EP2054510A2 - Préparation énantiosélective d'esters aliphatiques acycliques et de cétones aliphatiques acycliques - Google Patents

Préparation énantiosélective d'esters aliphatiques acycliques et de cétones aliphatiques acycliques

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Publication number
EP2054510A2
EP2054510A2 EP07785706A EP07785706A EP2054510A2 EP 2054510 A2 EP2054510 A2 EP 2054510A2 EP 07785706 A EP07785706 A EP 07785706A EP 07785706 A EP07785706 A EP 07785706A EP 2054510 A2 EP2054510 A2 EP 2054510A2
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Prior art keywords
baeyer
radical
villiger
ketones
aliphatic acyclic
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EP07785706A
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German (de)
English (en)
Inventor
Uwe T. Bornscheuer
Anett Kirschner
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Universitaet Greifswald
Original Assignee
Ernst Moritz Arndt Universitaet Greifswald
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Filing date
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Application filed by Ernst Moritz Arndt Universitaet Greifswald filed Critical Ernst Moritz Arndt Universitaet Greifswald
Publication of EP2054510A2 publication Critical patent/EP2054510A2/fr
Withdrawn legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
    • C12P41/003—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by ester formation, lactone formation or the inverse reactions
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters

Definitions

  • the present invention relates to a process for the enantioselective (stereoselective) preparation of aliphatic acyclic esters from racemic aliphatic acyclic ketones by Baeyer-Villiger oxidation.
  • the invention further relates to the kinetic enzymatic racemate cleavage of racemic aliphatic acyclic ketones.
  • Baeyer and Villiger reported a reaction of cyclic ketones with peroxymonosulfuric acid, in which lactones were displayed (Baeyer and Villiger 1899).
  • the Baeyer-Villiger reaction has since been used in organic syntheses.
  • the chemical Baeyer-Villiger oxidation is normally catalyzed by peracids and found in one two-step procedure, which was described by Criegee (Criegee 1948).
  • Baeyer-Villiger monooxygenases (BVMOs, E.C. 1.14.13.x) are enzymes that belong to the class of oxidoreductases and can convert aliphatic and cyclic ketones to esters and lactones using oxygen (Mihovilovic et al., 2002).
  • BVMOs are flavin-dependent (generally FAD) and require NAD (P) H to catalyze the oxidation reaction.
  • BVMOs can be formed from a variety of bacteria and fungi.
  • the best-studied BVMO to date is an enzyme from Acinetobacter calcoaceticus NCIMB 9871, which is also referred to as cyclohexanone monooxygenase (CHMO). This enzyme converts a variety of mono- and bicyclic ketones (Donoghue et al., 1976, Stewart 1998).
  • BVMOs in addition to metal based chiral catalysts (Strukul 1998) is one way to enantioselectively esters and lactones represent.
  • the present invention solves these problems. It provides a method for the production of aliphatic acyclic esters from racemic aliphatic acyclic ketones by Baeyer-Villiger oxidation, in which the Baeyer-Villiger oxidation proceeds enantioselectively and Baeyer-Villiger monooxygenases (BVMO) are used for the oxidation.
  • BVMO Baeyer-Villiger monooxygenases
  • enantioselective or stereoselective if an enantioselectivity E of more than 1, preferably more than 2, is reached, ie one enantiomer is present in excess over the other.
  • An industrial application is especially worthwhile if a high enantioselectivity E of at least 10, preferably of at least 40, is achieved.
  • enantioselectivity can be achieved in the enzymatic reaction of aliphatic acyclic ketones by means of BVMO.
  • Enantiomeric excess and enantioselectivity are determined as appropriate, e.g. Gas chromatographic analysis of the products of the reaction calculated according to the following formula (Chen 1982):
  • the Baeyer-Villiger monooxygenase used in the method of the invention is specific for aliphatic acyclic ketones.
  • the term aliphatic acyclic ketones indicates that the keto group is neither directly linked to an aromatic or cyclic group nor does the molecule contain aromatic or cyclic substituents.
