Effect Of Substrate Structure Research Articles

Ultrauniform Plating of Lithium on 10-nm-Scale Ordered Carbon Grids for Long Lifespan Lithium Metal Batteries.

Tailorable lithium (Li) nucleation and uniform early-stage plating is essential for long-lifespan Li metal batteries. Among factors influencing the early plating of Li anode, the substrate is critical, but a fine control of the substrate structure on a scale of ≈10nm has been rarely achieved. Herein, a carbon consisting of ordered grids is prepared, as a model to investigate the effect of substrate structure on the Li nucleation. In contrast to the individual spherical Li nuclei formed on the flat graphene, an ultrauniform and nuclei-free Li plating is obtained on the ordered carbon with a grid size smaller than the thermodynamical critical radius of Li nucleation (≈26nm). Simultaneously, an inorganic-rich solid-electrolyte-interphase is promoted by the cross-sectional carbon layers of such ordered grids which are exposed to the electrolyte. Consequently, the carbon grids with a grid size of ≈10nm show a favorable cycling stability for more than 1100 cycles measured at 2mA cm-2 in a half cell. With LiNi0.8Co0.1Mn0.1O2 as cathode, the assembled full cell with a cathode capacity of 3 mAh cm-2 and a negative/positive ratio of 1.67 demonstrates a stable cycling for over 130 cycles with a capacity retention of 88%.

A Comparative Study on the Effect of Substrate Structure on Electrochemical Performance and Stability of Electrodeposited Platinum and Iridium Oxide Coatings for Neural Electrodes.

Implantable electrodes are crucial for stimulation safety and recording quality of neuronal activity. To enhance their electrochemical performance, electrodeposited nanostructured platinum (nanoPt) and iridium oxide (IrOx) have been proposed due to their advantages of in situ deposition and ease of processing. However, their unstable adhesion has been a challenge in practical applications. This study investigated the electrochemical performance and stability of nanoPt and IrOx coatings on hierarchical platinum-iridium (Pt-Ir) substrates prepared by femtosecond laser, compared with the coatings on smooth Pt-Ir substrates. Ultrasonic testing, agarose gel testing, and cyclic voltammetry (CV) testing were used to evaluate the coatings' stability. Results showed that the hierarchical Pt-Ir substrate significantly enhanced the charge-storage capacity of electrodes with both coatings to more than 330 mC/cm2, which was over 75 times that of the smooth Pt-Ir electrode. The hierarchical substrate could also reduce the cracking of nanoPt coatings after ultrasonic, agarose gel and CV testing. Although some shedding was observed in the IrOx coating on the hierarchical substrate after one hour of sonication, it showed good stability in the agarose gel and CV tests. Stable nanoPt and IrOx coatings may not only improve the electrochemical performance but also benefit the function of neurobiochemical detection.

The 5′-phosphate enhances the DNA-binding and exonuclease activities of human mitochondrial genome maintenance exonuclease 1 (MGME1)

In higher eukaryotes, mitochondria play multiple roles in energy production, signaling, and biosynthesis. Mitochondria possess multiple copies of mitochondrial DNA (mtDNA), which encodes 37 genes that are essential for mitochondrial and cellular function. When mtDNA is challenged by endogenous and exogenous factors, mtDNA undergoes repair, degradation, and compensatory synthesis. mtDNA degradation is an emerging pathway in mtDNA damage response and maintenance. A key factor involved is the human mitochondrial genome maintenance exonuclease 1 (MGME1). Despite previous biochemical and functional studies, controversies exist regarding the polarity of MGME1-mediated DNA cleavage. Also, how DNA sequence may affect the activities of MGME1 remains elusive. Such information is not only fundamental to the understanding of MGME1 but critical for deciphering the mechanism of mtDNA degradation. Herein, we use quantitative assays to examine the effects of substrate structure and sequence on the DNA-binding and enzymatic activities of MGME1. We demonstrate that MGME1 binds to and cleaves from the 5′-end of single-stranded DNA substrates, especially in the presence of 5′-phosphate, which plays an important role in DNA binding and optimal cleavage by MGME1. In addition, MGME1 tolerates certain modifications at the terminal end, such as a 5′-deoxyribosephosphate intermediate formed in base excision repair. We show that MGME1 processes different sequences with varying efficiencies, with dT and dC sequences being the most and least efficiently digested, respectively. Our results provide insights into the enzymatic properties of MGME1 and a rationale for the coordination of MGME1 with the 3′–5′ exonuclease activity of DNA polymerase γ in mtDNA degradation.

