Strain selection and scale-up fermentation for FR901379 acylase production by Streptomyces sp. no. 6907

Micafungin (FK463) is a widely used treatment for life-threatening, deep-seated fungal infections. It is an echinocandin-like lipopeptide derived from the chemical modification of deacylated FR901379, a type of lipopeptide antibiotic produced by Coleophoma empetri F-11899. The palmitoyl moiety of FR901379 is deacylated by FR901379 acylase produced by Streptomyces sp. no. 6907. In this study, our goal was to generate an improved strain of Streptomyces sp. no. 6907 capable of hyperproducing the FR901379-acylase enzyme. To accomplish this goal, modified strains of Streptomyces sp. no. 6907 were generated using UV-irradiation mutagenesis, and strain selection was performed using an agar-plate screening method to efficiently select an acylase-hyperproducing strain. Three marker indices were shown to correlate with elevated acylase production: decreased candidacidal activity of FR901379, decreased proteolytic activity on skim milk, and phenotypic characteristics. Cloning and subsequent sequencing of the acylase gene from the hyperproducing mutant revealed no mutations in either the acylase structural gene or the 5'-flanking region required for gene expression. The growth medium was also modified to maximize acylase production. We successfully increased acylase activity approximately 65-fold, compared with the original growth conditions (wild strain cultured in the original unmodified medium). To minimize formation of excess foam during the fermentation process, we optimized the parameters of agitation speed, as calculated from the discharge flow rate. Using our improved strain and the optimized medium and growth conditions, we have developed an improved and highly reproducible method for stable large-scale production of FR901379-acylase.     

Molded micro- and mesoporous carbon/silica composite from rice husk and beet sugar

A molded carbon/silica composite with high micro- and mesoporosity, as well as a high bulk density, was fabricated by activating a disk-molded precursor made from carbonized rice husk (RH) and beet sugar (BS) at 875 °C in CO₂. The pore structure of the RH- and BS-based carbon/silica composite (RBC) was analysed in relation to the bulk density. An activation time of 2.0 h provided the largest BET specific surface area (1027 m²/g) and total pore volume (0.68 cm³/g) and a low bulk density (0.54 g/cm³). An RBC that was first activated for 1 h was immersed again in BS syrup and then activated in CO₂ for 1 h. This two-step activation process provided both a high bulk density (0.69 g/cm³) and a highly textured structure (BET specific surface area, 943 m²/g; total pore volume, 0.56 cm³/g). The immersion in BS syrup was useful for improving the texture without reducing the bulk density, in comparison to one-step activation for 1.0 h. The suspension of the RBCs was basic because of the residual inorganic compounds of potassium and calcium. However, the basicity of the suspension was alleviated by washing the RBCs with water.     

Structural and electrical characteristics of high-κ Er2O3 and Er2TiO5 gate dielectrics for a-IGZO thin-film transistors

In this letter, we investigated the structural and electrical characteristics of high-κ Er₂O₃ and Er₂TiO₅ gate dielectrics on the amorphous indium-gallium-zinc-oxide (a-IGZO) thin-film transistor (TFT) devices. Compared with the Er₂O₃ dielectric, the a-IGZO TFT device incorporating an Er₂TiO₅ gate dielectric exhibited a low threshold voltage of 0.39 V, a high field-effect mobility of 8.8 cm²/Vs, a small subthreshold swing of 143 mV/decade, and a high I_on/I_off current ratio of 4.23 × 10⁷, presumably because of the reduction in the oxygen vacancies and the formation of the smooth surface roughness as a result of the incorporation of Ti into the Er₂TiO₅ film. Furthermore, the reliability of voltage stress can be improved using an Er₂TiO₅ gate dielectric.     

Structure of silane layer formed on silica particle surfaces by treatment with silane coupling agents having various functional groups

The surface treatment of spherical silica particles with silane coupling agents having various organic functional groups was conducted and the effect of the alkoxy group number on the molecular flexibility of the silane chain with multilayer coverage was investigated using ¹H-pulse nuclear magnetic resonance spectroscopy. The silica particles were treated with 2-propanol solution and heated at 120?°C for 24?h after solvent evaporation to accelerate the polycondensation reaction of silanol groups. For multilayer coverage, flexible linear chain and rigid network structures were expected to form on the surface from the di- and trialkoxy structures, respectively. However, the rigid network structure was formed from both the di- and trialkoxy structures with glycidoxy, amino, and methacryloxy functional silanes. Ring opening of the epoxy group occurred, followed by reaction to form the network structure, even with the dialkoxy structure of the glycidoxy functionality. Ring opening of the epoxy group could be reduced by pH adjustment of the treatment solution and the linear chain structure was formed from the dialkoxy structure. In the case of amino and methacryloxy functional groups, hydrogen bonds were formed between the amino or methacryloxy groups and the silanol groups on the silica surface or silane molecules.     

