CN109486866A - The method that biocatalysis prepares (S) -4- decyl alcohol - Google Patents
The method that biocatalysis prepares (S) -4- decyl alcohol Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 38
- 238000006243 chemical reaction Methods 0.000 claims abstract description 70
- 108090000790 Enzymes Proteins 0.000 claims abstract description 28
- 102000004190 Enzymes Human genes 0.000 claims abstract description 28
- 238000006555 catalytic reaction Methods 0.000 claims abstract description 26
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims abstract description 19
- 101001110310 Lentilactobacillus kefiri NADP-dependent (R)-specific alcohol dehydrogenase Proteins 0.000 claims abstract description 15
- 239000002994 raw material Substances 0.000 claims abstract description 14
- XJLXINKUBYWONI-DQQFMEOOSA-N [[(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-3-hydroxy-4-phosphonooxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical compound NC(=O)C1=CC=C[N+]([C@@H]2[C@H]([C@@H](O)[C@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-DQQFMEOOSA-N 0.000 claims abstract description 13
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 claims abstract description 13
- MKJDUHZPLQYUCB-UHFFFAOYSA-N decan-4-one Chemical compound CCCCCCC(=O)CCC MKJDUHZPLQYUCB-UHFFFAOYSA-N 0.000 claims abstract description 12
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000001257 hydrogen Substances 0.000 claims abstract description 11
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 11
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 claims abstract description 10
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 9
- 239000008103 glucose Substances 0.000 claims abstract description 9
- 239000000203 mixture Substances 0.000 claims abstract description 9
- 239000005515 coenzyme Substances 0.000 claims abstract description 8
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 6
- 102000005751 Alcohol Oxidoreductases Human genes 0.000 claims abstract description 4
- 108010031132 Alcohol Oxidoreductases Proteins 0.000 claims abstract description 4
- 102000007698 Alcohol dehydrogenase Human genes 0.000 claims abstract description 3
- 108010021809 Alcohol dehydrogenase Proteins 0.000 claims abstract description 3
- 150000001299 aldehydes Chemical class 0.000 claims abstract description 3
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 claims abstract 4
- DTDMYWXTWWFLGJ-JTQLQIEISA-N (4s)-decan-4-ol Chemical compound CCCCCC[C@@H](O)CCC DTDMYWXTWWFLGJ-JTQLQIEISA-N 0.000 claims description 39
- 239000000243 solution Substances 0.000 claims description 26
- 239000000047 product Substances 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- 239000003153 chemical reaction reagent Substances 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 13
- 229910019142 PO4 Inorganic materials 0.000 claims description 11
- BZLVMXJERCGZMT-UHFFFAOYSA-N Methyl tert-butyl ether Chemical compound COC(C)(C)C BZLVMXJERCGZMT-UHFFFAOYSA-N 0.000 claims description 10
- 108010050375 Glucose 1-Dehydrogenase Proteins 0.000 claims description 9
- 238000000605 extraction Methods 0.000 claims description 9
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 8
- 239000007853 buffer solution Substances 0.000 claims description 5
- 239000012043 crude product Substances 0.000 claims description 5
- 239000003960 organic solvent Substances 0.000 claims description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 5
- 239000010452 phosphate Substances 0.000 claims description 5
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 4
- 238000006911 enzymatic reaction Methods 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 108090000854 Oxidoreductases Proteins 0.000 claims description 2
- 102000004316 Oxidoreductases Human genes 0.000 claims description 2
- 239000007864 aqueous solution Substances 0.000 claims description 2
- 238000004587 chromatography analysis Methods 0.000 claims description 2
- 239000012044 organic layer Substances 0.000 claims description 2
- 239000008363 phosphate buffer Substances 0.000 claims description 2
- 239000008366 buffered solution Substances 0.000 claims 1
- 239000003599 detergent Substances 0.000 claims 1
- 239000002904 solvent Substances 0.000 claims 1
- 238000010626 work up procedure Methods 0.000 claims 1
- 238000002360 preparation method Methods 0.000 abstract description 11
- 230000003287 optical effect Effects 0.000 abstract description 7
- 239000000126 substance Substances 0.000 abstract description 6
- 238000012805 post-processing Methods 0.000 abstract 1