  • ketones have a formula
  • R 1, R 2, R 3 and R 4 independently of one another are a substituted or unsubstituted, linear or branched alkyl radical, alkenyl radical, alkynyl radical, a hydroxyalkyl radical, carboxyl radical, alkoxycarboxyl radical, amino radical, aminoalkyl radical, aminocarbonyl radical, hydroxyl radical or a halogen radical,
  • R3 or R4 can also be hydrogen
  • R 1 is preferably a linear or branched alkyl radical having 1-6 C atoms, in particular methyl, ethyl, propyl, n-butyl or isobutyl.
  • R 2 is preferably a hydroxyl radical, amino radical or a halogen radical, ie, for example, chlorine, bromine or iodine. It is particularly preferred for the process according to the invention if R 2 is a hydroxyl radical, ie the ketone is a ⁇ -hydroxyketone.
  • the C atom attached to R2 is chiral, so R2, R3 and R4 are different.
  • racemic mixtures are used as starting materials.
  • R3 or R4 is hydrogen. It is particularly preferred that R 3 or R 4 is an alkyl radical having a chain length of 1 to 14 C atoms, in particular 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 C atoms, and that R3 or R4 - that is the other substituent - is hydrogen.
  • the alkyl radical is preferably linear and unsubstituted.
  • n 1, but n may be e.g. also be 2, 3, 4, 5, 6, 7, 8, 9 or 10.
  • the ⁇ -hydroxyketone has the following formula:
  • R3 is an alkyl radical having a chain length of 2-20 C atoms, in particular having a chain length of 2-10 C atoms.
  • the ⁇ -hydroxy ketone at the ⁇ -C atom instead of the methyl group can carry a radical Rl according to the above definition.
  • Non-limiting examples of a linear or branched alkenyl of the invention i. for a hydrocarbon radical containing one or more C-C double bonds, propenyl, 2-butenyl, 3-butenyl, 3-pentenyl, 1,3-hexadienyl, etc. are.
  • Alkynyl 1 represents , for example, propynyl, 2-butynyl, 3-butynyl, 4-hexynyl, 1-decynyl, etc., ie a hydrocarbon radical which contains one or more C-C triple bonds.
  • hydrocarbon radicals are also included which contain both double and triple bonds.
  • hydrocarbon radicals mentioned may optionally be monosubstituted or polysubstituted, e.g. by alkyl, halogen, hydroxyl and / or amino groups.
  • Hydroxyalkyl stands for an abovementioned alkyl radical which is hydroxylated. For example, hydroxyethyl may be mentioned here.
  • Aminoalkyl-, Alkylaminoalkyl- each represent an above-mentioned alkyl radical which carries an amino group, which may be mono- but also dialkylated. Examples are the aminoethyl and the dimethylaminoethyl radical.
  • Alkoxyalkyl represents an abovementioned alkyl radical to which an alkoxy group is bonded. Examples of these are ethoxyethyl and ethoxypropyl.
  • Alkoxycarbonyl is a carbonyl radical to which an alkoxy group is bonded. Examples are the methoxycarbonyl and the ethoxycarbonyl.
  • Alkoxycarbonylalkyl is an abovementioned alkyl radical to which an alkoxycarbonyl group is bonded. Examples of these are ethoxycarbonylmethyl and methoxycarbonylmethyl radicals.
  • the method of the invention uses a Baeyer-Villiger monooxygenase (BVMO) derived from Pseudomonas fluorescens (P. fluorescens) or Pseudomonas putida (P. putida) or a functional derivative thereof.
  • BVMO Baeyer-Villiger monooxygenase
  • the BVMO from P. fluorescens has a particularly high specificity for ketones in which R3 is a short-chain alkyl radical, e.g. with 4, 6, or 8 C atoms, so that this enzyme is preferably used for the reaction of these ketones.
  • R3 is a short-chain alkyl radical, e.g. with 4, 6, or 8 C atoms, so that this enzyme is preferably used for the reaction of these ketones.
  • the BVMO from P. putida has a similar specificity. In both enzymes ⁇ -hydroxy ketones are preferred substrates.
  • the Baeyer-Villiger monooxygenase preferably comprises a sequence which has a homology of at least 70%, at least 80%, at least 90% or at least 95% to SEQ ID NO: 1 (BVMO from P. fluorescens) or SEQ ID NO: 3 (BVMO from P. putida).
  • the Baeyer-Villiger monooxygenase used in the method according to the invention preferably has the sequence according to SEQ ID NO: 1 or SEQ ID NO: 3.
