Rie Takada

Microwave-assisted pretreatment of recalcitrant softwood in aqueous glycerol

Microwave-assisted pretreatment of recalcitrant softwood in aqueous glycerol containing a series of organic and inorganic acids with different pK(a) values was examined. The pulp obtained by organosolvolysis with 0.1% hydrochloric acid (pK(a) -6) at 180 degrees C for 6 min gave the highest sugar yield, 53.1%, based on the weight of original biomass. The pretreatment efficiency correlated linearly with the pK(a) of the acids, with the exception of malonic and phosphoric acids. Organosolvolysis with 1.0% phosphoric acid (pK(a) 2.15) gave a saccharification yield (50.6%) higher than that expected from its pK(a), while the catalytic effect of malonic acid (pK(a) 2.83) was negligible. Extensive exposure of crystalline and non-crystalline cellulose by the glycerolysis with strong inorganic acids was demonstrated by using fluorescent-labeled recombinant carbohydrate-binding modules (CBMs). Because of the low concentration of the acid catalysts and availability of glycerol as a by-product from biodiesel and fatty acid production, organosolvolysis in glycerol is an appealing process for pretreatment of recalcitrant softwood.

Analysis of exposed cellulose surfaces in pretreated wood biomass using carbohydrate-binding module (CBM)-cyan fluorescent protein (CFP).

In enzymatic saccharification of lignocellulosics, the access of the enzymes to exposed cellulose surfaces is a key initial step in triggering hydrolysis. However, knowledge of the structure–hydrolyzability relationship of the pretreated biomass is still limited. Here we used fluorescent‐labeled recombinant carbohydrate‐binding modules (CBMs) from Clostridium josui as specific markers for crystalline cellulose (CjCBM3) and non‐crystalline cellulose (CjCBM28) to analyze the complex surfaces of wood tissues pretreated with NaOH, NaOH–Na2S (kraft pulping), hydrothermolysis, ball‐milling, and organosolvolysis. Japanese cedar wood, one of the most recalcitrant softwood species was selected for the analysis. The binding analysis clarified the linear dependency of the exposure of crystalline and non‐crystalline cellulose surfaces for enzymatic saccharification yield by the organosolv and kraft delignification processes. Ball‐milling for 5–30 min increased saccharification yield up to 77%, but adsorption by the CjCBM–cyan fluorescent proteins (CFPs) was below 5%. Adsorption of CjCBM–CFPs on the hydrothermolysis pulp were less than half of those for organosolvolysis pulp, in coincidence with low saccharification yields. For all the pretreated wood, crystallinity index was not directly correlated with the overall saccharification yield. Fluorescent microscopy revealed that CjCBM3–CFP and CjCBM28–CFP were site‐specifically adsorbed on external fibrous structures and ruptured or distorted fiber surfaces. The assay system with CBM–CFPs is a powerful measure to estimate the initiation sites of hydrolysis and saccharification yields from chemically delignified wood pulps. Biotechnol. Bioeng. 2010; 105: 499–508.

Surface carbohydrate analysis and bioethanol production of sugarcane bagasse pretreated with the white rot fungus, Ceriporiopsis subvermispora and microwave hydrothermolysis

Effects of pretreatments with a white rot fungus, Ceriporiopsis subvermispora, and microwave hydrothermolysis of bagasse on enzymatic saccharification and fermentation were evaluated. The best sugar yield, 44.9 g per 100g of bagasse was obtained by fungal treatments followed by microwave hydrothermolysis at 180°C for 20 min. Fluorescent-labeled carbohydrate-binding modules which recognize crystalline cellulose (CjCBM3-GFP), non-crystalline cellulose (CjCBM28-GFP) and xylan (CtCBM22-GFP) were applied to characterize the exposed polysaccharides. The microwave pretreatments with and without the fungal cultivation resulted in similar levels of cellulose exposure, but the combined treatment caused more defibration and thinning of the plant tissues. Simultaneous saccharification and fermentation of the pulp fractions obtained by microwave hydrothermolysis with and without fungal treatment, gave ethanol yields of 35.8% and 27.0%, respectively, based on the holocellulose content in the pulp. These results suggest that C. subvermispora pretreatment could be beneficial part of the process to produce ethanol from bagasse.

Recognition of cellooligosaccharides by a family 28 carbohydrate-binding module.

The crystal structures of a carbohydrate‐binding module (CBM) family 28 domain of endoglucanase Cel5A from Clostridium josui have been determined in ligand‐free and complex forms with cellobiose, cellotetraose, and cellopentaose as the first complex structures of this family. In the cleft of a β‐sandwich fold, the ligands are recognized by stacking interactions and hydrogen bonds. Conformations of the bound cellooligosaccharides are similar to those in crystals and solution but clearly different from the cellulose structure. Interestingly, the glucan chain bound on CBM28 is in the opposite direction of that bound to CBM17, although these families share significant structural similarity.

Chemometric Analysis with Near-Infrared Spectroscopy for Chemically Pretreated Erianthus toward Efficient Bioethanol Production

In this paper, we report the combination of a near-infrared (NIR) spectroscopic method with multivariate analysis in order to develop a calibration model of the saccharification ratio of chemically pretreated Erianthus. The regression models clearly depend on the NIR spectral regions, and the information of CH and aromatic framework vibrations contributed most effectively to the alkaline dataset. From interpretations of the regression coefficient, lignin and cellulose were negatively and positively correlated with the saccharification ratio, respectively, and this result was supported by the data from wet chemical analysis. A more complex dataset was obtained from varied chemical pretreatments; here, the saccharification ratio was either small or had no linear correlation with each structural monocomponent. These results enabled the successful construction of the PLS regression model. NIR spectroscopy can be a rapid screening method for the saccharification ratio, and furthermore, can provide information of the key factors influencing the realization of more efficient enzymatic accessibility.

Near-Infrared Chemometric Approach to Exhaustive Analysis of Rice Straw Pretreated for Bioethanol Conversion

We report a simple analytical procedure combining near-infrared (NIR) spectroscopy with multivariate analysis to detect the saccharification efficiency of pretreated rice straw. Three types of sample preparation methods were tested to develop a powerful calibration model, with the disk sample used as the standard protocol. From the spectra dataset of NaOH-treated biomass, we obtained a good calibration for the saccharification ratio and some major structural components by partial least-squares regression. Adding dataset from hot water and dilute sulfuric acid pretreatments to NaOH sample dataset, an acceptable calibration model to predict the saccharification ratio as well as the glucose, xylose, and lignin contents was generated. NIR has a great potential for rapid screening of saccharification efficiency of pretreated biomass, which would allows us to control the quality of processing toward better bioethanol production.

22. EFFECTS OF WAVE RADIATION STRESS AND VERTICAL MIXING ON STORM SURGE

Since major driving forces of storm surge are pressure depression and wind stresse, a set of depth integrated equations is widely used for storm surge simulations to estimate anomaly sea level rise. However, there are several phenomena should be taken into consideration. This study estimates effects of wave radiation stress, vertical mixing models and boundary conditions at ocean upper layers on storm surge. The three dimensional hydro-static model including turbulence mixing and diffusion are used to predict the sea surface elevation in the storm surge. The numerical results show that the wave radiation stress and the turbulent flux significantly influence on sea levels and currents, respectively.