Aspergillus oryzae PrtR alters transcription of individual peptidase genes in response to the growth environment.

Aspergillus oryzae PrtR is an ortholog of the transcription factor PrtT, which positively regulates the transcription of extracellular peptidase genes in Aspergillus niger and Aspergillus fumigatus. To identify the genes under the control of PrtR and elucidate its regulatory mechanism in A. oryzae, prtR gene disruption mutants were generated. The control strain clearly showed a halo on media containing skim milk as the nitrogen source, whereas the ΔprtR strain formed a smaller halo. Measurement of acid peptidase activity revealed that approximately 84% of acidic endopeptidase and 86% of carboxypeptidase activities are positively regulated by PrtR. As the transcription of the prtR gene varied depending on culture conditions, especially with or without a protein substrate, it was considered that its transcription would be regulated in response to a nitrogen source. In addition, contrary to previous expectations, PrtR was found to act both in promoting and repressing the transcription of extracellular peptidase genes. The mode of regulation varied from gene to gene. Some genes were regulated in the same manner in both liquid and solid cultures, whereas others were regulated in different ways depending on the culture conditions. Furthermore, PrtR has been suggested to regulate the transcription of peptidase genes that are closely associated with other transcription factors. KEY POINTS: • Almost all peptidase genes in Aspergillus oryzae are positively regulated by PrtR • However, several genes are regulated negatively by PrtR • PrtR optimizes transcription of peptidase genes in response to culture conditions.

Voltammetric and In Situ Spectroscopic Investigations on the Redox Processes of Trioxotriangulene Neutral Radicals on Graphite Electrodes.

To investigate the microscopic electrochemical dynamics of a stable trioxotriangulene (TOT) organic neutral π-radical on a graphite electrode surface, voltammetric and in situ infrared (IR) spectroelectrochemical studies were conducted using electrolyte solutions containing TOT monoanions. Upright columnar crystals (face-on alignment) of the TOT neutral radical were preferentially formed and dissolved in a rather reversible manner in the electrolyte with a low concentration of TOT monoanion under electrochemical conditions; however, more flat-lying columnar crystals (edge-on alignment) were formed in a higher concentration electrolyte. The flat-lying crystals remained on the graphite surface even at a fully reduced potential, owing to the lack of direct π-π interactions between the molecules and the graphite electrode. In situ IR attenuated total reflectance spectroscopy analyses successfully characterized the alignment of the columnar crystals of the TOT neutral radicals and their electrochemical behaviors, including the possible origins of the irreversible redox reaction of TOT on the graphite electrode.

A Redox-Active Microporous Organosiloxane Containing a Stable Neutral Radical, Trioxotriangulene.

A new silyl-substituted trioxotriangulene (TOT) neutral radical and corresponding porous organosiloxanes (POSs) were synthesized. The neutral radical exhibited a peculiarly high stability and formed a diamagnetic π-dimer characteristic to TOT neutral radicals stabilized by the strong multiple SOMO-SOMO interaction in both solution and solid states. POSs including TOT units within the organosiloxane-wall were prepared by polycondensation of the silyl groups and formed microporous structures with ∼1 nm-size diameters. Redox ability of TOT units in the POS was demonstrated by the treatment of oxidant/reductant in heterogeneous suspension condition, where the TOT units were reversibly converted between reduced and neutral radical species. Furthermore, the solid-state electrochemical measurements of the POS revealed the reversible multi-stage redox ability of TOT units involving polyanionic species within the organosiloxane-wall.

High Capacity and Energy Density Organic Lithium-Ion Battery Based on Buckypaper with Stable π-Radical.

Owing to an increasing demand on high performance and rare-metal free energy storage systems, organic rechargeable battery has attracted much attention. To increase the capacity of the whole battery, we have fabricated coin-type buckypaper cells composed of a trioxotriangulene neutral radical derivative (H3 TOT) and single-walled carbon nanotubes as a cathode and lithium metal plate as an anode without current collector. The cells exhibited a stable charge-discharge behavior even at a 90 wt % H3 TOT content with a high-rate performance of 10 C originating from high electrical conductivity of H3 TOT. Furthermore, based on the four-stage redox ability of H3 TOT, the H3 TOT 90 wt % cathode showed a high capacity of approximately 260 mAh g-1 and a high energy density of 546 Wh g-1 . In view of the simple fabrication of the cathode and excellent performance, TOT-based buckypaper will open a new strategy for the flexible cells for next-generation energy storages.

2D Coordination Network of Trioxotriangulene with Multiple Redox Abilities and Its Rechargeable Battery Performance.

