Combining aldolases and transaminases for the synthesis of 2‑amino-4-hydroxybutanoic acid

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%.This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement no. 635595 (CarbaZymes), the Ministerio de Economía y Competitividad (MINECO), the Fondo Europeo de Desarrollo Regional (FEDER) (grant no. CTQ2015-63563-R to P.C.), and COST action CM1303 Systems Biocatalysis.We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI).Peer reviewe

Structure-guided redesign of D-fructose-6-phosphate aldolase from E. coli: remarkable activity and selectivity towards acceptor substrates by two-point mutation

Structure-guided re-design of the acceptor binding site of D-fructose-6-phosphate aldolase from E. coli leads to the construction of FSA A129S/A165G double mutant with an activity between 5- to >900-fold higher than that of wild-type towards N-Cbz-aminoaldehyde derivatives.Peer reviewe

Procedimiento quimioenzimático para la preparación de iminociclitoles

Traducción de patente europea publicada el 11JUN2010. Titulares: Consejo Superior de Investigaciones Científicas (CSIC) y Bioglane S.L.N.E.La presente invención se refiere a un procedimiento quimioenzimático para la preparación de iminociclitoles. Los productos sintetizados se pueden usar como complementos dietéticos e ingredientes funcionales en la industria alimenticia, así como agentes terapéuticos (p. ej., en el tratamiento de diabetes).Peer reviewedConsejo Superior de Investigaciones Científicas (España), Bioglane SLNET3 Traducción de patente europe

Derivados de amida de ácidos grasos con anfetaminas para el tratamiento de desórdenes alimenticios

La presente invención se refiere a nuevos derivados amidas de ácidos grasos conjugados con anfetaminas, que se comportan como ligandos duales de los receptores cannabionides tipo 1 (CB 1) Y del subtipo alfa de los receptores activados por el proliferador de peroxisomas (PPARalfa), y como potentes agentes inhibidores de la oxidación de la Lipoproteína de Baja Densidad (LDL), así como a su procedimiento de preparación, y su utilización como herramienta farmacológica y como fármacos para modular las acciones reguladas por los citados receptores, como la inducción de la saciedad y control de ingesta, la disminución de la grasa corporal y la regulación del metabolismo lipídico.Peer reviewedFundación IMIM, Fundación IMABIS-Instituto Mediterráneo para el Avance de la Biotecnología y la Investigación Sanitaria, Consejo Superior de Investigaciones Científicas (España) (CSIC) (Titular al 10%)A1 Solicitud de patente con informe sobre el estado de la técnic

Identification of new ozonation disinfection byproducts of 17β-estradiol and estrone in water

Estrogens are a class of micro-pollutants found in water at low concentrations (in the ng L−1 range), but often sufficient to exert estrogenic effects due to their high estrogenic potency. Disinfection of waters containing estrogens through oxidative processes has been shown to lead to the formation of disinfection byproducts, which may also be estrogenic. The present work investigates the formation of disinfection byproducts of 17β-estradiol (E2) and estrone (E1) in the treatment of water with ozone. Experiments have been carried out at two different concentrations of the estrogens in ground water (100 ng L−1 and 100 μg L−1) and at varying ozone dosages (0–30 mg L−1). Detection of the estrogens and their disinfection byproducts in the water samples has been performed by means of ultra performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) with a triple quadrupole (QqQ) and a quadrupole-time of flight (QqTOF) instrument. Both E2 and E1 have been found to form two main byproducts, with molecular mass (MM) 288 and 278 in the case of E2, and 286 and 276 in the case of E1, following presumably the same reaction pathways. The E2 byproduct with MM 288 has been identified as 10epsilon-17beta-dihydroxy-1,4-estradieno-3-one (DEO), in agreement with previously published results. The molecular structures and the formation pathways of the other three newly identified byproducts have been suggested. These byproducts have been found to be formed at both high and low concentrations of the estrogens and to be persistent even after application of high ozone dosages.The research leading to these results has received funding from the European Community’s Seventh Framework Programme ([FP7/2007-2013]) under grant agreement n. 265264” and from the Spanish Ministry of Science and Innovation through the projects SCARCE (Consolider-Ingenio 2010 CSD2009-00065) and CEMAGUA (CGL2007-64551/HID). It reflects only the author’s views. The Community is not liable for any use that may be made of the information contained therein. Renata Pereira thanks the CNPQ (Conselho Nacional de Desenvolvimento Cientifico e Tecnológico) for the doctorate scholarship (process 200100/2009-2) and gratefully acknowledges FAPESP for its support (process 2008/070439) in the development of the project in Brazil. Merck (Darmstadt, Germany) is also acknowledged for the gift of LC columns.Peer reviewe

Procedimiento quimo-enzimático para la síntesis de 1-deoxi-D-xilulosa

Procedimiento quimo-enzimático para la síntesis de 1- deoxi-D-xilulosa. Procedimiento para la obtención de 1-deoxi-D-xilulosa que comprende la adición aldólica de benciloxiacetaldehído a hidroxiacetona catalizado por FSA para obtener 5-Obencil- 1-deoxi-D-xilulosa; y posterior eliminación del grupo bencilo del aducto, 5-O-bencil-1-deoxi-D-xilulosa, mediante hidrogenolisis catalítica para dar lugar a 1-deoxi-Dxilulosa.Peer reviewedConsejo Superior de Investigaciones Científicas (España), Bioglane SLNEA1 Solicitud de patente con informe sobre el estado de la técnic