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 21
- 238000003756 stirring Methods 0.000 description 12
- 238000004817 gas chromatography Methods 0.000 description 11
- 239000012074 organic phase Substances 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 7
- 229950006238 nadide Drugs 0.000 description 7
- 238000000926 separation method Methods 0.000 description 7
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 238000004821 distillation Methods 0.000 description 5
- 239000011521 glass Substances 0.000 description 5
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000012159 carrier gas Substances 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- 239000007818 Grignard reagent Substances 0.000 description 3
- 239000003054 catalyst Substances 0.000 description 3
- DTDMYWXTWWFLGJ-UHFFFAOYSA-N decan-4-ol Chemical compound CCCCCCC(O)CCC DTDMYWXTWWFLGJ-UHFFFAOYSA-N 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 150000004795 grignard reagents Chemical class 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 239000007810 chemical reaction solvent Substances 0.000 description 2
- MWKFXSUHUHTGQN-UHFFFAOYSA-N decan-1-ol Chemical compound CCCCCCCCCCO MWKFXSUHUHTGQN-UHFFFAOYSA-N 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- KWGKDLIKAYFUFQ-UHFFFAOYSA-M lithium chloride Chemical compound [Li+].[Cl-] KWGKDLIKAYFUFQ-UHFFFAOYSA-M 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 210000001130 astrocyte Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010364 biochemical engineering Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- ORTQZVOHEJQUHG-UHFFFAOYSA-L copper(II) chloride Chemical compound Cl[Cu]Cl ORTQZVOHEJQUHG-UHFFFAOYSA-L 0.000 description 1
- DOBRDRYODQBAMW-UHFFFAOYSA-N copper(i) cyanide Chemical compound [Cu+].N#[C-] DOBRDRYODQBAMW-UHFFFAOYSA-N 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 150000002118 epoxides Chemical class 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000011261 inert gas Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- FRIJBUGBVQZNTB-UHFFFAOYSA-M magnesium;ethane;bromide Chemical compound [Mg+2].[Br-].[CH2-]C FRIJBUGBVQZNTB-UHFFFAOYSA-M 0.000 description 1
- YCCXQARVHOPWFJ-UHFFFAOYSA-M magnesium;ethane;chloride Chemical compound [Mg+2].[Cl-].[CH2-]C YCCXQARVHOPWFJ-UHFFFAOYSA-M 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000005292 vacuum distillation Methods 0.000 description 1
Classifications
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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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
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- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The present invention discloses a kind of method that biocatalysis prepares (S) -4- decyl alcohol, is under the conditions of existing for the hydrogen source and coenzyme, to prepare (S) -4- decyl alcohol by enzymic catalytic reaction using 4- decanone as raw material；Wherein, enzyme used in enzymic catalytic reaction is selected from least one of ketoreductase, aldehyde ketone reductase, carbonyl reductase or alcohol dehydrogenase；The pH of enzymic catalytic reaction is 6~8, and reaction temperature is 15~50 DEG C；Hydrogen source is selected from the mixture of isopropanol or glucose or isopropanol and glucose；Coenzyme is selected from least one of NAD, NADH, NADP or NADPH.(S) -4- decyl alcohol product of high-optical-purity can be obtained with high conversion in preparation method of the invention, then by conventional post-processing operation, can high yield chiral product (S) -4- decyl alcohol that chemical purity is high, optical purity is high is prepared.
Description
Technical Field
The invention belongs to the technical field of biological pharmacy and biochemical engineering, and particularly relates to a method for preparing enantiomerically pure (S) -4-decanol by enzymatic asymmetric reduction.
Background
(S) -4-decanol (shown as formula II) is a common and widely used chiral alcohol. As the chiral center of the chiral alcohol is connected with an active hydroxyl functional group, the substance becomes a key chiral intermediate for synthesizing a plurality of important chiral drugs, agricultural chemicals and natural products. For example, (S) -4-decanol is an important chiral intermediate for preparing a drug for inhibiting astrocyte activation, and its chemical and optical purity determines the final quality of the chiral drug, so that improvement of the preparation method thereof is very important.
