Heterogeneous Fe-N-C Catalyst for Aerobic Dehydrogenation of Hydrazones to Diazo Compounds Used for Carbene Transfer.

Organic diazo compounds are versatile reagents in chemical synthesis and would benefit from improved synthetic accessibility, especially for larger scale applications. Here, we report a mild method for the synthesis of diazo compounds from hydrazones using a heterogeneous Fe-N-C catalyst, which has Fe ions dispersed within a graphitic nitrogen-doped carbon support. The reactions proceed readily at room temperature using O2 (1 atm) as the oxidant. Aryl diazoesters, ketones, and amides are accessible, in addition to less stable diaryl diazo compounds. Initial-rate data show that the Fe-N-C catalyst achieves faster rates than a heterogeneous Pt/C catalyst. The oxidative dehydrogenation of hydrazones may be performed in tandem with Rh-catalyzed enantioselective C-H insertion and cyclopropanation of alkenes, without requiring isolation of the diazo intermediate. This sequence is showcased by using a flow reactor for continuous synthesis of diazo compounds.

Chemical Kinetic Method for Active-Site Quantification in Fe-N-C Catalysts and Correlation with Molecular Probe and Spectroscopic Site-Counting Methods.

Mononuclear Fe ions ligated by nitrogen (FeNx) dispersed on nitrogen-doped carbon (Fe-N-C) serve as active centers for electrocatalytic O2 reduction and thermocatalytic aerobic oxidations. Despite their promise as replacements for precious metals in a variety of practical applications, such as fuel cells, the discovery of new Fe-N-C catalysts has relied primarily on empirical approaches. In this context, the development of quantitative structure-reactivity relationships and benchmarking of catalysts prepared by different synthetic routes and by different laboratories would be facilitated by the broader adoption of methods to quantify atomically dispersed FeNx active centers. In this study, we develop a kinetic probe reaction method that uses the aerobic oxidation of a model hydroquinone substrate to quantify the density of FeNx centers in Fe-N-C catalysts. The kinetic method is compared with low-temperature Mössbauer spectroscopy, CO pulse chemisorption, and electrochemical reductive stripping of NO derived from NO2- on a suite of Fe-N-C catalysts prepared by diverse routes and featuring either the exclusive presence of Fe as FeNx sites or the coexistence of aggregated Fe species in addition to FeNx. The FeNx site densities derived from the kinetic method correlate well with those obtained from CO pulse chemisorption and Mössbauer spectroscopy. The broad survey of Fe-N-C materials also reveals the presence of outliers and challenges associated with each site quantification approach. The kinetic method developed here does not require pretreatments that may alter active-site distributions or specialized equipment beyond reaction vessels and standard analytical instrumentation.

Quinone-mediated hydrogen anode for non-aqueous reductive electrosynthesis.

Electrochemical synthesis can provide more sustainable routes to industrial chemicals1-3. Electrosynthetic oxidations may often be performed 'reagent-free', generating hydrogen (H2) derived from the substrate as the sole by-product at the counter electrode. Electrosynthetic reductions, however, require an external source of electrons. Sacrificial metal anodes are commonly used for small-scale applications4, but more sustainable options are needed at larger scale. Anodic water oxidation is an especially appealing option1,5,6, but many reductions require anhydrous, air-free reaction conditions. In such cases, H2 represents an ideal alternative, motivating the growing interest in the electrochemical hydrogen oxidation reaction (HOR) under non-aqueous conditions7-12. Here we report a mediated H2 anode that achieves indirect electrochemical oxidation of H2 by pairing thermal catalytic hydrogenation of an anthraquinone mediator with electrochemical oxidation of the anthrahydroquinone. This quinone-mediated H2 anode is used to support nickel-catalysed cross-electrophile coupling (XEC), a reaction class gaining widespread adoption in the pharmaceutical industry13-15. Initial validation of this method in small-scale batch reactions is followed by adaptation to a recirculating flow reactor that enables hectogram-scale synthesis of a pharmaceutical intermediate. The mediated H2 anode technology disclosed here offers a general strategy to support H2-driven electrosynthetic reductions.

