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We demonstrated the effects of substituents in fluorescein on the photoredox catalytic performance under visible light. For the systematic investigation, the phenyl ring of fluorescein was substituted with six different functional groups (i.e., amine, amide, isothiocyanate, aminomethyl, bromo, or nitro group) at the 5- or 6-position. The fluorescein derivatives were carefully characterized through photophysical and electrochemical analyses. The substituent effects were estimated by comparing the photopolymerization of poly(ethylene glycol) diacrylate (PEGDA) and N -vinylpyrrolidone (VP) in the presence of triethanolamine (TEOA) under aerobic conditions to that of intact fluorescein. As a result, the amine and nitro groups exhibited the lowest performances, presumably due to intramolecular photoinduced electron transfer (PET) promoted by the strong electron push-pull effect. The others, representative moderate or weak deactivators and activators, exhibited inferior performances than intact fluorescein, presumably owing to the more negative Δ G PET values, resulting in a decreased rate of intermolecular PET. These results are crucial for understanding the structure-performance relationship and the development of visible-light photoredox catalysts with improved performance and functionality., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

Anti-ambipolar switch (AAS) devices at a narrow bias region are necessary to solve the intrinsic leakage current problem of ternary logic circuits. In this study, an AAS device with a very high peak-to-valley ratio (∼10 6 ) and adjustable operating range characteristics was successfully demonstrated using a ZnO and dinaphtho[2,3- b :2',3'- f ]thieno[3,2- b ]thiophene heterojunction structure. The entire device integration was completed at a low thermal budget of less than 200 °C, which makes this AAS device compatible with monolithic 3D integration. A 1-trit ternary full adder designed with this AAS device exhibits excellent power-delay product performance (∼122 aJ) with extremely low power (∼0.15 μW, 7 times lower than the reference circuit) and lower device count than those of other ternary device candidates.

Thin polymeric films were developed for the vapor-phase sequential colorimetric detection of a nerve agent mimic and ammonia with high sensitivity. N -(4-Benzoylphenyl)acrylamide (BPAm), N , N -dimethylacrylamide (DMA), and ( E )-2-(methyl(4-(pyridine-4yldiazenyl)phenyl)amino)ethyl acrylate (MPDEA, M1) were copolymerized via free radical polymerization (FRP) to yield p(BPAm- co -DMA- co -MPDEA), hereafter referred to as P1. P1 exhibits selective sensing properties toward diethyl chlorophosphate (DCP), a nerve agent mimic, in pure aqueous media. Upon the addition of DCP, the pyridine groups of P1 were quaternized with DCP, accompanied by a color change from yellow to pink due to the enhancement of the intramolecular charge transfer (ICT) effect. In situ generated quaternized P1, hereafter referred to as P2, after DCP sensing was used to selectively detect ammonia via dequaternization in an aqueous medium. Ammonia detection was indicated by a color change in the solution from pink back to yellow. A surface-immobilized P1 film was prepared and employed for the vapor-phase detection of DCP, demonstrating that an amount of as low as 2 ppm was detectable. Ammonia vapor was also successfully detected by the P2 film via the ammonia-triggered removal of the quaternized phosphates. Alternating exposure of the film to DCP and ammonia resulted in the corresponding color changes, thereby demonstrating the reversibility of the system. The reusability of the polymeric film for detecting DCP and ammonia in the vapor phase was confirmed by performing four sequential colorimetric detection cycles.

Two-dimensional (2D) transition-metal dichalcogenides have attracted significant attention as gas-sensing materials owing to their superior responsivity at room temperature and their possible application as flexible electronic devices. Especially, reliable responsivity and selectivity for various environmentally harmful gases are the main requirements for the future chemiresistive-type gas sensor applications. In this study, we demonstrate improved sensitivity of a 2D MoS 2 -based gas sensor by controlling the Schottky barrier height. Chemical vapor deposition process was performed at low temperature to obtain layer-controlled 2D MoS 2 , and the NO 2 gas responsivity was confirmed by the fabricated gas sensor. Then, the number of MoS 2 layers was fixed and the types of electrode materials were varied for controlling the Schottky barrier height. As the Schottky barrier height increased, the NO 2 responsivity increased, and it was found to be effective for CO and CO 2 gases, which had little reactivity in 2D MoS 2 -based gas sensors.

A well-defined amphiphilic phenylthiosemicarbazone-based block copolymer was successfully synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization, followed by postpolymerization modification. Poly( N,N-dimethylacrylamide) (pDMA) was synthesized via RAFT polymerization of N,N-dimethylacrylamide (DMA). The resulting pDMA macrochain transfer agent was further extended using 3-vinylbenzaldehyde (VBA) to yield the poly[( N,N-dimethylacrylamide)- b-(3-vinylbenzaldehyde)] [p(DMA- b-VBA)] block copolymer. The aldehyde groups of p(DMA- b-VBA) were then made to react with 4-phenylthiosemicarbazide to yield the target block copolymer poly{ N,N-dimethylacrylamide- b-[ N-phenyl-2-(3-vinylbenzylidene)hydrazine carbothioamide]} [p(DMA- b-PVHC)]. p(DMA- b-PVHC) self-assembled in aqueous solution to yield polymeric micelles that comprise a pDMA block that forms a hydrophilic shell and a pPVHC block that forms a hydrophobic core. p(DMA- b-PVHC) micelles can detect Cu(II) ions which can be determined by a color change from colorless to yellow induced by the formation of coordination complexes between Cu(II) ions and the phenylthiosemicarbazone units of p(DMA- b-PVHC). As Cu(II) ions slowly penetrated the core of p(DMA- b-PVHC) micelles, these cores cross-linked with each other, which in turn resulted in the micelle particles swelling in water. Upon the addition of Cu(II) ions to a solution of p(DMA- b-PVHC) micelles encapsulating the hydrophobic model drug coumarin 102, this drug was released from the micelles in a sustained manner due to the gradual swelling of the cross-linked micelle cores caused by the slow penetration of Cu(II) ions.