  • the P. fluorescens BVMO sequence is published under the GenBank accession numbers AAC36351 (protein) and AF090329 (genome) and the BVMO sequence from P. putida under AAN68413 (protein) and AE015451 (genome), respectively.
  • the BVMO is able to convert aliphatic acyclic ketones to esters in a Baeyer-Villiger oxidation.
  • the Baeyer-Villiger monooxygenase sequence motif described by Fraaije et al. (2002, see also WO 03/020890) is also present in the protein sequence of the enzyme preferably used in the method according to the invention.
  • BVMO from P. fluorescens does not convert cyclic Baeyer-Villiger substrates, such as cycloketones or aromatic ketones, because the enzyme, when compared to characterized BVMOs with a known substrate spectrum, has the highest sequence similarity.
  • Hydroxyacetophenone monooxygenase HAPMO, 37% amino acid identity.
  • Phylogenetic analysis showed the same close affinity between HAPMO and the BVMO of P. fluorescens DSM50106.
  • BVMOs may also be used in the method of the invention, such as the tridecanone monooxygenase from P. cepacia (Britton and Markovetz 1977), the cyclohexanone monooxygenase (CHMO) from Acinetobacter calcoaceticus NCIMB 9871 (Donoghue et al., 1976) or the cyclopentanone monooxygenase (CPMO) from Comamonas sp. NCIMB 9872 (Griffin and Trudgill 1976).
  • CHMO can industrially rewarding be used for example for the implementation of a ß-hydroxyketone having a radical R3 with 1, 2, 3 or 4 C-atoms.
  • BVMO suitability of a BVMO for the implementation of a particular substrate or production of a specific ester for a specific, e.g. industrial purpose
  • a BVMO for the implementation of a particular substrate or production of a specific ester for a specific, e.g. industrial purpose
  • the ketones are preferably oxidized by incubating them with growing or quiescent cells containing the Baeyer-Villiger monooxygenases.
  • the enzyme it is not necessary to purify the enzyme prior to the reaction and to use it in isolation with cofactors such as NADPH isolated in the reaction.
  • cofactors such as NADPH isolated in the reaction.
  • this is also possible in principle, as shown, for example, in Britton and Markovetz, 1977.
  • Dormant cells are preferably used.
  • the cells also overexpress chaperones, for example, by expression of GroES / GroEL from plasmid vectors (Ikura et al., 2002, Lee et al., 2004, Nishihara et al., 1998, Nishihara et al., 2000).
  • the cells may be eukaryotic cells, such as yeasts, but are preferably bacterial cells, such as Escherichia coli, Pseudomonas sp. , Bacillus sp., Aspergillus sp. Most preferred is expression in E. coli, e.g. in E. coli DH5 ⁇ , E. coli JM109, E. coli BL21 (DE3) or E. coli Origami TM (DE3). Preferably, the expression of the enzyme is inducible.
  • Preferred expression vectors are based on pJOE or pET vectors and are described in detail in the examples, e.g. the expression vector pJOE4072.6 or pET22BVMO.
  • the inventive method is preferably carried out at about 20 to 37 0 C.
  • an incubation temperature of about 25 to 30 0 C, in particular about 30 0 C, since volatile volatiles and esters evaporate here only to a small extent.
  • the temperature can also be varied for different steps.
  • the esters produced by the Baeyer-Villiger oxidation can be isolated and purified.
  • esters can be carried out by extraction with suitable solvents, for example dichloromethane. However, it was found that extraction with ethyl acetate gave a better yield. Subsequently, the ester of simultaneously extracted unreacted ketone be separated, for example by column chromatography on silica gel, in particular using a suitable solvent mixture, for example, of hexane: ethyl acetate 5: 1.
  • the present invention in addition to the process for the enantioselective preparation of aliphatic acyclic esters, simultaneously provides a process for the enantioselective preparation of aliphatic acyclic ketones.
  • the enantiomer unreacted by the particular enzyme is obtained.
  • the starting material used is a racemic mixture of ketones, from which an enantiomer is enantioselectively converted into the ester by the enzyme.
  • isolating and purifying the unreacted ketones e.g. by extraction and subsequent separation by column chromatography, this gives the other enantiomer of the ketone.