Effect of substrate structure on medium chain fatty acids production and reactor microbiome

The medium chain fatty acids (MCFAs) produced from organic wastes can replace part fossil-fuel-based products to promote the sustainable development of economy and environment. However, the selection and collocation of feedstocks for MCFAs production are lack of reference basis. This study thereby aimed to investigate how the commonly used electron donor (ED) and substrate configuration affect MCFAs synthesis and then obtain the optimal substrate composition. It was found that the optimized ratios for ethanol/acetate, lactate/acetate, and ethanol/lactate/acetate were 3/1, 2/1, and 2/1/1, respectively, and the optimal substrate concentration was 400 mM C. Combining ethanol and lactate as co-EDs effectively concentrated substrate-carbon-flow (increased by 20–28% than sole ED) on MCFAs synthesis by promoting the elongation of butyrate and reutilization of by-products. As a result, the higher MCFAs yield of 646.22 mg COD/g COD and selectivity of 67.72% were obtained from co-EDs than those from sole ED. Moreover, the key functional bacteria enriched under different ED were also discrepant, which were Clostridium sensu stricto for ethanol, Corynebacterium for lactate, and Veillonella and Oscillibacter for ethanol-lactate, respectively. This study provided a basic but significant reference for the scale-up MCFAs production.

(Invited) Ga2O3 Phase Control and Heterojunctions Using Plasma-Enhanced Atomic Layer Epitaxy

