alkaline pH of 10.5.     

The immune privilege of the oral mucosa

Despite high bacterial colonization and frequent allergen contact, acute inflammatory and allergic reactions are rarely seen in the oral mucosa. Therefore we assert that immune tolerance predominates at this site and antigen presenting cells, such as dendritic cells and different T cell subtypes, serve as key players in oral mucosal tolerance induction. In this article we describe the mechanisms that lead to tolerance induced in the oral mucosa and how they differ from tolerance induced in the lower gastrointestinal tract. Furthermore we discuss ways in which novel nonparenteral approaches for immune intervention, such as allergen-specific immunotherapy applied by way of the sublingual route, might be improved to target the tolerogenic properties of the sophisticated oral mucosal immune network.     

Highly expressed cholera toxin B subunit in the fruit of a transgenic tomato (<Emphasis Type="Italic">Lycopersicon esculentum</Emphasis> L.)

The cholera toxin B subunit (CTB), a nontoxic molecule with potent biological properties, is a powerful mucosal and parenteral adjuvant that induces a strong immune response against co-administered or coupled antigens. A gene encoding CTB, which was modified based on the optimized codon usage in the plant, was synthesized and fused to the endoplasmic reticulum retention signal KDEL to enhance its expression level in plants. The synthetic CTB (sCTB) gene was introduced into a plant expression vector adjacent to the CaMV 35S promoter, and was transformed into tomato using an Agrobacterium-mediated transformation method. The integration of the sCTB gene into the genomic DNA of transgenic plants was confirmed by genomic DNA PCR amplification. The synthesis and assembly of CTB protein in transgenic plants was demonstrated through immunoblot analysis and G_M1-ELISA. The highest amount of CTB protein produced in transgenic tomatoes was approximately 0.9% of total soluble fruit protein which was 10-fold greater than the previously 0.081%. G_M1-ELISA indicated that plant-synthesized CTB protein bound specifically to G_M1-gangliosides, suggesting that the CTB subunits formed active pentamers.     

Chlorine transport in a small catchment in southeast Sweden during two years

Previous studies have revealed that chlorine participates in a complex biogeochemical cycle in soil, which suggests that the transport of chloride through catchments may also be influenced. The present study is based on field observations of organic carbon, chloride (Cl_in), and chlorinated organic carbon (Cl_org) in precipitation, soil, and runoff over a 2-year period from a small, forested catchment in southeast Sweden. The study reveals that (1) the soil pool is dominated by Cl_org, (2) the input via wet deposition and output of Cl_in via runoff is 30 times smaller than the total storage of chlorine (Cl_in + Cl_org) in soil, and (3) the transport is dominated by Cl_in. The organic matter that entered the outlet of the catchment was more chlorinated in the autumn than during the rest of the year, and rain events taking place in low-flow periods had a greater influence on TOC, Cl_org, and Cl_in than did rain events taking place in high-flow periods. The seasonal pattern in combination with the low-flow versus high-flow pattern and previous findings of increasing chlorine-to-carbon ratios with soil depth suggests that the chlorine-to-carbon ratio variation in the leached organic matter is due that water preferentially comes from deeper layers in low-flow conditions. This study provides well-founded estimates of Cl_org and Cl_in storage and fluxes for the studied catchment; however, the processes underlying the observed seasonal Cl_org variations and transportation processes need further study.