Heterogeneous M-N-C Catalysts for Aerobic Oxidation Reactions: Lessons from Oxygen Reduction Electrocatalysts.

Nonprecious metal heterogeneous catalysts composed of first-row transition metals incorporated into nitrogen-doped carbon matrices (M-N-Cs) have been studied for decades as leading alternatives to Pt for the electrocatalytic O2 reduction reaction (ORR). More recently, similar M-N-C catalysts have been shown to catalyze the aerobic oxidation of organic molecules. This Focus Review highlights mechanistic similarities and distinctions between these two reaction classes and then surveys the aerobic oxidation reactions catalyzed by M-N-Cs. As the active-site structures and kinetic properties of M-N-C aerobic oxidation catalysts have not been extensively studied, the array of tools and methods used to characterize ORR catalysts are presented with the goal of supporting further advances in the field of aerobic oxidation.

Molecular Catalyst Synthesis Strategies to Prepare Atomically Dispersed Fe-N-C Heterogeneous Catalysts.

We report a strategy to integrate atomically dispersed iron within a heterogeneous nitrogen-doped carbon (N-C) support, inspired by routes for metalation of molecular macrocyclic iron complexes. The N-C support, derived from pyrolysis of a ZIF-8 metal-organic framework, is metalated via solution-phase reaction with FeCl2 and tributyl amine, as a Brønsted base, at 150 °C. Fe active sites are characterized by 57Fe Mössbauer spectroscopy and aberration-corrected scanning transmission electron microscopy. The site density can be increased by selective removal of Zn2+ ions from the N-C support prior to metalation, resembling the transmetalation strategy commonly employed for the preparation of molecular Fe-macrocycles. The utility of this approach is validated by the higher catalytic rates (per total Fe) of these materials relative to established Fe-N-C catalysts, benchmarked using an aerobic oxidation reaction.

Chemical and Electrochemical O2 Reduction on Earth-Abundant M-N-C Catalysts and Implications for Mediated Electrolysis.

M-N-C catalysts, incorporating non-precious-metal ions (e.g. M = Fe, Co) within a nitrogen-doped carbon support, have been the focus of broad interest for electrochemical O2 reduction and aerobic oxidation reactions. The present study explores the mechanistic relationship between the O2 reduction mechanism under electrochemical and chemical conditions. Chemical O2 reduction is investigated via the aerobic oxidation of a hydroquinone, in which the O-H bonds supply the protons and electrons needed for O2 reduction to water. Mechanistic studies have been conducted to elucidate whether the M-N-C catalyst couples two independent half-reactions (IHR), similar to electrode-mediated processes, or mediates a direct inner-sphere reaction (ISR) between O2 and the organic molecule. Kinetic data support the latter ISR pathway. This conclusion is reinforced by rate/potential correlations that reveal significantly different Tafel slopes, implicating different mechanisms for chemical and electrochemical O2 reduction.

Anthraquinone-Mediated Fuel Cell Anode with an Off-Electrode Heterogeneous Catalyst Accessing High Power Density when Paired with a Mediated Cathode.

The development of processes for electrochemical energy conversion and chemical production could benefit from new strategies to interface chemical redox reactions with electrodes. Here, we employ a diffusible low-potential organic redox mediator, 9,10-anthraquinone-2,7-disulfonic acid (AQDS), to promote efficient electrochemical oxidation of H2 at an off-electrode heterogeneous catalyst. This unique approach to integrate chemical and electrochemical redox processes accesses power densities up to 228 mW/cm2 (528 mW/cm2 with iR-correction). These values are significantly higher than those observed in previous mediated electrochemical H2 oxidation methods, including those using enzymes or inorganic mediators. The approach described herein shows how traditional catalytic chemistry can be coupled to electrochemical devices.

