Yoshikuni Teramoto

Pretreatment of woody and herbaceous biomass for enzymatic saccharification using sulfuric acid-free ethanol cooking

A sulfuric acid-free ethanol cooking (SFEC) treatment was developed to achieve complete saccharification of the cellulosic component of eucalyptus and baggase flour, thereby avoiding the problems associated with the use of strong acid catalysts. Cutter-milled flours were exposed to an ethanol (EtOH)/water/acetic acid mixture in an autoclave. Enzymatic hydrolysis experiments of the pretreated samples demonstrated that almost complete conversion of the cellulosic components to glucose was achieved under optimal conditions. A large-scale trial revealed that there was little consumption of in-feed EtOH during SFEC; therefore, it is considered that most part EtOH used can be essentially recovered and reused. Field emission scanning electron microscopy showed that SFEC induced the formation of pores ranging in size from approximately 10 to several 100nm. It can be assumed that the porous surface was due to the partial removals of lignin and hemicellulose, which improved the accessibility of the enzyme onto the substrate.

Enzymatic saccharification of woody biomass micro/nanofibrillated by continuous extrusion process I--effect of additives with cellulose affinity.

Mechanical micro/nanofibrillation of Douglas fir was performed by a continuous extrusion process in an attempt to develop a cost-effective pretreatment method for enzymatic saccharification. Additives with cellulose affinity (ethylene glycol, glycerol, and dimethyl sulfoxide) were used to effectively fibrillate the wood cell wall into submicron- or nano-scale, thus opening up the cell wall structure for improving enzymatic accessibility, and lower the extrusion torque. Morphological characterization showed that ethylene glycol was the most effective additive for fibrillation. The fibrillated products were converted into glucose with a high yield by enzymatic saccharification. The maximum cellulose-to-glucose conversion was achieved when ethylene glycol was used; the value was 62.4%. The glucose yield was approximately 6 times higher than that of the untreated raw material.

Optimization of microwave-assisted extraction of carbohydrates from industrial waste of corn starch production using response surface methodology.

Microwave-assisted extraction (MAE) was applied for production of carbohydrates mainly consisting of arabinoxylan from corn pericarp which is an industrial waste of corn starch production by using hot compressed water as a solvent. The solubilization rate increased with increase in heating temperature and reached 75.2% at 220 °C. The main extracted materials were carbohydrates consist of glucose, xylose and arabinose indicating solubilization of starch and hemicellulose, while residues were composed of cellulose. Four independent variables (heating temperature, come-up time, heating time and solid to liquid ratio) were optimized for maximizing the carbohydrates yield using the response surface methodology including fractional factorial design, the path of steepest ascent and central composite design. The optimized condition was as follows; heating temperature 176.5 °C, come-up time 2 min, heating time 16 min and solid to liquid ratio 1/20 (g/mL), respectively. The maximal yield attained 70.8% of carbohydrates with predominant production of xylo-oligosaccharides.

Fabrication and properties of chitin/hydroxyapatite hybrid hydrogels as scaffold nano-materials.

Novel hybrid hydrogels were prepared by introducing nano-hydroxyapatite (nHAp) into chitin solution dissolved in NaOH/urea aqueous solution at low temperature, and then by cross-linking with epichlorohydrin (ECH). Their structure and morphology were characterized by FTIR spectra, wide-angle X-ray diffraction (WAXD), thermo-gravimetric analysis (TGA), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Our findings revealed that hydroxyapatite nano-particles were uniformly dispersed in chitin hydrogel networks. The chitin/nHAP hybrid hydrogel (Gel2) exhibited about 10 times higher mechanical properties (compressive strength: 274 kPa) than that of chitin hydrogel. Moreover, COS-7 cell culture experiment proved that cells could adhere and proliferate well on the chitin/nHAp hydrogels, suggesting good biocompatibility. All these results signified that these bio-materials could be potential candidates as scaffolds for tissue engineering.

Enzymatic saccharification of woody biomass micro/nanofibrillated by continuous extrusion process II: Effect of hot-compressed water treatment

An extrusion process involving a twin-screw extruder was used for the micro/nanofibrillation of Douglas fir and Eucalyptus treated with hot-compressed water (HCW). Partial removal of hemicellulose and lignin by HCW treatment effectively improved the fibrillation by extrusion. Only HCW treatment produced glucose less than 5 weight percent (wt.%) in Douglas fir in a temperature range of 140-180 degrees C by enzymatic hydrolysis. Glucose production yields of 18 and 26 wt.% were obtained by HCW treatment at 170 and 180 degrees C, respectively, in Eucalyptus. Use of extrusion after HCW treatment drastically improved monosaccharide production yield in both woods. In the case of Douglas fir, the obtained values were 5 times higher than those obtained by HCW treatment alone. Total monosaccharide production yields were higher in Eucalyptus than in Douglas fir. The extruded production had a fine fibrous morphology on a sub-micro/nanoscopic scale. This result shows the great potential of the extrusion process after HCW treatment as a cost-effective pretreatment for enzymatic saccharification of woody biomass.

