Yuka Miyoshi

Degradation and Synthesis of β-Glucans by a Magnaporthe oryzae Endotransglucosylase, a Member of the Glycoside Hydrolase 7 Family

Background: Plant pathogens secrete enzymes that degrade plant cell walls to enhance infection and nutrient acquisition. Results: A novel endotransglucosylase catalyzes cleavage and transfer of β-glucans and decreases the physical strength of plant cell walls. Conclusion: Endotransglucosylation causes depolymerization and polymerization of β-glucans, depending on substrate molecular size. Significance: Enzymatic degradation of plant cell walls is required for wall loosening, which enhances pathogen invasion. A Magnaporthe oryzae enzyme, which was encoded by the Mocel7B gene, was predicted to act on 1,3–1,4-β-glucan degradation and transglycosylation reaction of cellotriose after partial purification from a culture filtrate of M. oryzae cells, followed by liquid chromatography-tandem mass spectrometry. A recombinant MoCel7B prepared by overexpression in M. oryzae exhibited endo-typical depolymerization of polysaccharides containing β-1,4-linkages, in which 1,3–1,4-β-glucan was the best substrate. When cellooligosaccharides were used as the substrate, the recombinant enzyme generated reaction products with both shorter and longer chain lengths than the substrate. In addition, incorporation of glucose and various oligosaccharides including sulforhodamine-conjugated cellobiose, laminarioligosaccharides, gentiobiose, xylobiose, mannobiose, and xyloglucan nonasaccharide into β-1,4-linked glucans were observed after incubation with the enzyme. These results indicate that the recombinant enzyme acts as an endotransglucosylase (ETG) that cleaves the glycosidic bond of β-1,4-glucan as a donor substrate and transfers the cleaved glucan chain to another molecule functioning as an acceptor substrate. Furthermore, ETG treatment caused greater extension of heat-treated wheat coleoptiles. The result suggests that ETG functions to induce wall loosening by cleaving the 1,3–1,4-β-glucan tethers of plant cell walls. On the other hand, use of cellohexaose as a substrate for ETG resulted in the production of cellulose II with a maximum length (degree of polymerization) of 26 glucose units. Thus, ETG functions to depolymerize and polymerize β-glucans, depending on the size of the acceptor substrate.

Temperature dependences of the dynamic viscoelastic properties of wood and acetylated wood swollen by water or organic liquids

In this study, to summarize the changes of thermal-softening behaviors of wood and acetylated wood due to differences in the kinds of swelling liquids, the following measurements were conducted. Untreated and acetylated wood samples were swollen by various liquids and the temperature dependences of the dynamic viscoelastic properties were measured after the heating and cooling histories were unified among the samples. The results obtained are as follows. Untreated samples swollen by high-polarity liquid had lower peak temperature of tanδ, however acetylated samples had higher peak temperature of tanδ than those of untreated wood. On the other hand, untreated wood samples swollen by low-polarity liquid had higher peak temperature of tanδ, however acetylated samples had lower peak temperature of tanδ than those of untreated wood. The amount of swelling is determined by interaction between wood and liquid due to proton-accepting power and molar volumes of liquid and so on, therefore the peak temperature of tanδ and degree of reduction in dynamic elastic modulus (E´) with increasing temperature were corresponded to the amount of swelling.

Effects of density and anatomical feature on mechanical properties of various wood species in lateral tension

This study is focused on what factor mainly affects the mechanical properties of each wood species in the lateral direction. At first, the influence of the density which is closely related to mechanical properties in the longitudinal direction was also researched in the lateral direction. Thus, the elastic modulus, strength, and failure strain in the lateral tension were measured using thin cross-sectional samples of softwoods and hardwoods, having wide varieties in the density and anatomical features. The results obtained are as follows. The linear relationship between the density and the elastic modulus which has been verified in the longitudinal direction was not observed in the lateral tension for the samples with the annual ring inclination of 90°, which samples were influenced by ray arranged parallel to the tensile direction. However, samples with the annual ring inclination of 45° showed the high correlation between the density and the elastic modulus due to the shearing deformation of the cell shape. On the other hand, the proportional relationship between the elastic modulus and strength which has been verified in the longitudinal direction was not observed in the lateral tension except for the samples with the annual ring inclination of 90°. From the results obtained, it was revealed that the mechanical properties of wood in the lateral direction were significantly affected not only by the density but also by the structural features such as deformation of cell shapes, arrangement of ray or vessels, and the degree of the transition from the earlywood to the latewood.