Currently, in the methods for preparing (S) -4-decanol, including the conventional chemical method and the bio-enzyme catalytic method, only one method is used to obtain chiral decanol with a single configuration by the chemical synthesis method of the grignard reagent.
Related literature (Organic Process Research and Development,2005,9(6), 774-. Specifically, cuprous cyanide and lithium chloride are added into a Grignard reagent EtMgBr for reaction and are used as catalysts to react for 2 hours at the temperature of-10-0 ℃; the addition of the Grignard reagent EtMgCl and the use of copper chloride as a catalyst required a reaction for 3.5 hours at-40 ℃ in an inert gas atmosphere. The reaction conditions for preparing (S) -4-decanol by the chemical method are relatively severe and need to be carried out at low temperature.
There is another related literature (Tetrahedron Asymmetry,2012,23(8), 583-. The reaction was carried out at 30 ℃ for 16 hours, and the conversion of the final product was only 51%, with 33% of form S. Although lipase is used as a catalyst in the method, the reaction condition is mild, but the conversion rate and the stereoselectivity are not ideal.
In addition, the biological preparation method for preparing the (S) -4-decanol with high optical purity by carrying out enzyme-catalyzed reduction on the 4-decanone has no related literature report at present.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a preparation method of (S) -4-decanol with high yield and high chiral purity, and mainly solve the technical problems that the existing preparation method has complex process conditions, is difficult to obtain a chiral product with high optical purity, and is difficult to realize large-scale preparation of (S) -4-decanol with high chiral purity.
In order to solve the technical problems, the invention provides a method for preparing (S) -4-decanol by biological catalysis, which takes 4-decanone (a compound shown in a formula I) as a raw material and prepares (S) -4-decanol (a compound shown in a formula II) by enzyme catalytic reaction in the presence of a hydrogen source and coenzyme; wherein,
the enzyme used for the enzyme-catalyzed reaction is at least one selected from ketoreductase, aldehyde ketoreductase, carbonyl reductase or alcohol dehydrogenase;
the pH of the enzyme catalysis reaction is 6-8, and the reaction temperature is 15-50 ℃;
the hydrogen source is selected from isopropanol, or glucose, or a mixture of isopropanol and glucose;
the coenzyme is selected from at least one of NAD, NADH, NADP or NADPH.
The specific reaction formula is as follows:
preferably, the enzyme used in the enzyme-catalyzed reaction is a ketoreductase. More preferably, the ketoreductase is a reductase available from the general pharmaceutical industry as reagent number R308055, R130709, R113020, R103014, or R103022.
Preferably, the dosage of the enzyme used in the enzyme catalysis reaction is 0.002-10 times of the mass of the raw materials; more preferably, the amount of the enzyme is 0.02 to 1 time of the mass of the raw material.
Preferably, the amount of the hydrogen source is 0.5-10 times of the mass of the raw materials; more preferably, Glucose Dehydrogenase (GDH) may be further added to the hydrogen source; the dosage of the Glucose Dehydrogenase (GDH) is 0.01-1 time of the mass of the raw material.
Preferably, the coenzyme is a combination of NAD and NADP. Wherein the amount of NAD is 0.001-0.1 times of the mass of the raw material, and the amount of NADP is 0.001-0.1 times of the mass of the raw material.
Preferably, the pH of the enzyme catalysis reaction is 6.2-7.5, and the reaction temperature is 20-40 ℃.
Specifically, the reaction solvent for enzyme catalysis is water or a mixture of water and an organic solvent. Preferably, the water is pure water or a phosphate buffered water solution; the phosphate buffer aqueous solution is KH containing phosphate with a concentration of 7.0-9.0 in the range of 0.1-2 mmol/L, pH2PO4-K2HPO4Or NaH2PO4-Na2HPO4And (4) a buffer solution. More preferably, the phosphate concentration is 0.2mol/L, and the pH is 6.2-7.5.
