Kinetic Resolution Of Racemic Mixtures Research Articles

Kinetic Resolution as a General Approach to Enantioenrichment in Prebiotic Chemistry.

ConspectusThe origin of the single chirality of the chemical building blocks of life remains an intriguing topic of research, even after decades of experimental and theoretical work proposing processes that may break symmetry and induce chiral amplification, a term that may be defined as the enhancement of enantiomeric excess starting from prochiral substrates or from a racemic mixture or a small imbalance between enantiomers. Studies aimed at understanding prebiotically plausible pathways to these molecules have often neglected the issue of chirality, with a focus on the stereochemical direction of these reactions generally being pursued after reaction discovery. Our work has explored how the stereochemical outcome for the synthesis of amino acids and sugars might be guided to rationalize the origin of biological homochirality. The mechanistic interconnection between enantioenrichment in these two groups of molecules provides insights concerning the handedness extant in modern biology. In five separate examples involving the synthesis of life's building blocks, including sugars, RNA precursors, amino acids, and peptides, kinetic resolution emerges as a key protocol for enantioenrichment from racemic molecules directed by chiral source molecules. Several of these examples involve means not only for chiral amplification but also symmetry breaking and chirality transfer across a range of racemic monomer molecules. Several important implications emerge from these studies: one, kinetic resolution of the primordial chiral sugar, glyceraldehyde, plays a key role in a number of different prebiotically plausible reactions; two, the emergence of homochirality in sugars and amino acids is inherently intertwined, with clear synergy between the biological hand of each molecule class; three, the origin story for the homochirality of enzymes and modern metabolism points toward kinetic resolution of racemic amino acids in networks that later evolved to include sophisticated and complete catalytic and co-catalytic cycles; four, a preference for heterochiral ligation forming product molecules that cannot lead to biologically competent polymers can in fact be a driving force for a route to homochiral polymer chains; and five, enantioenrichment in complex mixtures need not be addressed one compound at a time, because kinetic resolution induces symmetry breaking and chirality transfer that may lead to general protocols rather than specific cases tailored to each individual molecule. Such chirality transfer mechanisms perhaps presage strategies utilized in modern biology.Our latest work extends the study of monomer enantioenrichment to the ligation of these molecules into the extended homochiral chains leading to the complex polymers of modern biology. A central theme in all of these reactions is the key role that kinetic resolution of a racemic mixture of amino acids or sugars plays in enabling enantioenrichment under prebiotically plausible conditions. This work has uncovered important trends in symmetry breaking, chirality transfer, and chiral amplification. Kinetic resolution of racemic mixtures emerges as a general solution for chiral amplification in prebiotic chemistry, leading to the single chirality of complex biological molecules and genetic polymers.

Kinetic Resolution of Racemic Mixtures via Enantioselective Photocatalysis.

Despite the increasing demand for enantiopure drugs in the pharmaceutical industry, currently available chiral separation technologies are still lagging behind, whether due to throughput or to operability considerations. This paper presents a new kinetic resolution method, based on the specific adsorption of a target enantiomer onto a molecularly imprinted surface of a photocatalyst and its subsequent degradation through a photocatalytic mechanism. The current model system is composed of an active TiO2 layer, on which the target enantiomer is adsorbed. A photocatalytic suppression layer of Al2O3 is then grown around the adsorbed target molecules by atomic layer deposition. Following the removal of the templating molecules, molecularly imprinted cavities that correspond to the adsorbed species are formed. The stereospecific nature of these pores encourages enantioselective degradation of the undesired species through its enhanced adsorption on the photocatalyst surface, while dampening nonselective photocatalytic activity around the imprinted sites. The method, demonstrated with the dipeptide leucylglycine as a model system, revealed a selectivity factor of up to 7 and an enrichment of a single enantiomer to 85% from an initially racemic mixture. The wide range of parameters that can be optimized (photocatalyst, concentration of imprinted sites, type of passivating layer, etc.) points to the great potential of this method for obtaining enantiomerically pure compounds, beginning from racemic mixtures.

