In Vitro Evaluation of Lignin-Containing Nanocellulose.

The increasing importance of environmental sustainability has led to the development of new materials that are environmentally friendly, functional, and cost-effective. Lignin-containing cellulose nanomaterials are a common example of these. The advantages of lignocelluloses include their renewability, sustainability, and functionality combined with molecular rigidity and enhanced hydrophobicity. In order to valorize these beneficial traits from lignin-containing nanocellulose, various approaches have been examined in industrial applications. However, the safety of these materials has not been tested or validated in humans. In this study, we tested 21 wt% lignin-containing nanocellulose (L-MFC) in vitro using the human lung and kidney cell lines, H460 and HEK293 cells, respectively. The cytotoxicity of cellulose, L-MFC, and lignin was compared using the water-soluble tetrazolium salt assays. In addition, the gene expressions of HSP70 and HSP90 as cellular stress markers treated with cellulose, L-MFC, and lignin were quantified using real-time polymerase chain reaction (PCR) and Western blotting. Our data indicated little cytotoxicity for cellulose and significant cytotoxicity for lignin and a relatively low level of cytotoxicity for L-MFC, providing the lethal median concentration (LC50) values of L-MFC and lignin. The gene expression of HSP70 and HSP90 was little affected by moderate concentrations of L-MFC. Interestingly, the lignin contained in L-MFC influenced the cell viability and the gene expression of HSP70 and HSP90 less than the same amount of lignin alone. These results indicate that L-MFC displays cell-type-dependent sensitivity and suggest that L-MFC could serve as a new eco-friendly material that is relatively safe for humans.

Impact of hardwood species on production cost of second generation ethanol.

The present work targeted the understanding of the influence of nine different hardwood species as feedstock on ethanol production yield and costs. It was found that the minimum ethanol revenue (MER) ($ per gallon to the producer) to achieve a 12% internal rate of return (IRR) on invested capital was smaller for low lignin content samples and the influence of species characteristics remained restricted to high residual lignin content. We show that if the pretreatment being applied to the feedstock targets or is limited to low lignin removal, one can expect the species to have a significant impact on overall economics, playing important role to project success. This study also showed a variation of up to 40% in relative MER among hardwood species, where maple, globulus and sweet gum varied the least. Sensitivity analysis showed ethanol yield per ton of feedstock had the largest influence in MER, followed by CAPEX.

Effects of hardwood structural and chemical characteristics on enzymatic hydrolysis for biofuel production.

This study investigated the influence of various hardwood characteristics on enzymatic hydrolysis. Important hardwood species, including three Eucalyptus species, were comprehensively characterized using quantitative (13)C NMR, image analysis and fiber quality analysis. Hydrolysis efficiency from all the hardwoods was correlated to the wood chemical composition and lignin characteristics. Among the key wood components that control enzymatic hydrolysis efficiency, lignin content, enzyme adsorption on substrate and, the ratio of syringyl/guaiacyl (S/G) of the pretreated feedstock were identified as the most important. No wood morphological feature was found to have a significant influence on enzymatic conversion of the pretreated samples.

A comparison of the autohydrolysis and ammonia fiber explosion (AFEX) pretreatments on the subsequent enzymatic hydrolysis of coastal Bermuda grass.

Two distinct pretreatment technologies, autohydrolysis and AFEX, have been applied to coastal Bermuda grass (CBG) followed by enzymatic hydrolysis in order to compare the effects of pretreatment on the subsequent sugar generation. Furthermore, the influence of structural features from each pretreatment on biomass digestibility was characterized with SEM, ATR-FTIR, and XRD. Enzymatic conversion of pretreated solids from the pretreatments increased with elevated temperature and longer residence times. AFEX pretreatment at 100 degrees C for 30 min produced a sugar yield of 94.8% of theoretical possible with 30 FPU/g enzymatic loading, the maximum achieved with AFEX. It was also shown that with autohydrolysis at 170 degrees C for 60 min that 55.4% sugar yield of the theoretical possible was produced with a 30 FPU/g enzymatic loading, the maximum with autohydrolysis. AFEX pretreatment does not change the chemical composition of CBG but autohydrolysis reduces hemicellulose content in the pretreated solids. Both pretreatments cause re-localization of lignin components. There was no observed correlation between crystallinity and enzyme digestibility of the pretreated solids. AFEX pretreatment developed more enzymatic accessibility to pretreated solids of CBG than did autohydrolysis pretreatment, leading to more sugar generation through the whole process. The total amount of sugars accounted for with autohydrolysis decreases with increasing temperature, consistent with increased byproduct generation via thermal degradation reactions.

Autohydrolysis pretreatment of coastal Bermuda grass for increased enzyme hydrolysis.

Coastal Bermuda grass (GBG) was pretreated using an autohydrolysis process with different temperatures and times, and the pretreated materials were enzymatically hydrolyzed using a mixture of cellulase, xylanase and beta-glucosidase with different enzyme loadings to evaluate sugar yields. Compared with untreated CBG, autohydrolysis pretreatments at all elevated temperatures and residence times tested enhanced enzymatic digestibility of both cellulose and hemicellulose. Increasing the temperature and residence time also helps to solubilize hemicelluloses, with 83.3% of the hemicelluloses solubilized at 170 degrees C for 60 min treatment. However, higher temperatures and longer times resulted in an overall lower sugar recovery when considering monosaccharides in the prehydrolyzate combined with the enzyme hydrolyzate. Autohydrolysis at 150 degrees C for 60 min provided the highest overall sugar yield for the entire process. A total of 43.3 g of sugars, 70% of the theoretical sugar yield, can be generated from 100g CBG, 15.0 g of monosaccharide in the prehydrolyzate and 28.3 g in the enzyme hydrolyzate. The conversion efficiency could be further improved by optimizing enzyme dosages and xylanases:cellulases ratio and pretreatment conditions to minimize sugar degradation.