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Cells regulate their intracellular mRNA levels by using specific ribonucleases. Oligoribonuclease (ORN) is a 3′–5′ exoribonuclease for small RNA molecules, important in RNA degradation and re-utilisation. However, there is no structural information on the ligand-binding form of ORNs. In this study, the crystal structures of oligoribonuclease from Colwellia psychrerythraea strain 34H (CpsORN) were determined in four different forms: unliganded-structure, thymidine 5′-monophosphate p-nitrophenyl ester (pNP-TMP)-bound, two separated uridine-bound, and two linked uridine (U-U)-bound forms. The crystal structures show that CpsORN is a tight dimer, with two separated active sites and one divalent metal cation ion in each active site. These structures represent several snapshots of the enzymatic reaction process, which allowed us to suggest a possible one-metal-dependent reaction mechanism for CpsORN. Moreover, the biochemical data support our suggested mechanism and identified the key residues responsible for enzymatic catalysis of CpsORN.

Dihydrodipicolinate reductase (DHDPR) is a key enzyme in the diaminopimelate- and lysine-synthesis pathways that reduces DHDP to tetrahydrodipicolinate. Although DHDPR uses both NADPH and NADH as a cofactor, the structural basis for cofactor specificity and preference remains unclear. Here, we report that Paenisporosarcina sp. TG-14 PaDHDPR has a strong preference for NADPH over NADH, as determined by isothermal titration calorimetry and enzymatic activity assays. We determined the crystal structures of PaDHDPR alone, with its competitive inhibitor (dipicolinate), and the ternary complex of the enzyme with dipicolinate and NADPH, with results showing that only the ternary complex had a fully closed conformation and suggesting that binding of both substrate and nucleotide cofactor is required for enzymatic activity. Moreover, NADPH binding induced local conformational changes in the N-terminal long loop (residues 34–59) of PaDHDPR, as the His35 and Lys36 residues in this loop interacted with the 2′-phosphate group of NADPH, possibly accounting for the strong preference of PaDHDPR for NADPH. Mutation of these residues revealed reduced NADPH binding and enzymatic activity, confirming their importance in NADPH binding. These findings provide insight into the mechanism of action and cofactor selectivity of this important bacterial enzyme.

Xylose isomerase (XI; E.C. 5.3.1.5) catalyzes the isomerization of xylose to xylulose, which can be used to produce bioethanol through fermentation. Therefore, XI has recently gained attention as a key catalyst in the bioenergy industry. Here, we identified, purified, and characterized a XI (

Additional file 1: Fig. S1. Phylogenetic analysis of SfSFGH. Fig. S2. Gene clustering analysis of SfSFGH. Fig. S3. Recombinant SfSFGH protein purification, crystallization, and X-ray diffraction data collection. Fig. S4. Freezeâ thaw cycles of SfSFGH.

Methionine sulfoxide reductase (Msr) is a family of enzymes that reduces oxidized methionine and plays an important role in the survival of bacteria under oxidative stress conditions. MsrA and MsrB exist in a fusion protein form (MsrAB) in some pathogenic bacteria, such as Helicobacter pylori (Hp), Streptococcus pneumoniae, and Treponema denticola. To understand the fused form instead of the separated enzyme at the molecular level, we determined the crystal structure of HpMsrABC44S/C318S at 2.2 Å, which showed that a linker region (Hpiloop, 193–205) between two domains interacted with each HpMsrA or HpMsrB domain via three salt bridges (E193-K107, D197-R103, and K200-D339). Two acetate molecules in the active site pocket showed an sp2 planar electron density map in the crystal structure, which interacted with the conserved residues in fusion MsrABs from the pathogen. Biochemical and kinetic analyses revealed that Hpiloop is required to increase the catalytic efficiency of HpMsrAB. Two salt bridge mutants (D193A and E199A) were located at the entrance or tailgate of Hpiloop. Therefore, the linker region of the MsrAB fusion enzyme plays a key role in the structural stability and catalytic efficiency and provides a better understanding of why MsrAB exists in a fused form.

