Chemoenzymatic Synthesis of Glutamic Acid Analogues:  Substrate Specificity and Synthetic Applications of Branched Chain Aminotransferase from <i>Escherichia coli</i>

Xian, Mo; Alaux, Sébastien; Sagot, Emmanuelle; Gefflaut, Thierry

doi:10.1021/jo070805q

Cited by 41 publications

(21 citation statements)

References 28 publications

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“…Such information would be of importance for evaluating process technology options [10]. Earlier studies [11][12][13][14][15] had alerted us to the potential of BCAT as a biocatalyst for production of high-value non-natural amino acids, such as L-tert-leucine and L-3-hydroxyadamantylglycine. Therefore, the overall substrate specificity and kinetic properties of E. coli BCAT have been revisited using the novel coupled assay, which delivers reliable initial-rate measurements.…”

Section: Introductionmentioning

confidence: 99%

The specificity and kinetic mechanism of branched‐chain amino acid aminotransferase from Escherichia coli studied with a new improved coupled assay procedure and the enzyme's potential for biocatalysis

Wang

Engel

2013

The FEBS Journal

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Branched-chain amino acid aminotransferase (BCAT) plays a key role in the biosynthesis of hydrophobic amino acids (such as leucine, isoleucine and valine), and its substrate spectrum has not been fully explored or exploited owing to the inescapable restrictions of previous assays, which were mainly based on following the formation/consumption of the specific branched-chain substrates rather than the common amino group donor/ acceptor. In our study, detailed measurements were made using a novel coupled assay, employing (R)-hydroxyglutarate dehydrogenase from Acidaminococcus fermentans as an auxiliary enzyme, to provide accurate and reliable kinetic constants. We show that Escherichia coli BCAT can be used for asymmetric synthesis of a range of non-natural amino acids such as L-norleucine, L-norvaline and L-neopentylglycine and compare the kinetic results with the results of molecular modelling. A full two-substrate steadystate kinetic study for several substrates yields results consistent with a bibi ping-pong mechanism, and detailed analysis of the kinetic constants indicates that, for good 2-oxoacid substrates, release of 2-oxoglutarate is much slower than release of the product amino acid during the transamination reaction. The latter is in fact rate-limiting under conditions of substrate saturation. DatabaseBranched-chain amino acid aminotransferase EC 2.6.1.42; (R)-2-hydroxyglutarate dehydrogenase EC 1.1.99.2

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Section: Introductionmentioning

confidence: 99%

The specificity and kinetic mechanism of branched‐chain amino acid aminotransferase from Escherichia coli studied with a new improved coupled assay procedure and the enzyme's potential for biocatalysis

Wang

Engel

2013

The FEBS Journal

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“…Enantioselective acetoacetate additions to acyclic α,β-unsaturated carbonyls await discovery, but the easily accessible adducts (±)- 8a 15 and (±)- 8b 16 provide their derived α-keto esters (±)- 9a and (±)- 9b in the presence of Cu(II)/air (Scheme 3). The utility of these racemic products in subsequent stereodynamic processes is under investigation.…”

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confidence: 99%

Asymmetric Synthesis of α-Keto Esters via Cu(II)-Catalyzed Aerobic Deacylation of Acetoacetate Alkylation Products: An Unusually Simple Synthetic Equivalent to the Glyoxylate Anion Synthon

Steward

Johnson

2011

Org. Lett.

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A simple and efficient method for the preparation of β-stereogenic α-keto esters is described using a copper(II)-catalyzed aerobic deacylation of substituted acetoacetate esters. The substrates for the title process arise from catalytic, enantioselective conjugate additions and alkylation reactions of acetoacetate esters. The mild conditions do not induce racemization of the incipient enolizable α-keto ester. The reaction is tolerant of esters, certain ketones, ketals, and nitro groups and utilizes inexpensive, readily available materials.

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“…In this case the amino acid racemase is of course omitted [Scheme 14 (b)]. [67][68][69] Interestingly, amino acid dehydrogenases and atransaminases have also been coupled in a linear way, realizing a system for deracemization of amino acids. [70] Branched-chain amino acid transaminase (BCAAT) from Sinorhizobium meliloti ATCC 51124 was cloned and overexpressed in E. coli.…”

Section: Multi-enzymatic Synthesis Of Amines and Amino Acids Employinmentioning

confidence: 99%

Multi‐Enzymatic Cascade Reactions: Overview and Perspectives

Brucher²,

2011

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Multi-enzymatic cascade reactions, i.e., the combination of several enzymatic transformations in concurrent one-pot processes, offer considerable advantages: the demand of time, costs and chemicals for product recovery may be reduced, reversible reactions can be driven to completion and the concentration of harmful or unstable compounds can be kept to a minimum. This review summarizes the developments in multi-enzymatic cascades employed for the asymmetric synthesis of chiral alcohols, amines and amino acids, as well as for C À C bond formation. In addition, a general classification of biocatalytic cascade systems is provided and bioprocess engineering aspects associated with the topic are discussed.

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Chemoenzymatic Synthesis of Glutamic Acid Analogues: Substrate Specificity and Synthetic Applications of Branched Chain Aminotransferase from Escherichia coli

Cited by 41 publications

References 28 publications

The specificity and kinetic mechanism of branched‐chain amino acid aminotransferase from Escherichia coli studied with a new improved coupled assay procedure and the enzyme's potential for biocatalysis

The specificity and kinetic mechanism of branched‐chain amino acid aminotransferase from Escherichia coli studied with a new improved coupled assay procedure and the enzyme's potential for biocatalysis

Asymmetric Synthesis of α-Keto Esters via Cu(II)-Catalyzed Aerobic Deacylation of Acetoacetate Alkylation Products: An Unusually Simple Synthetic Equivalent to the Glyoxylate Anion Synthon

Multi‐Enzymatic Cascade Reactions: Overview and Perspectives

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