Synthesis of the major metabolites of Paroxetine

11 pages, 3 figures.-- PMID: 12818234 [PubMed].-- Printed version published Jun 2003.Paroxetine is a well-known antidepressant, used worldwide in therapeutics. In comparison with other selective serotonin reuptake inhibitors, it exhibits the highest activity in serotonin reuptake inhibition. Paroxetine metabolism initially involves its demethylenation to the catechol intermediate, which is then O-methylated at positions C3 or C4. Herein, the chemistry resulting in the syntheses of these metabolites (3S,4R)-4-(4-fluorophenyl)-3-(hydroxymethyl)piperidine and (3S,4R)-4-(4-fluorophenyl)-3-(4-hydroxy-3-methoxyphenoxymethyl)piperidine is described starting from the common intermediate (3S,4R)-4-(4-fluorophenyl)-3-hydroxymethyl-1-methylpiperidine. Additionally, the common intermediate was used to synthesize paroxetine, which had the same structure and stereochemistry as commercial paroxetine, thereby confirming our synthetic route.This investigation was supported by: FIS 98/0181, CIRIT 99-SGR-00187, and PNSD (Spain).Peer reviewe

Synthesis of the major metabolites of Paroxetine

11 pages, 3 figures.-- PMID: 12818234 [PubMed].-- Printed version published Jun 2003.Paroxetine is a well-known antidepressant, used worldwide in therapeutics. In comparison with other selective serotonin reuptake inhibitors, it exhibits the highest activity in serotonin reuptake inhibition. Paroxetine metabolism initially involves its demethylenation to the catechol intermediate, which is then O-methylated at positions C3 or C4. Herein, the chemistry resulting in the syntheses of these metabolites (3S,4R)-4-(4-fluorophenyl)-3-(hydroxymethyl)piperidine and (3S,4R)-4-(4-fluorophenyl)-3-(4-hydroxy-3-methoxyphenoxymethyl)piperidine is described starting from the common intermediate (3S,4R)-4-(4-fluorophenyl)-3-hydroxymethyl-1-methylpiperidine. Additionally, the common intermediate was used to synthesize paroxetine, which had the same structure and stereochemistry as commercial paroxetine, thereby confirming our synthetic route.This investigation was supported by: FIS 98/0181, CIRIT 99-SGR-00187, and PNSD (Spain).Peer reviewe

Enzymatic desaturation of fatty acids: delta11 desaturase activity on cyclopropane acid probes

10 pages, 6 figures, 3 schemes.-- PMID: 12662058 [PubMed].-- Printed version published Apr 4, 2003.The formation of methylenecyclopropanes by enzymatic desaturation of 11-cyclopropylundecanoic acid (1) and its disubstituted derivatives cis- and trans-3−5 has been investigated using the Δ11 desaturase of Spodoptera littoralis as model enzyme. Gas chromatography coupled to mass spectrometry analyses of methanolyzed lipidic extracts from tissues incubated with each probe revealed that all the cyclopropyl fatty acids were transformed into the corresponding 11-cyclopropylidene acids, except for compound trans-5 (5b), which was not desaturated at C11. The formation of methylenecyclopropane 9 as the only reaction product from 1 indicates that a potential radical intermediate is too short-lived to allow rearrangement reactions. Information on the Δ11 desaturase substrate binding domain is provided considering the cyclopropyl probes 3−5 as conformationally restricted analogues of the straight-chain substrates.Financial support from CICYT (Grant No. AGF98-0844), MCYT (Grant No. AGL2001-0585), CIRIT (Grant Nos. 1999SGR-0187 and 2001SGR-0342), and FEDER fonds from EC.Peer reviewe

Chemoenzymatic Synthesis and Inhibitory Activities of Hyacinthacines A1 and A2 Stereoisomers.

6 páginas,1 figura, 2 esquemas, 2 tablas.A novel straightforward chemoenzymatic procedure for the synthesis of hyacinthacine stereoisomers based on the aldol addition of dihydroxyacetone phosphate (DHAP) to N-Cbz-prolinal under catalysis by l-rhamnulose 1-phosphate aldolase from E. coli is presented. The synthesis is complemented by a simple and effective purification protocol consisting of ion-exchange chromatography on CM-sepharose. As examples, ( )-hyacinthacine A2 [the enantiomer of (+)-hyacinthacine A2], 7-deoxy- 2-epialexine (the enantiomer of 3-epihyacinthacine A2), ent-7-deoxyalexine (the enantiomer of 7-deoxyalexine) and 2-epihyacinthacine A2 were synthesized by these procedures and characterized for the first time. These new isomers were assayed as inhibitors of glycosidases. As a result, ( )-hyacinthacine A2 demonstrated to be a good inhibitor of a-d-glucosidase from rice whereas the natural enantiomer, hyacinthacine A2, was not. Moreover, a new family of inhibitors of a-l-rhamnosidase was uncovered.Financial support from the Spanish MEC (CTQ2005–25182- E, CTQ2006–01080 and CTQ2006–01345/BQU), La Marat5 de TV3 foundation (Ref: 050931) and Generalitat de Catalunya DURSI 2005-SGR-00698 is acknowledged. J. Calveras acknowledges the CSIC UAs pre-doctoral scholarship programs.Peer reviewe