Developing a simple and cheap analytical method for the selective detection and quantitative separation of toxic ions present in aqueous media is the biggest challenge faced by the chemosensing research community. Here, a 5,5-difluoro-1,3,7,9-tetramethyl-10-phenyl-5 H-dipyrrolo-diazaborinine-derived water-soluble polymer integrated with thiosemicarbazone units was rationally designed and synthesized for the simultaneous detection and separation of Hg(II) ions in pure aqueous solution. The water-soluble polymer scaffold poly( N, N'-dimethyl acrylamide- co-5,5-difluoro-1,3,7,9-tetramethyl-10-phenyl-5 H-dipyrrolo-diazaborinine-2-carbaldehyde) was synthesized by reversible addition-fragmentation chain transfer polymerization, followed by post-polymerization modification with thiosemicarbazide, leading to the formation of the target probe, P1. The nonemitting P1 exhibited bright yellow emission upon exposure to Hg(II) ions, with a limit of detection as low as 0.37 μM. This turn-on emission behavior triggered by Hg(II) ions might originate from the suppression of isomerization around the C═N bond of the thiosemicarbazone moiety caused by the formation of a coordination complex between P1 and Hg(II) ions. In addition, P1 displayed excellent selectivity toward Hg(II) ions over other metal cations. Finally, the selective removal of Hg(II) ions from an aqueous solution containing various metal ions was achieved by precipitation, which is probably caused by the fact that coordination complexes whereby Hg(II) ions acted as bridgeheads between P1 molecules had formed.

Thin-film probes have been developed for the reversible detection and separation of picric acid (PA) with extreme sensitivity in aqueous media. The free radical copolymerization of dimethylacrylamide (DMA), benzophenone acrylamide (BPAM), and glycidyl methacrylate (GMA) with a feed ratio of 95:1:4 yielded [p(DMA- co-BPAM- co-GMA)] (P1). P1 was transformed to the final polymeric probe, P2, by a subsequent ring-opening reaction between N-(pyren-1-yl-methyl)propan-1-amine (Py-PA) with the epoxide unit of P1. P2 exhibited rapid and selective sensing properties toward PA in aqueous media via turn-off fluorescence emission. The detection sensitivity was tuned precisely by varying the pH of the solution. After the immobilization of P2 on a quartz slide by spin-coating, followed by exposure to UV light, the resulting film exhibited an attogram-level detection limit toward PA. The photoinduced electron transfer together with an energy-transfer process between PA and the pyrene units of P2 were maximized by the strong π-π stacking of pyrene units of P2, which, in turn, induced rapid exciton energy diffusion. Furthermore, the separation of PA from the mixture of the various nitroaromatic compounds by the P2 film was achieved. Whereas the detection process of PA was reversible and repeatable over multiple cycles, the P2 film could be recycled.

Polymeric micelles based on light-responsive block copolymers were prepared and used for the phototunable detection of mercury(II) ions. 2-Nitrobenzyl acrylate (NBA) and ( E)-2-((4-((4-formylphenyl)diazenyl)phenyl)(methyl)amino) ethyl acrylate (FPDEA) were copolymerized from a poly(ethylene oxide) (PEO) macroinitiator via atom transfer radical polymerization (ATRP), leading to a well-defined block copolymer of PEO 113 - b-[p(NBA 10 - co-FPDEA 3 )] with a low polydispersity index (PDI = 1.16). After polymerization, the aldehyde groups of PEO- b-[p(NBA- co-FPDEA)] were converted to aldoxime groups by reacting with hydroxylamine, leading to the formation of a final oxime-containing polymeric probe, PEO- b-[p(NBA- co-HPDEA)], P1. The resulting block copolymer, P1, was self-assembled in water to yield spherical micelles that consist of a PEO block forming a hydrophilic shell and a copolymer of light-responsive NBA and a mercury(II) ion-detecting HPDEA block forming a hydrophobic core. Upon the addition of mercury(II) ions to this micellar solution, no detection was observed since water-soluble mercury(II) ions have limited accessability to the oxime units of P1, which are located in the hydrophobic core. After UV light irradiation, however, the photolabile 2-nitrobenzyl moieties were cleaved, and hydrophobic PNBA was transformed to hydrophilic poly(acrylic acid) (PAA), leading to the photoinduced dissociation of micelles to unimers. As a result, the oxime units of P1 were exposed to a hydrophilic environment and could react with mercury(II) ions to form nitrile groups, resulting in the turn-on detection of mercury(II) ions by UV light irradiation.

