Extraction of lignins from aqueous-ionic liquid mixtures by organic solvents

The commercial development of ionic liquids (ILs) to pretreat lignocellulose by dissolution of whole biomass and cellulose precipitation by addition of water is hindered by the absence of an effective technique to recover the lignin content of the biomass from the IL. Three organic solvents [ethyl acetate, 1,4-dioxane, and tetrahydrofuran (THF)] were studied for their ability to form a two-liquid-phase system with water and 1-ethyl-3-methylimidazolium acetate ([C(2)mim][OAc]), and for partitioning model lignins and lignin monomers between the two liquid phases. Ternary diagrams were obtained for three [C(2)mim][OAc]/organic solvent/water systems at 22°C. Partition coefficients were measured for several types of lignin in these three systems. Partition coefficients increase with rising water content in the IL phase, and depend strongly on the type of lignin and on the organic solvent. Partition coefficients rise as the pH of the ionic-liquid-rich phase falls. Small molecule model lignin monomer compounds (guaiacol, syringaldehyde) are also readily extracted from the IL/water system by THF.     

Energy, wealth, and human development: Why and how biomass pretreatment research must improve

A high level of human development is dependent on energy consumption (roughly 4 kW per person), and most developed countries that have reached this level have done so through the extensive use of fossil energy. However, given that fossil resources are finite, in order for developed countries to maintain their level of development and simultaneously allow developing countries to reach their potential, it is essential to develop viable renewable energy alternatives. Of particular importance are liquid fuel replacements for petroleum, the fossil resource that primarily drives commerce and economic growth. The intent of this article is to remind our fellow biofuel researchers, particularly those involved in lignocellulosic pretreatment, of these global issues and the serious nature of our work. We hope that this will inspire us to generate and report higher quality and more thorough data than has been done in the past. Only in this way can accurate comparisons and technoeconomic evaluations be made for the many different pretreatment technologies that are currently being researched. The data that primarily influence biorefinery economics can be subdivided into three main categories: yield, concentration, and rate. For these three categories we detail the specific data that should be reported for pretreatment research. In addition, there is other information that is needed to allow for a thorough comparison of pretreatment technologies. An overview of these criteria and our comparison of the current state of a number of pretreatment technologies with respect to these criteria are covered in the last section. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 28: 893–898, 2012     

Biological pretreatment of sugarcane trash for its conversion to fermentable sugars

Pretreatment of lignocellulosic biomasses, the first step in their conversion to utilizable molecules requires very high energy (steam and electricity), corrosion resistant high-pressure reactors and high temperatures. These severe conditions not only add to the cost component of the entire process but also lead to the loss of sugars to the side reactions. Microbial pretreatments have been reported to be associated with reducing the cost factors as well as the severities of the reactions. Eight bioagents, including fungi and bacteria, were screened for their pretreatment effects on sugarcane trash. They narrowed down the C:N ratio of trash from 108:1 to a varying range of approximately 42:1 to 60:1.The maximum drop in C:N ratio of 61% was observed using Aspergillus terreus followed by Cellulomonas uda (52%) and Trichoderma reesei and Zymomonas mobilis (49%). The bioagents helped in degradation of sugarcane trash by production of cellulases, the maximum being produced by A. terreus, (12 fold) followed by C. uda (10 fold), Cellulomonas cartae (9 fold) and Bacillus macerans (8 fold). The microbial pretreatment of trash rendered the easy accessibility of sugars for enzymatic hydrolysis, which can be directed for production of alcohol.     

