Catalytic activity of Cu2O nanoparticles supported on cellulose beads prepared by emulsion-gelation using cellulose/LiBr solution and vegetable oil.

Nanocatalysts tend to aggregate and are difficult to recycle, limiting their practical applications. In this study, an environmentally friendly method was developed to produce cellulose beads for use as supporting materials for Cu-based nanocatalysts. Cellulose beads were synthesized from a water-in-oil emulsion using cellulose dissolved in an LiBr solution as the water phase and vegetable oil as the oil phase. Upon cooling, the gelation of the cellulose solution produced spherical cellulose beads, which were then oxidized to introduce surface carboxyl groups. These beads (diameter: 95-105 µm; specific surface area: 165-225 m2 g-1) have a three-dimensional network of nanofibers (width: 20-30 nm). Furthermore, the Cu2O nanoparticles were loaded onto oxidized cellulose beads before testing their catalytic activity in the reduction of 4-nitrophenol using NaBH4. The apparent reaction rate constant increased with increasing loading of Cu2O nanoparticles and the conversion efficiency was >90 %. The turnover frequency was 376.2 h-1 for the oxidized cellulose beads with the lowest Cu2O loading, indicating a higher catalytic activity compared to those of other Cu-based nanoparticle-loaded materials. In addition to their high catalytic activity, the cellulose beads are reusable and exhibit excellent stability.

Elucidation of the Specific Ion Effects and Intermediate Structures of Cellulose Fibers Swollen in Inorganic Salt Solutions via In Situ X-ray Diffraction.

The solubilities of many substances are significantly affected by specific ions, as demonstrated by the Hofmeister series of proteins. Cellulose has a resistant fibrillar structure that hinders its swelling and dissolution. Certain inorganic salt solutions are effective swelling agents and solvents for cellulose. However, the precise effects of these ions on cellulose are not fully understood. In this study, we studied the intermediate structures of cellulose fibers during their swelling process in ZnCl2 and LiBr solutions via in situ X-ray diffraction. Two swollen phases with characteristic morphologies were observed for both salt treatments. Only the surfaces of the fibers were swollen in ZnCl2, whereas the ions penetrated the fibers and formed complexes with cellulose while the morphology of the fibers was maintained in LiBr. Our findings clarify the reasons that ZnCl2 has been used as an excellent swelling agent, whereas LiBr has been used as a good solvent for cellulose.

α-d-(1 → 3)-graft-(1 → 6)-glucan: Comb-like polysaccharide synthesized in vitro with α-1,3/1,6-glucosyltransferase L from Streptococcus salivarius.

We demonstrated that a unique polysaccharide with extremely high molecular weight can be easily obtained via a low-cost, mild reaction in a water medium from sucrose, a photosynthetic product. α-1,3/1,6-Glucosyltransferase L (GtfL) from Streptococcus salivarius produced water-insoluble α-d-glucan from sucrose at 37 °C. Gel permeation chromatography revealed the molecular weight was extremely high; the weight-average molecular weight values were more than 1,000,000 irrespective of the substrate concentration. The Smith degradation of neat glucan and NMR spectroscopic analyses of the acetyl derivative revealed a structure similar to that of a comb-type graft copolymer, α-d-(1 â†’ 3)-graft-(1 â†’ 6)-glucan. The anhydroglucose units (AGUs) in the main-chain backbone are linked by (1 â†’ 3)-glycosidic bonds, whereas a side chain consisting of four AGUs via (1 â†’ 6)-glycosidic bonds alternately extends from C6 of the main chain.

Untangling the threads of cellulose mercerization.