Enzymes for pharmaceutical and therapeutic applications.

Enzymes are highly efficient and selective biocatalysts, present in the living beings. They exist in enormous varieties in terms of the types of reactions catalyzed by them for instance oxidation-reduction, group transfers within the molecules or between the molecules, hydrolysis, isomerization, ligation, bond cleavage, and bond formation. Besides, enzyme based catalyses are performed with much higher fidelity, under mild reaction conditions and are highly efficient in terms of number of steps, giving them an edge over their chemical counter parts. The unique characteristics of enzymes makes them highly applicable fora number of chemical transformation reactions in pharmaceutical industries, such as group protection and deprotection, selective acylation and deacylation, selective hydrolysis, deracemization, kinetic resolution of racemic mixtures, esterification, transesterification, and many others. In this review, an overview of the enzymes, their production and their applications in pharmaceutical syntheses and enzyme therapies are presented with diagrams, reaction schemes and table for easy understanding of the readers.

Making the Silylation of Alcohols Chiral: Asymmetric Protection of Hydroxy Groups.

The non-enzymatic kinetic resolution of racemic mixtures of alcohols by silylation had been unknown before the turn of the century. This stands in stark contrast to established acylation techniques. The same applies to the related desymmetrization of diols. This might come as a surprise, given the significance of silyl ethers as protecting groups in multistep synthesis of complex molecules. The situation changed after a seminal report by Ishikawa nearly twenty years ago. Since then, enantioselective silylation of alcohols has matured and grown into an independent research field with organocatalytic and transition-metal-catalyzed approaches providing powerful solutions. This Minireview summarizes these recent advances with particular emphasis on the stereoselective dehydrogenative coupling of alcohols and hydrosilanes.

Insights on the Origin of Regiodivergence in the Parallel Kinetic Resolution of rac-Aziridines Using a Chiral Lanthanum–Yttrium Bimetallic Catalyst

Parallel kinetic resolution of racemic mixtures is an important method used in asymmetric synthesis of chiral compounds. In a recent example, a rac-cis-2,3-substituted chiral N-benzoyl aziridine was reacted with dimethyl malonate, in the presence of a La–Y heterobimetallic chiral BINAM Schiff base (L) catalyst, to form enantiomerically pure (ee > 98%) γ-amino acid derivatives through a ring-opening reaction in near-quantitative yields from both the enantiomers (∼48%). High regio- and enantioselectivities even with a rac-aziridine, having C2 and C3 substituents as similar as ethyl and n-propyl. Through a comprehensive computational investigation, we delineate the origin of regio-divergent and enantioselective formation of γ-amino ester derivatives. The Gibbs free energy of the transition state for the ring-opening at the propyl substituted C2 carbon leading to 3-benzamidoheptan-4-yl malonate is found to be 7.2 kcal/mol lower than that at the ethyl substituted C3 carbon in the case of (2R,3S)-aziridine. A r...

Fungal synthesis of chiral phosphonic synthetic platform – Scope and limitations of the method

Chiral hydroxyphosphonates due to their wide range of biological properties are industrially important chemicals. Chemical synthesis of their optical isomers is expensive, time consuming and not friendly to the environment, so biotransformations are under consideration. Among others, these compounds act as enzymes inhibitors. This makes the bioconversions of phosphonates, especially scaling experiments, hard to perform. Biocatalysis is one of the methods that can be applied in synthesis of optically pure compounds. To increase the efficiency of the process with whole cell biocatalysts, it is essential to ensure optimal reaction conditions that minimize cellular stress and can enhance the metabolic activity of cells. The present investigation focuses on the scaling up of the kinetic resolution of racemic mixture of 2-butyryloxy-2-(ethoxy-P-phenylphosphinyl)acetic acid, applying free and immobilized form of the fungal biocatalysts and two operation systems: shake flask and recirculated fixed-bed batch reactor. Protocols of effective mycelium immobilization on polyurethane foams were set for T. purpurogenus IAFB 2512, F. oxysporum, P. commune. The best results of biotransformation were obtained with the immobilized P. commune in the column recirculated fixed-bed batch reactor. The conversion reaches 56% (maximal for the kinetic process) and the enantiomeric enrichment of the isomers mixture ranges between 82 and 93% (93% for ester of RP,R conformation). All biocatalysts exhibit SP-preference toward tested compound, what is essential because of importance of the phosphorus atom chirality for its biological activity.