This study aims to examine and analyze the current status of patent trend for housing modification related technologies for the elderly in order to understand the current situation of housing modification industry in Korea and other countries. Using KOMPASS patent search system as a searching tool, patents for housing modification related technologies in Korea, Japan, the U.S., and Europe were searched and 999 valid patents were found. The country with the most patents was Japan, which was followed by the U.S.. Valid patents were categorized based on 9 elements of criteria for technology classification (secondary lifting device, porch, kitchen, bathroom or toilet, moving assistive device, lighting or door, IT-related technology, reform plan and extra), and these were divided by country, year, and technology classification and analyzed. Through the yearly analysis of housing modification related technologies, it was found that the increase and decrease of the patent trend of each country was related to the duration of aging society and super-aged society, which is the index of the number of the elderly compared to the total population. In case of Korea, from the late 80s to the late 90s, there were intermittent applications for patents and since 2000, the year that Korea reached aging society, the number has continuously increased and patent applications for bathroom-related technology took the biggest portion. Another thing to note was that compared to other countries researched, Korea showed an increasing trend in the number of patent applications related to IT technologies such as emergency call system and abnormality detection monitoring. Still, the number of Korea's patent applications was smaller than that of other countries analyzed, and the area of technologies that were applied for patent was not diverse. In the future, Korea will have to focus on developing housing modification technologies related to the elderly, which is at a poor stage, and for this, it will have to put efforts for developing R&D technology and securing intellectual property. Especially, compared to other countries, Korea will be able to use its domestic industrial environment that can easily apply IT technologies.

Dye-sensitized solar cells (DSCs) are promising solar energy conversion devices with aesthetically favorable properties such as being colorful and having transparent features. They are also well-known for high and reliable performance even under ambient lighting, and these advantages distinguish DSCs for applications in window-type building-integrated photovoltaics (BIPVs) that utilize photons from both lamplight and sunlight. Therefore, investigations on bifacial DSCs have been done intensively, but further enhancement in performance under back-illumination is essential for practical window-BIPV applications. In this research, highly efficient bifacial DSCs were prepared by a combination of electropolymerized poly(3,4-ethylenedioxythiphene) (PEDOT) counter electrodes (CEs) and cobalt bipyridine redox ([Co(bpy)3]3+/2+) electrolyte, both of which manifested superior transparency when compared with conventional Pt and iodide counterparts, respectively. Keen electrochemical analyses of PEDOT films verified t...

Bacterial cytochrome P450 (CYP) steroid hydroxylases are effectively useful in the pharmaceutical industry for introducing hydroxyl groups to a wide range of steroids. We found a putative CYP steroid hydroxylase (

To find the catalytic activities of CYP191A1 from Mycobacterium smegmatis, in which functions of most P450s are unknown, by using a set of reductase systems, peroxides, and various substrates including fatty acids and human drugs. CYP191A1 was functionally expressed in Escherichia coli and purified. Its catalytic activities were examined with fatty acids, chromogenic and fluorogenic substrates, and several human P450 substrates, in the presence of six different types of electron transfer systems, such as rat NADPH-P450 reductase, Candida NADPH-P450 reductase, ferredoxin/ferredoxin reductase, putidaredoxin/putidaredoxin reductase, and peroxides (H2O2 and t-butyl hydroperoxide). The reactions catalyzed by CYP191A1 included the hydroxylation and O-dealkylation of several substrates. CYP191A1 preferentially catalyzes the peroxide-dependent oxidation of various substrates over the reductase-dependent reaction. Its peroxygenase activity may be used an effective biocatalytic tool to synthesize the metabolites of drugs.

