Preparation of (S)-N-Substituted 4-Hydroxy-pyrrolidin-2-ones by Regio- and Stereoselective Hydroxylation with Sphingomonas sp. HXN-200

Chang, Dongliang; Witholt, Bernard; Li, Zhi

doi:10.1021/ol006735q

Cited by 44 publications

(15 citation statements)

References 20 publications

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“…In subsequent work, Sphingomonas sp. strain HXN200 was shown to hydroxylate many 4-, 5,-and 6-ring alicyclic compounds with rates of up to 10-20 µmol min -1 g -1 cell dry weight [17,112,113]. The stereoselective oxidation of isopropyl moieties by alkane-grown Rhodococcus and Pseudomonas strains has also been studied in some detail [85,114,115].…”

Section: Applications Of Alkane Hydroxylase Systemsmentioning

confidence: 99%

Diversity of Alkane Hydroxylase Systems in the Environment

Beilen

Duetz

et al. 2003

Oil & Gas Science and Technology - Rev. IFP

326

227

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Résumé -Diversité des systèmes alcane hydroxylase dans l'environnement -La première étape dans la dégradation aérobie des alcanes par les bactéries, les levures et les champignons est catalysée par des oxygénases, une classe d'enzymes capables d'introduire des atomes d'oxygène issus de l'oxygène moléculaire dans le substrat alcane. Ces enzymes jouent un rôle important dans la biodégradation du pétrole et dans la biodégradation cométabolique de composés tels que le trichloroéthylène et les éthers-carburants. De plus, ce sont des biocatalyseurs très utiles qui peuvent également servir de modèles pour caractériser une réaction chimique difficile : l'activation régio-et stéréospécifique de la liaison C-H. Plusieurs autres classes d'enzymes catalysent l'oxydation des alcanes. Les souches de levures capables de dégrader les alcanes contiennent plusieurs alcanes hydroxylases appartenant à la superfamille des P450, alors que différentes bactéries contiennent des enzymes similaires au système alcane hydroxylase membranaire de Pseudomonas putida GPo1. Les alcanes à courte chaîne sont probablement oxydés par des alcanes hydroxylases solubles similaires aux méthanes monooxygénases. La présence dans l'environnement de ces oxygénases a été étudiée dans des échantillons de sols et aquifères uniquement pour les alcanes hydroxylases associés aux membranes. Abstract -Diversity of Alkane Hydroxylase Systems in the Environment

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Section: Applications Of Alkane Hydroxylase Systemsmentioning

confidence: 99%

Diversity of Alkane Hydroxylase Systems in the Environment

Beilen

Duetz

et al. 2003

Oil & Gas Science and Technology - Rev. IFP

326

227

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“…Five different CYP153 enzymes obtained from Sphingomonas sp. HXN-200 were also confirmed to convert piperidines, pyrrolidines, and azetidines to useful pharmaceutical intermediates [2,3].…”

Section: Introductionmentioning

confidence: 94%

Biotransformation of medium-chain alkanes using recombinant P450 monooxygenase from Alcanivorax borkumensis SK2 expressed in Escherichia coli

Baik

2010

Korean J. Chem. Eng.

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A bioconversion system for medium-chain alkanes was constructed by using a recombinant Escherichia coli whole-cell biocatalyst expressing P450 monooxygenase genes, ferredoxin, and ferredoxin reductase cloned from Alcanivorax borkumensis as an operon. The recombinant E. coli harboring the P450 gene and two related expression component enzymes, ferredoxin and ferredoxin reductase, was constructed in a single vector pET21(a) and successfully expressed in E. coli BL21(DE3) as a soluble form, showing a molecular weight of 53 kDa on 10% SDS-PAGE. When the cell-free extract of E. coli BL21 expressing p450 monooxygenase was subjected to reduced CO difference spectral analysis, a soret band near 450 nm appeared indicating that the cloned P450 was expressed as a functionally active enzyme. The E. coli cells harboring the expressed P450 gene were able to convert n-octane and 1-decene, producing approximately 450 µg/ml of n-octanol and 290 µg/ml of 1,2-epoxydecane, respectively, at pH 7.0 and 30 o C. However, the recombinant E. coli cells were not able to convert the branched alkane, 2,6,10,14-tetramethylpentadecane (C19).

