Cloning and characterization of a β-N-acetylglucosaminidase (BmFDL) from silkworm Bombyx mori

In insects, β-N-acetylglucosaminidase (GlcNAcase) participates in critical physiological processes such as fertilization, metamorphosis, and glycoconjugate degradation. Insects produce glycoproteins carrying paucimannosidic-type N-glycans, the terminal GlcNAc residue of which is cleaved by a GlcNAc-linkage specific GlcNAcase, also known as the fused lobes (FDL) protein. To obtain information on the structure of GlcNAcases and insight into their contribution to physiological processes, we cloned Bombyx mori FDL (BmFDL) from silkworm larvae. The full-length cDNA (1.9 kb) encoded a protein of 633 amino acids with 42% amino acid sequence identity to Drosophila melanogaster FDL (DmFDL). Recombinant BmFDL cleaved only β-1,2-linked GlcNAc residues from the α-1,3 branch of biantennary N-glycan. This substrate specificity was similar to that of DmFDL. Microsomal FDL activity was inhibited by anti-BmFDL antibodies. Taken together, our results suggest that BmFDL is a N-glycan-processing GlcNAcase in B. mori.     

Template-free hydrothermal synthesis of hollow hematite microspheres

Hollow hematite (α-Fe₂O₃) microspheres with an average diameter of 3-4 μm and a shell thickness of approximate 150 nm was synthesized by a simple hydrothermal route using FeCl₃·6H₂O solution and acetic acid without using any templates. The hollow microspheres were composed of α-Fe₂O₃ nanoparticles with the diameter range from 20 to 40 nm. The effects of reaction parameters such as reaction time, temperature, concentration of FeCl₃·6H₂O solution, and initial pH on the morphology of the final products were investigated. A possible formation mechanism of hollow α-Fe₂O₃ microspheres was also proposed, where the acetic acid played a role of etching in the formation of hollow structure.     

Involvement of cytochrome P450 in hydroxylation of propylbenzene by Fusarium moniliforme strain MS31

Fusarium moniliforme strain MS31 can oxidize propylbenzene to (R)-1-phenylpropanol with what may be a cytochrome P450. Hydroxylation of propylbenzene needed molecular oxygen, and NADPH as a coenzyme gave a higher yield than NADH. The hydroxylation proceeded further when FAD and FMN were added than in their absence, suggesting that the enzyme was a flavo-protein. Carbon monoxide inhibited the hydroxylation, as did other cytochrome P450 inhibitors such as SKF 525A and miconazole. These characteristics matched those of a microsomal cytochrome P450 monooxygenase system that contained NADPH-cytochrome P450 reductase.     

Updated data dissemination methods for updating old replicas in ad hoc networks

In this paper, we propose two updated data dissemination methods to not only reduce the number of accesses to old replicas, but also to improve the data accessibility in ad hoc networks where data items are updated regularly. In the first method, when a mobile host updates a data item, it disseminates the updated data item after the flooding of with invalidation reports. In the second method, two newly connected mobile hosts disseminate updated data items with each other after the flooding with invalidation reports.     

Properties of graphite prepared from boron-doped pitch as an anode for a rechargeable Li ion battery

Boron-doped graphites were derived from a naphthalene-based pitch mixed with para-xylene glycol (PXG) or dimethyl para-xylene glycol (DMPXG) as a cross-linking agent and three types of boron-containing compounds as a graphitization catalyst, and their anode performances were investigated. The structural analysis of the obtained graphites revealed that PXG functioned mainly as a two-dimensional cross-linking agent during the heat treatment process and DMPXG functioned partially as a three-dimensional. The average interlayer spacing decreased and lattice constant, a₀, and graphitizability increased with increasing the amount of boron atoms added. The result indicated that the carbon atoms were replaced by boron atoms. The anode performance was improved by the enhancement of graphitizability. The structural parameters and anode performance of boron-doped graphites did not depend on the kind of boron-containing compounds but the amount of boron atoms added in pitch and the kind of cross-linking agent.     

Reduction of germination frequency in Anabaena akinetes by sediment drying: a possible method by which to inhibit bloom formation

The effect of desiccation on germination frequency of Anabaena akinetes was investigated to evaluate the practical usefulness of sediment drying in reservoirs as a measure of inhibition of Anabaena bloom formation. Sediment samples taken from a small reservoir, where an abundance of Anabaena blooms occur every summer were desiccated in the outdoors and in an incubator at 5, 10, 15, and 25 °C. As a result, the germination frequency of Anabaena akinetes was strongly inhibited in comparison with that of Anabaena akinetes preserved in a refrigerator without desiccation as a control. Greater inhibition of Anabaena germination found at a higher temperature was examined, and almost all akinetes lost the ability of germination at 25 °C after 3 days of the desiccation treatment. Therefore, sediment drying is considered to be a useful measure to reduce periods and scales of Anabaena blooms, and its effect will be enhanced by performance during the warmer seasons.     

Efficient synthesis of nonnatural mutants in Escherichia coli S30 in vitro protein synthesizing system

Factors that affect the efficiency of in vitro synthesis of mutant proteins that contain nonnatural amino acids were investigated. The process of the nonnatural mutagenesis consists of chemical aminoacylation of a tRNA that contains a 4-base anticodon, followed by in vitro synthesis in the presence of an mRNA that contains the corresponding 4-base codon. Detailed studies on the time courses of the synthesis revealed two major factors that suppress the yield of nonnatural mutants compared with the wild-type protein. First, a cyclic tRNA that exists as a by-product of the chemical aminoacylation inhibits the protein synthesis. Second, the very short lifetime of a tRNA aminoacylated with a nonnatural amino acid limits the protein yield. As a simple and practical way of surmounting these factors, aminoacyl tRNA was added into the in vitro system at 5 min after the start of the synthesis. The addition increased the protein yield up to the level of conventional proteins in the in vitro system.     

An aqueous suspension system for phospholipase D-mediated synthesis of PS without toxic organic solvent

Enzymatic synthesis of PS by phospholipase D (PLD)-mediated transphosphatidylation in an aqueous media was investigated. The purpose of this study was to establish a novel synthetic method where no toxic organic solvents were used. An attempt to react soybean lecithin (simply dispersed in an aqueous buffer) with an aqueous solution of l-serine and PLD was unsuccessful, giving only 20% of PS. By contrast, a suspension of lecithin adsorbed on fine powders such as silica was effectively converted into PS in an aqueous solution of l-serine and PLD. After screening various powders for use as the lecithin adsorbent, calcium sulfate was found to be the best with respect to lecithin conversion. In addition, calcium sulfate did not require prior adsorption of lecithin (i.e., the reaction proceeded effectively simply by adding the powder to an aqueous mixture of lecithin, l-serine, and PLD). With this “aqueous suspension system” of calcium sulfate, up to 180 mg/mL lecithin was completely converted, resulting in more than 80% PS in 24 h. The synthesized PS could easily be recovered from the powder by extracting with a mixture of n-hexane, ethanol, and diluted HCl.