Escherichia coli is a representative microorganism that is frequently used for industrial biotechnology; thus its cellular robustness should be enhanced for the widespread application of E. coli in biotechnology. Stress response genes from the extremely radioresistant bacterium Deinococcus radiodurans have been used to enhance the stress tolerance of E. coli. In the present study, we introduced the cold shock domain-containing protein PprM from D. radiodurans into E. coli and observed that the tolerance to hydrogen peroxide (H2O2) was significantly increased in recombinant strains (Ec-PprM). The overexpression of PprM in E. coli elevated the expression of some OxyR-dependent genes, which play important roles in oxidative stress tolerance. Particularly, mntH (manganese transporter) was activated by 9-fold in Ec-PprM, even in the absence of H2O2 stress, which induced a more than 2-fold increase in the Mn/Fe ratio compared with wild type. The reduced production of highly reactive hydroxyl radicals (·OH) and low protein carbonylation levels (a marker of oxidative damage) in Ec-PprM indicate that the increase in the Mn/Fe ratio contributes to the protection of cells from H2O2 stress. PprM also conferred H2O2 tolerance to E. coli in the absence of OxyR. We confirmed that the H2O2 tolerance of oxyR mutants reflected the activation of the ycgZ-ymgABC operon, whose expression is activated by H2O2 in an OxyR-independent manner. Thus, the results of the present study showed that PprM could be exploited to improve the robustness of E. coli.

This study employs for the first time surface-nitrided Ni foam as a counter-electrode free of Pt and transparent conducting oxides (TCOs) to realize a cost-effective counter electrode for dye-sensitized solar cells (DSSCs). This electrode simultaneously features high catalytic activity for triiodide reduction and high conductivity in a single layer. The nitrided Ni foam is synthesized by nitridation treatment of open-cell Ni foam in an ammonia atmosphere. This electrode presents high catalytic activity on the nitrided surface and easy electron transport ability in the three-dimensional, interconnected metallic structure. This study provides a preliminary design concept for utilizing the nitrided Ni foam as a promising cost-effective counter electrode that does not require expensive Pt and TCO.

An inverse-opal poly(3,4-ethylene dioxythiophene) (PEDOT) electrode was fabricated by electrodeposition of EDOT onto a PEDOT:poly(styrene sulfonate) filler in a self-assembled polystyrene template. By this novel fabrication method, the inverse-opal electrode was directly assembled into a dye-sensitized solar cell (DSSC) as a Pt-free counter electrode without any additional processing. This inverse-opal PEDOT electrode exhibits superior electrocatalytic activity and easy electrolyte diffusion in the electrode, as the inverse-opal electrode provides a porous morphology. The improved reflectivity of the inverse-opal electrode was also observed in the incident-photon conversion-efficiency spectra at low wavelength, which was a characteristic of its well-ordered structure. Therefore, the inverse-opal PEDOT electrode could successfully act as an alternative counter electrode for DSSCs to replace Pt in the counter electrode.

A large set of Bacillus megaterium CYP102A1 mutants are known to metabolize various drugs to form human metabolites. Omeprazole (OMP), a proton pump inhibitor, has been widely used as an acid inhibitory agent for the treatment of gastric acid hypersecretion disorders. It is primarily metabolized by human CYP2C19 and CYP3A4 to 5'-OH OMP and a sulfone product, respectively. It was recently reported that several CYP102A1 mutants can oxidize racemic and S-OMP to 5'-OH OMP and that these mutants can further oxidize 5'-OH racemic OMP to 5'-COOH OMP. Here, we report that the S- and R-enantiomers of OMP are hydroxylated by 26 mutants of CYP102A1 to produce 1 major metabolite (5'-OH OMP) regardless of the chirality of the parent substrates. Although the binding of R-OMP to the CYP102A1 active site caused a more apparent change of heme environment compared with binding of S-OMP, there was no correlation between the spectral change upon substrate binding and catalytic activity of either enantiomer. The 5'-OH OMP produced from racemic, S-, and R-OMP could be obtained with a high conversion rate and high selectivity when the triple R47L/F87V/L188Q mutant was used. These results suggest that bacterial CYP102A1 mutants can be used to produce the human metabolite 5'-OH OMP from both the S- and R-enantiomers of OMP.