Light-optimized growth of cyanobacterial cultures: Growth phases and productivity of biomass and secreted molecules in light-limited batch growth.

Cyanobacteria are photosynthetic microorganisms whose metabolism can be modified through genetic engineering for production of a wide variety of molecules directly from CO2, light, and nutrients. Diverse molecules have been produced in small quantities by engineered cyanobacteria to demonstrate the feasibility of photosynthetic biorefineries. Consequently, there is interest in engineering these microorganisms to increase titer and productivity to meet industrial metrics. Unfortunately, differing experimental conditions and cultivation techniques confound comparisons of strains and metabolic engineering strategies. In this work, we discuss the factors governing photoautotrophic growth and demonstrate nutritionally replete conditions in which a model cyanobacterium can be grown to stationary phase with light as the sole limiting substrate. We introduce a mathematical framework for understanding the dynamics of growth and product secretion in light-limited cyanobacterial cultures. Using this framework, we demonstrate how cyanobacterial growth in differing experimental systems can be easily scaled by the volumetric photon delivery rate using the model organisms Synechococcus sp. strain PCC7002 and Synechococcus elongatus strain UTEX2973. We use this framework to predict scaled up growth and product secretion in 1L photobioreactors of two strains of Synechococcus PCC7002 engineered for production of l-lactate or L-lysine. The analytical framework developed in this work serves as a guide for future metabolic engineering studies of cyanobacteria to allow better comparison of experiments performed in different experimental systems and to further investigate the dynamics of growth and product secretion.

High-CO2 Requirement as a Mechanism for the Containment of Genetically Modified Cyanobacteria.

As researchers engineer cyanobacteria for biotechnological applications, we must consider potential environmental release of these organisms. Previous theoretical work has considered cyanobacterial containment through elimination of the CO2-concentrating mechanism (CCM) to impose a high-CO2 requirement (HCR), which could be provided in the cultivation environment but not in the surroundings. In this work, we experimentally implemented an HCR containment mechanism in Synechococcus sp. strain PCC7002 (PCC7002) through deletion of carboxysome shell proteins and showed that this mechanism contained cyanobacteria in a 5% CO2 environment. We considered escape through horizontal gene transfer (HGT) and reduced the risk of HGT escape by deleting competence genes. We showed that the HCR containment mechanism did not negatively impact the performance of a strain of PCC7002 engineered for L-lactate production. We showed through coculture experiments of HCR strains with ccm-containing strains that this HCR mechanism reduced the frequency of escape below the NIH recommended limit for recombinant organisms of one escape event in 108 CFU.

Platinum-Based Heterogeneous Catalysts for Nitrile Synthesis via Aerobic Oxidative Coupling of Alcohols and Ammonia.

Transition-metal-catalyzed aerobic oxidative coupling of alcohols and ammonia represents an attractive atom-economical synthetic route to prepare nitriles. Heterogeneous platinum catalysts have been widely used for aerobic alcohol oxidation to aldehydes and carboxylic acids but have not been applied to nitrile synthesis. In this work, we show that carbon-supported Pt catalysts are effective for this transformation. Unpromoted Pt is competent with benzylic substrates bearing either electron-donating or electron-withdrawing substituents. Use of both K2CO3 and Bi additives accelerates the rate of alcohol oxidation and affords high yields with challenging heterocyclic alcohols.

Discovery of Multicomponent Heterogeneous Catalysts via Admixture Screening: PdBiTe Catalysts for Aerobic Oxidative Esterification of Primary Alcohols.