Naturally occurring plant cellulose, our most abundant renewable resource, consists of fibers of long polymer chains that are tightly packed in parallel arrays in either of two crystal phases collectively referred to as cellulose I. During mercerization, a process that involves treatment with sodium hydroxide, cellulose goes through a conversion to another crystal form called cellulose II, within which every other chain has remarkably changed direction. We designed a neutron diffraction experiment with deuterium labelling in order to understand how this change of cellulose chain direction is possible. Here we show that during mercerization of bacterial cellulose, chains fold back on themselves in a zigzag pattern to form crystalline anti-parallel domains. This result provides a molecular level understanding of one of the most widely used industrial processes for improving cellulosic materials.

Time-Dependent Elastic Tensor of Cellulose Nanocrystal Probed by Hydrostatic Pressure and Uniaxial Stretching.

The elastic properties of crystals are fundamental for structural material. However, in the absence of macroscopic single crystals, the experimental determination of the elastic tensor is challenging because the measurement depends on the transmission of stress inside the material. To avoid arbitrary hypotheses about stress transfer, we combine hydrostatic pressure and uniaxial-stretching experiments to investigate the elastic properties of cellulose Iß. Three orthogonal compressibilities are 50.0, 6.6, and 1.71 TPa-1. Combining Poisson's ratios from a uniaxial stretching experiment directly gives the Young's modulus along the chain direction (E33). However, Poisson's ratio depends on the deformation rate leading to apparent modulus E33 = 113 GPa using a slow cycle (hours) and 161 GPa using a fast cycle (minutes). The lattice deformation along the chain is not time-dependent, so the off-diagonal elements are time-dependent on the scale of minutes to hours.

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.

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, M w: 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.

Cationic hydrogels prepared from regioselectively azidated (1→3)-α-d-glucan via crosslinking and amination: Physical and adsorption properties.

Cationic hydrogels with amino groups were successfully prepared using (1→3)-α-d-glucan synthesized by glucosyltransferase J (GtfJ) cloned from Streptococcus salivarius through a three-step reaction: (i) Azido groups were regioselectively introduced at the C6 position of (1→3)-α-d-glucan by a bromination-azidation process (degree of substitution 0.94), (ii) Azido groups were partially crosslinked with 1,8-nonadiyne via a copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction, (iii) Azido groups that were unused for crosslinking were reduced to amino groups by sodium borohydride (NaBH4). The introduction of amino groups was confirmed quantitatively and qualitatively by elemental, Fourier transform infrared (FT-IR), and nuclear magnetic resonance (NMR) analyses. These cationic hydrogels showed a specific adsorption ability for bovine serum albumin (BSA) over a wide pH range of 4.5-8.0 due to their high pH values at the point of zero charge (pHpzc 8.80-8.92).

Highly swellable hydrogel of regioselectively aminated (1→3)-α-d-glucan crosslinked with ethylene glycol diglycidyl ether.

(1→3)-α-d-glucan synthesized by glucosyltransferase J (GtfJ) cloned from Streptococcus salivarius was regioselectively aminated as 6-amino-6-deoxy-(1→3)-α-d-glucan (aminoglucan) through three steps: bromination, azidation, and reduction. The degree of substitution of the amino group was determined by elemental analysis to be 0.97 and the molecular weight was 3.74×104 as measured by size exclusion chromatography. The regioselective amination at the C6 position of every pyranose ring was confirmed by 1H/13C NMR and solid state 15N cross polarization/magic angle spinning NMR spectroscopy. Aminoglucan was characterized by FT-IR, X-ray diffraction and thermogravimetric analysis. Solubility of aminoglucan in various solvents was investigated and confirmed in aqueous solution at pH ≤ 11. Therefore, aminoglucan was crosslinked with ethylene glycol diglycidyl ether (EGDE) by an epoxy-ring opening reaction under alkaline conditions. The obtained EGDE-crosslinked aminoglucan hydrogels were highly swellable in water owing to a strong water-holding ability and no water was released on compression and breaking of the gels.

In situ solid-state NMR of a magnetically oriented microcrystal suspension.