Human drug metabolites produced by cytochrome P450 enzymes are critical for safety testing and may themselves act as drugs or leads in the drug discovery and development process. Here, highly active chimeric fusion proteins (chimeras) were obtained by reductase domain swapping of mutants at key catalytic residues of the heme domain with that of a natural variant (CYP102A1.2) of P450 BM3 (CYP102A1.1) from Bacillus megaterium. Random mutagenesis at the heme domain of the chimera was also used to generate chimeric mutants that were more active and diverse than the chimeras themselves. To determine whether the chimeras and several mutants of the highly active chimera displayed enhanced catalytic activity and, more importantly, whether they acquired activities of biotechnological importance, we measured the oxidation activities of the chimeras and chimeric mutants toward human P450 substrates, mainly drugs. Some of the chimeric mutants showed high activity toward typical human P450 substrates including drugs. Statin leads, especially chiral products, with inhibitory effects toward HMG-CoA reductase could be obtained from metabolites of statin drugs generated using these chimeric mutants. This study reveals the critical role of the reductase domain for the activity of P450 BM3 and shows that chimeras generated by domain swapping can be used to develop industrial enzymes for the synthesis of human metabolites from drugs and drug leads. Biotechnol. Bioeng. 2014;111: 1313–1322. © 2014 Wiley Periodicals, Inc.

Cytochromes P450 (CYP for isoforms) play a central role in biological processes especially metabolism of chiral molecules; thus, development of computational methods to predict parameters for chiral reactions is important for advancing this field. In this study, we identified the most optimal artificial neural networks using conformation-independent chirality codes to predict CYP2C19 catalytic parameters for enantioselective reactions. Optimization of the neural networks required identifying the most suitable representation of structure among a diverse array of training substrates, normalizing distribution of the corresponding catalytic parameters ( k cat , K m , and k cat / K m ), and determining the best topology for networks to make predictions. Among different structural descriptors, the use of partial atomic charges according to the CHelpG scheme and inclusion of hydrogens yielded the most optimal artificial neural networks. Their training also required resolution of poorly distributed output catalytic parameters using a Box–Cox transformation. End point leave-one-out cross correlations of the best neural networks revealed that predictions for individual catalytic parameters ( k cat and K m ) were more consistent with experimental values than those for catalytic efficiency ( k cat / K m ). Lastly, neural networks predicted correctly enantioselectivity and comparable catalytic parameters measured in this study for previously uncharacterized CYP2C19 substrates, R - and S -propranolol. Taken together, these seminal computational studies for CYP2C19 are the first to predict all catalytic parameters for enantioselective reactions using artificial neural networks and thus provide a foundation for expanding the prediction of cytochrome P450 reactions to chiral drugs, pollutants, and other biologically active compounds.

Coumadin (R/S-warfarin) is a highly efficacious and widely used anticoagulant; however, its highly variable metabolism remains an important contributor to uncertainties in therapeutic responses. Pharmacogenetic studies report conflicting findings on the clinical relevance of CYP2C19. A resolution to this controversy is impeded by a lack of detail on the potential role of CYP2C19 in warfarin metabolism. Consequently, we assessed the efficiency of CYP2C19 metabolism of R- and S-warfarin and explored possible contributions in the liver using in vitro methods. Recombinant CYP2C19 metabolized R- and S-warfarin mainly to 6-, 7-, and 8-hydroxywarfarin, while 4′-hydroxywarfarin was a minor metabolite. Overall R-warfarin metabolism was slightly more efficient than that for S-warfarin. Metabolic pathways that produce R-6-, 7-, and 8-hydroxywarfarin in human liver microsomal reactions correlated strongly with CYP2C19 S-mephenytoin hydroxylase activity. Similarly, CYP1A2 activity toward phenacetin correlated with formation of R-6 and 7-hydroxywarfarin such that R-8-hydroxywarfarin seems unique to CYP2C19 and possibly a biomarker. In following, CYP2C19 likely impacts R-warfarin metabolism and patient response to therapy. Intriguingly, CYP2C19 may contribute to S-warfarin metabolism in patients, especially when CYP2C9 activity is compromised due to drug interactions or genetic polymorphisms.