Biocatalyzed kinetic resolution of racemic mixtures of chiral α-aminophosphonic acids

Abstract Several fungal species namely: Aspergillus niger , Aspergillus parasiticus , Penicillium funiculosum , Trigonopsis variabilis and two different strains of Fusarium oxysporum were tested toward racemic mixtures of following phosphonic acids: 1-amino-2-methylpropanephosphonic acid, 1-aminophenylmethanephosphonic acid and 1-amino-2-phenylethanephosphonic acid. Application of F. oxysporum strain (UW1) allowed obtaining optically pure S -isomer of 1-aminophenylmethanephosphonic acid and R -isomer of 1-amino-2-phenylethanephosphonic acid, whereas biotransformation of racemic mixture of 1-amino-2-methylpropanephosphonic acid with F. oxysporum strain (DSM 12646) cells resulted in obtaining pure enantiomer of R -configuration.

Mutational and Structural Analysis ofl-N-Carbamoylase Reveals New Insights into a Peptidase M20/M25/M40 Family Member

N-Carbamoyl-L-amino acid amidohydrolases (L-carbamoylases) are important industrial enzymes used in kinetic resolution of racemic mixtures of N-carbamoyl-amino acids due to their strict enantiospecificity. In this work, we report the first L-carbamoylase structure belonging to Geobacillus stearothermophilus CECT43 (BsLcar), at a resolution of 2.7 Å. Structural analysis of BsLcar and several members of the peptidase M20/M25/M40 family confirmed the expected conserved residues at the active site in this family, and site-directed mutagenesis revealed their relevance to substrate binding. We also found an unexpectedly conserved arginine residue (Arg(234) in BsLcar), proven to be critical for dimerization of the enzyme. The mutation of this sole residue resulted in a total loss of activity and prevented the formation of the dimer in BsLcar. Comparative studies revealed that the dimerization domain of the peptidase M20/M25/M40 family is a "small-molecule binding domain," allowing further evolutionary considerations for this enzyme family.

Quantitative analysis of enzymatic fractionation of multiple substrate mixtures

The enzymatic conversion of mixtures of multiple substrates was studied quantitatively, based on established methodology used for the enzymatic kinetic resolution of racemic mixtures, involving the use of competitive factors: ratios of specificity constants (k(cat)/K(M)) of substrate pairs. The competitive factors of the substrates were defined in relation to a reference substrate. These competitive factors were used to predict the composition of the reaction mixture as a function of the degree of conversion of the reaction. The methodology was evaluated using three different lipases to hydrolyze a model mixture of four fatty acid methyl esters and for the esterification of a mixture of the same fatty acids in free form with ethanol. In most cases, the competitive factors determined from the initial phase of the reactions predicted the product composition during the rest of the reaction very well. The slowest reacting fatty acid was erucic acid (both in free form and as methyl ester), which was thus enriched in the remaining substrate fraction, while the other fatty acids: lauric acid, palmitic acid and oleic acid were converted faster. Simulations of the compositions of reaction mixtures with different values of the competitive factors were carried out to provide an overview of what could be achieved using enzymatic enrichment. Possible applications include reactions involving homologous substrates and mixtures of multiple isomers. The analysis presented provides guidelines that can be useful in the screening and development of enzymes for enzymatic enrichment applications.