To synthesize a thermally robust Ni 1-x Pt x Si film suitable for ultrashallow junctions in advanced metal-oxide-semiconductor field-effect transistors, we used a continuous laser beam to carry out millisecond annealing (MSA) on a preformed Ni-rich silicide film at a local surface temperature above 1000 °C while heating the substrate to initiate a phase transition. The melting and quenching process by this unique high-temperature MSA process formed a Ni 1-x Pt x Si film with homogeneous Pt distribution across the entire film thickness. After additional substantial thermal treatment up to 800 °C, the noble Ni 1-x Pt x Si film maintained a low-resistive phase without agglomeration and even exhibited interface flattening with the underlying Si substrate.

Nanostructured silicon (Si) is useful in many applications and has typically been synthesized by bottom-up colloid-based solution processes or top-down gas phase reactions at high temperatures. These methods, however, suffer from toxic precursors, low yields, and impractical processing conditions (i.e., high pressure). The magnesiothermic reduction of silicon oxide (SiO 2 ) has also been introduced as an alternative method. Here, we demonstrate the reduction of SiO 2 by a simple milling process using a lab-scale planetary-ball mill and industry-scale attrition-mill. Moreover, an ignition point where the reduction begins was consistently observed for the milling processes, which could be used to accurately monitor and control the reaction. The complete conversion of rice husk SiO 2 to high purity Si was demonstrated, taking advantage of the rice husk's uniform nanoporosity and global availability, using a 5L-scale attrition-mill. The resulting porous Si showed excellent performance as a Li-ion battery anode, retaining 82.8% of the initial capacity of 1466 mAh g -1 after 200 cycles.

Recently, hybrid supercapacitors (HSCs), which combine the use of battery and supercapacitor, have been extensively studied in order to satisfy increasing demands for large energy density and high power capability in energy-storage devices. For this purpose, the requirement for anode materials that provide enhanced charge storage sites (high capacity) and accommodate fast charge transport (high rate capability) has increased. Herein, therefore, a preparation of nanocomposite as anode material is presented and an advanced HSC using it is thoroughly analyzed. The HSC comprises a mesoporous Nb2O5/carbon (m-Nb2O5-C) nanocomposite anode synthesized by a simple one-pot method using a block copolymer assisted self-assembly and commercial activated carbon (MSP-20) cathode under organic electrolyte. The m-Nb2O5-C anode provides high specific capacity with outstanding rate performance and cyclability, mainly stemming from its enhanced pseudocapacitive behavior through introduction of a carbon-coated mesostructure within a voltage range from 3.0 to 1.1 V (vs Li/Li(+)). The HSC using the m-Nb2O5-C anode and MSP-20 cathode exhibits excellent energy and power densities (74 W h kg(-1) and 18,510 W kg(-1)), with advanced cycle life (capacity retention: ∼90% at 1000 mA g(-1) after 1000 cycles) within potential range from 1.0 to 3.5 V. In particular, we note that the highest power density (18,510 W kg(-1)) of HSC is achieved at 15 W h kg(-1), which is the highest level among similar HSC systems previously reported. With further study, the HSCs developed in this work could be a next-generation energy-storage device, bridging the performance gap between conventional batteries and supercapacitors.

Carbon nanofiber/nanotube (CNF/CNT) composite catalysts grown on carbon felt (CF), prepared from a simple way involving the thermal decomposition of acetylene gas over Ni catalysts, are studied as electrode materials in a vanadium redox flow battery. The electrode with the composite catalyst prepared at 700 °C (denoted as CNF/CNT-700) demonstrates the best electrocatalytic properties toward the V(2+)/V(3+) and VO(2+)/VO2(+) redox couples among the samples prepared at 500, 600, 700, and 800 °C. Moreover, this composite electrode in the full cell exhibits substantially improved discharge capacity and energy efficiency by ~64% and by ~25% at 40 mA·cm(-2) and 100 mA·cm(-2), respectively, compared to untreated CF electrode. This outstanding performance is due to the enhanced surface defect sites of exposed edge plane in CNF and a fast electron transfer rate of in-plane side wall of the CNT.

A composite air electrode consisting of Ketjenblack carbon (KB) supported amorphous manganese oxide (MnOx) nanowires, synthesized via a polyol method, is highly efficient for the oxygen reduction reaction (ORR) in a Zn-air battery. The low-cost and highly conductive KB in this composite electrode overcomes the limitations due to low electrical conductivity of MnOx while acting as a supporting matrix for the catalyst. The large surface area of the amorphous MnOx nanowires, together with other microscopic features (e.g., high density of surface defects), potentially offers more active sites for oxygen adsorption, thus significantly enhancing ORR activity. In particular, a Zn-air battery based on this composite air electrode exhibits a peak power density of ∼190 mW/cm2, which is far superior to those based on a commercial air cathode with Mn3O4 catalysts.