Specifically, the enzyme catalysis reaction is carried out by adding buffer solution or alkali liquor into the reaction system for pH control. The buffer solution comprises KH2PO4-K2HPO4Or NaH2PO4-Na2HPO4The buffer solution, commonly used lye comprises KOH or NaOH solution.
Specifically, the preparation method also comprises the step of carrying out post-treatment by a conventional method to separate the enantiomer of the product formula II. Conventional separation methods include extraction with organic solvents, ion exchange or chromatography. Among them, the enantiomer of formula II is preferably separated using a solvent extraction method. For example, after the reaction is finished, organic solvents such as dichloromethane or methyl tert-butyl ether are used for extraction, and after layering, an organic layer is obtained by separation, namely a crude product solution; drying the crude product solution with anhydrous sodium sulfate, filtering, directly concentrating under reduced pressure at a temperature of not higher than 45 deg.C, and finally obtaining colorless to light yellow liquid, i.e. the product (S) -4-decanol with enantiomeric excess value not less than 97% and purity not less than 95%.
In the preparation method, the reaction is carried out under the condition that the pH range is 6-8. When the pH is less than 6, the catalytic activity of the enzyme is significantly reduced. When the pH value is more than 8, the enzyme activity is also reduced. The preferable reaction pH is 6.2-7.5. The pH control is carried out by adding a buffer or an alkali solution to the reaction solvent.
In the preparation method, the reaction is carried out at the temperature of 15-60 ℃. When the temperature is lower than 15 ℃, the reaction speed is slow. However, above 60 ℃ the enzyme is irreversibly inactivated. The optimal reaction temperature is 20-40 ℃ for ensuring the stable and efficient reaction.
In the preparation method of the invention, (S) -4-decanol is prepared by carrying out asymmetric reduction on 4-decanone through an enzyme catalysis reaction. In the preparation method of the present invention, (S) -4-decanol product of high optical purity can be obtained at high conversion rate by enzymatic reaction. Then, the chiral product (S) -4-decanol with high chemical purity and high optical purity can be prepared with high yield by conventional post-treatment operation.
The invention has the beneficial effects that the enantiomeric excess value of the (S) -4-decanol prepared by the enzyme catalysis method is not less than 97 percent, and the purity is not less than 95 percent. Compared with the traditional method which uses ester or epoxide as raw materials and has relatively harsh reaction conditions, the method for efficiently producing the (S) -4-decanol with enantiomerically pure by a novel enzyme catalysis technology has important significance. The method is simple and easy to operate, the product obtained by post-treatment has high yield and good selectivity, and is suitable for large-scale industrial production.
Drawings
FIG. 1 is a high performance gas chromatography spectrum of a racemate of 4-decanol.
FIG. 2 is a chiral high performance gas chromatography spectrum of 4-decanol converted according to the method of example 6 of the present invention, in which the peak at 12.286min is the starting material 4-decanone and the peak at 19.299min is the target compound (S) -4-decanol.
FIG. 3 is a chiral high performance gas chromatography spectrum of 4-decanol converted according to the method of example 2 of the present invention, in which the peak at 19.821min is the target compound (S) -4-decanol.
FIG. 4 is a hydrogen nuclear magnetic spectrum of (S) -4-decanol after conversion according to the method of example 1 of the present invention. 1H NMR (400MHz, CDCl3) delta ppm 0.82-0.96(m,6H)1.21-1.51(m,15H)3.54-3.64(m, 1H).
Detailed Description
The technical solutions of the present invention will be described clearly and completely below, and it should be apparent that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
The reagents and starting materials used in the present invention are commercially available.
Example 1
In a 50mL glass jacketed reaction flask, 20mL water, 0.212g KH was added2PO4And 0.557g K2HPO4Stirring until the solid is dissolved; adding potassium hydroxide to adjust the pH value to 6.2-7.5; is turned to the reverse directionAdding 3g glucose, 1g ketoreductase (commercial reagent, available from general pharmaceutical industry, reagent No. R308055), 0.02g NADP and 0.2g GDH into the reaction solution, stirring, and adding 2g 4-decanone; the reaction solution was adjusted and controlled to have a temperature of 25 ℃ and a pH of 7.0, and stirred continuously for 24 hours.
