Coexistence of Urbach-Tail-Like Localized States and Metallic Conduction Channels in Nitrogen-Doped 3D Curved Graphene.

Nitrogen (N) doping is one of the most effective approaches to tailor the chemical and physical properties of graphene. By the interplay between N dopants and 3D curvature of graphene lattices, N-doped 3D graphene displays superior performance in electrocatalysis and solar-energy harvesting for energy and environmental applications. However, the electrical transport properties and the electronic states, which are the key factors to understand the origins of the N-doping effect in 3D graphene, are still missing. The electronic properties of N-doped 3D graphene are systematically investigated by an electric-double-layer transistor method. It is demonstrated that Urbach-tail-like localized states are located around the neutral point of N-doped 3D graphene with the background metallic transport channels. The dual nature of electronic states, generated by the synergistic effect of N dopants and 3D curvature of graphene, can be the electronic origin of the high electrocatalysis, enhanced molecular adsorption, and light absorption of N-doped 3D graphene.

Dirac Fermion Kinetics in 3D Curved Graphene.

3D integration of graphene has attracted attention for realizing carbon-based electronic devices. While the 3D integration can amplify various excellent properties of graphene, the influence of 3D curved surfaces on the fundamental physical properties of graphene has not been clarified. The electronic properties of 3D nanoporous graphene with a curvature radius down to 25-50 nm are systematically investigated and the ambipolar electronic states of Dirac fermions are essentially preserved in the 3D graphene nanoarchitectures, while the 3D curvature can effectively suppress the slope of the linear density of states of Dirac fermion near the Fermi level are demonstrated. Importantly, the 3D curvature can be utilized to tune the back-scattering-suppressed electrical transport of Dirac fermions and enhance both electron localization and electron-electron interaction. As a result, nanoscale curvature provides a new degree of freedom to manipulate 3D graphene electrical properties, which may pave a new way to design new 3D graphene devices with preserved 2D electronic properties and novel functionalities.

Understanding the Detection Mechanisms and Ability of Molecular Hydrogen on Three-Dimensional Bicontinuous Nanoporous Reduced Graphene Oxide.

Environmental safety has become increasingly important with respect to hydrogen use in society. Monitoring techniques for explosive gaseous hydrogen are essential to ensure safety in sustainable hydrogen utilization. Here, we reveal molecular hydrogen detection mechanisms with monolithic three-dimensional nanoporous reduced graphene oxide under gaseous hydrogen flow and at room temperature. Nanoporous reduced graphene oxide significantly increased molecular hydrogen physisorption without the need to employ catalytic metals or heating. This can be explained by the significantly increased surface area in comparison to two-dimensional graphene sheets and conventional reduced graphene oxide flakes. Using this large surface area, molecular hydrogen adsorption behaviors were accurately observed. In particular, we found that the electrical resistance firstly decreased and then gradually increased with higher gaseous hydrogen concentrations. The resistance decrease was due to charge transfer from the molecular hydrogen to the reduced graphene oxide at adsorbed molecular hydrogen concentrations lower than 2.8 ppm; conversely, the resistance increase was a result of Coulomb scattering effects at adsorbed molecular hydrogen concentrations exceeding 5.0 ppm, as supported by density functional theory. These findings not only provide the detailed adsorption mechanisms of molecular hydrogen, but also advance the development of catalyst-free non-heated physisorption-type molecular detection devices.

Three-dimensional porous graphene networks expand graphene-based electronic device applications.

In recent years, there has been increasing demand for 3D porous graphene structures with excellent 2D characteristics and great potential. As one avenue, several approaches for fabricating 3D porous graphene network structures have been proposed to realize multi-functional graphene materials with 2D graphene structures. Herein, we overview characteristics of 3D porous graphene for applications in future electronic devices along with physical insights into "2D to 3D graphene", in which the characters of 2D graphene such as massless Dirac fermions are well preserved. The present review thus summarizes recent 3D porous graphene studies with a perspective for providing new and board applications of graphene in electronic devices.