A magnesiothermic reduction synthesis approach is reported for direct conversion of SiO(2)/C composite nanostructures to corresponding SiC materials without losing their nanostructure morphologies. Crystalline SiC materials can be obtained by this approach at a temperature as low as 600 degrees C, only approximately half of that applied in the generally used carbothermal reduction and preceramic polymer pyrolysis methods. An ordered hierarchical macro-mesoporous SiC material was synthesized for the first time as a demonstration. This pseudomorphic transformation can be regarded as a general synthesis method for different kinds of SiC nanostructures, and it can also be readily extended to other metal carbide materials as well as TiC.

An efficient stereocontrolled route to the azatricyclic core of an advanced halichlorine intermediate is described. Reaction of the oxime derived from 2-(oxo-cyclopentyl)acetic acid ethyl ester with 2,3-bis(phenylsulfonyl)-1,3-butadiene gives rise to a 7-oxa-1-azanorbornane cycloadduct in high yield. The formation of the bicyclic isoxazolidine arises from conjugate addition of the oxime onto the diene to afford a transient nitrone that then undergoes an intramolecular dipolar cycloaddition. Treatment of the cycloadduct with 5% Na/Hg results in reductive nitrogen-oxygen bond cleavage to furnish a spirocyclic piperidinone, which was further elaborated to an advanced intermediate employed in an earlier synthesis of halichlorine.

Ordered mesoporous materials (OMMs) with well-defined pore sizes (>2 nm) and pore geometries are important in various applications that require fast mass transfer or deal with large molecules. Extensive research has resulted in the discovery of OMMs with three types of mesostructures: (i) bi- or multicontinuous, (ii) columnar, and (iii) discontinuous (cagelike). However, another type, the pillared lamellar structure, which has long been sought and has been mathematically computed and known to exist in the research fields of surfactant and multiblock-copolymer mesophases, still remains a mesostructure that has not been observed in real OMMs for any specific symmetry. Herein, we report an unprecedented type of ordered mesoporous material with a hexagonally pillared lamellar (HPL) structure (P6(3)/mmc) that can be synthesized via a phase transformation from a lamellar mesophase by hydrothermal reaction in the presence of an organosilica precursor and a high concentration of a designed Gemini surfactant (Gem(16-3-16)) that has a large g value. The present GMO-HPL, which has an unique three-dimensional periodic structure with two-dimensionally connected pore channels running between the framework layers, provides a fascinating topological link between the lamellar and columnar (2D hexagonal) mesophases. It is unique in its application potential by making possible selective 2D diffusion in different directions.

An efficient stereocontrolled route to the isoschizozygane alkaloid core has been developed utilizing an intramolecular 1,4-dipolar cycloaddition of a cross-conjugated heteroaromatic betaine. The resulting cycloadduct undergoes loss of COS, and further reduction delivers a 5a-azaacenaphthylene intermediate that was transformed into the isoschizozygane skeleton upon treatment with acid. A variation of this tactic was then employed for a synthesis of the hexacyclic framework of the shizozygane alkaloid (+/-)-strempeliopine. The key step of the synthesis corresponds to an intramolecular 1,4-dipolar cycloaddition of a heteroaromatic betaine across a tethered 4-((2-nitrophenyl)but-3-enyl) side chain. Catalytic reduction of the nitro group followed by reaction with NBS resulted in the formation of the required pentacyclic indoline framework of the target alkaloid. Closure of the final ring of the shizozygane skeleton was carried using an oxidative cyclization.

This work reports on repeatable adhesive materials prepared by controlled grafting of dangling hetero chains from polymer elastomers. The dangling chain elastomer system was prepared by grafting poly(n-butyl acrylate) (PBA) chains from prefunctionalized polydimethylsiloxane (PDMS) elastomer networks using atom transfer radical polymerization. To study the effects of chain growth and network strain as they relate to network adhesion mechanics, various lengths of PBA chains with degree of polymerizations (DP) of 65, 281, 508, and 1200 were incorporated into the PDMS matrix. PBA chains with a DP value of 281 grafted from a flat PDMS substrate showed the highest (approximately 3.5-fold) enhancement of nano- and macroscale adhesion relative to a flat raw (ungrafted and not prefunctionalized) PDMS substrate. Moreover, to study the effect of PBA dangling chains on adhesion in fibrillar elastomer structures inspired by gecko foot hairs, a dip-transfer fabrication method was used to graft PBA chains with a DP value of 296 from the tip endings of mushroom-shaped PDMS micropillars. A PBA chain covered micropillar array showed macroscale adhesion enhancement up to approximately 7 times relative to the flat ungrafted prefunctionalized PDMS control substrate, showing additional nonoptimized approximately 2-fold adhesion enhancement due to fibrillar structuring and mushroom-shaped tip ending. These dangling hetero chains on elastomer micro-/nanofibrillar structures may provide a novel fabrication platform for multilength scale, repeatable, and high-strength fibrillar adhesives inspired by gecko foot hairs.

Adsorption-induced degradation of brushlike macromolecules was monitored through molecular imaging by atomic force microscopy. The rate constant for C-C bond cleavage was shown to be extremely sensitive to the substrate surface energy. A few percent increase in the surface energy from 69.2 to 71.2 mN/m led to an order of magnitude increase of the scission rate. The absolute values of the rupture forces ranging from 2.57 to 2.47 nN are in agreement with previously calculated and measured values for stretching surface-tethered molecules.

