Asymmetric Epoxidation Of Styrene Research Articles

Enantioselectivity and key residue of Herbaspirillum huttiense monooxygenase in asymmetric epoxidation of styrenes.

Styrene monooxygenases (SMOs) are powerful enzymes for the synthesis of enantiopure epoxides, but these SMOs have narrow substrate spectra, and the residues in controlling enantioselectivity of SMOs remains unclear. A monooxygenase from Herbaspirillum huttiense (HhMO) was found to have excellent enantioselectivities and diastereoselectivities in the epoxidation of unconjugated terminal alkenes. Here we found that HhMO could also transfer styrene into styrene epoxide with 75% ee, and it could also catalyze the epoxidation of styrene derivatives into the corresponding epoxides with enantioselectivities up to 99% ee. Meanwhile, site 199 in the substrate access channel of HhMO was found to play an important role in the controlling enantioselectivity of the epoxidation. The E199L variant catalyzed the epoxidation of styrene with > 99% ee. The identification of critical residue that affects the enantioselectivity of SMOs would thus be valuable for creating efficient monooxygenases for the preparation of essential enantiopure epoxides. KEY POINTS: • Bioexpoxidation of both conjugated and unconjugated alkenes by HhMO with excellent enantioselectivities. • Gating residue 199 played an essential role in controlling the enantioselectivity of SMO. • HhMO E199L catalyzed the epoxidation of styrenes with up to > 99% ee.

Synthesis of new chiral Mn(iii)–salen complexes as recoverable and reusable homogeneous catalysts for the asymmetric epoxidation of styrenes and chromenes

New chiral Mn(iii)–salen complexes 1a–e and 2a–e were synthesized from the reaction of C2-symmetric chiral salen ligands and Mn(CH3COO)2·4H2O under an inert atmosphere followed by aerobic oxidation.

Chiral Phosphotungstate Functionalized with (S)-1-Phenylethylamine: Synthesis, Characterization, and Asymmetric Epoxidation of Styrene.

In the present work, an attempt has been made to induce chirality in copper-substituted phosphotungstate (PW11Cu) by functionalization with (S)-(+)-1-phenylethylamine (S-PEA) via a ligand substitution approach. The formation of a N→Cu dative bond was confirmed by 13C NMR, while 1H NMR, circular dichroism spectroscopy and optical rotation studies confirmed the introduction of chirality to the Keggin structure. The synthesized material was used as the heterogeneous catalyst for the asymmetric epoxidation of styrene using various green oxidants to obtain high enantiomeric excess (ee), and the reaction with molecular oxygen was found to give the best ee. Regeneration studies were carried out, and the catalyst was found to be suitable for the same. A probable mechanism is also proposed. A comparison with other copper-based polyoxometalate catalysts clearly demonstrate the superiority and novelty of the present catalyst in terms of the reaction conditions as well as the obtained ee.

Recent Developments in Heterogeneous Catalyzed Epoxidation of Styrene to Styrene Oxide

AbstractEpoxidation of styrene under heterogeneous catalyzed conditions provides environmentally friendly routes to produce styrene oxide, an important synthetic intermediate for synthesis of perfumes, epoxy resins, plasticizers, drugs, sweeteners and fine chemicals. The present review explores epoxidation of styrene under heterogeneous catalysed conditions using a variety of catalysts such as metal nanoparticles, metal oxides (of group 12 and 13 metals, transition metals, rare earth metals), transition metals and complexes, polyoxometallates, microporous and mesoporous materials (zeolites, modified zeolites, organic/inorganic porous hybrid materials, porous metal oxides, SBA 15) and polymers using hydrogen peroxide (H2O2), tert‐butyl hydroperoxide (TBHP), air or oxygen as oxidants.The use of salen based transition metal complexes and of chiral porphyrins as catalysts in asymmetric epoxidation of styrene is also reviewed. The reaction mechanism studies of most of these reactions suggest an intermediate formation of a reactive metal‐oxygen species, by interactions between the catalyst and oxidant.