Cold-storage of mixed inoculum of Glomus intraradices enhances root colonization,phosphorus status and growth of hot pepper

Growth response of hot pepper (Capsicum annuum L.) inoculated with the arbuscular mycorrhizal (AM) fungus, Glomus intraradices Schenck and Smith was evaluated in a greenhouse study. Three treatments in a soil-based medium amended with rock phosphate were: (1) control (CON), (2) inoculation of G. intraradices as a freshly prepared soil mixture of spores, hyphae and colonized roots of Sorghum vulgare (FM), and (3) inoculation of the fungus as cold-stored mixed inoculum (CM). Colonization at 14 weeks after inoculation with CM was 42.5%, but was significantly lower with FM (14.5%). Inoculation with G. intraradices as FM and CM increased growth of pepper, and total phosphorus and nitrogen uptake in shoots and roots compared with the CON treatment. Inoculation with CM resulted in significant increases in plant dry weight and chlorophyll concentration compared to the FM and CON treatments. Acid phosphatase activity in the rhizosphere was generally increased by AM fungal treatments. Highest acid phosphatase activity occurred at 14 weeks after inoculation with CM. Alkaline phosphatase activity in the CM treatment was significantly higher compared to that in CON and FM treatments throughout the growth period. Thus, cold storage of mixed inoculum enhanced colonization and growth-promoting activity of G. intraradices compared to freshly prepared inoculum.     

Effects of colchicine on anther and microspore culture of bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

The aim of this work was to study the effects of colchicine application on chromosome doubling and androgenic response in anther and microspore culture of different bread wheat genotypes. Colchicine was applied during a mannitol stress pretreatment or during the first 48 h of culture at concentrations of 0, 150 and 300 mg l⁻¹. When colchicine was applied during stress pretreatment, the percentage of doubling depended on genotype and concentration. A significant increase in doubling was observed with 300 mg l⁻¹ in the low androgenic responding cv. Caramba. Colchicine incorporation during the first hours of culture improved percentage of doubling in all genotypes, in both anther and microspore culture. Application of 300 mg l⁻¹ colchicine improved the percentage of doubling in the two low responding genotypes, to 118% of control in DH24033, and 75% in Caramba in microspore and anther culture, respectively. Concerning the androgenic response, the effect of colchicine on embryo formation and percentage of green plants depended on the genotype and on the culture method. In cv. Pavon, a 2- and a 3-fold increase in percentage of embryogenesis and green plants, respectively, were obtained with 300 mg l⁻¹ colchicine in microspore culture. However, no significant differences in these two variables were observed in anther culture. The number of green doubled haploid (DH) plants reflects the index of success of the procedure. Regardless of the culture method, when colchicine was incorporated during the first hours of culture, the number of green DH plants increased significantly in three of four genotypes. These results confirm the usefulness of colchicine application during the first hours of culture in wheat breeding programs.     

Isolation of polyphenols from spent coffee grounds and silverskin by mild hydrothermal pretreatment

In this study, a new method for isolation of polyphenols (PP) from spent coffee grounds (SCG) and coffee silverskin (CS) is described. The method consisted of a mild hydrothermal pretreatment at 120°C, for 20 min, using a liquid-to-solid ratio of 20 mL/g. PP (determined as gallic acid equivalents, GAE) were the most abundant components in the extracts produced by this method, corresponding to 32.92 mg_GAE/g_SCG and 19.17 mg_GAE/g_CS, among which flavonoids corresponded to 8.29 and 2.73 mg quercetin equivalents/g of SCG and CS, respectively. Both extracts presented antioxidant activity but the results were higher for SCG extract, probably due to the highest content of PP present. Negligible effects (less than 1% solubilization) were caused by the hydrothermal pretreatment on cellulose, hemicellulose, and protein fractions of these materials. Some mineral elements were present in the extracts, with potassium being the most abundant. Hydrothermal pretreatment under mild conditions was demonstrated to be an efficient method to recover antioxidant PP from coffee residues.     

