Biocatalytic Aldol Addition of Simple Aliphatic Nucleophiles to Hydroxyaldehydes

This ACS article is provided to You under the terms of this Standard ACS AuthorChoice/Editors' Choice usage agreement between You and the American Chemical Society ("ACS")(https://pubs.acs.org/page/policy/authorchoice_termsofuse.html)Asymmetric aldol addition of simple aldehydes and ketones to electrophiles is a cornerstone reaction for the synthesis of unusual sugars and chiral building blocks. We investigated -fructose-6-phosphate aldolase from E. coli (FSA) D6X variants as catalysts for the aldol additions of ethanal and nonfunctionalized linear and cyclic aliphatic ketones as nucleophiles to nonphosphorylated hydroxyaldehydes. Thus, addition of propanone, cyclobutanone, cyclopentanone, or ethanal to 3-hydroxypropanal or (S)- or (R)-3-hydroxybutanal catalyzed by FSA D6H and D6Q variants furnished rare deoxysugars in 8-77% isolated yields with high stereoselectivity (97:3 dr and >95% ee)

Recent Advances in the Substrate Selectivity of Aldolases

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Efficient biocatalytic processes for highly valuable terminally phosphorylated C5 to C9 D-ketoses.

International audienceA green enzymatic strategy for the synthesis of terminally phosphorylated C5 to C9 naturally occurring D-ketose phosphates and analogues was developed using D-fructose-6-phosphate aldolase (FSA) as a catalyst. This enzyme has stereoselectively catalysed aldol reactions between glycolaldehyde phosphate or ribose-5-phosphate as an acceptor substrate and dihydroxyacetone, hydroxyacetone or hydroxybutanone as a donor. Furthermore, D-glycero-D-altro-2-octulose 8-phosphate was obtained using a straightforward one-pot domino biocatalytic system involving FSA, ribulose-5-phosphate epimerase and ribose-5-phosphate isomerase controlling five contiguous asymmetric centres and starting from achiral material

Aldolases from biodiversity: exploration of their substrates promiscuity

International audienceFor many years, aldolases catalysing stereoselective C-C bond formation have been considered essential for synthetic applications.[1] Biocatalysed aldolisation reactions are performed under mild conditions, without any protections and are therefore highly valuable for the development of green synthetic processes. In addition, there is room for new C-C bond forming enzymes to construct more complex molecules since this category of enzymes is underused when compared with other biocatalysts.[2] The classification of aldolases is based on the structure of natural nucleophiles leading to five main classes: dihydroxyacetone phosphate- (DHAP), pyruvate-, ethanal-, dihydroxyacetone- (DHA), and glycine- aldolases.[1a] Concerning their substrate specificity, one generally admitted that if they accept a broad range of aldehydes as electrophiles, most of them are strictly dependent on a sole nucleophile substrate.Our work highlight that aldolases are not always dependent on aldehydes as electrophile or on a sole nucleophile substrates. They complete the recent discoveries reported by us and others on their higher nucleophile tolerance.[3] Recent results in exploring nucleophile and electrophile substrates promiscuity among aldolases from biodiversity (see scheme) will be described.In particular, we have revisited DHAP-dependent aldolases with ketones as electrophiles.[4] We have demonstrated that rhamnulose-1-phosphate aldolases display an unprecedented versatility for activated ketones. We selected and characterized a rhamnulose aldolase from Bacteroides thetaio-taomicron as a proof of concept. DHAP was added to several hydroxylated ketones used as electro-philes. This aldol addition was stereoselective and produced branched-chain monosaccharide adducts with a tertiary alcohol moiety, which is rather difficult to prepare optically pure. Other nucleophiles [5] or electrophiles, with different aldolase classes are currently under investigation in our lab, which would confirmed the unprecedented substrate tolerance among aldolases.References:1. (a) P. Clapés, X. Garrabou, Adv. Synth. Catal. 2011, 353, 2263-2283; (b) P. Clapés, W. D. Fessner, G. A. Sprenger, A. K. Samland, Curr. Opin. Chem. Biol. 2010, 14, 154-167; (c) M. Müller, Adv. Synth. Catal. 2012, 354, 3161-3174; (d) M. Brovetto, D. Gamenara, P. Mendez, G. Seoane, Chem. Rev. 2011, 111, 4346-4403; (e) A. Bolt, A. Berry, A. Nelson, Arch. Biochem. Biophys. 2008, 474, 318-330; (f) A. K. Samland, G. A. Sprenger, Appl. Microbiol. Biotechnol. 2006, 71, 253-264.2. N. J. Turner, E. O'Reilly Nature Chem. Biol. 2013, 9, 285–288.3 (a) R. Roldaú n, K. Hernandez, J. Joglar, J. Bujons, T. Parella, I. Saú nchez-Moreno, V. Hélaine, M. Lemaire, C. Gueú rard-Heú laine, W.-D. Fessner, and P. Clapeú s ACS Catal. 2018, 8, 8804−880. (b) I. Sanchez-Moreno, T. Scheidt, V. Hélaine, M. Lemaire, T. Parella, P. Clapés, W.-D. Fessner, C. Guérard-Hélaine Chem. Eur. J., 2017, 23, 2005-2009. (c) V. de Berardinis, C. Guérard-Hélaine, E. Darii, K. Bastard, V. Hélaine, A. Mariage, J.-L. Petit, N. Poupard, I. Sanchez-Moreno, M. Stam, T. Gefflaut, M. Salanoubat, M. Lemaire Green Chem., 2017, 19, 519-526.4 (a) V. Laurent, E. Darii, A. Aujon, M. Debacker, J.-L. Petit, V. Hélaine, T. Liptaj, M. Breza, L. Nauton, M. Traïkia, M. Salanoubat, M. Lemaire, C. Guérard-Hélaine, V. de Berardinis Angew. Chem., Int. Ed. Engl., 2018, 57, 5467-5471. (b) M. Salanoubat, M. Lemaire, C. Guérard-Hélaine, V. de Berardinis WO 2018/215476 A1.5 V. Laurent, A. Uzel, V. Hélaine, L. Nauton, M. Traïkia, T. Gefflaut, M. Salanoubat, V. de Berardinis, M. Lemaire and C. Guérard-Hélaine Adv. Synth. Catal. 2019, accepted (special Biotrans 2019 issue