Enhanced Turnover for the P450 119 Peroxygenase-Catalyzed Asymmetric Epoxidation of Styrenes by Random Mutagenesis.

A randomized library is constructed based on pET30a-CYP119-T214V plasmid. This library of random mutants of CYP119-T214V was screened by means of the reduced CO difference spectra and epoxidation of styrene. By using directed evolution, a new CYP119 quadruple mutant S148P/I161T/K199E/T214V is constructed, expressed, and purified. This quadruple mutant significantly increases the turnover rate and conversion for the asymmetric epoxidation of styrene and its derivatives. The kcat. value of cis-β-methylstyrene epoxidation catalyzed by the quadruple mutant exhibits an approximately 10-fold increase, relative to the previously reported T213M mutant under the same conditions. This is the first engineered CYP119 peroxygenase for the epoxidation of cis-β-methylstyrene with a high turnover rate. The proposed mechanism, on the basis of a molecular docking study, for the asymmetric epoxidation suggests that the introduction of an acidic amino acid side chain into the active site and a hydrophobic amino acid into the substrate channels of CYP119 peroxygenase might result in high efficiency for the formation of compound I, and its subsequent peroxygenation by reconstructing the hydrogen-bonding interaction and increasing the substrate affinity and access.

New chiral catalytic membrane reactor created by immobilizing salen-Mn(III) onto APTES modified ceramic membrane and its performances in epoxidation of styrene

New chiral catalytic ceramic membrane (CCCM) reactor was constructed by immobilizing mono-CH2Cl-salen-Mn(III) catalysts onto 3-aminopropyltriethoxysilane modified ceramic ultrafiltration membranes through chemical bonding. The performance of CCCM reactor was investigated by catalyzing the asymmetric epoxidation of styrene and was comparable to the homogeneous salen-Mn(III) catalyst under the same reaction conditions. Furthermore, the obtained reactor showed relatively stable catalytic performances in four runs of epoxidation reaction. And in this reactor, the catalyst can be reused directly without any separation stage.

Enhanced turnover rate and enantioselectivity in the asymmetric epoxidation of styrene by new T213G mutants of CYP 119

New CYP 119 T213G mutants were constructed and characterized. Introduction of T213G mutation into the wild-type CYP 119 enhances the turnover rate for the styrene epoxidation to 346.2 min−1, and the double T213G/T214V mutant improves the ratio of the S- and R-enantiomers of the epoxide products to 5.8. The molecular docking results support our initial design and experimental data.

Gas chromatography-mass spectrometry/selected ion monitoring for screening asymmetric epoxidation of styrene.

A screening method by gas chromatography-mass spectrometry (GC-MS) with selected ion monitoring (SIM) has been developed for asymmetric epoxidation of styrene. All samples were analyzed by GC-MS with the mode of SIM by using ethylbenzene as the internal standard, by which styrene and styrene epoxide enantiomers were completely separated and their concentrations were accurately determined. Furthermore, this method was used to evaluate the epoxidation reaction of styrene, which was optimized by orthogonal experiment. The advantages of the method are as follows: excellent precision (RSD in the range of 1. 2% - 5. 2%, n= 5), good linearity (the correlation coefficients for styrene, (R)/(S)-styrene oxide were 0. 9997, 0. 9932 and 0. 9963, respectively) , low limits of quantitation (LOQs of styrene, (R)/(S)-styrene oxide were 1. 3, 1. 1 and 0. 7 mg/L, respectively) and reasonable recoveries (98. 2% - 108. 2%). All in all, the method provided a new way to determine the asymmetric epoxidation mixture of styrene and its homologs.