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“…HXN‐200, a class I P450 requiring ferredoxin (Fdx) and ferredoxin reductase (FdR) for electron transfer, for the hydroxylation at non‐activated carbon atom with unique substrate specificity, broad substrate range, and good to excellent regio‐ and stereoselectivity (Chang et al, 2000, 2002a, b; Li et al, 1999, 2001). It represents the best hydroxylation system known thus far for the hydroxylation of a series of alicycles such as N ‐substituted pyrrolidines (Li et al, 1999, 2001), pyrrolidinones (Chang et al, 2000), piperidines (Chang et al, 2002b), piperidinones (Chang et al, 2002a), and azetidines (Chang et al, 2002b) to prepare the corresponding alcohols as useful and valuable pharmaceutical intermediates (Fromtling and Castaner, 1995; Nagahara et al, 1994; Wallbaum et al, 1994; Weber and Gmeiner, 1998). To improve the enantioselectivity in the hydroxylation of N ‐benzyl pyrrolidine, we performed the directed evolution of P450pyr hydroxylase.…”

Section: Introductionmentioning

confidence: 99%

Engineering of recombinant E. coli cells co‐expressing P450pyrTM monooxygenase and glucose dehydrogenase for highly regio‐ and stereoselective hydroxylation of alicycles with cofactor recycling

Pham

Gao

2012

Biotech & Bioengineering

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E. coli (P450pyrTM-GDH) with dual plasmids, pETDuet containing P450pyr triple mutant I83H/M305Q/A77S (P450pyrTM) and ferredoxin reductase (FdR) genes and pRSFDuet containing glucose dehydrogenase (GDH) and ferredoxin (Fdx) genes, was engineered to show a high activity (12.7 U g⁻¹ cdw) for the biohydroxylation of N-benzylpyrrolidine 1 and a GDH activity of 106 U g⁻¹ protein. The E. coli cells were used as efficient biocatalysts for highly regio- and stereoselective hydroxylation of alicyclic substrates at non-activated carbon atom with enhanced productivity via intracellular recycling of NAD(P)H. Hydroxylation of N-benzylpyrrolidine 1 with resting cells in the presence of glucose showed excellent regio- and stereoselectivity, giving (S)-N-benzyl-3-hydroxypyrrolidine 2 in 98% ee as the sole product in 9.8 mM. The productivity is much higher than that of the same biohydroxylation using E. coli (P450pyrTM)b without expressing GDH. E. coli (P450pyrTM-GDH) was found to be highly regio- and stereoselective for the hydroxylation of N-benzylpyrrolidin-2-one 3, improving the regioselectivity from 90% of the wild-type P450pyr to 100% and giving (S)-N-benzyl-4-hydroxylpyrrolidin-2-one 4 in 99% ee as the sole product. A high activity of 15.5 U g⁻¹ cdw was achieved and (S)-4 was obtained in 19.4 mM. E. coli (P450pyrTM-GDH) was also found to be highly regio- and stereoselective for the hydroxylation of N-benzylpiperidin-2-one 5, increasing the ee of the product (S)-N-benzyl-4-hydroxy-piperidin-2-one 6 to 94% from 33% of the wild-type P450pyr. A high activity of 15.8 U g⁻¹ cdw was obtained and (S)-6 was produced in 3.3 mM as the sole product. E. coli (P450pyrTM-GDH) represents the most productive system known thus far for P450-catalyzed hydroxylations with cofactor recycling, and the hydroxylations with E. coli (P450pyrTM-GDH) provide with simple and useful syntheses of (S)-2, (S)-4, and (S)-6 that are valuable pharmaceutical intermediates and difficult to prepare.

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Preparation of (S)-N-Substituted 4-Hydroxy-pyrrolidin-2-ones by Regio- and Stereoselective Hydroxylation with Sphingomonas sp. HXN-200

Cited by 44 publications

References 20 publications

Diversity of Alkane Hydroxylase Systems in the Environment

Diversity of Alkane Hydroxylase Systems in the Environment

Biotransformation of medium-chain alkanes using recombinant P450 monooxygenase from Alcanivorax borkumensis SK2 expressed in Escherichia coli

Engineering of recombinant E. coli cells co‐expressing P450pyrTM monooxygenase and glucose dehydrogenase for highly regio‐ and stereoselective hydroxylation of alicycles with cofactor recycling

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