Stable biodegradable nanogels cross-linked with disulfide linkages were prepared by inverse miniemulsion atom transfer radical polymerization (ATRP). These nanogels could be used for targeted drug delivery scaffolds for biomedical applications. The nanogels had a uniformly cross-linked network, which can improve control over the release of encapsulated agents, and the nanogels biodegraded into water-soluble polymers in the presence of a biocompatible glutathione tripeptide, which is commonly found in cells. The biodegradation of nanogels can trigger the release of encapsulated molecules including rhodamine 6G, a fluorescent dye, and Doxorubicin (Dox), an anticancer drug, as well as facilitate the removal of empty vehicles. Results obtained from optical fluorescence microscope images and live/dead cytotoxicity assays of HeLa cancer cells suggested that the released Dox molecules penetrated cell membranes and therefore could suppress the growth of cancer cells. Further, OH-functionalized nanogels were prepared to demonstrate facile applicability toward bioconjugation with biotin. The number of biotin molecules in each nanogel was determined to be 142,000, and the formation of bioconjugates of nanogels with avidin was confirmed using optical fluorescence microscopy.

Improved 1H ENDOR data from the S(EPR1) intermediate formed during turnover of the nitrogenase alpha-195Gln MoFe protein with C2(1,2)H2 in (1,2)H2O buffers, taken in context with the recent study of the intermediate formed from propargyl alcohol, indicate that S(EPR1) is a product complex, likely with C2H4 bound as a ferracycle to a single Fe of the FeMo-cofactor active site. 35 GHz CW and Mims pulsed 57Fe ENDOR of 57Fe-enriched S(EPR1) cofactor indicates that it exhibits the same valencies as those of the CO-bound cofactor of the lo-CO intermediate formed during turnover with CO, [Mo4+, Fe3+, Fe6(2+), S9(2-)(d43)](+1), reduced by m = 2 electrons relative to the resting-state cofactor. Consideration of 57Fe hyperfine coupling in S(EPR1) and lo-CO leads to a picture in which CO bridges two Fe of lo-CO, while the C2H4 of S(EPR1) binds to one of these. To correlate these and other intermediates with Lowe-Thorneley (LT) kinetic schemes for substrate reduction, we introduce the concept of an "electron inventory". It partitions the number of electrons a MoFe protein intermediate has accepted from the Fe protein (n) into the number transmitted to the substrate (s), the number that remain on the intermediate cofactor (m), and the additional number delivered to the cofactor from the P clusters (p): n = m + s - p (with p = 0 here). The cofactors of lo-CO and S(EPR1) both are reduced by m = 2 electrons, but the intermediates are not at the same LT reduction stage (E(n)): (n = 2; m = 2, s = 0) for lo-CO; (n = 4; s = 2, m = 2) for S(EPR1). This is the first proposed correlation of an LT E(n) kinetic state with a well-defined chemical state of the enzyme.

A high population intermediate has been trapped on the nitrogenase active site FeMo cofactor during reduction of N2. In addition, intermediates have been trapped during reduction of CH3-N=NH by the alpha-195Gln variant and during reduction of H2N-NH2 by the alpha-70Ala/alpha-195Gln variant. Each of these trapped states shows an EPR signal arising from an S = 1/2 state of the FeMo cofactor. 15N ENDOR shows that each intermediate has a nitrogenous species bound to the FeMo cofactor, with a single type of N seen for each bound intermediate. The g tensors are unique to each intermediate, g(e) = [2.084, 1.993, 1.969], g(m) = [2.083, 2.021, 1.993], g(l) = [2.082, 2.015, 1.987], as are the 15N hyperfine couplings at g1, which suggests that three distinct stages of NN reduction may have been trapped. The 1H ENDOR spectra show that the N2 intermediate is at a distinct and earlier stage of reduction from the other two, so at least two stages of NN reduction have been trapped. Some possible structures of the hydrazine intermediate are presented.

X-ray crystallographic study of the nitrogenase MoFe protein revealed electron density from an atom (denoted X) inside the active-site metal cluster, the [MoFe7S9:homocitrate] FeMo-cofactor. The electron density associated with X is consistent with a single N, O, or C atom. We now have tested whether X is an N or not by comparing the Q-band ENDOR and ESEEM signals from resting-state (S = 3/2) MoFe protein and NMF-extracted FeMo-co from bacteria grown with either 14N or 15N as the exclusive N source. All of the 14N or 15N signals associated with the protein are lost upon extraction of the FeMo-co. We interpret this as strong evidence that X is not an N.

[reaction: see text] A new strategy for the synthesis of the isoschizozygane alkaloid core has been developed that is based on a 1,4-dipolar cycloaddition reaction of a cross-conjugated heteroaromatic betaine. The resulting cycloadduct undergoes loss of COS, and further reduction delivers a 5a-aza-acenaphthylene intermediate that was transformed into the isoschizozygane skeleton upon treatment with acid.