Kinetic resolution of racemic mixtures in multiphase enzyme membrane reactors: Influence of ester type and oil/water interface on process performance

Kinetic resolution of racemic mixtures in multiphase enzyme membrane reactors: Influence of ester type and oil/water interface on process performance

Dynamic kinetic resolution of racemic tropic acid ethyl ester and its derivatives

The dynamic kinetic resolution of racemic mixtures of tropic acid ethyl ester under substrate racemizing conditions was studied using lipase PS with a ruthenium catalyst. Isopropenyl acetate was used as an acyl donor, since it was found to be compatible with both catalysts; this resulted in an efficient dynamic kinetic resolution. With this process, a variety of racemic tropic acid ethyl esters were transformed to optically active acetoxy-2-arylpropionic acid ethyl esters with 60–88% yields and 53–92% ee.

Influence of [sbnd]OR ester group length on the catalytic activity and enantioselectivity of free lipase and immobilized in membrane used for the kinetic resolution of naproxen esters

Lipases are suitable catalysts for the kinetic resolution of racemic mixtures due to their ability to discriminate between enantiomers. For this reason, they have been studied largely to develop reactors for the production of optically pure enantiomers. The main problem in these productive systems is enzyme stability and enantiocatalytic selectivity as a function of time. In this work, the enantiocatalytic properties of lipase as a function of the OR group length were studied. The methyl, butyl, and octyl esters of naproxen were synthesized and used as reagents. The lipase was used as a free agent and immobilized in a polymeric membrane reactor. The results show that selectivity and stability of the enzyme improved while catalytic activity decreased with the OR length group. The immobilized enzyme had higher activity compared with the free enzyme.

Conformational rigidity of silicon-stereogenic silanes in asymmetric catalysis: A comparative study.

In recent years, cyclic silicon-stereogenic silanes were successfully employed as stereoinducers in transition metal-catalyzed asymmetric transformations as exemplified by (1) the hydrosilylation of alkenes constituting a chirality transfer from silicon to carbon and (2) the kinetic resolution of racemic mixtures of alcohols by dehydrogenative silicon-oxygen coupling. In this investigation, a cyclic and a structurally related acyclic silane with silicon-centered chirality were compared using the above-mentioned model reactions. The stereochemical outcome of these pairs of reactions was correlated with and rationalized by the current mechanistic pictures. An acyclic silicon-stereogenic silane is also capable of inducing excellent chirality transfer (ct) in a palladium-catalyzed intermolecular carbon-silicon bond formation yet silicon incorporated into a cyclic framework is required in the copper-catalyzed silicon-oxygen bond forming reaction.

Bio-resolution of a chiral epoxide using whole cells of Bacillus megaterium ECU1001 in a biphasic system

Optically active epoxides can be prepared by kinetic resolution of racemic mixtures using stereospecific epoxide hydrolases. To increase the bio-resolution efficiency of a sparingly water-soluble epoxide (glycidyl phenyl ether, GPE), we investigated the use of organic/aqueous two-phase system. Various conditions were systematically examined and optimized in shake flasks. Isooctane was found to be the most suitable solvent as the organic phase. The phase volume ratio ( ϕ o/w) and biocatalyst concentration were shown to be sensitive parameters affecting both the reaction rate and the enzyme enantiospecificity in the biphase system. An isooctane/aqueous system was developed to overcome the low solubility and instability of GPE in the aqueous phase, resulting in a significant improvement of enatiomeric ratio ( E-value) from 39.5 to 94.0 and an average productivity of 18.8 mg GPE/(h g) biocatalyst to 48.9 mg GPE/(h g) biocatalyst, respectively. Resolution of a 90.1 g/l solution of racemic glycidyl phenyl ether in isooctane phase was successfully carried out in a mechanically stirred reactor (120 ml), affording ( S)-glycidyl phenyl ether in high (100%) enantiomeric excess with a yield of 44.5%.

Enzymatic resolution of chiral phosphinate esters.