  • the process according to the invention is also a novel process for the racemate resolution of aliphatic acyclic ketones, ie for the separation of racemates into the enantiomers.
  • the process is referred to as catalytic kinetic resolution because the enzyme reacts one enantiomer of the reactant faster than the mirror image enantiomer of the ketone.
  • esters and / or ketones obtained by the process according to the invention preferably have a purity of the enantiomers of at least 70%, preferably at least 80%, 90% or 95% or 99%.
  • the process according to the invention is therefore also particularly advantageous since both enantioselective preparation of aliphatic acyclic ketones and aliphatic acyclic esters is possible with only one reaction step.
  • the present invention furthermore relates to the use of Baeyer-Villiger monooxygenases for carrying out the process according to the invention.
  • Particularly preferred here too is the use of the Baeyer-Villiger monooxygenases described above, in particular the BVMOs which originate from Pseudomonas fluorescens (P. fluorescens) or P. putida or are functional derivatives thereof.
  • the invention thus also relates to the use of Baeyer-Villiger monooxygenases for the steroe-selective Baeyer-Villiger oxidation of racemic aliphatic acyclic ketones.
  • E. coli JM109 was obtained from New England Biolabs (Beverly, MA, USA).
  • E. coli BL21 (DE3), Origami TM (DE3) and pET22b (+) were purchased from Novagen (Darmstadt, Germany).
  • P. fluorescens DSM 50106 is commonly available (for example, Khalameyzer et al., 1999) and may be obtained, for example, from DSMZ (Braunschweig, Germany).
  • the chaperone plasmid set containing plasmids pG-KJE8, pGro7, pKJE7, pG-Tf2 and pTfl6 was purchased from TaKaRa Bio Inc. (Otsu, Japan).
  • the BVMO gene was used without stop codon by means of the polymerase chain reaction (PCR) using the oligonucleotides S3089 (5'-AAA ACA, TAT GAA TGC CCA CAG TGA TT-3 1 (SEQ ID NO: 4) and S3090 (5 ').
  • PCR polymerase chain reaction
  • S3089 5'-AAA ACA, TAT GAA TGC CCA CAG TGA TT-3 1 (SEQ ID NO: 4) and S3090 (5 ').
  • Taql DNA polymerase Biozym Diagnostik GmbH, Oldendorf, Germany
  • pJOE4072.6 was digested with Ndel and Hindl.II and the resulting small fragment (about 1.6 kb) carrying the gene and the His tag was digested with pET22b (+). which was digested with the same restriction enzymes (pET22BVMO).
  • Constructs without C-terminal His tag were prepared by converting the codon GGA from the BamHI site inserted between gene and His tag into the stop codon TGA using QuikChang TM Site Directed Mutagenesis (Stratagene, La Jolla, USA).
  • E. coli JM109 was transformed with pJOE constructs. Expression was performed in 30 ml LB medium (10 g tryptone, 10 g NaCl, 5 g yeast extract / L) containing 100 ⁇ g per ml ampicillin (LB amp ) at 37, 30 and 25 ° C. The cells were cultured to an optical density of 0.6 to 0.7 at 600 nm, at which time BVMO expression was induced by the addition of L-rhamnose (0.2% (w / v) final concentration). To After further cultivation for up to 20 hours, the cells were harvested by centrifugation and disrupted by sonication (60 seconds, 50% intensity) in sodium phosphate buffer (50 mM, pH 7.5).
  • Inclusion bodies were separated from the cell lysate by centrifugation for 10 minutes at 800 g, treated with 0.1% (v / v) Triton X-100 (37 ° C, 10 minutes) and washed twice with sodium phosphate buffer. Cell debris was removed from the cell lysate by additional centrifugation at high speed (10 minutes, 16000 g). Supernatants and fractions with inclusion bodies were analyzed by SDS-PAGE and Western blot.
  • E. coli BL21 (DE3) and Origami TM (DE3) were transformed with pET22b (+) constructs.
  • the cells were cultured in 30 ml LB ara p at 37, 30, 25 and 20 ° C. to an optical density of 0.7 to 0.8 at 600 nm. Thereafter, BVMO expression was induced by the addition of IPTG at a final concentration of 0.1 mM. After further expression for 20 hours, the cells were harvested by centrifugation and worked up as previously described.