Ga2O3 has emerged as a promising material for next generation power electronics. While β-Ga2O3 (monoclinic) is the most stable and studied of six Ga2O3 polymorphs, the slightly less energetically favorable α-, ε-, and κ-Ga2O3 phases have unique characteristics that can be exploited such as larger bandgaps, alloying for dopant control, or polarization beneficial to the formation of two-dimensional electron gas (2DEG) channels. Specifically, α-Ga2O3 (rhombohedral, corundum) has the largest bandgap of ~5.3 eV and can be alloyed with α-Al2O3 (8.8 eV) and α-In2O3 (3.7 eV) for bandgap engineering. Both ε-Ga2O3 (hexagonal, P63mc) and κ-Ga2O3 (orthorhombic, Pna21 ) phases are polar, with a predicted spontaneous polarization strength up to 10 times larger than GaN and 3 times larger than AlN. Like the III-N system, polarization induced charges can lead to higher charge densities and mobilities in 2DEGs formed at heterojunctions, which would improve the viability of Ga2O3 electronic devices. Plasma-enhanced atomic layer deposition (PEALD) is a popular, conformal, energy-enhanced synthesis method for thin films due to its many advantages, including: deposition at reduced growth temperatures, access to metastable phases, improved crystallinity, and increased growth rates. In this work, we use PEALD to produce high-quality heteroepitaxial Ga2O3 and (AlxGa1-x)2O3 (AlGO) films and investigate materials properties such as phase selectivity, ternary solubility limits, and electrical and optical performance.All Ga2O3 films were deposited in a Veeco Fiji G2 reactor equipped with a load lock and turbo pump using trimethygallium, trimethylaluminum, and O2 plasma. Initial studies on c-plane sapphire substrates at 350°C and 8 mTorr show the phase could be altered from β to α by a varying the pure O2 flow during plasma pulse from 5-40 sccm [1]. Optical emission spectroscopy indicate that the changes in the relative concentration of atomic oxygen is crucial for phase selectivity while the high ion flux to the surface can contribute to the crystallinity at low Tg [2]. To grow ε(κ)-Ga2O3 on c-plane sapphire required going to a much higher temperature (500°C), pressure (100’s mTorr), and O2 flow (100sccm) [1]. Without modifications to the current ALD system, pure ε(κ)-Ga2O3 on sapphire was not achieved under any conditions. Using optimum growth conditions for the three phases on sapphire, films were deposited on GaN and diamond to determine the effect of substrate structure. Transmission electron microscopy was conducted to determine the specific phases (β, ε, and κ) present in each case, and showed the amount of each phase varied with PEALD parameters. While films on diamond resulted in mixed β/ε(κ) phases, pure ε(κ)-phase films were attained on GaN and the strain varied with pressure and Tg. Vertical breakdown measurements were taken for both β- and ε(κ)-Ga2O3/n+ GaN substrates. Breakdown fields varied between 3.8-7.0 MV/cm dependent on the phase and strained state of the Ga2O3 films. β-Ga2O3 films showed less variability in breakdown field from device to device than ε(κ)-Ga2O3 films, but neither showed a dependence on device size.While PEALD is beneficial for depositing thin films of metastable phases, practical devices often require much thicker barrier and active layers. For this reason, we investigated integrating PEALD metastable Ga2O3 films with traditional semiconductor deposition techniques, such as molecular beam epitaxy (MBE), capable of extending these layers beyond 100 nm in thickness. The same MBE conditions were used to deposit Ga2O3 films on GaN substrates with and without PEALD ε(κ)-Ga2O3 nucleation layers. Those deposited without the PEALD metastable nucleation layer resulted in stable β-phase films, while those with nucleation layers resulted in pure ε(κ)-phase films. This shows importance of PEALD for realizing practical device structures using metastable phases.Finally, to investigate heterojunctions for 2DEG formation, AlxGa1-xO films were developed. While the full stoichiometric range could be reached using a PEALD digital alloying method, crystallinity was lost above x = 0.2 for the β phase, x = 0.35 for the ε(κ) phase, and x=0.6 for the α phase. Initial device structures will be shown in order to establish the feasibility of these films in device applications.[1] Wheeler, et al. Chem. Mater. 2020, 32, 1140-1152[2] Boris, et al. JVST A 2019, 37(6), 060909 Figure 1

Structure-based design, synthesis of novel probes for cytochrome P450 OleT

Cytochrome P450 OleTSA, a new cytochrome P450 enzyme from Staphylococcus aureus, catalyzes the oxidative decarboxylation and hydroxylation of fatty acids to generate terminal alkenes and fatty alcohols. The mechanism of this bifurcative chemistry remains largely unknown. Herein, a class of derivatized fatty acids were synthesized as probes to investigate the effects of substrate structure on the product type of P450 OleTSA. The results demonstrate that the fine-tuned structure of substrates, even in a remote distance from the carboxyl group, significantly regulates OleT catalyzed decarboxylation/hydroxylation reactions. Molecular docking analysis indicated the potential interactions between the carboxylate groups of different probes and the enzyme active center which was attributed to the bifurcative chemistry.

Cellulose hydrolysis by Trichoderma viride enzyme complex in the presence of sodium fluoride: effect of substrate structure and cellulase sorption activity

The dependence of cellulase activity of the Trichoderma viride fungus on three factors – substrate type, enzyme sorption properties and sodium fluoride (NaF) concentration – was studied. The four studied water-insoluble substrates were based on two native (birch wood and wheat straw) and two model (microcrystalline cellulose and filter paper) materials. In addition, a water-soluble Na-carboxymethyl-cellulose substrate specific for endoglucanase activity was studied. The T. viride cellulase complex from the “Celloviridin G3x” preparation was separated into fractions differing in sorption properties using the affinity chromatography method. The cellulase activity of the initial complex and obtained fractions was measured with respect to all substrates at different concentrations of NaF ranging from 0 to 1000 mg/L. Qualitative differences in the protein composition of the preparation and fractions were studied by SDS electrophoresis. With the introduction of NaF (100–500 mg/L), the initial cellulase complex activity increased with respect to all substrates. The hydrolysis rate of model samples was observed to increase by up to 100 %. Comparison of chromatographic fraction activity relative to model substrates demonstrated the NaF to have the opposite effect on the catalytic and sorption properties of cellulases. The activity of high affinity fractions containing cellobiohydrolases and low molecular weight endoglucanases in the presence of fluoride decreased by 35–90 % in relation to all substrates. Although weakly adsorbing fractions (endoglucanases I, II + cellobiasis) under the same conditions were inhibited by 15–20 % with respect to cellulose with a high degree of crystallinity (MCC), these were activated 1.3–3 times slower with respect to a water-soluble substrate (Na-CMC) and a substrate with less crystallinity (filter paper). Thus, the total effect of fluorides on the enzymatic hydrolysis of the plant substrate is determined by the crystallinity degree of the cellulose substrate, as well as by the ratio of stronglyand weakly-sorbed components in the cellulase complex. Possible molecular mechanisms of these effects are additionally proposed.