Chemical Synthesis of a Synthetic Analogue of the Sialic Acid‐Binding Lectin Siglec‐7

As a basis for the development of an artificial carbohydrate‐binding lectin, we chemically synthesized a domain of siglec‐7, a well‐characterized sialic‐acid‐binding lectin. The full polypeptide (127 amino acids) was constructed by sequential native chemical ligation (NCL) of five peptide segments. Because of poor cysteine availability for NCL, cysteine residues were introduced at suitable ligation sites; these cysteine residues were alkylated in order to mimic native glutamine or asparagine residues, or converted to an alanine residue by desulfurization after NCL. After folding the full‐length polypeptide, the sialic‐acid‐binding activity of the synthetic siglec‐7 was clearly demonstrated by STD NMR and ELISA experiments. We succeeded in the synthesis of siglec‐7 by installing three extra cysteine residues with side‐chain modifications and found that these modifications did not affect the binding activity.     

Preparation and characterization of sulfonated block‐graft copolyimide/sulfonated polybenzimidazole blend membranes for fuel cell application

Polymer electrolyte blend membranes composed of sulfonated block‐graft polyimide (S‐bg‐PI) and sulfonated polybenzimidazole (sPBI) were prepared and characterized. The proton conductivity and oxygen permeability coefficient of the novel blend membrane S‐bg‐PI/sPBI (7 wt%) were 0.38 S cm^?1 at 90 °C and 98% relative humidity and 7.2 × 10^?13 cm³(STP) cm (cm² s cmHg)^?1 at 35 °C and 76 cmHg, respectively, while those of Nafion^® were 0.15 S cm^?1 and 1.1 × 10^?10 cm³(STP) cm (cm² s cmHg)^?1 under the same conditions. The apparent (proton/oxygen transport) selectivity calculated from the proton conductivity and the oxygen permeability coefficient in the S‐bg‐PI/sPBI (7 wt%) membrane was 300 times larger than that determined in the Nafion membrane. Besides, the excellent gas barrier properties based on an acid ? base interaction in the blend membranes are expected to suppress the generation of hydrogen peroxide and reactive oxygen species, which will degrade fuel cells during operation. The excellent proton conductivity and gas barrier properties of the novel membranes promise their application for future fuel cell membranes. © 2015 Society of Chemical Industry     

Organofunctionalized catalyst surfaces highly active and selective for carbon–carbon bond-forming reactions

This article illustrates two types of organofunctionalized heterogeneous catalysts for variety of organic carbon–carbon bond-forming reactions, summarizing our previous reports and also presenting new data. Organic amines with an alkoxysilane moiety were immobilized on inorganic silica-alumina surfaces (SA-NR₂) by simple silane-coupling reactions between the silica-alumina surface (SA) and the alkoxysilane. This SA-NR₂ acted as acid–base bifunctional heterogeneous catalysts for carbon–carbon bond-forming reactions, such as cyano-ethoxycarbonylation, Michael reaction of nitriles, and nitro-aldol reaction. These reactions did not occur with either SA or homogeneous amine compounds. In addition, the mixture of SA and homogeneous amine showed low catalytic activity due to undesirable acid–base neutralization reaction. Achiral organic silane-coupling reagents with a variety of functional groups were also immobilized on a SiO₂ surface that had been immobilized with chiral bis(oxazoline) (BOX), to which Cu ions were coordinated to make chiral Cu–BOX complexes on the SiO₂ surface. The SiO₂-supported Cu–BOX complex catalyst functionalized with achiral 3-methacryloxypropyltrimethoxysilane dramatically increased enantioselectivity in the asymmetric Diels–Alder reaction of cyclopentadiene and 3-acryloyl-2-oxazolidinone. The organofunctionalized catalysts showed much better performances for the C–C bond-forming reactions compared to the corresponding homogeneous systems. The heterogeneous catalysts thus obtained were characterized by solid-state ¹³C and ²⁹Si MAS NMR, FT-IR, UV/vis, XAFS, ESR, XRF, and elemental analysis.     

A method to modify coordinates of detectors in positron emission tomography systems

A new method to modify coordinates of detectors in any positron emission tomography (PET) system using a radioactive point source is developed. This method is based on selecting a centered detector in each detector block of PET and measuring coincidence counts between the two centered detectors in opposite detector blocks to find the coordinates of their LOR (Line of Response). Due to slight misalignment of detector positions, measured LORs may not intersect at a single point. Based on the proposed method, the coordinates of detectors can be measured with very high accuracy and the coordinate of the center of the gantry (which is normally the same as the center of field of view) can be defined correctly. The results of the application of our method to a small animal PET system (FinePET), which was recently developed at Tohoku University, Japan, are shown here. The method is expected to contribute to the design and development of PET systems which can realize a very high spatial resolution of less than 1 mm FWHM.     

Phthalocyanine molecular nanowires that were prepared using porous alumina as a template: Development in the sample preparation procedure to evaluate electronic properties

We have proposed the synthesis of organic molecular nanowires using porous alumina as a template. We also proposed the use of a magnetic field to control the molecular packing structure in the nanowires. In this paper, we developed the method to evaluate the electronic properties of the nanowire of a phthalocyanine derivative that was synthesized using porous alumina as a template. The developed method facilitates the study in the organic molecular nanowires that were synthesized using templates and helps their use in future electronic devices.