Large-Area and Transferred High-Quality Three-Dimensional Topological Insulator Bi2-xSbxTe3-ySey Ultrathin Film by Catalyst-Free Physical Vapor Deposition.

Uniform and large-area synthesis of bulk insulating ultrathin films is an important subject toward applications of a surface of three-dimensional topological insulators (3D-TIs) in various electronic devices. Here we report epitaxial growth of bulk insulating three-dimensional topological insulator (3D-TI) Bi2-xSbxTe3-ySey (BSTS) ultrathin films, ranging from a few quintuple to several hundreds of layers, on mica in a large-area (1 cm2) via catalyst-free physical vapor deposition. These films can nondestructively be exfoliated using deionized water and transferred to various kinds of substrates as desired. The transferred BSTS thin films show good ambipolar characteristics as well as well-defined quantum oscillations arising from the topological surface states. The carrier mobility of 2500-5100 cm2/(V s) is comparable to the high-quality bulk BSTS single crystal. Moreover, tunable electronic states from the massless to the massive Dirac fermion were observed with a decrease in the film thickness. Both the feasible large-area synthesis and the reliable film transfer process can promise that BSTS ultrathin films will pave a route to many applications of 3D-TIs.

In-plane topological p-n junction in the three-dimensional topological insulator Bi2-xSbxTe3-ySey.

A topological p-n junction (TPNJ) is an important concept to control spin and charge transport on a surface of three-dimensional topological insulators (3D-TIs). Here we report successful fabrication of such TPNJ on a surface of 3D-TI Bi2-xSbxTe3-ySey thin films and experimental observation of the electrical transport. By tuning the chemical potential of n-type topological Dirac surface of Bi2-xSbxTe3-ySey on its top half by using tetrafluoro-7,7,8,8-tetracyanoquinodimethane as an organic acceptor molecule, a half surface can be converted to p-type with leaving the other half side as the opposite n-type, and consequently TPNJ can be created. By sweeping the back-gate voltage in the field effect transistor structure, the TPNJ was controlled both on the bottom and the top surfaces. A dramatic change in electrical transport observed at the TPNJ on 3D-TI thin films promises novel spin and charge transport of 3D-TIs for future spintronics.

Electric Properties of Dirac Fermions Captured into 3D Nanoporous Graphene Networks.

Nanoporous graphene- based electric-double-layer transistors (EDLTs) are successfully fabricated. Transport measurements of the EDLTs demonstrate that the ambipolar electronic states of massless Dirac fermions with a high carrier mobility are well preserved in 3D nanoporous graphene along with anomalous nonlinear Hall resistance and exceptional transistor on/off ratio. This study may open a new avenue for device applications of graphene.

Structural and binding properties of laminarin revealed by analytical ultracentrifugation and calorimetric analyses.

One of the ß-1,3-glucans, laminarin, has been widely used as a substrate for enzymes including endo-1,3-ß-glucanase. To obtain quantitative information about the molecular interaction between laminarin and endo-1,3-ß-glucanase, the structural properties of laminarin should be determined. The results from pioneering work using analytical ultracentrifugation for carbohydrate analysis showed that laminarin from Laminaria digitata predominantly exists as a single-chain species with approximately 5% of triple-helical species. Differential scanning calorimetry experiments did not show a peak assignable to the transition from triple-helix to single-chain, supporting the notion that a large proportion of laminarin is the single-chain species. The interaction of laminarin with an inactive variant of endo-1,3-ß-glucanase from Cellulosimicrobium cellulans, E119A, was quantitatively analyzed using isothermal titration calorimetry. The binding was enthalpically driven and the binding affinity was approximately 10(6) M(-1). The results from binding stoichiometric analysis indicated that on average, E119A binds to laminarin in a 2:1 ratio. This seems to be reasonable, because laminarin mainly exists as a monomer, the apparent molecular mass of laminarin is 3.6 kDa, and E119A would have substrate-binding subsites corresponding to 6 glucose units. The analytical ultracentrifugation experiments could detect different complex species of laminarin and endo-1,3-ß-glucanase.