Novel chiral (salen)Mn(iii) complexes containing a calix[4]arene unit in 1,3-alternate conformation as catalysts for enantioselective epoxidation reactions of (Z)-aryl alkenes

Two new chiral calix[4]arene-salen ligands 1a,b, based on calix[4]arene platforms in 1,3-alternate conformation, have been prepared by a new general synthetic pathway. Their Mn(III) complexes, 3a,b have shown fairly good efficiency in the asymmetric epoxidation of styrene and substituted styrenes, whereas excellent catalytic activity and selectivity were observed with rigid bicyclic alkenes, namely 1,2-dihydro-naphthalene and substituted 2,2'-dimethyl-chromene. The higher catalytic properties of 3a may be ascribed to the more rigid and inherently chiral structure as proved by molecular modelling, NMR spectroscopy and X-ray data of the similarly structured UO2 complexes 2a,b.

Synthesis of new perfluorinated binaphthyl Mn(III) complexes and epoxidation of styrene in supercritical carbon dioxide

In this article, new perfluorinated binaphthyl Mn(III) complexes have been synthesized by template effect. The synthesized complexes were characterized by elemental analysis, FT-IR, LC/MS, UV–vis spectroscopy. After the solubility of the catalysts in scCO2 was observed through the high pressure reactor, catalytic activities of the catalysts have been tested in homogeneous asymmetric epoxidation of styrene in both dichloromethane (organic solvent) and supercritical carbon dioxide (scCO2). In order to determine and compare to complexes, the effect of synthesized catalysts to conversion and selectivity was investigated. Reaction parameters such as; reaction time, solvent type and position of substituents were also assessed during the analysis.

The elpaN-salen series: multifunctional ligands based on d-glucose for the Mn(III)-catalyzed enantioselective epoxidation of styrenes

It is described the simple design of a series of N,N′,O,O′-chiral ligands (elpaN-salen-H2), which represent a subset of the elpaN-type library of ligands based on β-1,2-d-glucodiamine. The corresponding Mn(III) complexes were examined in the asymmetric epoxidation of styrenes with satisfactory results, e.g. cis-β-methylstyrene was functionalised in ees up to 88%.

Engineered catalytic biofilms for continuous large scale production of n‐octanol and (S)‐styrene oxide

This study evaluates the technical feasibility of biofilm-based biotransformations at an industrial scale by theoretically designing a process employing membrane fiber modules as being used in the chemical industry and compares the respective process parameters to classical stirred-tank studies. To our knowledge, catalytic biofilm processes for fine chemicals production have so far not been reported on a technical scale. As model reactions, we applied the previously studied asymmetric styrene epoxidation employing Pseudomonas sp. strain VLB120ΔC biofilms and the here-described selective alkane hydroxylation. Using the non-heme iron containing alkane hydroxylase system (AlkBGT) from P. putida Gpo1 in the recombinant P. putida PpS81 pBT10 biofilm, we were able to continuously produce 1-octanol from octane with a maximal productivity of 1.3 g L ⁻¹(aq) day⁻¹ in a single tube micro reactor. For a possible industrial application, a cylindrical membrane fiber module packed with 84,000 polypropylene fibers is proposed. Based on the here presented calculations, 59 membrane fiber modules (of 0.9 m diameter and 2 m length) would be feasible to realize a production process of 1,000 tons/year for styrene oxide. Moreover, the product yield on carbon can at least be doubled and over 400-fold less biomass waste would be generated compared to classical stirred-tank reactor processes. For the octanol process, instead, further intensification in biological activity and/or surface membrane enlargement is required to reach production scale. By taking into consideration challenges such as biomass growth control and maintaining a constant biological activity, this study shows that a biofilm process at an industrial scale for the production of fine chemicals is a sustainable alternative in terms of product yield and biomass waste production.