A major challenge in understanding the mechanism of nitrogenase, the enzyme responsible for the biological fixation of N(2) to two ammonias, is to trap a nitrogenous substrate at the enzyme active site in a state that is amenable to further characterization. In the present work, a strategy is described that results in the trapping of the substrate hydrazine (H(2)N-NH(2)) as an adduct bound to the active site metal cluster of nitrogenase, and this bound adduct is characterized by EPR and ENDOR spectroscopies. Earlier work has been interpreted to indicate that nitrogenous (e.g., N(2) and hydrazine) as well as alkyne (e.g., acetylene) substrates can bind at a common FeS face of the FeMo-cofactor composed of Fe atoms 2, 3, 6, and 7. Substitution of alpha-70(Val) that resides over this FeS face by the smaller amino acid alanine was also previously shown to improve the affinity and reduction rate for hydrazine. We now show that when alpha-195(His), a putative proton donor near the active site, is substituted by glutamine in combination with substitution of alpha-70(Val) by alanine, and the resulting doubly substituted MoFe protein (alpha-70(Ala)/alpha-195(Gln)) is turned over with hydrazine as substrate, the FeMo-cofactor can be freeze-trapped in a S = (1)/(2) state in high yield ( approximately 70%). The presumed hydrazine-FeMo-cofactor adduct displays a rhombic EPR signal with g = [2.09, 2.01, 1.93]. The optimal pH for the population of this state was found to be 7.4. The EPR signal showed a Curie law temperature dependence similar to the resting state EPR signal. Mims pulsed ENDOR spectroscopy at 35 GHz using (15)N-labeled hydrazine reveals that the trapped intermediate incorporates a hydrazine-derived species bound to the FeMo-cofactor; in spectra taken at g(1) this species gives a single observed (15)N signal, A(g(1)) = 1.5 MHz.

We here show that the iron-molybdenum (FeMo)-cofactor of the nitrogenase alpha-70(Ile) molybdenum-iron (MoFe) protein variant accumulates a novel S = (1)/(2) state that can be trapped during the reduction of protons to H(2). (1,2)H-ENDOR measurements disclose the presence of two protons/hydrides (H(+/)(-)) whose hyperfine tensors have been determined from two-dimensional field-frequency (1)H ENDOR plots. The two H(+/)(-) have large isotropic hyperfine couplings, A(iso)( )() approximately 23 MHz, which shows they are bound to the cofactor. The favored analysis for these plots indicates that the two H(+/)(-) have the same principal values, which indicates that they are chemically equivalent. The tensors are further related to each other by a permutation of the tensor components, which indicates an underlying symmetry of binding relative to the cofactor. At present, no model for the structure of the iron-molybdenum (FeMo)-cofactor in the S = (1)/(2) state trapped during the reduction of H(+) can be shown unequivocally to satisfy all of the constraints generated by the ENDOR analysis. The data disfavors any model that involves protonation of sulfides, and thus suggests that the intermediate instead contains two chemically equivalent bound hydrides; it appears unlikely that these are terminal monohydrides.

The chemical mechanism for biological cleavage of the N(2) triple bond at ambient pressure and temperature has been the subject of intense study for many years. The site of substrate activation and reduction has been localized to a complex cofactor, called FeMo cofactor, yet until now the complexity of the system has denied information concerning exactly where and how substrates interact with the metal-sulfur framework of the active site. In this Account, we describe a combined genetic, biophysical, and biochemical approach that was used to provide direct and detailed information concerning where alternative alkyne substrates interact with FeMo cofactor during catalysis. The relevance and limitations of this work with respect to N(2) binding and reduction also are discussed.

A convenient synthesis of variously substituted octahydroindolo[7a,1a]-isoquinolinones has been achieved by an acid-induced cyclization of hexahydroindolinones bearing tethered phenethyl groups. The formation of a single lactam diastereomer is the result of the stereoelectronic preference for axial attack by the aromatic ring onto the initially formed N-acyliminium ion from the least hindered side. Additional experiments showed that a variety of hexahydroindolinones containing tethered pi-bonds undergo a related acid-induced cyclization reaction. Treatment of the 3-methylbut-3-enyl-substituted hexahydroindolinone with acid furnished a 3:1 mixture of isomeric octahydropyrido[2,1-i]indolinones in near-quantitative yield. Interestingly, cyclization of the closely related 1-(3-methoxybut-3-enyl)-substituted hexahydroindolin-one afforded a pyrrolo[3,2,1-ij]quinolinone as the exclusive product. With this system, initial protonation takes place on the more nucleophilic enol ether pi-bond and the resulting carbonium ion undergoes a subsequent cyclization with the enamido pi-bond to give the observed product. The electrophilic promoted cyclizations were extended to include the related hexahydro[1]pyrindinone and 1H-quinolinone systems. An NBS-promoted intramolecular electrophilic aromatic substitution reaction of 1-[2-(3,4-dimethoxyphenyl)ethyl]-1,4,5,6-tetrahydroindolinone was used to assemble the tetracyclic core of the erythrinone skeleton. The resulting cyclized product was transformed into (+/-)-erysotramidine in three additional steps.

