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.     

Simultaneous three-dimensional step-height measurement and high-resolution tomographic imaging with a spectral interferometric microscope

A noncontact, nonmechanical scanning, wide-field spectral interference microscope is developed for simultaneous measurement of three-dimensional step-height of discontinuous objects and tomographic imaging. A superluminescent diode (SLD) is used as a broadband light source and a liquid-crystal Fabry-Perot interferometer (LC-FPI) as a frequency-scanning device. By means of changing the injection current to the SLD, the spectral profile of the SLD is equalized, and a constant light input to the interferometer is achieved over the entire frequency-scan range. The Fourier-transform technique is used to determine both the amplitude and the phase of spectral fringe signals. Three-dimensional height distribution of a discontinuous object is obtained from the phase information, whereas optically sectioned images of the object are obtained either from the amplitude information alone or from the combination of both the amplitude and phase information. Experimental results with submicrometer resolution are presented for both step-height measurement and tomographic sectioning.     

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.     

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.     

红外相机CASCAM的电动系统设计

介绍了红外相机CASCAM的电动系统设计。系统采用AT90S8515单片机和TA8435集成芯片组成驱动电路的核心,通过电机选择电路和位置反馈电路实现对步进电机的选择驱动和定位、复位。系统采用MAXIM232芯片实现单片机和上位机之间的串行通讯。系统软件以汇编语言编制,并给出了系统程序框图。经实验验证,该系统运行可靠,达到了设计要求。     

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.     

Spectral interferometric microscope with tandem liquid-crystal Fabry-Perot interferometers for extension of the dynamic range in three-dimensional step-height measurement

The maximum measurable range of a spectral interference microscope depends on the coherence length of the light transmitted by its tunable spectral filter. To achieve a large range in step-height measurement we have developed a new tunable spectral filter that uses tandem liquid-crystal Fabry-Perot interferometers (LC-FPIs), which can simultaneously attain both a high spectral resolution and a large tuning range. Fringe visibility measurements were carried out, and it was found that the coherence length of the light transmitted through tandem LC-FPIs is two times larger than that transmitted through a single LC-FPI. Using this novel tunable spectral filter, we developed a new spectral interference microscope for the measurement of three-dimensional shapes of discontinuous objects. Experimental results of step-height measurements both with a single LC-FPI and with tandem LC-FPIs are presented for a combination of standard steel gauge block sets with 1-, 99-, and 100-microm steps. A large range (1-100 microm) of measurement with submicrometer resolution was achieved with tandem LC-FPIs that was not possible with our previous system in which a single LC-FPI was used.     

Cloning of a fatty acid synthase component FAS1 gene from Saccharomyces kluyveri and its functional complementation of S. cerevisiae fas1 mutant

A gene encoding a fatty acid synthase component, FAS1, has been cloned from a genomic library of the polyunsaturated fatty acid (PUFA)-producing yeast Saccharomyces kluyveri. This gene (named Sk-FAS1) was found to contain an open reading frame of 6150 bp, coding for 2049 amino acids. The deduced Sk-FAS1 protein showed significant (75-59%) homology with FAS proteins from the other yeasts, including S. cerevisiae, Candida albicans and Yarrowia lipolytica. The substrate-binding sites of the acetyl transferase and malonyl/palmitoyl transferase domains, and the FMN- and NADPH-binding sites of the enoyl reductase domain, were all highly conserved. Expression of the Sk-FAS1 gene in S. cerevisiae complemented genetic disruption of the S. cerevisiae FAS1 gene (Sc-FAS1), suggesting the formation of a heterogeneous complex of Sk-FAS1 (beta) and Sc-FAS2 (alpha), which is able to function to synthesize fatty acids. Compared with the isogenic wild-type of S. cerevisiae, as well as S. kluyveri, the S. cerevisiae fas1 mutant carrying the Sk-FAS1 gene showed an increase in the relative amount of 16-carbon fatty acids and a decrease in 18-carbon fatty acids.