Multifunctional Porous Graphene for High-Efficiency Steam Generation by Heat Localization.

Multifunctional nanoporous graphene is realized as a heat generator to convert solar illumination into high-energy steam. The novel 3D nanoporous graphene demonstrates a highly energy-effective steam generation with an energy conversation of 80%.

A Receptor-Like Kinase, Related to Cell Wall Sensor of Higher Plants, is Required for Sexual Reproduction in the Unicellular Charophycean Alga, Closterium peracerosum-strigosum-littorale Complex.

Here, we cloned the CpRLK1 gene, which encodes a receptor-like protein kinase expressed during sexual reproduction, from the heterothallic Closterium peracerosum-strigosum-littorale complex, one of the closest unicellular alga to land plants. Mating-type plus (mt(+)) cells with knockdown of CpRLK1 showed reduced competence for sexual reproduction and formed an abnormally enlarged conjugation papilla after pairing with mt(-) cells. The knockdown cells were unable to release a naked gamete, which is indispensable for zygote formation. We suggest that the CpRLK1 protein is an ancient cell wall sensor that now functions to regulate osmotic pressure in the cell to allow proper gamete release.

Tuning of the ground state in electron doped anthracene.

High quality bulk samples of anthracene (AN) doped with potassium (K) in 1 : 1 and 2 : 1 stoichiometries were successfully prepared by a method involving a room temperature solid-state mechanical diffusion process prior to intercalation reactions during heat treatment, and their physical properties were studied using both magnetic and optical measurements. The transfer of almost one electron from K to AN in K1(AN) was confirmed by SQUID and ESR measurements. A pronounced magnetic hump centered at 150 K associated with antiferromagnetic interactions was observed, which can most likely be interpreted in terms of on-site Coulomb repulsions of the Mott insulating states. Optical spectra of K1(AN) clearly showed the insulating states, as well as the electron occupation of the LUMO-derived band of AN. Our results demonstrated tuning of the ground state of a typical bulk hydrocarbon by alkali metal intercalation.

High-quality three-dimensional nanoporous graphene.

We report three-dimensional (3D) nanoporous graphene with preserved 2D electronic properties, tunable pore sizes, and high electron mobility for electronic applications. The complex 3D network comprised of interconnected graphene retains a 2D coherent electron system of massless Dirac fermions. The transport properties of the nanoporous graphene show a semiconducting behavior and strong pore-size dependence, together with unique angular independence. The free-standing, large-scale nanoporous graphene with 2D electronic properties and high electron mobility holds great promise for practical applications in 3D electronic devices.

Bicontinuous nanoporous N-doped graphene for the oxygen reduction reaction.

Bicontinuous nanoporous N-doped graphene with tunable pore size is synthesized by nanoporous Ni-based chemical vapor deposition. The novel 3D graphene material shows an outstanding catalytic activity towards the oxygen reduction reaction with a low onset potential of -0.08 V and a high kinetic current density of 8.2 mA cm(-2) at -0.4 V.

Critical roles of Asp270 and Trp273 in the α-repeat of the carbohydrate-binding module of endo-1,3-ß-glucanase for laminarin-binding avidity.

A carbohydrate-binding module from family 13 (CBM13), appended to the catalytic domain of endo-1,3-ß-glucanase from Cellulosimicrobium cellulans, was overexpressed in E. coli, and its interactions with ß-glucans, laminarin and laminarioligosaccharides, were analyzed using surface plasmon resonance biosensor and isothermal titration calorimetry. The association constants for laminarin and laminarioligosaccharides were determined to be approximately 10(6) M(-1) and 10(4) M(-1), respectively, indicating that 2 or 3 binding sites in the α-, ß-, and γ-repeats of CBM13 are involved in laminarin binding in a cooperative manner. The binding avidity is approximately 2-orders higher than the monovalent binding affinity. Mutational analysis of the conserved Asp residues in the respective repeats showed that the α-repeat primarily contributes to ß-glucan binding. A Trp residue is predicted to be exposed to the solvent only in the α-repeat and would contribute to ß-glucan binding. The α-repeat bound ß-glucan with an affinity of approximately 10(4) M(-1), and the other repeats additionally bound laminarin, resulting in the increased binding avidity. This binding is unique compared to the recognition mode of another CBM13 from Streptomyces lividans xylanase.

