Lipase catalysed Michael addition of secondary amines to acrylonitrile

Torre, Oliver; Alfonso, Ignacio; Gotor, Vicente

doi:10.1039/b402244k

Cited by 164 publications

(67 citation statements)

References 8 publications

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“…This proton helps to direct the ester oxygen atom to His 224 by providing a hydrogen-bonding interaction. Of course, the Michael addition reaction can proceed in the wild-type enzyme, as shown in previous reports, [12][13][14][15][16][17][18][19][20] but in the presence of an ester group and water, native hydrolysis is favored. As the nucleophilic Ser 105 residue is removed, the hydrolysis reaction is suppressed and the promiscuous Michael addition activity is revealed.…”

Section: Introductionmentioning

confidence: 71%

Suppressed Native Hydrolytic Activity of a Lipase to Reveal Promiscuous Michael Addition Activity in Water

et al. 2009

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Suppression of the native hydrolytic activity of Pseudozyma antarctica lipase B (PalB) (formerly Candida antarctica lipase B) in water is demonstrated. By replacing the catalytic Ser 105 residue with an alanine unit, promiscuous Michael addition activity is favored. A Michael addition reaction between methyl acrylate and acetylacetone was explored as a model system. For the PalB Ser 105 Ala mutant, the hydrolytic activity was suppressed more than 1000 times and, at the same time, the Michael addition activity was increased by a factor of 100. Docking studies and molecular dynamics simulations revealed an increased ability of the PalB Ser 105 Ala mutant to harbor the substrates close to a catalytically competent conformation.

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Section: Introductionmentioning

confidence: 71%

Suppressed Native Hydrolytic Activity of a Lipase to Reveal Promiscuous Michael Addition Activity in Water

et al. 2009

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“…the ability of an enzyme to exhibit catalysis of other than its native reaction) towards e.g. aldol condensations, [19,20] epoxidations [21] and Michael additions, [22,23,24,25,26] not seldom enhanced by point mutations. [27] An often-seen feature in promiscuous catalysis is that only parts of the catalytic machinery are used, so that removal of one or more of For Production's Reference Only Click here to download Manuscript: Brinck_2010-DielsAlder_revised.pdf 2 …”

Section: Introductionmentioning

confidence: 99%

Computational design of a lipase for catalysis of the Diels-Alder reaction

et al. 2010

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Combined molecular docking, molecular dynamics (MD) and density functional theory (DFT) studies have been employed to study catalysis of the Diels-Alder reaction by a modified lipase. Six variants of the versatile enzyme {\em Candida Antarctica} lipase B (CALB) have been rationally engineered {\em in silico} based on the specific characteristics of the pericyclic addition. A kinetic analysis reveals that hydrogen bond stabilization of the transition state and substrate binding are key components of the catalytic process. In the case of substrate binding, which has the greater potential for optimization, both binding strength and positioning of the substrates are important for catalytic efficiency. The binding strength is determined by hydrophobic interactions and can be tuned by careful selection of solvent and substrates. The MD simulations show that substrate positioning is sensitive to cavity shape and size, and can be controlled by a few rational mutations. The well-documented S105A mutation is essential to enable sufficient space in the vicinity of the oxyanion hole. Moreover, bulky residues on the edge of the active site hinders the formation of a sandwich-like near-attack conformer (NAC), and the I189A mutation is needed to obtain enough space above the face of the $\alpha$,$\beta$-double bond on the dienophile. The double mutant S105A/I189A performs quite well for two of three dienophiles. Based on binding constants and NAC energies obtained from MD simulations combined with activation energies from DFT computations, relative catalytic rates ($v_{cat}/v_{uncat}$) of up to $10^3$ are predicted.Response to Reviewers: We are happy with the favorable comments by the reviewers. We have have corrected the manuscript as suggested by reviewer 1 1. A sentence explaining catalytic promiscuity has been added. 2. A paragraph discussing the relationship between the NAC concept and the Reaction Force has been added.We have shortened the manuscript as suggested by reviewer 2. In particular, we have moved information regarding the structural relaxation of protein structure in MD simulations to the supplementary material. Abstract Combined molecular docking, molecular dynamics (MD) and density functional theory (DFT) studies have been employed to study catalysis of the DielsAlder reaction by a modified lipase. Six variants of the versatile enzyme Candida Antarctica lipase B (CALB) have been rationally engineered in silico based on the specific characteristics of the pericyclic addition. A kinetic analysis reveals that hydrogen bond stabilization of the transition state and substrate binding are key components of the catalytic process. In the case of substrate binding, which has the greater potential for optimization, both binding strength and positioning of the substrates are important for catalytic efficiency. The binding strength is determined by hydrophobic interactions and can be tuned by careful selection of solvent and substrates. The MD simulations show that substrate positioning is sensitive to cavity s...

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“…The conversion to give an optically active product was ~60% in 144 hours with vinyl acetate. Earlier, Torre et al, (2004) [79] reported Michael addition of secondary amines to acrylonitrile in a solvent free system using different preparations of CAL B. Both Wu et al, (2005) [78] and Torre et al, (2004) [79] discussed the possible mechanism for the doi: 10.7243/2053-7670-1-1 enzyme catalyzed reaction which does involve active site of the enzymes.…”

Section: Catalytic Promiscuity In Nearly Anhydrous Organic Solventsmentioning

confidence: 99%

“…Earlier, Torre et al, (2004) [79] reported Michael addition of secondary amines to acrylonitrile in a solvent free system using different preparations of CAL B. Both Wu et al, (2005) [78] and Torre et al, (2004) [79] discussed the possible mechanism for the doi: 10.7243/2053-7670-1-1 enzyme catalyzed reaction which does involve active site of the enzymes. Lou et al, (2008) [80] showed that selectivity between Markonikov addition and anti-Markonikov addition of thiols to vinyl esters catalyzed by CAL B can be altered by changing the reaction medium.…”

Section: Catalytic Promiscuity In Nearly Anhydrous Organic Solventsmentioning

confidence: 99%

Use of high activity enzyme preparations in neat organic solvents for organic synthesis

Gupta¹,

Mukherjee²,

Malhotra³

2013

Univers Org Chem

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The use of enzymes in nearly anhydrous organic solvents generally results in low initial rates/percentage conversions. The current review focuses on some biocatalyst designs like immobilization on nanomaterials, enzyme precipitated and rinsed with organic solvents (EPROS), crosslinked enzyme crystals (CLEC), crosslinked enzyme aggregates (CLEA), protein coated microcrystals (PCMC) and crosslinked protein coated microcrystals (CLPCMC) which show much better catalytic efficiency in such media as compared to other kinds of biocatalyst preparations. The basic methodology and principle behind these efficient designs are described. The relatively recent results on catalytic promiscuity as seen in low water media have also been covered. It is hoped that this will further encourage wider applications of enzymes in neat organic solvents in organic synthesis.

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Lipase catalysed Michael addition of secondary amines to acrylonitrile

Abstract: A new enzymatic process is described. Different preparations of lipase B from Candida antarctica are able to catalyse Michaeltype addition of secondary amines to acrylonitrile. This new reaction widens the applicability of these biocatalysts in organic synthesis.

Cited by 164 publications

References 8 publications

Suppressed Native Hydrolytic Activity of a Lipase to Reveal Promiscuous Michael Addition Activity in Water

Suppressed Native Hydrolytic Activity of a Lipase to Reveal Promiscuous Michael Addition Activity in Water

Computational design of a lipase for catalysis of the Diels-Alder reaction

Use of high activity enzyme preparations in neat organic solvents for organic synthesis

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