In this study, we investigated the potential of using nickel oxide (NiO)-lithium phosphorous oxynitride (LiPON) composite as an anode electrode in electrochromic devices and evaluated the effect of Li+ insertion (or extraction) kinetics and diffusion of Li+. NiO–LiPON composite thin films were fabricated using the RF magnetron co-sputtering method. Here, we observed, for the first time, the bimodal colored states of the NiO–LiPON composite. This was caused by reacting both Ni metal and LiO under extremely cathodic condition, which enhanced the electrochromic properties – electrochromic response time and optical modulations – of the device. We also found that the NiO–LiPON composite thin films displayed long term stability.

Cellular robustness is an important trait for industrial microbes, because the microbial strains are exposed to a multitude of different stresses during industrial processes, such as fermentation. Thus, engineering robustness in an organism in order to push the strains toward maximizing yield has become a significant topic of research. We introduced the deinococcal response regulator DR1558 into Escherichia coli (strain Ec -1558), thereby conferring tolerance to hydrogen peroxide (H 2 O 2 ). The reactive oxygen species (ROS) level in strain Ec -1558 was reduced due to the increased KatE catalase activity. Among four regulators of the oxidative-stress response, OxyR, RpoS, SoxS, and Fur, we found that the expression of rpoS increased in Ec -1558, and we confirmed this increase by Western blot analysis. Electrophoretic mobility shift assays showed that DR1558 bound to the rpoS promoter. Because the alternative sigma factor RpoS regulates various stress resistance-related genes, we performed stress survival analysis using an rpoS mutant strain. Ec -1558 was able to tolerate a low pH, a high temperature, and high NaCl concentrations in addition to H 2 O 2 , and the multistress tolerance phenotype disappeared in the absence of rpoS . Microarray analysis clearly showed that a variety of stress-responsive genes that are directly or indirectly controlled by RpoS were upregulated in strain Ec -1558. These findings, taken together, indicate that the multistress tolerance conferred by DR1558 is likely routed through RpoS. In the present study, we propose a novel strategy of employing an exogenous response regulator from polyextremophiles for strain improvement.

New nickel oxide thin films with a micro-scale thickness and porous structure were fabricated using a sol–gel method in a mixed solvent containing N,N-dimethylaminoethanol (dmaeH) and distilled water at a 1:1 ratio. After annealing, the deposited film transformed to a well oriented, porous nickel oxide thin film. This nickel oxide thin film showed enhanced charge capacity and greater optical transmittance changes in a lithium-based electrolyte, and may be applied to WO3/Li+-based electrolyte/NiO electrochromic devices requiring long cycles.

Thin film electrodes have been intensively studied for active materials and current collectors toenhance the electrochemical performance. Here, porous structures of nickel-based oxide films, con-sisting of nickel oxide and copper (II) oxide, which was derived from the copper substrate duringthe annealing process, were deposited on metallic copper foils. The half-cell tests revealed excellentcapacity retention after 80 th charge/discharge cycles. Some films showed an excess of the theoret-ical capacity of nickel oxides, which mainly originate from partially oxidized copper substrates dur-ing annealing. These results exhibit that both a preparation method of an active materials andpartially oxidized current collectors could be important roles to apply thin film electrodes.Keywords : Nickel Oxides, Copper Oxides, Current Collectors, Anode, Lithium Ion Batteries Received December 3, 2011 : Accepted December 27, 2011 1. Introduction The decreasing size of electronic devices has focusedincreasing attention on thin film batteries as an on-chippower supplier. Like the general types of lithium ionbatteries, thin film batteries use an anode and cathode.In particular, the anode is composed of a current collec-tor and an active material that reacts with lithium ionsdirectly. Most preliminary approaches focused on cur-rent collectors