In situ solid-state NMR measurements of a magnetically oriented microcrystal suspension (MOMS) were demonstrated. Under modulated rotation of the static field, or equivalently, of the sample tube, randomly oriented microcrystals in a viscous liquid medium feel a torque arising from the anisotropic bulk susceptibility and eventually aligned in the same direction. In this way, a three-dimensional MOMS (3D-MOMS) was obtained. To apply an elliptically rotating magnetic field to microcrystals in suspension, a probe to rotate the sample tube around an axis perpendicular to the static magnetic field was developed. Single-crystal (SC) rotation patterns were obtained from the 3D-MOMS by solid-state CP measurements triggered in synchronous with the sample-tube rotation. Unlike the traditional SC method, the 3D-MOMS approach presented here does not require the elaborate adjustment of the direction of the reference frame. The process of three-dimensional magnetic alignment was also studied by monitoring the spectral changes during continuous application of the modulated sample rotation.

Facile preparation of cellulose-SiO2 composite aerogels with high SiO2 contents using a LiBr aqueous solution.

Cellulose-SiO2 composite aerogels (CSGs) with high SiO2 content up to 90% were successfully developed via the mixing of SiO2 particles and a cellulose solution dissolved in a LiBr aqueous solution. The characteristic properties resulting from the direct gelation of a dissolved cellulose solution could facilitate the preparation of a variety of forms of SiO2-embedded gels. The introduction of SiO2 particles reduced the pore sizes while maintaining the three-dimensional porous structure of the regenerated cellulose matrix, resulting in increased density and decreased porosity. CSGs exhibited excellent mechanical properties, large surface areas, and thermal stability up to 250 °C in an air atmosphere. Furthermore, CSGs showed low thermal conductivity ranging from 0.038 to 0.048 W/m K despite the influence of higher density and lower porosity with increased SiO2 contents. These results suggested that CSGs could provide a new option with regard to advanced porous organic-inorganic composite materials.

Highly enhanced adsorption of Congo red onto dialdehyde cellulose-crosslinked cellulose-chitosan foam.

The adsorption of Congo red (CR) was evaluated using cellulose-chitosan foam crosslinked via dialdehyde cellulose (DAC). DAC-crosslinked cellulose-chitosan foam (CCLBD) was obtained by dissolution/regeneration using a LiBr aqueous solution, followed by crosslinking between chitosan and DAC. CCLBD possessed a three-dimensional structure with 40-200 nm wide pores composed of nanofibrils with a width of 10-20 nm, resulting in a high specific surface area of 230 m2/g. CCLBD was highly stable even acidic conditions in spite of a low crosslinking degree of 10.3%, which induced a slight reduction in the amino groups that interact with CR. CCLBD showed a CR adsorption capacity of 1548.2 mg/g and the adsorption process followed the Sips isotherm and pseudo-second-order models.

Purification of Branched Dextrin from Nägeli Amylodextrin by Ethanol Precipitation and Characterization of Its Aggregation Property in Methanol-Water.

Ethanol precipitation process for purification of branched dextrin (BD) in Nägeli amylodextrin from waxy rice starch was developed. Temperature and ethanol concentration for precipitation were main parameters affecting the recovery and purity of BD, and the purification condition at 4 °C and 10 % (v/v) ethanol in water was adopted. After four-time precipitation, the BD recovery was 34.6 %, whereas the purity improved from 78.5 % at the initial to 94.5 % at the four-time purified BD (BD4). BD4 mainly showed a chain length distribution between 18 to 35 with a mode length of 25, which shifted after enzymatic debranching with isoamylase to that between 9 and 20 with a mode length of 14. Each purified BD was solubilized in water, and each solution was mixed with methanol-water at 25 °C to a final methanol concentration of 16 M. The flakes of BD precipitated with 16 M methanol exhibited an A-type crystal structure by an X-ray diffraction analysis, and the speed generation of white flakes in 16 M methanol dramatically increased as the purification time increased. The effect of addition of highly branched cyclic dextrin (HBCD) or sodium tetraborate on BD aggregation in 16 M methanol was also investigated, where the former retarded aggregation but the latter had no effect on the velocity. Thus, the purified BD enables rapid characterization of aggregation of double helix structures of A-type crystal structure, and screening of compounds which could affect the phenomena for prediction of potentials in starch modification as well.

