Yu Ogawa

Crystal analysis and high-resolution imaging of microfibrillar α-chitin from Phaeocystis.

The ultrastructure of α-chitin microfibril produced by marine alga Phaeocystis was investigated by FT-IR spectroscopy, X-ray diffraction and electron microscopy. The average size of the microfibril was 17.1±1.8 μm in length and 39.8±8.8 nm in width. The FT-IR spectrum shows typical α-chitin pattern, and each band was sharper than crustacean chitins, indicating higher crystallinity of the Phaeocystis chitin. The X-ray diffraction gave crystallite size more than twice of crustacean tendons. The fiber diffraction pattern is consistent with previous studies with two-chains orthorhombic unit cell (Minke and Blackwell, 1978; Sikorski et al., 2009), and refined unit cell dimensions are a=4.742 Å, b=18.871 Å, c=10.338Å. High-resolution electron microscopy of ultrathin sections gave the cross-sectional shape of microfibril as hexagon. The lattice images of (020) plane (d=0.94 nm) were frequently observed extending the entire cross-section of microfibril, indicating its single crystalline nature. These results allowed construction of a molecular packing model for α-chitin crystal.

Electron diffraction and high-resolution imaging on highly-crystalline β-chitin microfibril.

The ultrastructure of β-chitin microfibrils from a centric diatom, Thalassiosira, and a tubeworm, Lamellibrachia, was studied using electron diffraction and high-resolution electron microscopy. Electron microdiffraction diagrams corresponding to each projection of the β-chitin crystals were obtained, and all the data support the structure model of anhydrous β-chitin crystals proposed by X-ray diffraction experiments. From high-resolution electron microscopy on ultrathin sections, the cross-sectional shapes of the microfibrils from Thalassiosira and Lamellibrachia were observed as a rectangular and parallelogram, respectively. The lattice fringes corresponding to the (010) plane of anhydrous β-chitin crystals were clearly observed in both cross-sections. Based on these observations, we have constructed a molecular packing model for β-chitin microfibrils.

Comprehensive morphological and structural investigation of cellulose I and II nanocrystals prepared by sulphuric acid hydrolysis

Cellulose nanocrystals (CNCs) were produced from eucalyptus wood pulp using three different methods: (i) classical sulphuric acid hydrolysis (CN-I), (ii) acid hydrolysis of cellulose previously mercerized by alkaline treatment (MCN-II), and (iii) solubilization of cellulose in sulphuric acid and subsequent recrystallization in water (RCN-II). The three types of CNCs exhibited different morphologies and crystalline structures that were characterized using complementary imaging, diffraction and spectroscopic techniques. CN-I corresponded to the type I allomorph of cellulose while MCN-II and RCN-II corresponded to cellulose II. CN-I and MCN-II CNCs were acicular particles composed of a few laterally-bound elementary crystallites. In both cases, the cellulose chains were oriented parallel to the long axis of the particle, although they were parallel in CN-I and antiparallel in MCN-II. RCN-II particles exhibited a slightly tortuous ribbon-like shape and it was shown that the chains lay perpendicular to the particle long axis and parallel to their basal plane. The unique molecular and crystal structure of the RCN-II particles implies that a higher number of reducing chain ends are located at the surface of the particles, which may be important for subsequent chemical modification. While other authors have described nanoparticles prepared by regeneration of short-chain cellulose solutions, no detailed description was proposed in terms of particle morphology, crystal structure and chain orientation. We provide such a description in the present paper.

Alternative hydrogen bond models of cellulose II and IIII based on molecular force-fields and density functional theory

Alternative hydrogen-bond structures were found for cellulose II and IIII based on molecular dynamics simulations using four force fields and energy optimization based on density functional theory. All the modeling results were in support to the new hydrogen-bonding network. The revised structures of cellulose II and IIII differ with the fiber diffraction models mainly in the orientation of two hydroxyl groups, namely, OH2 and OH6 forming hydrogen-bond chains perpendicular to the cellulose molecule. In the alternative structures, the sense of hydrogen bond is inversed but little difference can be seen in hydrogen bond geometries. The preference of these alternative hydrogen bond structures comes from the local stabilization of hydroxyl groups with respect to the β carbon. On the other hand when simulated fiber diffraction patterns were compared with experimental ones, the current structure of cellulose II with higher energy and the alternative structure of cellulose IIII with lower energy were in better agreement.

Facile preparation of highly crystalline lamellae of (1 → 3)-β-D-glucan using an extract of Euglena gracilis.

In vitro synthesis of (1 → 3)-β-D-glucan was performed using laminaribiose phosphorylase obtained by an extraction of Euglena gracilis with sucrose phosphorylase. The synthetic product was a linear (1 → 3)-β-D-glucan with a narrow distribution of degree of polymerization (DP) centered on DP=30. X-ray diffraction and electron microscopy revealed that the glucan molecules obtained were self-organized as highly crystalline hexagonal lamellae. This synthetic product has quite high structural homogeneity at every level from primary to higher-order structure, which is a great advantage for the detailed analyses of physiological functions of (1 → 3)-β-D-glucan.