Human cytochrome P450 (P450) enzymes metabolize a variety of endogenous and xenobiotic compounds, including steroids, drugs, and environmental chemicals. In this study, we examine the possibility that bacterial P450 BM3 (CYP102A1) mutants with indole oxidation activity have the catalytic activities of human P450 enzymes. Error-prone polymerase chain reaction was carried out on the heme domain-coding region of the wild-type gene to generate a CYP102A1 DNA library. The library was transformed into Escherichia coli for expression of the P450 mutants. A colorimetric colony-based method was adopted for primary screening of the mutants. When the P450 activities were measured at the whole-cell level, some of the blue colonies, but not the white colonies, possessed apparent oxidation activity toward coumarin and 7-ethoxycoumarin, which are typical human P450 substrates that produce fluorescent products. Coumarin is oxidized by the CYP102A1 mutants to produce two metabolites, 7-hydroxycoumarin and 3-hydroxycoumarin. In addition, 7-ethoxycoumarin is simultaneously oxidized to 7-hydroxycoumarin by O-deethylation reaction and to 3-hydroxy,7-ethoxycoumarin by 3-hydroxylation reactions. Highly active mutants are also able to metabolize several other human P450 substrates, including phenacetin, ethoxyresorufin, and chlorzoxazone. These results indicate that indigo formation provides a simple assay for identifying CYP102A1 mutants with a greater potential for human P450 activity. Furthermore, our computational findings suggest a correlation between the stabilization of the binding site and the catalytic efficiency of CYP102A1 mutants toward coumarin: the more stable the structure in the binding site, the lower the energy barrier and the higher the catalytic efficiency.

We prepared polyaniline-poly(4-styrenesulfonate) nanoparticles (PANI/PSS-NPs) by chemical oxidation polymerization in aqueous solution. We investigated the potential of the PANI/PSS-NPs to be used as an anode electrode for electrochromic devices and the effect of Li+ insertion (or deinsertion) kinetics and diffusion of Li+. A uniform electrochromic layer of PANI/PSS-NPs with a size of ca. 28 nm could be obtained by a solution process, specifically spin coating. The PANI/PSS-NPs film has a high Li+ diffusion coefficient (∼7.7 × 10−9 cm2 s−1) and low charge transfer resistance (∼99 Ω), which result in its having a fast electrochromic response time (coloring time 108 cm2 C−1). Keywords: Electrochromic, Polyaniline, Polymer nanoparticle, Conjugated polymer, Diffusion coefficient, Response time

This study examined the material and electrochromic properties of vanadium–titanium oxides (V–Ti oxides) as a counter electrode material in electrochromic devices. These oxides were deposited on an ITO substrate using a co-sputtering method at different levels of RF power. Electrochemical experiments of these oxides were carried out using half-cell and semi full-cell tests which are good methods for measuring the potential applied to each electrode. The change in electrochromic properties after 1000 cycles of a semi full-cell test was examined. Reversibility and durability of an electrochromic device were improved by increasing the titanium content in V–Ti oxides.

In this study, we fabricated nickel oxide thin film for lithium based electrolyte using sol-gel method. This film was deposited by dip-coating method with mixed solvent of DameH (N,N-dimethylaminoethanol) and DI water. As changing the ratio between DmaeH and DI water, nickel oxide thin film was presented in different charge density and optical transmittance because they were shown various thickness. It was accounted for changing viscosity and density by the ratio of DmaeH and DI water. The thin film synthesized with 1 : 1 ratio of DmaeH and DI water was expressed best electrochromic performance in lithium based electrolyte, because of thick thickness but porous structures.