In the present study, we demonstrate the utility of "admixture screening" for the discovery of new multicomponent heterogeneous Pd catalyst compositions that are highly effective for aerobic oxidative methyl esterification of primary alcohols. The identification of possible catalysts for this reaction was initiated by the screening of simple binary and ternary admixtures of Pd/charcoal in combination with one or two metal and/or metalloid components as the catalyst. This approach permitted rapid evaluation of over 400 admixture combinations for the oxidative methyl esterification of 1-octanol at 60 °C in methanol. Product yields from these reactions varied widely, ranging from 2% to 88%. The highest yields were observed with Bi-, Te-, and Pb-based additives, and particularly from those containing both Bi and Te. Validation of the results was achieved by preparing specific PdBiTe catalyst formulations via a wet-impregnation method, followed by application of response surface methodology to identify the optimal Pd-Bi-Te catalyst stoichiometry. This approach revealed two very effective catalyst compositions: PdBi0.47Te0.09/C (PBT-1) and PdBi0.35Te0.23/C (PBT-2). The former catalyst was used in batch aerobic oxidation reactions with different primary alcohols and shown to be compatible with substrates bearing heterocycle and halide substituents. The methyl ester products were obtained in >90% yield in nearly all cases. Implementation of the PBT-2 catalyst in a continuous-flow packed-bed reactor achieved nearly 60 000 turnovers with no apparent loss of catalytic activity.

A roadmap for the synthesis of separation networks for the recovery of bio-based chemicals: Matching biological and process feasibility.

Microbial conversion of renewable feedstocks to high-value chemicals is an attractive alternative to current petrochemical processes because it offers the potential to reduce net CO2 emissions and integrate with bioremediation objectives. Microbes have been genetically engineered to produce a growing number of high-value chemicals in sufficient titer, rate, and yield from renewable feedstocks. However, high-yield bioconversion is only one aspect of an economically viable process. Separation of biologically synthesized chemicals from process streams is a major challenge that can contribute to >70% of the total production costs. Thus, process feasibility is dependent upon the efficient selection of separation technologies. This selection is dependent on upstream processing or biological parameters, such as microbial species, product titer and yield, and localization. Our goal is to present a roadmap for selection of appropriate technologies and generation of separation schemes for efficient recovery of bio-based chemicals by utilizing information from upstream processing, separation science and commercial requirements. To achieve this, we use a separation system comprising of three stages: (I) cell and product isolation, (II) product concentration, and (III) product purification and refinement. In each stage, we review the technology alternatives available for different tasks in terms of separation principles, important operating conditions, performance parameters, advantages and disadvantages. We generate separation schemes based on product localization and its solubility in water, the two most distinguishing properties. Subsequently, we present ideas for simplification of these schemes based on additional properties, such as physical state, density, volatility, and intended use. This simplification selectively narrows down the technology options and can be used for systematic process synthesis and optimal recovery of bio-based chemicals.

Experimental Limiting Oxygen Concentrations for Nine Organic Solvents at Temperatures and Pressures Relevant to Aerobic Oxidations in the Pharmaceutical Industry.

Applications of aerobic oxidation methods in pharmaceutical manufacturing are limited in part because mixtures of oxygen gas and organic solvents often create the potential for a flammable atmosphere. To address this issue, limiting oxygen concentration (LOC) values, which define the minimum partial pressure of oxygen that supports a combustible mixture, have been measured for nine commonly used organic solvents at elevated temperatures and pressures. The solvents include acetic acid, N-methylpyrrolidone, dimethyl sulfoxide, tert-amyl alcohol, ethyl acetate, 2-methyltetrahydrofuran, methanol, acetonitrile, and toluene. The data obtained from these studies help define safe operating conditions for the use of oxygen with organic solvents.

Continuous Flow Aerobic Alcohol Oxidation Reactions Using a Heterogeneous Ru(OH) x /Al2O3 Catalyst.

