74 research outputs found

    In Vitro Synthesis of Branchless Linear (1 → 6)-α-d-Glucan by Glucosyltransferase K: Mechanical and Swelling Properties of Its Hydrogels Crosslinked with Diglycidyl Ethers

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    A hydrogel was prepared from a polysaccharide, enzymatically synthesized through a one-pot reaction in aqueous solution, and its properties as a functional material were evaluated. Enzymatic synthesis using glucosyltransferase K obtained from Streptococcus salivarius ATCC 25975 was performed with sucrose as a substrate. The synthetic product was unbranched linear (1 → 6)-α-d-glucan with a high molecular weight, Mw: 1.0–3.0 × 105. The synthesized (1 → 6)-α-d-glucan was insoluble in water and crystallized in a monoclinic unit cell, which is consistent with the hydrated form of dextran. Transparent and highly swellable (1 → 6)-α-d-glucan hydrogels were obtained by crosslinking with diglycidyl ethers. The hydrogels showed no syneresis and no volume change during compression, resulting in the retention of shape under repeated compression. The elastic moduli of these hydrogels (<60 kPa) are smaller than those of other polysaccharide-based hydrogels having the same solid contents. The oven-dried gels could be restored to the hydrogel state with the original transparency and a recovery ratio greater than 98%. The mechanism of water diffusion into the hydrogel was investigated using the kinetic equation of Peppas. The properties of the hydrogel are impressive relative to those of other polysaccharide-based hydrogels, suggesting its potential as a functional biomaterial

    Traffic Jams Reduce Hydrolytic Efficiency of Cellulase on Cellulose Surface

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    Abstract A deeper mechanistic understanding of the saccharification of cellulosic biomass could enhance the efficiency of biofuels development. We report here the real-time visualization of crystalline cellulose degradation by individual cellulase enzymes using an advanced version of high-speed atomic force microscopy. Trichoderma reesei 2 cellobiohydrolase I (TrCel7A) molecules were observed to slide unidirectionally along the crystalline cellulose surface, but at one point exhibited collective halting analogous to a traffic jam. Changing the crystalline polymorphic form of cellulose by an ammonia treatment increased the apparent number of accessible lanes on the crystalline surface and consequently the number of moving cellulase molecules. Treatment of this bulky crystalline cellulose simultaneously or separately with T. reesei cellobiohydrolase II (TrCel6A) resulted in a significant increase in the proportion of mobile enzyme molecules on the surface. Cellulose was completely degraded by the synergistic action between the two enzymes

    Emerging concepts in biomarker discovery; The US-Japan workshop on immunological molecular markers in oncology

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    Supported by the Office of International Affairs, National Cancer Institute (NCI), the "US-Japan Workshop on Immunological Biomarkers in Oncology" was held in March 2009. The workshop was related to a task force launched by the International Society for the Biological Therapy of Cancer (iSBTc) and the United States Food and Drug Administration (FDA) to identify strategies for biomarker discovery and validation in the field of biotherapy. The effort will culminate on October 28th 2009 in the "iSBTc-FDA-NCI Workshop on Prognostic and Predictive Immunologic Biomarkers in Cancer", which will be held in Washington DC in association with the Annual Meeting. The purposes of the US-Japan workshop were a) to discuss novel approaches to enhance the discovery of predictive and/or prognostic markers in cancer immunotherapy; b) to define the state of the science in biomarker discovery and validation. The participation of Japanese and US scientists provided the opportunity to identify shared or discordant themes across the distinct immune genetic background and the diverse prevalence of disease between the two Nations

    Extraction of cellulose-synthesizing activity of Gluconacetobacter xylinus by alkylmaltoside.

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    This study reinvestigated the synthesis of cellulose in vitro with a well-known cellulose-producing bacterium, Gluconacetobacter xylinus. Alkylmaltoside detergents, which are more frequently used in recent structural biological researches, are uniquely used in this study to solubilize cellulose-synthesizing activity from the cell membrane of G. xylinus. Activity comparable to that previously reported is obtained, while the synthesized cellulose is crystallized into a non-native polymorph of cellulose (cellulose II) as well as the previous studies. In spite of this failure to recover the native activity to synthesize cellulose I microfibril in vitro, the product is a polymer with a degree of polymerization greater than 45 as determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). It was thus concluded that the established protocol can solubilize cellulose-synthesizing activity of G. xylinus with polymerizing activity