Nickel nitride electrodes were prepared by reactive sputtering of nickel under a N2 atmosphere at room temperature for application in mesoscopic dye- or quantum dot- sensitized solar cells. This facile and reliable method led to the formation of a Ni2N film with a cauliflower-like nanostructure and tetrahedral crystal lattice. The prepared nickel nitride electrodes exhibited an excellent chemical stability toward both iodide and polysulfide redox electrolytes. Compared to conventional Pt electrodes, the nickel nitride electrodes showed an inferior electrocatalytic activity for the iodide redox electrolyte; however, it displayed a considerably superior electrocatalytic activity for the polysulfide redox electrolyte. As a result, compared to dye-sensitized solar cells (DSCs), with a conversion efficiency (η) = 7.62% and CdSe-based quantum dot-sensitized solar cells (QDSCs, η = 2.01%) employing Pt counter electrodes (CEs), the nickel nitride CEs exhibited a lower conversion efficiency (η = 3.75%) when applied to DSCs, but an enhanced conversion efficiency (η = 2.80%) when applied to CdSe-based QDSCs.

Mesoscopic solar cells based on nanostructured oxide semiconductors are considered as a promising candidates to replace conventional photovoltaics employing costly materials. However, their overall performances are below the sufficient level required for practical usages. Herein, this study proposes an anodized Ti foam (ATF) with multidimensional and hierarchical architecture as a highly efficient photoelectrode for the generation of a large photocurrent. ATF photoelectrodes prepared by electrochemical anodization of freeze-cast Ti foams have three favorable characteristics: (i) large surface area for enhanced light harvesting, (ii) 1D semiconductor structure for facilitated charge collection, and (iii) 3D highly conductive metallic current collector that enables exclusion of transparent conducting oxide substrate. Based on these advantages, when ATF is utilized in dye-sensitized solar cells, short-circuit photocurrent density up to 22.0 mA cm(-2) is achieved in the conventional N719 dye-I-3(-)/I- redox electrolyte system even with an intrinsically inferior quasi-solid electrolyte.

Human drug metabolites produced by cytochrome P450 enzymes are critical for safety testing and may themselves act as drugs or leads in the drug discovery and development process. Here, highly active chimeric fusion proteins (chimeras) were obtained by reductase domain swapping of mutants at key catalytic residues of the heme domain with that of a natural variant (CYP102A1.2) of P450 BM3 (CYP102A1.1) from Bacillus megaterium. Random mutagenesis at the heme domain of the chimera was also used to generate chimeric mutants that were more active and diverse than the chimeras themselves. To determine whether the chimeras and several mutants of the highly active chimera displayed enhanced catalytic activity and, more importantly, whether they acquired activities of biotechnological importance, we measured the oxidation activities of the chimeras and chimeric mutants toward human P450 substrates, mainly drugs. Some of the chimeric mutants showed high activity toward typical human P450 substrates including drugs. Statin leads, especially chiral products, with inhibitory effects toward HMG-CoA reductase could be obtained from metabolites of statin drugs generated using these chimeric mutants. This study reveals the critical role of the reductase domain for the activity of P450 BM3 and shows that chimeras generated by domain swapping can be used to develop industrial enzymes for the synthesis of human metabolites from drugs and drug leads.

An extreme diversity of substrates and catalytic reactions of cytochrome P450 (P450) enzymes is considered to be the consequence of evolutionary adaptation driven by different metabolic or environmental demands. Here we report the presence of numerous natural variants of P450 BM3 (CYP102A1) within a species of Bacillus megaterium. Extensive amino acid substitutions (up to 5% of the total 1049 amino acid residues) were identified from the variants. Phylogenetic analyses suggest that this P450 gene evolve more rapidly than the rRNA gene locus. It was found that key catalytic residues in the substrate channel and active site are retained. Although there were no apparent variations in hydroxylation activity towards myristic acid (C14) and palmitic acid (C16), the hydroxylation rates of lauric acid (C12) by the variants varied in the range of >25-fold. Interestingly, catalytic activities of the variants are promiscuous towards non-natural substrates including human P450 substrates. It can be suggested that CYP102A1 variants can acquire new catalytic activities through site-specific mutations distal to the active site.