Nitrogenase is the metalloenzyme that catalyzes the nucleotide-dependent reduction of N(2), as well as reduction of a variety of other triply bonded substrates, including the alkyne, acetylene. Substitution of the alpha-70(Val) residue in the nitrogenase MoFe protein by alanine expands the range of substrates to include short-chain alkynes not reduced by the unaltered protein. Rapid freezing of the alpha-70(Ala) nitrogenase MoFe protein during reduction of the alkyne propargyl alcohol (HC triple bond CH(2)OH; PA) traps an S = (1)/(2) intermediate state of the active-site metal cluster, the FeMo-cofactor. We have combined CW and pulsed (13)C ENDOR (electron-nuclear double resonance) with two quantitative 35 GHz (1,2)H ENDOR techniques, Mims pulsed ENDOR and the newly devised "stochastic field-modulated" ENDOR, to study this intermediate prepared with isotopically substituted ((13)C, (1,2)H) propargyl alcohol in H(2)O and D(2)O buffers. These measurements allow the first description of a trapped nitrogenase reduction intermediate. The S = (1)/(2) turnover intermediate generated during the reduction of PA contains the 3-carbon chain of PA and exhibits resolved (1,2)H ENDOR signals from three protons, two strongly coupled (H(a)) and one weakly coupled (H(b)); H(a)(c) originates as the C3 proton of PA, while H(a)(s) and H(b) are solvent-derived. The two H(a) protons have identical hyperfine tensors, despite having different origins. The equality of the (H(a)(s), H(a)(c)) hyperfine tensors strongly constrains proposals for the structure of the cluster-bound reduced PA. Through consideration of model structures found in the Cambridge Structural Database, we propose that the intermediate contains a novel bio-organometallic complex in which a reduction product of propargyl alcohol binds as a metalla-cyclopropane ring to a single Fe atom of the Fe-S face of the FeMo-cofactor that is composed of Fe atoms 2, 3, 6, and 7. Of the two most attractive structures, one singly reduced at C3 (4), the other being the doubly reduced allyl alcohol product (6), we tentatively favor 6 because of the "natural" assignment it affords for H(b).

An NBS-promoted intramolecular electrophilic aromatic substitution reaction of a hexahydroindolinone derivative was used to assemble the tetracyclic core of the erythrinane skeleton. The resulting cyclized product was transformed into (+/-)-erysotramidine in three additional steps. The cyclization reaction is also successful using variously substituted aryl and furanyl bicyclic lactams under acidic conditions. [reaction: see text]

A recent high-resolution X-ray crystallographic study (1.16 A) of the Azotobacter vinelandii nitrogenase MoFe protein revealed a previously undetected electron density associated with the active site FeMo-cofactor. The density is located inside the cluster at the center of the "trigonal prism" of six irons and is assigned to a species "X". The identity of species X was not resolved, although the electron density is consistent with a single N, O, or C atom. One proposal is that X is an N atom that derives from and exchanges with N from N2 during catalysis. In the present study, we have examined this possibility by employing 14N and 15N isotopes of N2 along with ENDOR and ESEEM spectroscopies. The WT MoFe protein and alpha-359Arg-->Lys and alpha-381Phe-->Leu variants were allowed to turn over in the presence of 14N2 or 15N2, and then were examined as resting enzymes by ENDOR and ESEEM at X- and Q-bands to look for all 14N and 15N signals coupled to the electron spin of the FeMo-cofactor and to determine if any exchanged during turnover. We have found five peaks in Q-band pulsed ENDOR spectra that appear to arise not only from previously reported N1/N2, which give rise to the ESEEM, but also from one or two additional coupled nitrogens. None of the ENDOR and ESEEM signals vanish or are altered by catalytic turnover with 15N2, and no new 15N signal is detected, leading to the conclusion that if species X is a nitrogen atom, it does not exchange during dinitrogen reduction.

Starting with the tricyclic core 2b, annulation to form the 13-membered western ring of sarain A has been achieved to afford the macrocycle 30a by initial construction of the sterically congested quaternary center at C-3, followed by elaboration of the C-3 side-chain and ring-closing olefin metathesis. Also included is a parallel conversion of tricycle 2c to macrocycle 30b containing a functionalized side-chain at N-1 suitable for attachment of the eastern macrocyclic ring.

[reaction: see text] An eight-membered ring was conveniently appended onto a cycloalkene carboxylate by employing a facile radical cyclization-fragmentation reaction of an omega-bromoalkyl spirocyclobutanone, which was readily accessible by the Kulinkovich cyclopropanation and subsequent electrophilic addition to a 3-bromoalkyl acetal.

[reaction: see text]. Intermolecular and intramolecular [4 + 3] cycloaddition reactions of readily available cyclopropanone hemiacetals with furans are described.

This study investigated an application of TiO2 photocatalyst sensitized with tris(4,4'-dicarboxy-2,2'-bipyridyl)ruthenium-(II) complex to CCl4 degradation under visible light irradiation. By injecting electrons from the photoexcited sensitizer to the conduction band, the sensitized TiO2 degraded CCl4 under the irradiation of lambda > 420 nm. The quantum yield of CCl4 dechlorination was about 10(-3). The dechlorination rate of CCl4 was reduced in the presence of dissolved O2 due to its competition for conduction band electrons. The photolysis rate was dependent on pH due to the strong pH dependence of the sensitizer adsorption on TiO2 surface with a maximum degradation rate achieved at pH approximately 3. A two-site Langmurian model successfully described the adsorption of the sensitizer on TiO2 particles. The monolayer coverage was achieved at the added sensitizer concentration of 10 microM at [TiO2] = 0.5 g/L. However, the photolysis rate of CCl4 showed a maximum at a sensitizer surface coverage of 0.3 monolayer. Since the photoinduced electron injection gradually depleted active sensitizer molecules on TiO2, sacrificial electron donors to regenerate the sensitizer were sought. 2-Propanol as an electron donor was efficient in the present RuIIL3/TiO2/CCl4 system, which showed no sign of deceleration in the dechlorination rate up to 6 h of irradiation.