Aldolases from biodiversity: exploration of their substrates promiscuity

For many years, aldolases catalysing stereoselective C-C bond formation have been considered essential for synthetic applications.[1] Biocatalysed aldolisation reactions are performed under mild conditions, without any protections and are therefore highly valuable for the development of green synthetic processes. In addition, there is room for new C-C bond forming enzymes to construct more complex molecules since this category of enzymes is underused when compared with other biocatalysts.[2] The classification of aldolases is based on the structure of natural nucleophiles leading to five main classes: dihydroxyacetone phosphate- (DHAP), pyruvate-, ethanal-, dihydroxyacetone- (DHA), and glycine- aldolases.[1a] Concerning their substrate specificity, one generally admitted that if they accept a broad range of aldehydes as electrophiles, most of them are strictly dependent on a sole nucleophile substrate.Our work highlight that aldolases are not always dependent on aldehydes as electrophile or on a sole nucleophile substrates. They complete the recent discoveries reported by us and others on their higher nucleophile tolerance.[3] Recent results in exploring nucleophile and electrophile substrates promiscuity among aldolases from biodiversity (see scheme) will be described.In particular, we have revisited DHAP-dependent aldolases with ketones as electrophiles.[4] We have demonstrated that rhamnulose-1-phosphate aldolases display an unprecedented versatility for activated ketones. We selected and characterized a rhamnulose aldolase from Bacteroides thetaio-taomicron as a proof of concept. DHAP was added to several hydroxylated ketones used as electro-philes. This aldol addition was stereoselective and produced branched-chain monosaccharide adducts with a tertiary alcohol moiety, which is rather difficult to prepare optically pure. Other nucleophiles [5] or electrophiles, with different aldolase classes are currently under investigation in our lab, which would confirmed the unprecedented substrate tolerance among aldolases.References:1. (a) P. Clapés, X. Garrabou, Adv. Synth. Catal. 2011, 353, 2263-2283; (b) P. Clapés, W. D. Fessner, G. A. Sprenger, A. K. Samland, Curr. Opin. Chem. Biol. 2010, 14, 154-167; (c) M. Müller, Adv. Synth. Catal. 2012, 354, 3161-3174; (d) M. Brovetto, D. Gamenara, P. Mendez, G. Seoane, Chem. Rev. 2011, 111, 4346-4403; (e) A. Bolt, A. Berry, A. Nelson, Arch. Biochem. Biophys. 2008, 474, 318-330; (f) A. K. Samland, G. A. Sprenger, Appl. Microbiol. Biotechnol. 2006, 71, 253-264.2. N. J. Turner, E. O'Reilly Nature Chem. Biol. 2013, 9, 285–288.3 (a) R. Roldaú n, K. Hernandez, J. Joglar, J. Bujons, T. Parella, I. Saú nchez-Moreno, V. Hélaine, M. Lemaire, C. Gueú rard-Heú laine, W.-D. Fessner, and P. Clapeú s ACS Catal. 2018, 8, 8804−880. (b) I. Sanchez-Moreno, T. Scheidt, V. Hélaine, M. Lemaire, T. Parella, P. Clapés, W.-D. Fessner, C. Guérard-Hélaine Chem. Eur. J., 2017, 23, 2005-2009. (c) V. de Berardinis, C. Guérard-Hélaine, E. Darii, K. Bastard, V. Hélaine, A. Mariage, J.-L. Petit, N. Poupard, I. Sanchez-Moreno, M. Stam, T. Gefflaut, M. Salanoubat, M. Lemaire Green Chem., 2017, 19, 519-526.4 (a) V. Laurent, E. Darii, A. Aujon, M. Debacker, J.-L. Petit, V. Hélaine, T. Liptaj, M. Breza, L. Nauton, M. Traïkia, M. Salanoubat, M. Lemaire, C. Guérard-Hélaine, V. de Berardinis Angew. Chem., Int. Ed. Engl., 2018, 57, 5467-5471. (b) M. Salanoubat, M. Lemaire, C. Guérard-Hélaine, V. de Berardinis WO 2018/215476 A1.5 V. Laurent, A. Uzel, V. Hélaine, L. Nauton, M. Traïkia, T. Gefflaut, M. Salanoubat, V. de Berardinis, M. Lemaire and C. Guérard-Hélaine Adv. Synth. Catal. 2019, accepted (special Biotrans 2019 issue