Determination of the Anisotropic Rotational Diffusion Constant of Microcrystals Dispersed in Liquid Medium.

Microcrystals of ErBa2Cu4O8 suspended in a liquid medium were triaxially aligned by a frequency-modulated magnetic field and allowed a free rotational relaxation after the magnetic field was turned off. In situ X-ray diffraction measurements of the suspension were performed during relaxation, and the temporal change of the orientation fluctuation was monitored via broadening of the diffraction spots. The rotational diffusion constants were determined using the plot of the orientation fluctuation versus the elapsed time of rotational relaxation. The diffusion constants thus determined were in close agreement with those evaluated by the Stokes law but showed slight anisotropy, indicating that the microcrystals studied had shape anisotropy. The present method can provide a useful means for experimentally determining rotational diffusion constants of microcrystals suspended in viscous media. This paper shows that, due to the combination of the initial triaxial alignment and the subsequent monitoring of the relaxation process by means of X-ray diffraction, the diffusion constants along arbitrary crystallographic axes are determined separately.

Cellulose hydrogel with tunable shape and mechanical properties: From rigid cylinder to soft scaffold.

Cellulose hydrogel from aqueous solution of lithium bromide demonstrated excellent tunability of mechanical property and shape. A series of compression tests showed that cellulose hydrogel covered a wide range of mechanical property, where the compressive Young's modulus was controllable from 30 kPa to 1.3 MPa by changing the initial concentration of cellulose solution. Meanwhile, the diameter of the building block of gel, namely nano-fibrous cellulose, was constant at 15-20 nm irrelevant of the initial concentration of cellulose solution. Moreover, thanks to the biocompatibility of cellulose, the cultivation of cartilage tissue was successful in the micro-porous sponge-like cellulose hydrogel prepared by salt-leaching process. These findings show that this environmentally-benign versatile gel offers a new substrate for the biomaterial-based nanomaterial in biomedical applications.

Crystal Orientation of Poly(l-Lactic Acid) Induced by Magnetic Alignment of a Nucleating Agent.

The orientation of poly(l-lactic acid) (PLLA) crystals was controlled through crystal growth from a magnetically oriented nucleating agent, phenylphosphonic acid zinc (PPAZn). The one-dimensional magnetically oriented microcrystal array of PPAZn microcrystals revealed the relationship between the magnetization and crystallographic axes in the PPAZn crystal. The PPAZn microcrystals were homogeneously dispersed in PLLA via melt mixing, which decreased the molecular weight of the PLLA component due to degradation. The PPAZn microcrystals in the molten PLLA were uniaxially aligned under an 8-T static or rotating magnetic field. The wide-angle X-ray diffraction and small-angle X-ray scattering patterns of the PPAZn/PLLA composite films crystallized under each magnetic field showed that the PLLA lamellae grew from the surface of the PPAZn microcrystals, which were uniaxially oriented along the easy- or hard-magnetization axis, with the c-axis of PLLA parallel to the bc-plane of PPAZn. It was also suggested that the greater nucleating effect of PPAZn on PLLA was derived not from geometrical matching, but from factors such as favorable interactions and/or the plate-like shape of the microcrystal.

Characterization of crystalline linear (1→3)-α-d-glucan synthesized in vitro.