Structure-activity relationship of coptis alkaloid bitterness inhibited by mPEG-PLLA

Inhibition of bitterness is a significant measure to improve the compliance and clinical efficacy of traditional Chinese medicine(TCM) decoction. According to the characteristics of TCM decoction, such as high dispersion of bitterness components, multi-component bitterness superposition and strong instantaneous stimulation, the research group put forward a new strategy to inhibit bitterness in the early stage based on the self-assembly characteristics of amphiphilic substances in aqueous solution, in order to reduce the distribution of bitterness components in real solution and achieve the purpose of bitter-masking. It was found that the bitter-masking effect of amphiphilic substances was different on the bitter compounds of various structures. Therefore, it was speculated that there might be a certain relationship between the bitter inhibition effect and the substrate structure. In this paper, the interaction between mPEG-PLLA and five bitter alkaloids(bamatine, jatrorrhizine, berberine, epiberberine and coptisine) in Coptidis Rhizoma was studied to explore the effect of substrate structure on the inhibition of bitterness. The sensory test of volunteers was used to determine the bitter-masking effect of mPEG-PLLA on the decoction of Coptidis Rhizoma and its main bitter alkaloids. The molecular docking and molecular force field were applied to locate the bitter groups and the bitter-masking parts. The relationship between the bitter strength and the structure was analyzed by the surface electrostatic potential of the bitter alkaloids, and the correlation between the bitter-masking effect and the structural parameters of the bitter components was explored by factor analysis, so as to clarify the structure-activity relationship of mPEG-PLLA in masking the bitterness of coptis alkaloids. It was found that mPEG-PLLA had significant taste masking effect on the decoction of Coptidis Rhizoma and five alkaloids. The masking effect was obviously related to the structure of different alkaloids: the effect increased with the increase of the number of hydrogen donors, rotatable bonds, molecular weight, and hydrophobicity, and decreased with the increase of surface electrostatic potential, electrophilicity and binding energy with bitter receptors. In this study, the influence of alkaloid structure of Coptidis Rhizoma on the butter-masking effect of mPEG-PLLA was preliminarily elucidated, providing a scientific basis for better exerting the bitter-masking effect of amphiphilic block copolymers.

Adhesion and Running Speed of a Tropical Arboreal Ant (Cephalotes atratus) on Rough, Narrow, and Inclined Substrates.

Arboreal ants must navigate variably sized and inclined linear structures across a range of substrate roughness when foraging tens of meters above the ground. To achieve this, arboreal ants use specialized adhesive pads and claws to maintain effective attachment to canopy substrates. Here, we explored the effect of substrate structure, including small and large-scale substrate roughness, substrate diameter, and substrate orientation (inclination), on adhesion and running speed of workers of one common, intermediately-sized, arboreal ant species. Normal (orthogonal) and shear (parallel) adhesive performance varied on sandpaper and natural leaf substrates, particularly at small size scales, but running speed on these substrates remained relatively constant. Running speed also varied minimally when running up and down inclined substrates, except when the substrate was positioned completely vertical. On vertical surfaces, ants ran significantly faster down than up. Ant running speed was slower on relatively narrow substrates. The results of this study show that variation in the physical properties of tree surfaces differentially affects arboreal ant adhesive and locomotor performance. Specifically, locomotor performance was much more robust to surface roughness than was adhesive performance. The results provide a basis for understanding how performance correlates of functional morphology contribute to determining local ant distributions and foraging decisions in the tropical rainforest canopy.

