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Manipulating the stereoselectivity of limonene epoxide hydrolase by directed evolution based on iterative saturation mutagenesis

Limonene epoxide hydrolase from Rhodococcus erythropolis DCL 14 (LEH) is known to be an exceptional epoxide hydrolase (EH) because it has an unusual secondary structure and catalyzes the hydrolysis of epoxides by a rare one-step mechanism in contrast to the usual two-step sequence. From a synthetic organic viewpoint it is unfortunate that LEH shows acceptable stereoselectivity essentially only in the hydrolysis of the natural substrate limonene epoxide, which means that this EH cannot be exploited as a catalyst in asymmetric transformations of other substrates. In the present study, directed evolution using iterative saturation mutagenesis (ISM) has been tested as a means to engineer LEH mutants showing broad substrate scope with high stereoselectivity. By grouping individual residues aligning the binding pocket correctly into randomization sites and performing saturation mutagenesis iteratively using a reduced amino acid alphabet, mutants were obtained which catalyze the desymmetrization of cyclopentene-oxide with stereoselective formation of either the (R,R)- or the (S,S)-diol on an optional basis. The mutants prove to be excellent catalysts for the desymmetrization of other meso-epoxides and for the hydrolytic kinetic resolution of racemic substrates, without performing new mutagenesis experiments. Since less than 5000 tranformants had to be screened for achieving these results, this study contributes to the generalization of ISM as a fast and reliable method for protein engineering. In order to explain some of the stereoselective consequences of the observed mutations, a simple model based on molecular dynamics simulations has been proposed.

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Reference£º
Synthesis and Crystal Structure of a Chiral?C3-Symmetric Oxygen Tripodal Ligand and Its Applications to Asymmetric Catalysis,
Chiral lanthanide(III) complexes of sulphur¨Cnitrogen¨Coxygen ligand derived from aminothiourea and sodium?D-camphor-¦Â-sulfonate

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Palladium allylic complexes with enantiopure bis(diamidophosphite) ligands bearing a cyclohexane-1,2-diamine skeleton as catalysts in the allylic substitution reaction

A series of cationic allyl palladium complexes [Pd(eta3-CH3-C3H5)(P-P)]X (X = PF6, 2a-c, 2e; and X = BPh4, 3a, 3b, 3d, 3e) and [Pd(eta3-1,3-Ph2-C3H3)(P-P)]X (X = PF6, 6b; and X = BPh4, 7a) have been prepared. The bis(diamidophosphite) ligands (P-P) contain a diazaphospholidine terminal fragment derived from (R,R)- and (S,S)-N,N?-dibenzyl- and (R,R)-N,N?-dimethyl-cyclohexane-1,2-diamines and dialcoxy bridging fragment derived from (R,R)- and (S,S)-butanediol, (R,R)-cyclohexanediol, (4R,5R)- and (4S,5S)-4,5-di(hydroxymethyl)-2,2-dimethyl-1,3-dioxolane and (R)- and (S)-binaphthol. Complexes [Pd(eta3-CH3-C3H5l)P2]X (X = PF6, 4f, 4g; and X = BPh4, 5f), where P are monodentate diamidophosphite ligands with diazaphospholidine heterocyclic backbone obtained from (R,R)- and (S,S)-N,N?-dibenzylcyclohexane-1,2-diamine and alcoxy groups coming from (R)-phenyl-ethanol and (S)-borneol have been also prepared. Neutral palladium complexes [PdCl2(P-P)] (1a, 1c) were synthesized to prove the C2symmetry of the P-P ligand. The new compounds were fully characterized in solution by NMR spectroscopy. The X-ray crystal structure determination for 2e-(R,R,Ral,Ral;R,R) and 1a-(S,S;Sal,Sal;S,S) has been achieved. The new allyl-palladium complexes were applied in the asymmetric allylic substitution reaction of the benchmark substrate rac-3-acetoxy-1,3-diphenyl-1-propene with dimethyl malonate and benzylamine as nucleophiles in order to test their catalytic potential. The best results were obtained with the 3a-(R,R;Ral,Ral;R,R) precursor (up to 84% ee) while complexes with the e ligand derived from the (R,R)-N,N?-dimethylcyclohexane-1,2-diamine terminal fragment resulted inactive in the process. The influence of the nature and the absolute configuration of both the bridging and the terminal fragments of the bis(diamidophosphite) ligand on the asymmetric induction is discussed. A preliminary study of the anion effect (PF6?vs. BPh4-) on the activity and the enantioselectivity of the Pd-catalysed allylic substitution has also been performed.