Recently, the wild-type and mutant forms of cytochrome P450 BM3 (CYP102A1) from Bacillus megaterium were found to oxidize various xenobiotic substrates, including pharmaceuticals, of human P450 enzymes. Simvastatin and lovastatin, which are used to treat hyperlipidemia and hypercholesterolemia, are oxidized by human CYP3A4/5 to produce several metabolites, including 6'β-hydroxy (OH), 3″-OH, and exomethylene products. In this report, we show that the oxidation of simvastatin and lovastatin was catalyzed by wild-type CYP102A1 and a set of its mutants, which were generated by site-directed and random mutagenesis. One major hydroxylated product (6'β-OH) and one minor product (6'-exomethylene), but not other products, were produced by CYP102A1 mutants. Formation of the metabolites was confirmed by high-performance liquid chromatography, liquid chromatography-mass spectroscopy, and NMR. Chemical methods to synthesize the metabolites of simvastatin and lovastatin have not been reported. These results demonstrate that CYP102A1 mutants can be used to produce human metabolites, especially chiral metabolites, of simvastatin and lovastatin. Our computational findings suggest that a conformational change in the cavity of the mutant active sites is related to the activity change. The modeling results also suggest that the activity change results from the movement of several specific residues in the active sites of the mutants. Furthermore, our computational findings suggest a correlation between the stabilization of the binding site and the catalytic efficiency of CYP102A1 mutants toward simvastatin and lovastatin.

The surface of long TiO2 nanotube (NT) electrodes in dye-sensitized solar cells (DSSCs) was modified without post-annealing by using atomic layer deposition (ALD) for the enhancement of photovoltage. Vertically oriented TiO2 NT electrodes with highly ordered and crack-free surface structures over large areas were prepared by a two-step anodization method. The prepared TiO2 NTs had a pore size of 80 nm, and a length of 23 μm. Onto these TiO2 NTs, an Al2O3 shell of a precisely controlled thickness was deposited by ALD. The conformally coated shell layer was confirmed by high-resolution transmission electron microscopy, energy-dispersive x-ray spectroscopy, and x-ray photoelectron spectroscopy. The open-circuit voltage (V(oc)) of the DSSCs was gradually enhanced as the thickness of the Al2O3 shell of the TiO2/Al2O3 NT electrodes was increased, which resulted from the enhanced electron lifetime. The enhanced electron lifetime caused by the energy barrier effect of the shell layer was measured quantitatively by the open-circuit voltage decay technique. As a result, 1- and 2-cycle-coated samples showed enhanced conversion efficiencies compared to the bare sample.

Influences of Psychological Resilience and Self-control on Suicidal Ideation of Undergraduate Students Im, SookBin · Heo, MyoungLyun · Kim, Se Young · Ahn, Hye Young · Kang, Saet Byul · Kim Mi, Song · Mun, Hea Mee · Park, Sun Ha · Won, A Rang · Lee, Ji Hye · Cho, Mi So College of Nursing, Eulji University, Daejeon Kunsan College of Nursing, Gunsan Department of Nursing, Changwon University, Changwon, Chungnam Mational University Hospital, Daejeon Korean Red Cross Chungbuk Blood Center, Cheongju, National Cancer Center, Goyang Seoul National University Hospital, Seoul, Dankook University Hospital, Cheonan Korean Armed Forces Capital Hospital, Seoul, Eulji University Hospital, Daejeon, Korea

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The authors deposited iridium tantalum oxide thin films using the reactive cosputtering system. The prepared IrTaO x thin films were characterized using X-ray photoelectron spectroscopy, transmission electron microscopy, chronocoloumetry, in situ transmittance measurements, and electrochemical impedance spectroscopy. The oxidized iridium in IrTaO x was increased by increasing the composition of tantalum. Most of IrTaO x thin films have high transmittance modulation, which is attributed by the proton conductivity of tantalum. As a result, the Ir 33 Ta 67 oxide thin film shows a performance of 1.4 s of response time, 5 × 10 -9 cm 2 /s of ion diffusion coefficient, and 20 cm 2 /C of coloration coefficient. The enhanced IrTaO x thin films are expected to be candidate of electrochromic materials for fast response time.