Monoterpenoid Oximes Hydrogenation Over Platinum Catalysts

Platinum nanoparticles supported on MgO, Al2O3, ZrO2, TiO2 were utilized for monoterpenoid oximes hydrogenation. Monocyclic menthone and carvone oximes synthesized from bio-derived monoterpenoids with a different structure were used to explore the effect of substrate structure on the reaction regularities. The oximes hydrogenation was carried out under hydrogen atmosphere at 100 °C using methanol as a solvent. Platinum catalysts were prepared by the impregnation methods. The catalysts were characterized by transmission electron microscopy, X-ray photoelectron spectroscopy, X-ray fluorescence spectroscopy, nitrogen physisorption. Hydrogenation of carvone oxime containing a conjugated oxime group and several reducible functional groups resulted in both hydrogenation and deoximation giving 5-isopropyl-2-methylcyclohexanamine and carvomenthone over Pt/Al2O3 and Pt/ZrO2 catalysts. Menthone oxime hydrogenation over Pt/Al2O3 catalysts with an average particle size of 0.8 nm provided the desired menthylamine formation with the selectivity of 90% at complete oxime conversion. Platinum catalysts based on MgO, ZrO2 and TiO2 enhanced menthone oxime deoximation to menthone.

ChemInform Abstract: Synthesis of Hydantoins and Dihydrouracils via Thermally‐Promoted Cyclization of Ureidoacetamides.

Abstract The synthesis of hydantoins and dihydrouracils from ureidoacetamides has been carried out at high temperature in glycol solvents. A series of substrates were prepared and examined to determine the effect of substrate structure, N -acyl substitution (X), and solvent on the course of the reaction. A dramatic effect was observed when using ureidoacetamides (e.g., X= N -methyl- N -phenyl), which led to higher yields, faster reaction times, and lower racemization of chiral substrates. The rate of racemization of a chiral hydantoin in the presence of dibenzylamine and N -methyl aniline has also been determined. The thermal cyclization methodology has been applied to the preparation of a complex hydantoin.

Synthesis of hydantoins and dihydrouracils via thermally-promoted cyclization of ureidoacetamides

The synthesis of hydantoins and dihydrouracils from ureidoacetamides has been carried out at high temperature in glycol solvents. A series of substrates were prepared and examined to determine the effect of substrate structure, N-acyl substitution (X), and solvent on the course of the reaction. A dramatic effect was observed when using ureidoacetamides (e.g., X=N-methyl-N-phenyl), which led to higher yields, faster reaction times, and lower racemization of chiral substrates. The rate of racemization of a chiral hydantoin in the presence of dibenzylamine and N-methyl aniline has also been determined. The thermal cyclization methodology has been applied to the preparation of a complex hydantoin.

Tertiary amine-promoted enone aziridination: investigations into factors influencing enantioselective induction

trans-N-Unsubstituted aziridines were synthesised (up to 77% ee) via a chiral tertiary amine-promoted nucleophilic aziridination of α,β-unsaturated ketones utilising in situ generated N–N ylides (aminimines). A wide range of chiral tertiary amines were synthesised and evaluated, allowing structure–activity relationships to be drawn. The most efficient promoter for asymmetric aziridination, quinine, was assessed with several enones to ascertain the effect of substrate structure on product ee, while the intermediate hydrazinium salt was characterised by X-ray crystallography.