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Reference£º
Synthesis and Crystal Structure of a Chiral?C3-Symmetric Oxygen Tripodal Ligand and Its Applications to Asymmetric Catalysis,
Chiral lanthanide(III) complexes of sulphur¨Cnitrogen¨Coxygen ligand derived from aminothiourea and sodium?D-camphor-¦Â-sulfonate

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Indolyl-oxazaphosphorine Precursors for Stereoselective Synthesis of Phosphite Triesters and Dithymidinyl Phosphorothioates

Several novel chiral indolyl-oxazaphosphorines 7 were synthesized, and their potential as precursors to chiral phosphorothioates was evaluated. Reaction of 7 with a thymidine derivative gave phosphite triester 8 with a large degree of stereoselectivity. Sulfurization with Beaucage’s reagent provided phosphorothioate triesters 9. The chiral auxiliary 9b containing a cyano group could be easily removed with aqueous ammonia to form dithymidinyl phosphorothioate in more than 97% diastereomeric excess. The chiral indolyl-oxazaphosphorines 7 are a new class of precursors for stereoselective synthesis of phosphorothioates.

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Reference£º
Synthesis and Crystal Structure of a Chiral?C3-Symmetric Oxygen Tripodal Ligand and Its Applications to Asymmetric Catalysis,
Chiral lanthanide(III) complexes of sulphur¨Cnitrogen¨Coxygen ligand derived from aminothiourea and sodium?D-camphor-¦Â-sulfonate

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The result showed that such a combination of chemo- and biocatalysis improved the catalytic yield more than two times compared with that of sole metal catalysis.HPLC of Formula: C4H10O2. I hope my blog about 19132-06-0 is helpful to your research.

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Preparation of Optically Active 1,2-Diols and alpha-Hydroxy Ketones Using Glycerol Dehydrogenase as Catalyst: Limits to Enzyme-Catalyzed Synthesis due to Noncompetitive and Mixed Inhibition by Product

Glycerol dehydrogenase (GDH, EC 1.1.1.6, from Enterobacter aerogenes or Cellulomonas sp.) catalyzes the interconversion of analogues of glycerol and dihydroxyacetone.Its substrate specificity is quite different from than of horse liver alcohol dehydrogenase (HLADH), yeast alcohol dehydrogenase, and other alcohol dehydrogenases used in enzyme-catalyzed organic synthesis and is thus a useful new enzymic catalyst for the synthesis of enantiomerically enriched and isotopically labeled organic molecules.This paper illustrates synthetic applications of GDH as a reduction catalyst by the enantioselective reduction of 1-hydroxy-2-propanone and 1-hydroxy-2-butanone to the corresponding R 1,2-diols (ee = 95-98percent). (R)-1,2-Butanediol-2-d1 was prepared by using formate-d1 as the ultimate reducing agent.Comparison of (R)-1,2-butanediol prepared by reduction of 1-hydroxy-2-butanone enzymatically and with actively fermenting bakers’ yeast indicated than yield and enantiomeric purity were similar by the two procedures.Reactions proceeding in the direction of substrate oxidation usually suffer from slow rates and incomplete conversions due to product inhibition.The kinetic consequences of product inhibition (competitive, noncompetitive, and mixed) for practical synthetic applications of GDH, HLADH, and other oxidoreductases are analyzed.In general, product inhibition seems the most serious limitation to the use of these enzymes as oxidation catalysts in organic synthesis.

The result showed that such a combination of chemo- and biocatalysis improved the catalytic yield more than two times compared with that of sole metal catalysis.HPLC of Formula: C4H10O2. I hope my blog about 19132-06-0 is helpful to your research.

Reference£º
Synthesis and Crystal Structure of a Chiral?C3-Symmetric Oxygen Tripodal Ligand and Its Applications to Asymmetric Catalysis,
Chiral lanthanide(III) complexes of sulphur¨Cnitrogen¨Coxygen ligand derived from aminothiourea and sodium?D-camphor-¦Â-sulfonate

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Enthalpy of vaporisation of butanediol isomers

The enthalpies of vaporisation of isomers of butanediol were determined by calorimetric measurements. A Knudsen effusion cell was used for this purpose. The values of the standard enthalpy of vaporisation obtained for the different isomers were compared and significant differences were found between them.

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Reference£º
Synthesis and Crystal Structure of a Chiral?C3-Symmetric Oxygen Tripodal Ligand and Its Applications to Asymmetric Catalysis,
Chiral lanthanide(III) complexes of sulphur¨Cnitrogen¨Coxygen ligand derived from aminothiourea and sodium?D-camphor-¦Â-sulfonate

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X=Y-ZH Systems as potential 1,3-dipoles. Part 38. 1,5-Electrocyclisation of vinyl-and iminyl-azomethine ylides. 2-Azaindolizines and pyrrolo-dihydro-isoquinolines

Azomethine ylides generated by the decarboxylation of imines of a- amino acids and 2,2?- dipyridyl ketone undergo 1,5-electrocyclisation and subsequent aromatisation to generate 1,3- disubstituted-2-azaindolizines. Azomethine ylides generated from 1,2,3,4- tetrahydroisoquinoline and diarylidene acetone undergo 1,5-electrocyclisation and subsequent prototropic rearrangement, to give pyrrolo-5,6-dihydroisoquinolines.