Subtoxic product levels limit the epoxidation capacity of recombinant E. coli by increasing microbial energy demands

The utilization of the cellular metabolism for cofactor regeneration is a common motivation for the application of whole cells in redox biocatalysis. Introduction of an active oxidoreductase into a microorganism has profound consequences on metabolism, potentially affecting metabolic and biotransformation efficiency. An ambitious goal of systems biotechnology is to design process-relevant and knowledge-based engineering strategies to improve biocatalyst performance.Metabolic flux analysis (MFA) has shown that the competition for NAD(P)H between redox biocatalysis and the energy metabolism becomes critical during asymmetric styrene epoxidation catalyzed by growing Escherichia coli containing recombinant styrene monooxygenase. Engineering TCA-cycle regulation allowed increased TCA-cycle activities, a delay of acetate formation, and enhanced NAD(P)H yields during batch cultivation. However, at low biomass and product concentrations, the cellular metabolism of both the mutants as well as the native host strains could cope with increased NADH demands during continuous two-liquid phase biotransformations, whereas elevated but still subtoxic product concentrations were found to cause a significantly increased NAD(P)H demand and a compromised efficiency of metabolic operation. In conclusion, operational conditions determine cellular energy and NAD(P)H demands and thus the biocatalytic efficiency of whole-cell redox biocatalysts.

Production host selection for asymmetric styrene epoxidation: Escherichia coli vs. solvent-tolerant Pseudomonas

Selection of the ideal microbe is crucial for whole-cell biotransformations, especially if the target reaction intensively interacts with host cell functions. Asymmetric styrene epoxidation is an example of a reaction which is strongly dependent on the host cell owing to its requirement for efficient cofactor regeneration and stable expression of the styrene monooxygenase genes styAB. On the other hand, styrene epoxidation affects the whole-cell biocatalyst, because it involves toxic substrate and products besides the burden of additional (recombinant) enzyme synthesis. With the aim to compare two fundamentally different strain engineering strategies, asymmetric styrene epoxidation by StyAB was investigated using the engineered wild-type strain Pseudomonas sp. strain VLB120ΔC, a styrene oxide isomerase (StyC) knockout strain able to accumulate (S)-styrene oxide, and recombinant E. coli JM101 carrying styAB on the plasmid pSPZ10. Their performance was analyzed during fed-batch cultivation in two-liquid phase biotransformations with respect to specific activity, volumetric productivity, product titer, tolerance of toxic substrate and products, by-product formation, and product yield on glucose. Thereby, Pseudomonas sp. strain VLB120ΔC proved its great potential by tolerating high styrene oxide concentrations and by the absence of by-product formation. The E. coli-based catalyst, however, showed higher specific activities and better yields on glucose. The results not only show the importance but also the complexity of host cell selection and engineering. Finding the optimal strain engineering strategy requires profound understanding of bioprocess and biocatalyst operation. In this respect, a possible negative influence of solvent tolerance on yield and activity is discussed.

Reusable chiral macrocyclic Mn(III) salen complexes for enantioselective epoxidation of nonfunctionalized alkenes

A series of new chiral monomeric and dimeric macrocyclic Mn(III) salen complexes 1–4 with trigol linker were synthesized, characterized (by microanalysis, IR spectroscopy, UV–vis. spectroscopy, optical rotation, and mass spectrometry), and used as catalysts in the enantioselective epoxidation of styrene, cis β-methyl styrene, indene, and chromenes in the presence of several N-oxides as an axial base and NaOCl as an oxidant at 0°C. With the use of chiral dimeric macrocyclic catalyst 3 (2.5mol%), enantio-pure epoxides were achieved in excellent yields (>99%) and enantioselectivities (ee up to 98% in selected cases). The recycling was demonstrated with complex 4 (recyclable up to six cycles studied with retention of enantioselectivity) in the asymmetric epoxidation of styrene. The kinetic investigation with complex 4 for the epoxidation of styrene as the representative substrate showed the first-order dependence on the catalyst and the oxidant but independent on the initial concentration of the substrate.