Freeze-quenching of nitrogenase during reduction of carbon disulfide (CS(2)) was previously shown to result in the appearance of a novel EPR signal (g = 2.21, 1.99, and 1.97) not previously associated with any of the oxidation states of the nitrogenase metal clusters. In the present work, freeze-quench X- and Q-band EPR and Q-band (13)C electron nuclear double resonance (ENDOR) spectroscopic studies of nitrogenase during CS(2) reduction disclose the sequential formation of three distinct intermediates with a carbon-containing fragment of CS(2) bound to a metal cluster inferred to be the molybdenum-iron cofactor. Modeling of the Q-band (35 GHz) EPR spectrum of freeze-trapped samples of nitrogenase during turnover with CS(2) allowed assignment of three signals designated "a" (g = 2.035, 1.982, 1.973), "b" (g = 2.111, 2.002, and 1.956), and "c" (g = 2.211, 1. 996, and 1.978). Freezing samples at varying times after initiation of the reaction reveals that signals "a", "b", and "c" appear and disappear in sequential order. Signal "a" reaches a maximal intensity at 25 s; signal "b" achieves maximal intensity at 60 s; and signal "c" shows maximal intensity at 100 s. To characterize the intermediates, (13)CS(2) was used as a substrate, and freeze-trapped turnover samples were examined by Q-band (13)C ENDOR spectroscopy. Each EPR signal ("a", "b", and "c") gave rise to a distinct (13)C signal, with hyperfine coupling constants of 4.9 MHz for (13)C(a), 1. 8 MHz for (13)C(b), and 2.7 MHz for (13)C(c). Models for the sequential formation of intermediates during nitrogenase reduction of CS(2) are discussed.

[formula: see text] A new approach to a suitably functionalized tricyclic core of sarains has been developed by means of Katritzky's cycloaddition using 3-oxidopyridinium betaines. A key step was the regioselective differentiation of the two nearly identical hydroxy groups derived from oxidative cleavage of the double bond in 8 to afford 14. A stereocontrolled construction of the tricyclic core 20 of sarains containing the requisite side chain at C-3' was achieved by an intramolecular conjugate addition.

Wild-type nitrogenase MoFe protein shows a deep 14N electron spin-echo envelope modulation (ESEEM) arising from a nitrogen nucleus (N1) coupled to the S = 3/2 spin system of the FeMo-cofactor of the MoFe protein. A previous ESEEM study on altered MoFe proteins generated by substitutions at the alpha-195-histidine position suggested that alpha-195-histidine provides a hydrogen bond to the FeMo-cofactor but is not the source of the 14N1 modulation [DeRose et al. (1995) Biochemistry 34, 2809-2814]. This study also raised the possibility of a correlation between ESEEM spectroscopic properties and the nitrogenase phenotype. We now report ESEEM studies on altered MoFe proteins with substitutions at residues alpha-96-arginine, alpha-359-arginine, and alpha-381-phenylalanine to (i) assign the first-shell hydrogen bonding as revealed by the 14N modulation; (ii) explore the mechanistic relevance of the ESEEM signatures to nitrogenase activity; and (iii) study microscopic changes within the polypeptide environment of the FeMo-cofactor. Present ESEEM data reveals that two kinds of 14N modulations are present in wild-type MoFe protein. A new 2-dimensional procedure for high-precision analysis of the ESEEM data of the MoFe proteins shows that the deep wild-type ESEEM modulation (denoted N1) has a hyperfine-coupling constant of Aiso = 1.05 MHz and nuclear quadrupole coupling parameters of e2qQ = 2.17 MHz, eta = 0.59; the other (denoted N2) has a smaller hyperfine coupling of Aiso = approximately 0.5 MHz and e2qQ = approximately 3.5 MHz, eta = approximately 0.4. The N2 ESEEM pattern is more obvious when unmasked by substitutions that result in the loss of the deep N1 modulation. Correlations of the ESEEM properties and catalytic activities of the altered MoFe proteins suggest that (i) the side chain of the alpha-359-arginine is the source of the deep ESEEM N1 modulation; (ii) one or both of the amide nitrogens of alpha-356-glycine/alpha-357-glycine are responsible for the weak N2 modulation; (iii) substitution of the nonpolar alpha-381-phenylalanine residue, as well as substitution of either the alpha-195-histidine or alpha-359-arginine residues, can eliminate the N1 interaction with FeMo-cofactor; and (iv) ESEEM can be used to detect slight reorientations of FeMo-cofactor within its polypeptide pocket, although the mechanistic relevance of the loss or perturbation of the hydrogen-bonding interactions between FeMo-cofactor and polypeptide environment has not yet been established.