L-Rhamnulose-1-phosphate and L-fuculose-1-phosphate aldolase mediated multi-enzyme cascade systems for nitrocyclitol synthesis

One-pot multistep stereoselective cascade reactions were implemented for the straightforward synthe-sis of various nitrocyclitols. Two kinases, an aldolase and a phosphatase were involved in this process,together with a spontaneous intramolecular Henry reaction to provide the nitrocyclitol moiety. The C Cbond formation catalysed by the aldolase and the nitroaldol reactions were key steps to build the carbocy-cle stereoselectively. The aldolase acceptor substrates were all 4-nitrobutanal structurally based, eitherhydroxylated or unsubstituted at the C2 and/or C3 positions. l-Fuculose-1-phosphate aldolase (FucA)catalysed the formation of the expected (R,R)- or d-erythro aldol, except in the case of 4-nitrobutanal,from which the epimeric (R,S)- or l-threo aldol was also formed. l-Rhamnulose-1-phosphate aldolase con-sistently formed the expected (R,S)- or l-threo aldol together with a minor amount of (R,R)- or d-erythroaldol. The intramolecular Henry reaction was also found to be stereoselective, occurring spontaneouslyonce the aldol was formed due to the presence of both ketone and a terminally positioned nitro group.The combination of this set of reactions successfully furnished 11 nitrocyclitols which have not beendescribed previously in the literature.CNRS International Relationship service is thanked for mobilityfinancial support to F. Camps Bres. This work was also supportedin part, by the CNRS GDR RDR2 “Aller vers une chimie éco-compatible” Grant (to F.C.B.). E G-J. has been supported by theSpanish Ministerio de Ciencia e Innovación (Grants PI11/01436 andCTQ2010-19073/BQU).Peer reviewe