Transient Kinetics and Rate-Limiting Steps for the Processive Cellobiohydrolase Cel7A: Effects of Substrate Structure and Carbohydrate Binding Domain

Cellobiohydrolases are exoacting, processive enzymes that effectively hydrolyze crystalline cellulose. They have attracted considerable interest because of their role in both natural carbon cycling and industrial enzyme cocktails used for the deconstruction of cellulosic biomass, but many mechanistic and regulatory aspects of their heterogeneous catalysis remain poorly understood. Here, we address this by applying a deterministic model to real-time kinetic data with high temporal resolution. We used two variants of the cellobiohydrolase Cel7A from Hypocrea jecorina , and three types of cellulose as substrate. Analysis of the pre-steady-state regime allowed delineation rate constants for both fast and slow steps in the enzymatic cycle and assessment of how these constants influenced the rate of hydrolysis at quasi-steady state. Processive movement on the cellulose strand advanced with characteristic times of 0.15-0.7 s per step at 25 °C, and the rate was highest on amorphous substrate. The cellulose binding module was found to raise this rate on crystalline, but not on amorphous, substrate. The rapid processive movement signified high intrinsic reactivity, but this parameter had marginal influence on the steady-state rate. This was because dissociation and association were slower and, hence, rate limiting. Specifically, the dissociation from the strand was found to occur with characteristic times of 45-100 s. This meant that dissociation was the bottleneck, except at very low substrate loads (0.5-1 g/L), where association became slower.

Synthesis of oxygenated cineole derivatives from cineole: utility of cytochrome P450cin as an enantioselective catalyst

The oxidation of cineole to (1R)-6β-hydroxycineole by cytochrome P450cin (CYP176A) is utilised as the initial step in the synthesis of a range of compounds, both novel and ones previously known only as racemates. The potential of P450cin to provide useful, enantiopure building blocks is thus demonstrated. In particular, hydroxylation by this enzyme was used as the initial step in the synthesis of a range of functionalised cineole derivatives that could be used as mechanistic probes to elucidate the effect of substrate structure on the transformations mediated by P450cin.

Au Clusters on TiO2(110) (1 × 1) and (1 × 2) Surfaces Examined by Polarization-Dependent Total Reflection Fluorescence XAFS

The structure of Au clusters on the TiO2(110) surface and the cluster-substrate interaction were investigated by polarization-dependent total reflection fluorescence X-ray absorption fine structure (PTRF-XAFS). The TiO2(110) (1 × 1) and (1 × 2) surfaces were compared in terms of substrate structural effects on Au cluster structure. We could not find evidence for the direct Au-substrate bonding and observed that the substrate morphology did not significantly affect the size and structure of Au clusters. Finally, we precisely determined that the Au clusters favored a geometry with icosahedral Au55 symmetry. We discuss the possible interaction mechanism between Au and TiO2(110) that leads to these results.

Substrate Structural Effects in Yttrium(III)-Catalyzed Hydroamination/Cyclizations of 1,2-Disubstituted and 1,1,2-Trisubstituted Aminoalkenes ­Terminated by 2-(Phenyl) and 2-(2-Heteroarenyl) Groups

A series of 2-phenyl- and 2-(2-heteroarenyl)-bearing amines possessing 1,2-disubstituted and 1,1,2-trisubststuted ­alkenes have been evaluated in intramolecular hydroaminations catalyzed by Y[N(TMS)2]3 (1Y ). Aminoalkenes possessing a terminal 2-(5-trimethylsilyl)thienyl group exhibited substantially enhanced reactivity compared to their 2-(phenyl)-containing counterparts. Cyclization efficiencies imparted by the 2-[(5-trimethylsilyl)furanyl] substituent were comparable or only slightly better than those obtained with the simple the 2-(phenyl) group.

Effect of substrate structure on properties of nitrided layers formed on pure iron

PurposeThis paper aims to investigate the structure and scratch resistance properties of gas nitrided pure iron samples.Design/methodology/approachThe effects of material strain hardening and amount of grain boundaries exposed on nitriding surface were evaluated by cold rolling the starting samples to different reduction levels before gas nitriding.FindingsThe study finds that nitriding without any prior cold rolling produced a comparatively wide compound layer with a large fraction of porous zone featuring low scratch hardness values but no evidence of damage. On the contrary, cold rolling before nitriding led to a more irregular and thinner compound layer with reduced amount of porous zone and much finer nitrides in the diffusion zone. Scratch hardness was increased but failure mechanism changed by generation of conformal cracks within the track groove and the appearance of discontinuous spallation at high loads.Originality/valueOne of the issues of great industrial importance concerning nitriding of steels is the need to predict the extent of the nitrided layer in products showing small variations in microstructure or in extent of cold working due to complex manufacturing cycles. Despite the practical importance, relatively little information is available in literature about these issues. The present paper is therefore aimed at investigating the structure and mechanical properties of pure iron samples, gas nitrided with different amounts of cold working and microstructural conditions.