Infrared study on deuteration of highly-crystalline chitin

We report an infrared study of the deuteration of highly crystalline α- and β-chitin. With exposure to D2O vapor, the deuteration of α-chitin progressed on surface molecules, in contrast with β-chitin, which is partially deuterated on the inner part of the crystal. The intracrystalline deuteration occurred with high-temperature annealing in liquid D2O for α- and β-chitin. In α-chitin, the deuteration was similar between all the three functional groups. The ratio of deuterium/hydrogen increased with treatment temperature, and all the hydrogen in the functional groups was accessible above 200 °C. In β-chitin, the deuteration was more specific for each functional group. The deuteration of O3-H groups progressed rapidly, even below 100 °C, and, by contrast, that of O6-H and N-H progressed relatively slowly. These differences of deuteration between α- and β-chitin presumably arises from the particular ability of β-chitin to form a complex with water molecules.

Ensemble evaluation of polydisperse nanocellulose dimensions: rheology, electron microscopy, X-ray scattering and turbidimetry

Six types of CNCs with different sizes were prepared from tunicins by sulfuric acid hydrolysis and subsequent sonication in water. The size distributions of CNCs were comprehensively evaluated by turbidimetry, small angle X-ray scattering, and microscopy to predict their intrinsic viscosities. Experimental intrinsic viscosities [η] of the CNC dispersions were evaluated by shear viscosity measurement, and then compared with their theoretical [η] values based on theories for rotational motions of rigid rods. The experimental [η] values for the straight CNCs were in good agreement with their theoretical [η] values, irrespective of the size and distributions. On the other hand, the experimental [η] value of the kinked CNC was higher than the theoretical [η] value, in agreement with a theoretical calculation giving higher intrinsic viscosities for bent fibers.

Estimating the Strength of Single Chitin Nanofibrils via Sonication-Induced Fragmentation

We report the mechanical strength of native chitin nanofibrils. Highly crystalline α-chitin nanofibrils were purified from filaments produced by a microalgae Phaeocystis globosa, and two types of β-chitin nanofibrils were purified from pens of a squid Loligo bleekeri and tubes of a tubeworm Lamellibrachia satsuma, with relatively low and high crystallinity, respectively. These chitin nanofibrils were fully dispersed in water. The strength of individualized nanofibrils was estimated using cavitation-induced tensile fracture of nanoscale filaments in a liquid medium. Both types of β-chitin nanofibrils exhibited similar strength values of approximately 3 GP; in contrast, the α-chitin nanofibrils exhibited a much lower strength value of 1.6 GPa. These strength estimates suggest that the tensile strength of chitin nanofibrils is governed by the molecular packing modes of chitin rather than their crystallinity.

X-ray crystal structure of anhydrous chitosan at atomic resolution.

We determined the crystal structure of anhydrous chitosan at atomic resolution, using X‐ray fiber diffraction data extending to 1.17 Å resolution. The unit cell [a = 8.129(7) Å, b = 8.347(6) Å, c = 10.311(7) Å, space group P212121] of anhydrous chitosan contains two chains having one glucosamine residue in the asymmetric unit with the primary hydroxyl group in the gt conformation, that could be directly located in the Fourier omit map. The molecular arrangement of chitosan is very similar to the corner chains of cellulose II implying similar intermolecular hydrogen bonding between O6 and the amine nitrogen atom, and an intramolecular bifurcated hydrogen bond from O3 to O5 and O6. In addition to the classical hydrogen bonds, all the aliphatic hydrogens were involved in one or two weak hydrogen bonds, mostly helping to stabilize cohesion between antiparallel chains.

Solid–solvent molecular interactions observed in crystal structures of β-chitin complexes

Abstract Three β-chitin structures [anhydrous, di-hydrate, mono-ethylenediamine (EDA)] recently determined by synchrotron X-ray and neutron fiber diffraction were reviewed from the viewpoint of molecular interactions. Both water and EDA molecules interact with the chitin chains through multiple hydrogen bonds. When water complexes with chitin, the hydrogen bonding pattern rearranges with the replacement of an intrachain chitin hydrogen bond by a stronger hydrogen bond between chitin and water, with an associated reduction in the degrees of freedom; the water oxygen is a much stronger acceptor than the O5 ring atom. The behavior of hydrogen exchange by deuterium supports this interpretation. EDA-molecules change the conformation of hydroxymethyl group from gg to gt, accompanied by changes in hydrogen bonds due to the strong accepting ability of the EDA nitrogen atoms. Some important interactions are in common with experimental crystallographic results of cellulosic crystals and of molecular dynamics studies. These new insights into solid–solvent interactions are valuable in understanding molecular interactions in other polysaccharides-solvents system in solution or on surface.