Enhanced poly(L-malic acid) production from pretreated cane molasses by Aureobasidium pullulans in fed-batch fermentation

Poly(L-malic acid) (PMA) is a natural polyester with many attractive properties for biomedical application. However, the cost of PMA production is high when glucose is used as a carbon source. To solve this problem, cane molasses as a low-cost feedstock was applied for the production of PMA. Six pretreatment methods were applied to cane molasses before fermentation. Pretreatment with combined tricalcium phosphate, potassium ferrocyanide, and sulfuric acid (TPFSA) removed significant amounts of metal ions from cane molasses. The PMA concentration increased from 5.4?g/L (untreated molasses) to 36.9?g/L (TPFSA-pretreated molasses) after fermentation in shake flasks. A fed-batch fermentation strategy was then developed. In this method, TPFSA-pretreated cane molasses solution was continuously fed into the fermentor to maintain the total sugar concentration at 20?g/L. This technique generated approximately 95.4?g/L PMA with a productivity of 0.57?g/L/hr. The present study indicated that fed-batch fermentation using pretreated cane molasses is a feasible technique for producing high amounts of PMA.     

Circulating muscle-specific microRNA, miR-206, as a potential diagnostic marker for rhabdomyosarcoma

Presently there is no serum biomarker of rhabdomyosarcoma (RMS). Several studies have shown that profiles of microRNA (miRNA) expression differ among tumor types. Here we evaluated the feasibility of using muscle-specific miRNAs (miR-1, -133a, -133b and -206) as biomarkers of RMS. Expression of muscle-specific miRNAs, especially miR-206, was significantly higher in RMS cell lines than in other tumor cell lines, as well as in RMS tumor specimens. Further, serum levels of muscle-specific miRNAs were significantly higher in patients with RMS tumors than in patients with non-RMS tumors. Normalized serum miR-206 expression level could be used to differentiate between RMS and non-RMS tumors, with sensitivity of 1.0 and specificity of 0.913. These results raise the possibility of using circulating muscle-specific miRNAs, especially miR-206, as landmark biomarkers for RMS.     

Transient β-glucuronidase activity after infiltration of Arabidopsis thaliana by Agrobacterium tumefaciens

Transient expression of the β-glucuronidase (GUS) gene introduced into Arabidopsis thaliana intact plants by T-DNA after vacuum infiltration of Agrobacterium tumefaciens was followed. The first incidence of GUS activity was found 2 - 3 d after treatment and a peak of activity one week after treatment in both A. thaliana races, Columbia and C24. GUS activity was sharply increased by cultivation of Arabidopsis plants at elevated temperature (29 °C) compared to cultivation at 25 °C. The density of inocula also influenced the GUS activity. This revised version was published online in July 2006 with corrections to the Cover Date.     

The plant growth-promoting fungus Penicillium simplicissimum GP17-2 induces resistance in Arabidopsis thaliana by activation of multiple defense signals

Arabidopsis thaliana grown in soil amended with barley grain inocula of Penicillium simplicissimum GP17-2 or receiving root treatment with its culture filtrate (CF) exhibited clear resistance to Pseudomonas syringae pv. tomato DC3000 (Pst). To assess the contribution of different defense pathways, Arabidopsis genotypes implicated in salicylic acid (SA) signaling expressing the NahG transgene or carrying disruption in NPR1 (npr1), jasmonic acid (JA) signaling (jar1) and ethylene (ET) signaling (ein2) were tested. All genotypes screened were protected by GP17-2 or its CF. However, the level of protection was significantly lower in NahG and npr1 plants than it was in similarly treated wild-type plants, indicating that the SA signaling pathway makes a minor contribution to the GP17-2-mediated resistance and is insufficient for a full response. Examination of local and systemic gene expression revealed that GP17-2 and its CF modulate the expression of genes involved in both the SA and JA/ET signaling pathways. Subsequent challenge of GP17-2-colonized plants with Pst was accompanied by direct activation of SA-inducible PR-2 and PR-5 genes as well as potentiated expression of the JA-inducible Vsp gene. In contrast, CF-treated plants infected with Pst exhibited elevated expression of most defense-related genes (PR-1, PR-2, PR-5, PDF1.2 and Hel) studied. Moreover, an initial elevation of SA responses was followed by late induction of JA responses during Pst infection of induced systemic resistance (ISR)-expressing plants. In conclusion, we hypothesize the involvement of multiple defense mechanisms leading to an ISR of Arabidopsis by GP17-2.