Aldolases from biodiversity: exploration of their substrates promiscuity

For many years, aldolases catalysing stereoselective C-C bond formation have been considered essential for synthetic applications.[1] Biocatalysed aldolisation reactions are performed under mild conditions, without any protections and are therefore highly valuable for the development of green synthetic processes. In addition, there is room for new C-C bond forming enzymes to construct more complex molecules since this category of enzymes is underused when compared with other biocatalysts.[2] The classification of aldolases is based on the structure of natural nucleophiles leading to five main classes: dihydroxyacetone phosphate- (DHAP), pyruvate-, ethanal-, dihydroxyacetone- (DHA), and glycine- aldolases.[1a] Concerning their substrate specificity, one generally admitted that if they accept a broad range of aldehydes as electrophiles, most of them are strictly dependent on a sole nucleophile substrate.Our work highlight that aldolases are not always dependent on aldehydes as electrophile or on a sole nucleophile substrates. They complete the recent discoveries reported by us and others on their higher nucleophile tolerance.[3] Recent results in exploring nucleophile and electrophile substrates promiscuity among aldolases from biodiversity (see scheme) will be described.In particular, we have revisited DHAP-dependent aldolases with ketones as electrophiles.[4] We have demonstrated that rhamnulose-1-phosphate aldolases display an unprecedented versatility for activated ketones. We selected and characterized a rhamnulose aldolase from Bacteroides thetaio-taomicron as a proof of concept. DHAP was added to several hydroxylated ketones used as electro-philes. This aldol addition was stereoselective and produced branched-chain monosaccharide adducts with a tertiary alcohol moiety, which is rather difficult to prepare optically pure. Other nucleophiles [5] or electrophiles, with different aldolase classes are currently under investigation in our lab, which would confirmed the unprecedented substrate tolerance among aldolases.References:1. (a) P. Clapés, X. Garrabou, Adv. Synth. Catal. 2011, 353, 2263-2283; (b) P. Clapés, W. D. Fessner, G. A. Sprenger, A. K. Samland, Curr. Opin. Chem. Biol. 2010, 14, 154-167; (c) M. Müller, Adv. Synth. Catal. 2012, 354, 3161-3174; (d) M. Brovetto, D. Gamenara, P. Mendez, G. Seoane, Chem. Rev. 2011, 111, 4346-4403; (e) A. Bolt, A. Berry, A. Nelson, Arch. Biochem. Biophys. 2008, 474, 318-330; (f) A. K. Samland, G. A. Sprenger, Appl. Microbiol. Biotechnol. 2006, 71, 253-264.2. N. J. Turner, E. O'Reilly Nature Chem. Biol. 2013, 9, 285–288.3 (a) R. Roldaú n, K. Hernandez, J. Joglar, J. Bujons, T. Parella, I. Saú nchez-Moreno, V. Hélaine, M. Lemaire, C. Gueú rard-Heú laine, W.-D. Fessner, and P. Clapeú s ACS Catal. 2018, 8, 8804−880. (b) I. Sanchez-Moreno, T. Scheidt, V. Hélaine, M. Lemaire, T. Parella, P. Clapés, W.-D. Fessner, C. Guérard-Hélaine Chem. Eur. J., 2017, 23, 2005-2009. (c) V. de Berardinis, C. Guérard-Hélaine, E. Darii, K. Bastard, V. Hélaine, A. Mariage, J.-L. Petit, N. Poupard, I. Sanchez-Moreno, M. Stam, T. Gefflaut, M. Salanoubat, M. Lemaire Green Chem., 2017, 19, 519-526.4 (a) V. Laurent, E. Darii, A. Aujon, M. Debacker, J.-L. Petit, V. Hélaine, T. Liptaj, M. Breza, L. Nauton, M. Traïkia, M. Salanoubat, M. Lemaire, C. Guérard-Hélaine, V. de Berardinis Angew. Chem., Int. Ed. Engl., 2018, 57, 5467-5471. (b) M. Salanoubat, M. Lemaire, C. Guérard-Hélaine, V. de Berardinis WO 2018/215476 A1.5 V. Laurent, A. Uzel, V. Hélaine, L. Nauton, M. Traïkia, T. Gefflaut, M. Salanoubat, V. de Berardinis, M. Lemaire and C. Guérard-Hélaine Adv. Synth. Catal. 2019, accepted (special Biotrans 2019 issue

2-Ketogluconate Kinase from Cupriavidus necator H16: Purification, Characterization, and Exploration of Its Substrate Specificity

International audienceWe have cloned, overexpressed, purified, and characterized a 2-ketogluconate kinase (2-dehydrogluconokinase, EC 2.7.1.13) from Cupriavidus necator (Ralstonia eutropha) H16. Exploration of its substrate specificity revealed that three ketoacids (2-keto-3-deoxy-d-gluconate, 2-keto-d-gulonate, and 2-keto-3-deoxy-d-gulonate) with structures close to the natural substrate (2-keto-d-gluconate) were successfully phosphorylated at an efficiency lower than or comparable to 2-ketogluconate, as depicted by the measured kinetic constant values. Eleven aldo and keto monosaccharides of different chain lengths and stereochemistries were also assayed but not found to be substrates. 2-ketogluconate-6-phosphate was synthesized at a preparative scale and was fully characterized for the first time

Mixing chemo- and biocatalysis for rare monosaccharide production by combining aldolase and N-heterocyclic carbene gold catalysts

International audienceChemo- and biocatalysis for multistep reactions were successfully combined to synthesize polyhydroxylated monosaccharides. Gold N-heterocyclic carbene, catalysing green hydration, was coupled to an aldolase to create two stereocenters via C–C bond formation in stereoselective and total atom economy fashions starting from simple and achiral compounds

Etude de l'immobilisation de la fructose-6-phosphate aldolase support hydroxyde double lamellaire

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