Cost reduction and feedstock diversity for sulfuric acid-free ethanol cooking of lignocellulosic biomass as a pretreatment to enzymatic saccharification

We have previously demonstrated that a sulfuric acid-free ethanol (EtOH) cooking treatment enhances the enzymatic digestibility of eucalyptus wood and bagasse flour. In the present study, a reconfigured process that achieves similar performance was developed by identifying possible cost-competitive pretreatments that provide high cellulose-to-glucose conversion during subsequent enzymatic hydrolysis. The series of reconfigurations reduced EtOH usage in the pretreatment by more than 80% in comparison with our previous research. Higher initial pressures and intensive size reduction of the starting material are not required. The reconfigured process was applied to rice straw and Douglas fir, in order to confirm the feasibility of feedstock diversity.

Enhancement of enzymatic accessibility by fibrillation of woody biomass using batch-type kneader with twin-screw elements.

Fibrillation of wood cell walls into submicron and/or nanoscale fibers was successfully carried out in the presence of water by using a batch-type kneader with combination-available twin-screw elements. The results obtained are expected to be used for the application of a twin-screw extruder. Two types of screw combinations were used for applying different shearing and distribution forces. Most of the fibers of the fibrillated products had diameters less than 1 microm, and some of them had diameters less than 100 nm. The maximum glucose yield by enzymatic saccharification was found to be 54.2% in the fibrillated products kneaded for 20 min after ball milling for 20 min using a screw combination for applying high shearing force. The fibrillation increased the surface area of cellulose. The glucose yield was improved by cooking the fibrillated products with water at 135 degrees C under 0.25 MPa, revealing that only mechanical kneading appears to have some limitation to expose cellulose for complete enzymatic saccharification.

Cuticular membrane of Fuyu persimmon fruit is strengthened by triterpenoid nano-fillers.

The mechanical defensive performance of fruit cuticular membranes (CMs) is largely dependent on the molecular arrangement of their constituents. Here, we elucidated nano-sized interactions between cutin and triterpenoids in the cuticular matrix of Fuyu persimmon fruits ( Diospyros kaki Thunb. cv. Fuyu), focusing on the mechanical properties using a combination of polymer analyses. The fruit CMs of Fuyu were primarily composed of wax (34.7%), which was predominantly triterpenoids followed by higher aliphatic compounds, and cutin (48.4%), primarily consisting of 9,10-epoxy-18-hydroxyoctadecanoic acid and 9,10,18-trihydroxyoctadecanoic acid. Based on the tensile tests of the CM, the removal of wax lead to a considerable decrease in the maximum stress and elastic modulus accompanied by an increase in the maximum strain, indicating that wax is of significant importance for maintaining the mechanical strength of the CM. Wide-angle X-ray diffraction and relaxation time measurements using solid-state 13C nuclear magnetic resonance indicated that the triterpenoids in the cuticular matrix construct a nanocomposite at a mixing scale below 20-24 nm; however, the higher aliphatic compounds did not exhibit clear interactions with cutin. The results indicated that the triterpenoids in the cuticular matrix endow toughness to the CM by functioning as a nanofiller.

Functional Thermoplastic Materials from Derivatives of Cellulose and Related Structural Polysaccharides

This review surveys advances in the development of various material functionalities based on thermoplastic cellulose and related structural polysaccharide derivatives. First, the dependence of thermal (phase) transition behavior on the molecular composition of simple derivatives is rationalized. Next, approaches enabling effective thermoplasticization and further incorporation of material functionalities into structural polysaccharides are discussed. These approaches include: (a) single-substituent derivatization, (b) derivatization with multi-substituents, (c) blending of simple derivatives with synthetic polymers, and (d) graft copolymerization. Some examples addressing the control of supramolecular structures and the regulation of molecular and segmental orientations for functional materials fabrication, which have especially progressed over the past decade, are also addressed. Attractive material functions include improved mechanical performance, controlled biodegradability, cytocompatiblity, and optical functions.

Anisotropic polymer composites synthesized by immobilizing cellulose nanocrystal suspensions specifically oriented under magnetic fields.

Novel polymer composites reinforced with an oriented cellulose nanocrystal (CNC) assembly were prepared from suspensions of CNC in aqueous 2-hydroxyethyl methacrylate (HEMA) via magnetic field application to the suspensions followed by polymerization treatment. The starting suspensions used at ∼6 wt % CNC separated into an upper isotropic phase and a lower anisotropic (chiral nematic) one in the course of quiescent standing. A static or rotational magnetic field was applied to the isolated isotropic and anisotropic phases. UV-induced polymerization of HEMA perpetuated the respective states of magnetic orientation invested for the CNC dispersions to yield variously oriented CNC/poly(2-hydroxyethyl methacrylate) composites. The structural characterization was carried out by use of X-ray diffractometry and optical and scanning electron microscopy. The result indicated that CNCs were aligned in the composites distinctively according to the static or rotational magnetic application when the anisotropic phases were used, whereas such a specific CNC orientation was not appreciable when the isotropic phases were sampled. This marks out effectiveness of a coherent response of CNCs in the mesomorphic assembly. In dynamic mechanical experiments in tensile or compressive mode, we observed a clear mechanical anisotropy for the polymer composites synthesized from wholly anisotropic suspensions under static or rotational magnetization. The higher modulus (in compression) was detected for a composite reinforced by locking-in the uniaxial CNC alignment attainable through conversion of the initial chiral nematic phase into a nematic phase in the rotational magnetic field.