The bacterial phosphotriesterase has been shown to catalyze the stereoselective hydrolysis of phosphinate esters. The wild-type enzyme preferentially hydrolyzes the SP-enantiomers of methyl phenyl p-X-phenylphosphinate esters by 3 orders of magnitude. The mutant enzyme, I106T/F132A/H254G/H257W, exhibits the opposite stereoselectivity and hydrolyzes the RP-enantiomer up to 30 times faster than the corresponding SP-enantiomer. The enantiomerically pure phosphinate esters, prepared from the kinetic resolution of racemic mixtures, can serve as the entry point for the chemoenzymatic preparation of P-chiral phosphines and phosphine oxides.

Crystal Structure of Escherichia coli Thioesterase I/Protease I/Lysophospholipase L 1: Consensus Sequence Blocks Constitute the Catalytic Center of SGNH-hydrolases through a Conserved Hydrogen Bond Network

Escherichia coli thioesterase I (TAP) is a multifunctional enzyme possessing activities of thioesterase, esterase, arylesterase, protease, and lysophospholipase. In particular, TAP has stereoselectivity for amino acid derivative substrates, hence it is useful for the kinetic resolution of racemic mixtures of industrial chemicals. In the present work, the crystal structure of native TAP was determined at 1.9 Å, revealing a minimal SGNH-hydrolase fold. The structure of TAP in complex with a diethyl phosphono moiety (DEP) identified its catalytic triad, Ser10-Asp154-His157, and oxyanion hole, Ser10-Gly44-Asn73. The oxyanion hole of TAP consists of three residues each separated from the other by more than 3.5 Å, implying that all of them are highly polarized when substrate bound. The catalytic (His)C ε1–H⋯OC hydrogen bond usually plays a role in the catalytic mechanisms of most serine hydrolases, however, there were none present in SGNH-hydrolases. We propose that the existence of the highly polarized tri-residue-constituted oxyanion hole compensates for the lack of a (His)C ε1–H⋯OC hydrogen bond. This suggests that members of the SGNH-hydrolase family may employ a unique catalytic mechanism. In addition, most SGNH-hydrolases have low sequence identities and presently there is no clear criterion to define consensus sequence blocks. Through comparison of TAP and the three SGNH-hydrolase structures currently known, we have identified a unique hydrogen bond network which stabilizes the catalytic center: a newly discovered structural feature of SGNH-hydrolases. We have defined these consensus sequence blocks providing a basis for the sub-classification of SGNH-hydrolases.

Parallel Kinetic Resolution of Racemic Mixtures: A New Strategy for the Preparation of Enantiopure Compounds?

AbstractFor Abstract see ChemInform Abstract in Full Text.

Lipase‐Catalyzed Acetylation of tert‐Butyl N‐(2‐Hydroxy‐3‐phenoxypropoxy)carbamates — Preparation of Optically Active Aminooxypropanols.

AbstractFor Abstract see ChemInform Abstract in Full Text.

Parallel kinetic resolution of racemic mixtures: a new strategy for the preparation of enantiopure compounds?

Kinetic resolution of racemic mixtures is a well-established methodology for the preparation of optically active compounds. However, excellent enantioselectivities are required to obtain them in enantiopure form, due to the decrease in ee when conversion values are close to 50%. To overcome this limitation, a parallel (asymmetric) reaction can remove the disfavored enantiomer. In this review, several examples of this strategy showing its wide range of applicability are described, as well as their mathematical treatment and some new applications in combinatorial chemistry.

LIPASE-CATALYZED ACETYLATION OF tert-BUTYL N-(2-HYDROXY-3-PHENOXYPROPOXY)CARBAMATES—PREPARATION OF OPTICALLY ACTIVE AMINOOXYPROPANOLS

ABSTRACT A simple method was developed for the preparation of optically active-3-aryloxy-1-aminooxy-2-propanols by a lipase-catalyzed kinetic resolution of racemic mixtures of their N-tert-butylcarbamate-protected derivatives.