Molecular characterization of endo-1,3-ß-glucanase from Cellulosimicrobium cellulans: effects of carbohydrate-binding module on enzymatic function and stability.

An endo-1,3-ß-glucanase was purified from Tunicase®, a crude enzyme preparation from Cellulosimicrobium cellulans DK-1, and determined to be a 383-residue protein (Ala1-Leu383), comprising a catalytic domain of the glycoside hydrolase family 16 and a C-terminal carbohydrate-binding module family 13. The Escherichia coli expression system of the catalytic domain (Ala1-Thr256) was constructed, and the protein with N-terminal polyhistidine tag was purified using a Ni-nitrilotriacetic acid column. We analyzed enzymatic properties of the recombinant catalytic domain, its variants, and the Tunicase®-derived full-length endo-1,3-ß-glucanase. Substitution of Glu119 with Ala and deletion of Met123, both of the residues are located in the catalytic motif, resulted in the loss of hydrolytic activity. In comparison between the full-length enzyme and isolated catalytic domain, their hydrolytic activities for soluble substrates such as laminarin and laminarioligosaccharides were similar. In contrast, the hydrolytic activity of the full-length enzyme for insoluble substrates such as curdlan and yeast-glucan was significantly higher than that of the catalytic domain. It should be noted that the acid stabilities for the hydrolysis of laminarin were clearly different. Secondary structure analysis using circular dichroism showed that the full-length enzyme was more acid stable than was the catalytic domain, possibly because of domain interactions between the catalytic domain and the carbohydrate-binding module.

Both electron and hole Dirac cone states in Ba(FeAs)2 confirmed by magnetoresistance.

Quantum transport of Dirac cone states in the iron pnictide Ba(FeAs)(2) with a d-multiband system is studied by using single crystal samples. Transverse magnetoresistance develops linearly against the magnetic field at low temperatures. The transport phenomena are interpreted in terms of the zeroth Landau level by applying the theory predicted by Abrikosov. The results of the semiclassical analyses of a two carrier system in a low magnetic field limit show that both the electron and hole reside as the high mobility states. Our results show that pairs of electron and hole Dirac cone states must be taken into account for an accurate interpretation in iron pnictides, which is in contrast with previous studies.

Organic ligand binding by a hydrophobic cavity in a designed tetrameric coiled-coil protein.

The design and characterization of a hydrophobic cavity in de novo designed proteins provides a wide range of information about the functions of de novo proteins. We designed a de novo tetrameric coiled-coil protein with a hydrophobic pocketlike cavity. Tetrameric coiled coils with hydrophobic cavities have previously been reported. By replacing one Leu residue at the a position with Ala, hydrophobic cavities that did not flatten out due to loose peptide chains were reliably created. To perform a detailed examination of the ligand-binding characteristics of the cavities, we originally designed two other coiled-coil proteins: AM2, with eight Ala substitutions at the adjacent a and d positions at the center of a bundled structure, and AM2W, with one Trp and seven Ala substitutions at the same positions. To increase the association of the helical peptides, each helical peptide was connected with flexible linkers, which resulted in a single peptide chain. These proteins exhibited CD spectra corresponding to superhelical structures, despite weakened hydrophobic packing. AM2W exhibited binding affinity for size-complementary organic compounds. The dissociation constants, K(d), of AM2W were 220 nM for adamantane, 81 microM for 1-adamantanol, and 294 microM for 1-adamantaneacetic acid, as measured by fluorescence titration analyses. Although it was contrary to expectations, AM2 did not exhibit any binding affinity, probably due to structural defects around the designed hydrophobic cavity. Interestingly, AM2W exhibited incremental structure stability through ligand binding. Plugging of structural defects with organic ligands would be expected to facilitate protein folding.