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Reference£º
Synthesis and Crystal Structure of a Chiral?C3-Symmetric Oxygen Tripodal Ligand and Its Applications to Asymmetric Catalysis,
Chiral lanthanide(III) complexes of sulphur¨Cnitrogen¨Coxygen ligand derived from aminothiourea and sodium?D-camphor-¦Â-sulfonate

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Alkali Metals Dissolved in Optically Active Solvents

Alkali metals dissolve in amines and ethers to give visible and infrared absorption bands.The visible band is believed to be due to absorption by the alkali metal anion and the infrared band to absorption of light by the solvated electrons.The visible and circular dichroism spectra of sodium-potassium alloy dissolved in several optically active ethers and amines were examined.In all cases, no circular dichroism could be detected corresponding to the alkali anion transition.An excellent synthetic route for the formation of a variety of chiral polyethers and cyclic polyethers is described.The method uses the readily available, chiral ethyl (S)-(+)-lactate.

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Reference£º
Synthesis and Crystal Structure of a Chiral?C3-Symmetric Oxygen Tripodal Ligand and Its Applications to Asymmetric Catalysis,
Chiral lanthanide(III) complexes of sulphur¨Cnitrogen¨Coxygen ligand derived from aminothiourea and sodium?D-camphor-¦Â-sulfonate

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Lewis pairs for ring-opening alternating copolymerization of cyclic anhydrides and epoxides

A simple and highly active catalytic process for ring-opening alternating copolymerization (ROAC) of cyclic anhydrides and epoxides still remains a key challenge. Herein, we have described an effective group of versatile and low-toxic zinc dicarbyl/amine Lewis pairs for the ROAC. The facile route showed a high catalytic activity (TOF ? 210 h-1 at 110 C) and perfectly alternating selectivity (>99%). An unexpected highly regioselective ring-opening of asymmetric epoxides (PO, ECH and SO) was also achieved by the combination of zinc alkyls (or aryls) and amines. Of note, deprotonation side reaction of alpha-H of anhydrides with organic bases was uncovered, and subsequently was inhibited by using nonpolar solvents and Lewis acid/base pairs. Thus, an array of polyesters was synthesized by the coupling of various anhydrides (PA, CHA, SA and NA) and epoxides (CHO, PO, ECH and SO) using the same Lewis pairs. Furthermore, variable temperature 1H NMR spectral and MALDI TOF MS analyses were performed to understand the possible mechanism and microstructure. The experimental results indicated that zwitterionic alkoxide and carboxylate intermediates alternately formed to enhance the ester repeat units in chain initiation and propagation. This work provides a simple and green catalytic strategy to prepare diversified polyesters from the ROAC process of cyclic anhydrides and epoxides with considerable catalytic activity and alternating selectivity.

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Reference£º
Synthesis and Crystal Structure of a Chiral?C3-Symmetric Oxygen Tripodal Ligand and Its Applications to Asymmetric Catalysis,
Chiral lanthanide(III) complexes of sulphur¨Cnitrogen¨Coxygen ligand derived from aminothiourea and sodium?D-camphor-¦Â-sulfonate

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PRODRUG COMPOUNDS USEFUL AS CANNABINOID LIGANDS

The present invention provides for compounds of formula (I) wherein A2, L2, R1g, R2A, R3A, R4A, R1a, R1b, q1, and z are as defined in the specification, are prodrugs of CB2 receptors ligands and as such are useful in the prevention and treatment of various diseases and conditions including, but not limited to, pain.

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Reference£º
Synthesis and Crystal Structure of a Chiral?C3-Symmetric Oxygen Tripodal Ligand and Its Applications to Asymmetric Catalysis,
Chiral lanthanide(III) complexes of sulphur¨Cnitrogen¨Coxygen ligand derived from aminothiourea and sodium?D-camphor-¦Â-sulfonate

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PHARMACEUTICAL FORMULATION CONTAINING AN SGLT2 INHIBITOR

Pharmaceutical formulations are provided which are in the form of capsules or tablets for oral use and which include a medicament dapagliflozin or its propylene glycol hydrate and a pharmaceutical acceptable carrier therefor, which formulation is designed for immediate release.

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Reference£º
Synthesis and Crystal Structure of a Chiral?C3-Symmetric Oxygen Tripodal Ligand and Its Applications to Asymmetric Catalysis,
Chiral lanthanide(III) complexes of sulphur¨Cnitrogen¨Coxygen ligand derived from aminothiourea and sodium?D-camphor-¦Â-sulfonate