  • E. coli JM109 and BL21 (DE3) cells were transformed with chaperone-encoding plasmids.
  • the cells were cultured in 20 ml LB containing 34 ⁇ g / ml chloramphenicol (LB cm ) at 37 ° C and competent cells were prepared. These were transformed with pJOE constructs (for JM109) or pET22b (+) constructs (for BL21 (DE3)) and selected on LB cm + a mp.
  • Racemic ⁇ -hydroxy ketones were synthesized via aldol condensation according to the method described by Smith and Levenberg (1981).
  • Hydroxyalkyl acetates were synthesized enzymatically using Candida antarctica lipase B. 5 mg of immobilized enzyme (Chirazyme L-2, C-2, Roche, Penzberg, Germany) were mixed with 300 ⁇ l of isooctane and 300 ⁇ l of vinyl acetate in a 2 ml glass jar. The reactions were started by addition of 10 ⁇ l of 1,2-diol and incubated at 25 ° C. in a thermoshaker (Eppendorf, Hamburg, Germany) for about two hours until the conversion was complete. The samples were used directly for GC analysis.
  • immobilized enzyme Chorazyme L-2, C-2, Roche, Penzberg, Germany
  • Hydroxyklaremethylester were synthesized from the corresponding ß-Keto acid methyl esters via ADH-catalyzed reduction, using an alcohol dehydrogenase from P. fluorescens DSM50106 was used (Hildebrand et al 2002). 5 mg of enzyme lyophilisate (produced recombinantly) were mixed with 800 ⁇ l of phosphate buffer (50 mM, pH 7.5), 200 ⁇ l of isopropanol and 2 ⁇ l of methyl ⁇ -keto acid ester. The reactions were incubated at 20 ° C. for 24 hours in a thermoshaker (Eppendorf, Hamburg, Germany). The reaction mixture was then extracted twice with 500 ⁇ l of ethyl acetate.
  • Methyl ⁇ -keto acid which was not commercially available, was prepared according to the method described by Oikawa et al. (1978).
  • E. coli JM109 pJOE4072.6 cells were cultured in 30 ml LB amp at 30 0 C to an optical density of 0.6 to 0.7 at 600 nm. Thereafter, BVMO expression was induced by addition of L-rhamnose at a final concentration of 0.2% (w / v). At the same time, 0.1 mmol of ⁇ -hydroxyketone was added and the culture was further incubated at 30 ° C. and 220 rpm. After certain periods, 500 ⁇ l samples were taken, extracted twice with dichloromethane and dried over anhydrous sodium sulfate. Excess solvent was removed in a nitrogen stream and the samples analyzed by gas chromatography.
  • BVMO in E. coli JM109 pGro7 pJOE4072.6 was determined in 200 ml LB c ⁇ H . at p / which contained 0.5 mg per ml of L-arabinose, performed at 30 ° C.
  • the cells were cultured to an optical density of 0.5 to 0.6 at 600 nm, at which time BVMO expression was induced by the addition of L-rhamnose at a final concentration of 0.2% (w / v) , After further growth for four hours, the cells were harvested by centrifugation and washed once with sterile phosphate buffer (50 mM, pH 7.5).
  • the cells were resuspended in the same buffer to an optical density of about 40, and 1 ml aliquots of this cell suspension were placed in 2 ml tubes containing 5 ⁇ mol ⁇ -hydroxyket . and 10 ⁇ l of a sterile 1 M glucose solution.
  • the tubes were sealed with air-permeable lids (Lid Ba c / Eppendorf, Hamburg, Germany) and incubated at 30 0 C in a thermoshaker (Eppendorf, Hamburg, Germany). At certain times, 300 ⁇ l samples were taken, extracted twice with ethyl acetate and dried over anhydrous sodium sulfate. Excess solvent was removed in a nitrogen stream and the samples were analyzed by gas chromatography. ,
  • the cell pellet was resuspended in 60 ml of phosphate buffer (OD of about 10), divided into 3 times 20 ml and transferred to 250 ml shake flasks. Then, in each case 0.1 mmol substrate and 200 ul sterile 1 M glucose solution were added and incubated at 30 0 C and 200 rpm on an orbital shaker for 15 to 16 hours. Each reaction was extracted once with 10 ml of ethyl acetate, dried over anhydrous sodium sulfate, and excess solvent was removed under reduced pressure. The samples were analyzed by gas chromatography.