A three-fold symmetric trioxotriangulene derivative with three pyridyl groups as coordinating sites was designed and synthesized. In a cyclic voltammetry measurement, the trioxotriangulene skeleton exhibited a multi-stage redox ability from neutral radical to radical tetra-anion species. In the zinc complex of monoanion species, three pyridyl groups coordinated to the zinc ion to build up a two-dimensional coordination network with a cavity larger than 12 Å in diameter. This complex was utilized as a cathode active material of a lithium ion battery, and it exhibited a capacity of ca. 60 mAh g-1 per the weight of the active material with a stable cycling performance up to 1000 cycles. This work shows that the coordination network formed by the trioxotriangulene-based ligand was effective in the improvement of cycle performance of the organic rechargeable battery.

Pyrolysis of Iron-Containing Polyanilines under Micropore Generation Control: Electrocatalytic Performance in the Oxygen Reduction Reaction.

Pyrolyzed iron-containing polyaniline (C-Fe-PANI) is one of the most promising candidates as a non-precious metal based electrocatalyst for oxygen reduction reaction (ORR). Although the ORR activity depends on the surface area arisen from pyrolysis-generated micropores on C-Fe-PANI particles, the micropore generation is hindered by pyrolysis-formed iron nanoparticles (Fe NPs) embedded inside C-Fe-PANI particles. Here, we demonstrate the pyrolysis of iron-containing PANIs under suppression of micropore-generation hindrance by blocking the Fe NPs formation. The higher-molecular-weight (MW: 100,000) PANI was dispersed in an FeCl3 solution before pyrolysis for preventing FeCl3 penetration inside PANI particles. As a result, as compared to the case of lower-MW (5,000) PANI, the Fe NPs formation was more suppressed inside catalyst particles to give 1.9 (1.8) times micropore volume (specific surface area), leading to a 11 % higher current density in ORR electrocatalytic performance test in acidic media.

Air-Stable Thin Films with High and Anisotropic Electrical Conductivities Composed of a Carbon-Centered Neutral π-Radical.

Air-stable thin films (50-720 nm thickness) composed of a carbon-centered neutral π-radical with high and anisotropic electrical conductivities were fabricated by vapor deposition of 4,8,12-trioxotriangulene (TOT). The thin films were air-stable over 15 months and were the aggregate of TOT microcrystals, in which a one-dimensional π-stacking column was formed through the strong singly occupied molecular orbital (SOMO)-SOMO interaction with two-electron-multicenter bond among the spin-delocalized π-planes. The orientations of the one-dimensional column of TOT were changed depending on the deposition rate and substrates, where face-on-oriented thin films were epitaxially grown on the graphite 0001 surface, and edge-on-oriented thin films were grown on glass, SiO2, and indium tin oxide substrates under a high-deposition rate condition. The films showed high electrical conductivities of 2.5 × 10-2 and 5.9 × 10-5 S cm-1 along and perpendicular to the π-stacking column, respectively, for an edge-on oriented thin film.

H-DROP: an SVM based helical domain linker predictor trained with features optimized by combining random forest and stepwise selection.

Domain linker prediction is attracting much interest as it can help identifying novel domains suitable for high throughput proteomics analysis. Here, we report H-DROP, an SVM-based Helical Domain linker pRediction using OPtimal features. H-DROP is, to the best of our knowledge, the first predictor for specifically and effectively identifying helical linkers. This was made possible first because a large training dataset became available from IS-Dom, and second because we selected a small number of optimal features from a huge number of potential ones. The training helical linker dataset, which included 261 helical linkers, was constructed by detecting helical residues at the boundary regions of two independent structural domains listed in our previously reported IS-Dom dataset. 45 optimal feature candidates were selected from 3,000 features by random forest, which were further reduced to 26 optimal features by stepwise selection. The prediction sensitivity and precision of H-DROP were 35.2 and 38.8%, respectively. These values were over 10.7% higher than those of control methods including our previously developed DROP, which is a coil linker predictor, and PPRODO, which is trained with un-differentiated domain boundary sequences. Overall, these results indicated that helical linkers can be predicted from sequence information alone by using a strictly curated training data set for helical linkers and carefully selected set of optimal features. H-DROP is available at http://domserv.lab.tuat.ac.jp.

Sumanenemonoone imines bridged by redox-active π-conjugated unit: synthesis, stepwise coordination to palladium(II), and laser-induced formation of nitrogen-doped graphitic carbon.

Sumanenemonoone imine compounds bridged by a redox-active π-conjugated unit on the basis of the conversion between 1,4-phenylenediamine and 1,4-benzoquinonediimine were synthesized and characterized. The stepwise coordination of the imino groups to Pd(II) in the sumanenemonoone imine compound bridged by 1,4-benzoquinonediimine was indicated by the titration experiment. Laser irradiation of a film of the metal-free quionediimine gave nitrogen-doped graphitic carbon, which was supported by an increase in conductivity and by Raman spectroscopy. The obtained graphitic carbon corresponds to carbonous compounds thermally treated at approximately 700-1000 °C. The ratio of nitrogen and carbon relative to that in the starting compound was nearly completely retained (5.4% decrease).