We investigated the crystal structure and molecular arrangement of the linear (1→3)-α-d-glucan synthesized by glucosyltransferase GtfJ cloned from Streptococcus salivarius using sucrose as a substrate. The synthetic products had two morphologies: wavy fibril-like crystals as major and thin lamellae as minor products. Their structures were analyzed using electron microdiffraction, synchrotron X-ray powder diffraction, and solid-state 13C NMR spectroscopy. The fibrils and lamellae had the same allomorphic form but different molecular arrangements. The wet crystals were in a hydrated form, which converted into an anhydrous form with a significant decrease in crystallinity on drying. The hydrated and anhydrous forms had an extended-chain conformation with 2/1 helix, and the hydrated form was estimated to contain one water molecule per glucose residue. The long glucan chains were folded in the fibril crystals, while the short, extended chains were arranged perpendicular to the base plane of the lamellae.

Protein adsorption of dialdehyde cellulose-crosslinked chitosan with high amino group contents.

Crosslinked chitosan was prepared by Schiff base formation between the aldehyde groups of dialdehyde cellulose (DAC) and the amino groups of chitosan and a subsequent reduction. DAC was obtained through periodate oxidation of cellulose and solubilization in hot water at 100°C for 1h. Three grades of DAC-crosslinked chitosan were prepared by adding various amounts DAC. The degrees of crosslinking as determined by amino group content were 3.8, 8.3, and 12.1%, respectively. DAC-crosslinked chitosan showed higher stability in the pH 2-9 range and no cytotoxicity was identified over the course of a 21-day long-term stability test. Also, DAC-crosslinked chitosan showed remarkably high bovine serum albumin (BSA) adsorption capacity at pH 5.5 as a result of the increased amino group content, due to the reaction between DAC and chitosan molecular chains occurring at multiple points even though DAC-crosslinked chitosan showed a lower degree of crosslinking.

Site-directed mutagenesis of bacterial cellulose synthase highlights sulfur-arene interaction as key to catalysis.

Cellulose is one of the most abundant biological polymers on Earth, and is synthesized by the cellulose synthase complex in cell membranes. Although many cellulose synthase genes have been identified over the past 25 years, functional studies of cellulose synthase using recombinant proteins have rarely been conducted. In this study, we conducted a functional analysis of cellulose synthase with site-directed mutagenesis, by using recombinant cellulose synthase reconstituted in living Escherichia coli cells that we recently constructed (cellulose-synthesizing E. coli, CESEC). We demonstrated that inactivating mutations at an important amino acid residue reduced cellulose production. In this study, an interesting loss-of-function mutation occurred on Cys308, whose main chain carbonyl plays an important role for locating the cellulose terminus. Mutating this cysteine to serine, thus changing sulfur to oxygen in the side chain, abolished cellulose production in addition to other apparent detrimental mutations. This unexpected result highlights that the thiol side-chain of this cysteine plays an active role in catalysis, and additional mutation experiments indicated that the sulfur-arene interaction around Cys308 is a key in cellulose-synthesizing activity. Data obtained by CESEC shed light on the function of cellulose synthase in living cells, and will deepen our understanding of the mechanism of cellulose synthase.

In vitro synthesis of linear α-1,3-glucan and chemical modification to ester derivatives exhibiting outstanding thermal properties.

Bio-based polymer is considered as one of potentially renewable materials to reduce the consumption of petroleum resources. We report herein on the one-pot synthesis and development of unnatural-type bio-based polysaccharide, α-1,3-glucan. The synthesis can be achieved by in vitro enzymatic polymerization with GtfJ enzyme, one type of glucosyltransferase, cloned from Streptococcus salivarius ATCC 25975 utilizing sucrose, a renewable feedstock, as a glucose monomer source, via environmentally friendly one-pot water-based reaction. The structure of α-1,3-glucan is completely linear without branches with weight-average molecular weight (Mw) of 700 kDa. Furthermore, acetate and propionate esters of α-1,3-glucan were synthesized and characterized. Interestingly, α-1,3-glucan acetate showed a comparatively high melting temperature at 339 °C, higher than that of commercially available thermoplastics such as PET (265 °C) and Nylon 6 (220 °C). Thus, the discovery of crystalline α-1,3-glucan esters without branches with high thermal stability and melting temperature opens the gate for further researches in the application of thermoplastic materials.