Mesoporous Silica Supported Unsymmetric Chiral Mn(III) Salen Complex: Synthesis, Characterization and Effect of Pore Size on Catalytic Performance

An unsymmetric chiral Mn(III) salen complex was immobilized onto a series of MCM-41 and MCM-48 mesoporous materials with different pore sizes. The as-synthesized catalysts were characterized by XRD, FT-IR, DR UV-Vis, N2 sorption and XPS. The results indicated that chiral Mn(III) salen complex was immobilized into the nanopores of supports and the long-range mesoporous ordering of parent supports was maintained after the immobilization. The heterogeneous catalysts were evaluated in the asymmetric epoxidation of styrene, indene, and 1-phenylcyclohexene, and the effect of 3-mercaptopropyltrimethoxysilane dosage and fine-tuning of pore size on the catalytic performance was studied. It was found that the activity and enantioselectivity were closely correlated with the pore sizes of heterogeneous catalysts. The larger pore sizes of supports could lead to higher conversions and ee values.

Highly Enantioselective Epoxidation of Unfunctionalized Olefins Catalyzed by Chiral Jacobsen’s Catalyst Immobilized on Phenoxy-Modified Zirconium Poly(syrene-phenylvinylphos- phonate)phosphate

Chiral Jacobsen’s catalyst was axially immobilized onto phenoxy-modified zirconium poly(styrene-phenylvinylphosphonate)phosphate (ZPS-PVPA). The immobilized catalysts show comparable ee values for asymmetric epoxidation of styrene and much higher ee values for α-methylstyrene (73.7% vs. 54.0%) and indene (99.9% vs. 65.0%) than the homogeneous Jacobsen’s catalyst. Moreover, the as-synthesized catalysts are relatively stable and can be recycled at least five times without significant loss of activity and enantioselectivity. A point worth emphasizing is that the heterogeneous catalysts afforded remarkable increases of conversion and ee values in the absence of expensive O-coordinating axial bases for the asymmetric epoxidation of olefins, especially for the epoxidation of α-methylstyrene (conversion: from 24.3% to 99.9%; ee: from 29.4% to 73.7%), which may overcome the last obstacle for the potential industry application of chiral Jacobsen’s catalyst.

Productive Asymmetric Styrene Epoxidation Based on a Next Generation Electroenzymatic Methodology

AbstractWe have established a novel and scalable methodology for the productive coupling of redox enzymes to reductive electrochemical cofactor regeneration relying on efficient mass transfer of the cofactor to the electron‐delivering cathode. Proof of concept is provided by styrene monooxygenase (StyA) catalyzing the asymmetric (S)‐epoxidation of styrene with high enantiomeric excess, space‐time yields, and current efficiencies. Highly porous reticulated vitreous carbon electrodes, maximized in volumetric surface area, were employed in a flow‐through mode to rapidly regenerate the consumed FADH2 cofactor required for StyA activity. A systematic investigation of the parameters determining cofactor mass transfer revealed that low FAD concentrations and high flow rates enabled the continuous synthesis of the product (S)‐styrene oxide at high rates, while at the same time the accumulation of the side‐products acetophenone and phenylacetaldehyde was minimized. At 10 μM FAD and a flow rate of 150 mL min−1, an average space‐time yield of 0.35 g L−1 h−1 could be achieved during 2 h with a final (S)‐styrene oxide yield of 75.2%. At two‐fold lower aeration rates, the electroenzymatic reaction could be sustained for 12 h, albeit at the expense of lower (59%) overall space‐time yields. Under these conditions, as much as 20.5% of the utilized current could be channeled into (S)‐styrene oxide formation. In comparison with state‐of‐the‐art electroenzymatic methodologies for the same conversion, (S)‐styrene oxide synthesis could be improved up to 150‐fold with respect to both reaction time and space‐time yield. These productivities constitute the most efficient reaction reported for asymmetric in vitro epoxidations of styrene.

ChemInform Abstract: Asymmetric Epoxidation of Styrenes Catalyzed by Molybdenum Complexes with Amino Alcohols Ligands.

AbstractChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 200 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

New chiral (salen)manganese(III) systems for asymmetric epoxidation of styrene

Highly enantioselelctive and repeatable epoxidation of styrene was performed by using new chiral (salen)Mn(III) catalysts, which were derived from the initial immobilization of a homogeneous (salen)Mn(III) complex on solid carriers and subsequent dispersion into ionic liquids.