Metal-ion-dependent GFP emission in vivo by combining a circularly permutated green fluorescent protein with an engineered metal-ion-binding coiled-coil.

Coordination of metal ions significantly contributes to protein structures and functions. Here we constructed a fusion protein, consisting of a de novo designed, metal-ion-binding, trimeric coiled-coil and a circularly permutated green fluorescent protein (cpGFP), where the fluorescent emission from cpGFP was induced by metal ion coordination to the coiled-coil. A circularly permutated GFP, (191)cpGFP(190), was constructed by connecting the original N- and C-termini of GFP(UV) by a GGSGG linker and cleaving it between Asp(190) and Gly(191). The metal-ion-binding coiled-coil, IZ-HH, was designed to have three alpha-helical structures, with 12 His residues in the hydrophobic core of the coiled-coil structure. IZ-HH exhibited an unfolded structure, whereas it formed the trimeric coiled-coil structure in the presence of divalent metal ions, such as Cu(2+), Ni(2+), or Zn(2+). The fusion protein (191)cpGFP(190)-IZ-HH was constructed, in which (191)cpGFP(190) was inserted between the second and third alpha-helices of IZ-HH. Escherichia coli cells, expressing (191)cpGFP(190)-IZ-HH, exhibited strong fluorescence when the Cu(2+) and Zn(2+) ions were present in the medium, indicating that they passed through the cell membrane and induced the proper folding of the (191)cpGFP(190) domain. This strategy, in which protein function is regulated by a metal-ion-responsive coiled-coil, should be applicable to the design of various metal-ion-responsive, nonnatural proteins that work both in vitro and in vivo.

Gene expression profiling using cDNA microarray analysis of the sexual reproduction stage of the unicellular charophycean alga Closterium peracerosum-strigosum-littorale complex.

The desmid Closterium peracerosum-strigosum-littorale complex, which is the closest unicellular sister to land plants, is the best characterized of the charophycean green algae with respect to the process of sexual reproduction. To elucidate the molecular mechanism of intercellular communication during sexual reproduction, we created a normalized cDNA library from mixed cells of the sexual and the vegetative phases and generated a cDNA microarray. In total, 3,236 expressed sequence tags, which were classified into 1,615 nonredundant groups, were generated for cDNA microarray construction. Candidate genes for key factors involved in fertilization, such as those that encode putative receptor-like protein kinase, leucine-rich-repeat receptor-like protein, and sex pheromone homologs, were up-regulated during sexual reproduction and/or by the addition of the purified sex pheromones, and the expression patterns of these genes were confirmed by quantitative real-time polymerase chain reaction analysis. This first transcriptome profile of Closterium will provide critical clues as to the mechanism and evolution of intercellular communication between the egg and sperm cells of land plants.

Isolation of myosin XI genes from the Closterium peracerosum-strigosum-littorale complex and analysis of their expression during sexual reproduction.

Myosins comprise a large superfamily of molecular motors that generate mechanical force in ATP-dependent interactions with actin filaments. On the basis of their conserved motor domain sequences, myosins can be divided into at least 17 classes, 3 of which (VIII, XI, XIII) are found in plants. Although full sequences of myosins are available from several species of green plants, little is known about the functions of these proteins. Additionally, sequence information for algal myosin is incomplete, and little attention has been given to the molecular evolution of myosin from green plants. In the present study, the Closterium peracerosum-strigosum-littorale complex was used as a model system for investigating a unicellular basal charophycean alga. This organism has been well studied with respect to sexual reproduction between its two mating types. Three types of partial sequences belonging to class XI myosins were obtained using degenerate primers designed to amplify motor domain sequences. Real-time polymerase chain reaction analysis of the respective myosin genes during various stages of the algal life cycle showed that one of the genes was more highly expressed during sexual reproduction, and that expression was cell-cycle-dependent in vegetatively grown cells.