  • the BVMO gene from P. fluorescens DSM50106 was PCR amplified and inserted into the L-rhamnose inducible expression vector pJOE3075. Thereby, the gene without its stop codon was linked in-frame with six histidine codons and a stop codon already present in the vector (His-tag) to give plasmid pJOE4072.6.
  • BVMO expression was first performed in E. coli JM109 with pJOE4072.6. different Temperatures were tested and the amount of BVMO in the soluble fraction and in inclusion bodies was examined by SDS-PAGE and Western blot. Western blot analysis was used to identify the correct protein band of approximately 56 kDa corresponding to the BVMO. The best results were obtained at 30 0 C because less enzyme was found in inclusion bodies here, compared to 37 ° C, and the entire expression level was higher than at 25 ° C. Nevertheless, the amount of soluble BVMO was relatively low. Therefore, the BVMO gene was subcloned into the multi-copy vector pET22b (+) and expressed in BL21 (DE3).
  • Racemic ⁇ -hydroxy ketones la-c (Reaction Scheme 1) synthesized by the method described by Smith and Levenberg (1981) were subjected to kinetic resolution by Baeyer-Villiger enzymatic oxidation to yield hydroxyalkyl acetates 2a-c (Reaction Scheme 1). Small amounts of the other possible products, methyl hydroxyacid 3a-c, were also formed (Scheme 1).
  • Table 3 Results of the kinetic resolution of hydroxy ketones la-c (see Reaction Scheme 1) using resting cells of E. coli JM109 pGro7 pJOE4072.6 at 30 0 C in shake flasks.
  • the BVMO from P. fluorescens preferably converts the (S) - ⁇ -hydroxy ketones to the corresponding (S) -hydroxyalkyl acetates.
  • reaction times could be significantly shortened by carrying out kinetic resolutions with a higher ratio of enzyme to substrate.
  • Reactions were performed with quiescent cells in small volumes of 1 ml of concentrated cell suspension in 2 ml tubes with air-permeable lids. When the same substrate concentrations were used as in shake flasks, high conversions could already be achieved after shorter reaction times (Table 4). In addition, the E values obtained were significantly higher.
  • Table 4 Results of the kinetic resolution of the hydroxyketones la-c (see reaction scheme 1) with resting cells of E. coli JM109 pGro7 pJOE4072.6 at 30 0 C in 2 ml tubes with air-permeable lids.
  • BVMO cyclohexanone monooxygenase
  • CPMO cyclopentanone monooxygenase
  • the CHMO shows a higher enantioselectivity in the oxidation of Ia (Scheme 1) compared to the BVMO from P. fluorescens DSM 50106, while in the conversion of Ib (Scheme 1) the E value is lower (Table 5).
  • This enzyme is likely to have a substrate inhibition in ⁇ -hydroxy ketone Ic (Reaction Scheme 1), but the enantioselectivity of the kinetic resolution of this compound is estimated to be relatively low.
  • the CPMO shows no economically acceptable E values for the Baeyer-Villiger oxidation of ⁇ -hydroxy ketones (Table 6).

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Abstract

L'invention concerne un procédé de préparation énantiosélective d'esters aliphatiques acycliques à partir de cétones aliphatiques acycliques racémiques par oxydation selon Baeyer-Villiger. L'invention porte également sur la séparation racémique enzymatique cinétique de cétones aliphatiques acycliques racémiques.
EP07785706A 2006-08-21 2007-08-20 Préparation énantiosélective d'esters aliphatiques acycliques et de cétones aliphatiques acycliques Withdrawn EP2054510A2 (fr)

Applications Claiming Priority (2)

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DE200610039189 DE102006039189B4 (de) 2006-08-21 2006-08-21 Enantioselektive Darstellung von aliphatischen azyklischen Estern und Ketonen
PCT/DE2007/001473 WO2008022627A2 (fr) 2006-08-21 2007-08-20 Préparation énantiosélective d'esters aliphatiques acycliques et de cétones aliphatiques acycliques

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CN110257348B (zh) * 2018-02-07 2021-03-30 凯莱英医药集团（天津）股份有限公司 一种单加氧酶突变体及其制备方法和应用
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