The effect of substrate structure on the chemoselectivity of Candida antarctica lipase B-catalyzed acylation of amino-alcohols

► A nucleophilic group in β of an amino group enhances rates of N-acylation. ► The more nucleophilic group in β the higher rates of N-acylation. ► Rates of N-acylation were improved for the short chain amino-alcohol. ► Rates of O-acylation were improved for the long chains amino-alcohols. The selective acylation of multifunctional compounds exhibiting both alcohol and amino groups gives interesting products with many applications in food, cosmetic and pharmaceutical industries, but it is real challenge. The current work describes the different behavior shown by Candida antarctica lipase B (Novozym 435) when catalyzing the O-acylation and N-acylation of bifunctional acyl acceptors. The acylation of three amino-alcohols (alaninol, 4-amino-1-pentanol and 6-amino-1-hexanol) was studied using myristic acid as an acyl donor. To achieve this, a structure-reactivity study was performed in tert -amyl alcohol as a solvent, comparing the three amino-alcohols as acyl acceptors and a series of structurally related amines, namely (R)- sec -butylamine, 1-methoxy-2-propylamine and 1,2-diaminopropane. These substrates were designed to investigate the effect of the group located in β-position of the amino group on the acyl acceptor: the more nucleophilic the group, the more the apparent maximal velocity ( V max,app ) of N-acylation increases. Moreover, the crucial role of the carbon chain length between the alcohol and amino groups on the chemoselectivity was also demonstrated. The chemoselectivity for the N-acylation was improved when the carbon chain included two carbons (alaninol) whereas the chemoselectivity for the O-acylation was improved when the carbon chain included four carbons or more (4-amino-1-pentanol and 6-amino-1-hexanol). These results provided new insights for the selective synthesis of amides or esters produced from the acylation of bifunctional substrates.

Complexity Generation in Fungal Peptidyl Alkaloid Biosynthesis: Oxidation of Fumiquinazoline A to the Heptacyclic Hemiaminal Fumiquinazoline C by the Flavoenzyme Af12070 from Aspergillus fumigatus

The human pathogen Aspergillus fumigatus makes a series of fumiquinazoline (FQ) peptidyl alkaloids of increasing scaffold complexity using L-Trp, 2 equiv of L-Ala, and the non-proteinogenic amino acid anthranilate as building blocks. The FQ gene cluster encodes two non-ribosomal peptide synthetases (NRPS) and two flavoproteins. The trimodular NRPS Af12080 assembles FQF (the first level of complexity) while the next two enzymes, Af12060 and Af12050, act in tandem in an oxidative annulation sequence to couple alanine to the indole side chain of FQF to yield the imidazolindolone-containing FQA. In this study we show that the fourth enzyme, the monocovalent flavoprotein Af12070, introduces a third layer of scaffold complexity by converting FQA to the spirohemiaminal FQC, presumably by catalyzing the formation of a transient imine within the pyrazinone ring (and therefore acting in an unprecedented manner as an FAD-dependent amide oxidase). FQC subsequently converts nonenzymatically to the known cyclic aminal FQD. We also investigated the effect of substrate structure on Af12070 activity and subsequent cyclization with a variety of FQA analogues, including an FQA diastereomer (2'-epi-FQA), which is an intermediate in the fungal biosynthesis of the tremorgenic tryptoquialanine. 2'-epi-FQA is processed by Af12070 to epi-FQD, not epi-FQC, illustrating that the delicate balance in product cyclization regiochemistry can be perturbed by a remote stereochemical center.