Ru(OH) x /Al2O3 is among the more versatile catalysts for aerobic alcohol oxidation and dehydrogenation of nitrogen heterocycles. Here, we describe the translation of batch reactions to a continuous-flow method that enables high steady-state conversion and single-pass yields in the oxidation of benzylic alcohols and dehydrogenation of indoline. A dilute source of O2 (8% in N2) was used to ensure that the reaction mixture, which employs toluene as the solvent, is nonflammable throughout the process. A packed bed reactor was operated isothermally in an up-flow orientation, allowing good liquid-solid contact. Deactivation of the catalyst during the reaction was modeled empirically, and this model was used to achieve high conversion and yield during extended operation in the aerobic oxidation of 2-thiophene methanol (99+% continuous yield over 72 h).

The effect of high intensity mixing on the enzymatic hydrolysis of concentrated cellulose fiber suspensions.

Enzymatic hydrolysis of lignocellulosic biomass in a high shear environment was examined. The conversion of cellulose to glucose in samples mixed in a torque rheometer producing shear flows similar to those found in twin screw extruders was greater than that of unmixed samples. In addition, there is a synergistic effect of mixing and enzymatic hydrolysis; mixing increases the rate of cellulose conversion while the increased conversion facilitates mixing. The synergy appears to result in part from particle size reduction, which is more significant when hydrolysis occurs during intense mixing.

Emergence of ideal membrane cascades for downstream processing.

An algorithm is developed for describing ideal membrane cascades for fractionation of binary and pseudo-binary mixtures. It is shown that solvent management plays a key role in determining both purification and yield. Development of efficient diafilters is needed if membrane cascades are to achieve their full potential in competing with both chromatography and simulated moving bed operations in downstream processing of proteins. Such a replacement will also be important for fractionation of higher titers and larger substrates, such as plasmids, viruses, and even whole cells.

Identification of non-idealities in gel electrophoresis.

The molecular weight separation, which is the second dimension of two-dimensional gel electrophoresis, is studied quantitatively with the goal of improving positional predictability and reproducibility. Mathematical modeling of carrier electrolyte dynamics is used to track the progress of a stacking front as a function of coulombs passed. In all test cases, the front moves more slowly than predicted and shows both curvature and tilt. These systematic deviations from the model are found to be influenced by a variety of factors, including both design and operating features. These factors are largely explained, and suggestions are made for improvements.

Adsorption and desorption behavior of plasmid DNA on ion-exchange membranes. Effect of salt valence and compaction agents.

In this study, we report the effect of salt type and compaction agents on adsorption and desorption behavior of plasmid DNA on strong anion-exchange membranes. Both divalent cations and compaction agents are known to reduce the effective charge density of plasmid DNA in solution, and compaction agents decrease the radius of gyration of plasmids. Differences in the batch uptake adsorption of a 6.1 kilo base pair plasmid in solution with sodium and magnesium salts were observed at low ionic strengths. Recoveries at high salt conditions, however, were independent of the cation, and measured only 63-76%. Similarly, no improvement in recoveries were observed when using sulfate rather than chloride anions as displacers. The compaction agents, spermine and spermidine, showed no strong effect on the uptake adsorption, capacity, or recovery of three different-sized plasmids on membrane sheets. It is recommended that further efforts to improve plasmid recoveries from anion-exchange membranes focus on properties of the adsorbent surface.

Performance and scale-up of adsorptive membrane chromatography.

Separation efficiency and scalability of Pall Corporation's new Mustang stacked membrane chromatographic devices were investigated, using both the 10-ml and l(-1) models and comparing the responses of tracer pulses obtained for conventional and reverse-flow operation. Tracers included AMP, lysozyme, and thyroglobulin, which vary in relative molecular mass from less than 1000 up to 650000. Both devices showed marked insensitivity to tracer size and flow-rate and gave sharper peaks than would have been expected from conventional 15-microm bead packings. However, reverse-flow peaks were always significantly sharper than those for conventional operation, and the differences were ascribed primarily to non-uniform header residence times. Numerical simulations of the macroscopic flow confirmed that this was indeed the case. This problem was much less pronounced for the l(-1) device so scale-up is conservative.