Enzyme‐Catalyzed Aldol Additions

Clapés, Pere; Joglar, Jesús

doi:10.1002/9783527656714.ch8

Cited by 13 publications

(4 citation statements)

References 154 publications

(197 reference statements)

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“…However, such enzymes easily become inactivated by nonspecific reactions involving the essential lysine residue in the presence of strong electrophiles, such as formaldehyde, even at low concentrations . In contrast, class II pyruvate aldolases utilize a divalent metal ion as an essential cofactor to promote the enolization of the pyruvate nucleophile and do not possess a sensitive active-site lysine and thus appear potentially more suitable for the target reaction . Among them, YfaU (EC 4.1.2.53) was selected from the genome of E.…”

Section: Resultsmentioning

confidence: 99%

Combining Aldolases and Transaminases for the Synthesis of 2-Amino-4-hydroxybutanoic Acid

et al. 2017

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Amino acids are of paramount importance as chiral building blocks of life, for drug development in modern medicinal chemistry, and for the manufacture of industrial products. In this work, the stereoselective synthesis of (S)-and (R)-2-amino-4-hydroxybutanoic acid was accomplished using a systems biocatalysis approach comprising a biocatalytic one-pot cyclic cascade by coupling of an aldol reaction with an ensuing stereoselective transamination. A class II pyruvate aldolase from E. coli, expressed as a soluble fusion protein, in tandem with either an S-or R-selective, pyridoxal phosphate dependent transaminase was used as a catalyst to realize the conversion, with formaldehyde and alanine being the sole starting materials. Interestingly, the class II pyruvate aldolase was found to tolerate formaldehyde concentrations of up to 1.4 M. The cascade system was found to reach product concentrations for (S)-or (R)-2-amino-4-hydroxybutanoic acid of at least 0.4 M, rendering yields between 86% and >95%, respectively, productivities of >80 g L −1 d −1 , and ee values of >99%.

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Section: Resultsmentioning

confidence: 99%

Combining Aldolases and Transaminases for the Synthesis of 2-Amino-4-hydroxybutanoic Acid

et al. 2017

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“…Aldol addition reaction is one of the most useful tools that have the synthetic chemist for the construction of new C–C bonds [ 2 , 3 , 4 ]. Dihydroxyacetone phosphate (DHAP)-dependent aldolases are among the most important biocatalysts for C–C bond formation [ 5 , 6 , 7 , 8 , 9 , 10 ]. Their major synthetic advantage is that the stereochemistry of the two newly formed stereogenic centers is controlled by the enzymes and, moreover, the four DHAP-dependent aldolases are stereocomplementary; so, from two given substrates, it is possible to obtain the four diastereoisomers.…”

Section: Introductionmentioning

confidence: 99%

Tuning the Phosphoryl Donor Specificity of Dihydroxyacetone Kinase from ATP to Inorganic Polyphosphate. An Insight from Computational Studies

Sánchez-Moreno

Bordes

Castillo

et al. 2015

IJMS

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Dihydroxyacetone (DHA) kinase from Citrobacter freundii provides an easy entry for the preparation of DHA phosphate; a very important C3 building block in nature. To modify the phosphoryl donor specificity of this enzyme from ATP to inorganic polyphosphate (poly-P); a directed evolution program has been initiated. In the first cycle of evolution, the native enzyme was subjected to one round of error-prone PCR (EP-PCR) followed directly (without selection) by a round of DNA shuffling. Although the wild-type DHAK did not show activity with poly-P, after screening, sixteen mutant clones showed an activity with poly-phosphate as phosphoryl donor statistically significant. The most active mutant presented a single mutation (Glu526Lys) located in a flexible loop near of the active center. Interestingly, our theoretical studies, based on molecular dynamics simulations and hybrid Quantum Mechanics/Molecular Mechanics (QM/MM) optimizations, suggest that this mutation has an effect on the binding of the poly-P favoring a more adequate position in the active center for the reaction to take place.

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“…Nature builds up carbohydrate, aminoacids, αhydroxy acids and other molecules by use of aldol reactions catalyzed by aldolase enzymes. Aldolases, mainly dihydroxyacetone phosphate (DHAP) dependent aldolases, have been extensively used in chemoenzymatic syntheses and many of these applications have been compiled in several superb reviews over the last years (Clapés and Joglar 2013;Fesko and Gruber-Khadjawi 2013;Müller 2012;Brovetto et al 2011;Clapés and Fessner 2011;Clapés et al 2010;Iturrate and García-Junceda 2008). Despite this well proven synthetic utility of aldolases, there are still few processes that have been developed at the industrial scale and none of them -to the best of our knowledge-have used DHAP-dependent aldolases (Patel 2011;Baik and Yoshioka 2009;Wolberg et al 2008;Castillo et al 2006;Müller 2005;Greenberg et al 2004).…”

Section: Introductionmentioning

confidence: 99%

Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima

Oroz‐Guinea

Sánchez-Moreno

Mena³

et al. 2014

Appl Microbiol Biotechnol

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The TM1072 gene from Thermotoga maritima codifies for a putative form of a rhamnulose-1-phosphate aldolase (Rha-1PA Tm). To investigate this enzyme further, its gene was cloned and expressed in Escherichia coli. The purified enzyme was activated by Co(2+) as a divalent metal ion cofactor, instead of Zn(2+) as its E. coli homologue, and exhibited a maximum of activity at 95 °C. Furthermore, the enzyme displayed a high stability against extreme reaction conditions, retaining 90 % of its activity in the presence of 40 % of acetonitrile and showing a half-life greater than 3 h at 115 °C. The kinetic parameters at room temperature (R/T) were also studied; the K M was calculated to be 3.6 ± 0.33 mM, while k cat/K M was found to be 0.7 × 10(3) s(-1) M(-1). Given these characteristics, Rha-1PA Tm is an attractive enzyme for use as a biocatalyst for industrial applications, offering intriguing possibilities for practical biocatalysis.

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Enzyme‐Catalyzed Aldol Additions

Cited by 13 publications

References 154 publications

Combining Aldolases and Transaminases for the Synthesis of 2-Amino-4-hydroxybutanoic Acid

Combining Aldolases and Transaminases for the Synthesis of 2-Amino-4-hydroxybutanoic Acid

Tuning the Phosphoryl Donor Specificity of Dihydroxyacetone Kinase from ATP to Inorganic Polyphosphate. An Insight from Computational Studies

Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima

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