The reaction solution was extracted with 16mL of methyl t-butyl ether, the organic phase was collected, a small amount of anhydrous sodium sulfate was added to remove water, and the mixture was filtered, and concentrated by distillation at 30 ℃ under reduced pressure. The obtained light yellow liquid, namely the product (S) -4-decanol, has the crude yield of 92.7 percent, and the conversion rate of 99.4 percent and the enantiomeric excess value of 97.6 percent by analyzing through a high performance gas chromatography. Analysis conditions were as follows: shimadzu gas chromatograph, AgilentCycyclosil-B (30X 0.25mm, 0.25 μm) chiral chromatographic column, constant temperature 110 deg.C for 25min, carrier gas is nitrogen, carrier gas flow rate is 30mL/min, and diluent is methyl tert-butyl ether. The NMR spectrum of the product is shown in FIG. 4.
After the enzyme catalysis reaction is finished, the (S) -4-decanol product with high yield and high purity can be obtained through simple extraction, liquid separation and concentration operations.
Analysis of the extracted organic phase revealed that (S) -4-decanol having high optical purity was produced at a high conversion rate after completion of the enzymatic reaction.
Example 2
In a 50mL glass jacketed reaction flask, 20mL water, 0.212g KH was added2PO4And 0.557g K2HPO4Stirring until the solid is dissolved; adding potassium hydroxide to adjust the pH value to 6.2-7.5; to the reaction solution were added 2g of glucose, 0.02g of ketoreductase (commercial reagent, available from Heyu Kagaku Kogyo, reagent No. R130709), 0.02g of NADP, and 0.1g of GDH, and after stirring, 1g of 4-decanone was added; the reaction solution was adjusted and controlled to have a temperature of 25 ℃ and a pH of 7.0, and stirred continuously for 24 hours.
The reaction solution was extracted with 16mL of methyl t-butyl ether, the organic phase was collected, a small amount of anhydrous sodium sulfate was added to remove water, and the mixture was filtered, and concentrated by distillation at 30 ℃ under reduced pressure. The obtained light yellow liquid, namely the product (S) -4-decanol, has the crude yield of 94.5 percent, and the conversion rate of 100 percent and the enantiomeric excess value of 100 percent are analyzed by high performance gas chromatography. The analysis conditions were the same as in example 1. The product has a gas chromatography pattern shown in FIG. 2.
After the enzyme catalysis reaction is finished, the (S) -4-decanol product with high yield and high purity can be obtained through simple extraction, liquid separation and concentration operations.
Example 3
In a 50mL glass jacketed reaction flask, 20mL water, 0.212g KH was added2PO4And 0.557g K2HPO4Stirring until the solid is dissolved; adding potassium hydroxide to adjust the pH value to 7.5; 3g of isopropyl alcohol, 1g of ketoreductase (commercial reagent, available from the general pharmaceutical industry, reagent No. R130709) and 0.02g of NAD were added to the reaction solution, and 2g of 4-decanone was added thereto after stirring; the temperature of the reaction solution was adjusted and controlled to 25 ℃ and pH 7.0, and the reaction solution was stirred continuously for 24 hours.
The reaction solution was extracted with 16mL of methyl t-butyl ether, the organic phase was collected, a small amount of anhydrous sodium sulfate was added to remove water, and the mixture was filtered, and concentrated by distillation at 30 ℃ under reduced pressure. The obtained light yellow liquid, namely the product (S) -4-decanol, has the crude yield of 96.2 percent, and the conversion rate of 100 percent and the enantiomeric excess value of 100 percent are analyzed by a high performance gas chromatography. The analysis conditions were the same as in example 1.
After the enzyme catalysis reaction is finished, the (S) -4-decanol product with high yield and high purity can be obtained through simple extraction, liquid separation and concentration operations.
Example 4
In a 50mL glass-jacketed reaction flask, 20mL of water, 0.212g of NaH was added2PO4And 0.557g Na2HPO4Stirring until the solid is dissolved; adding potassium hydroxide to adjust the pH value to 7; adding 3g of isopropanol, 1g of ketoreductase (commercial reagent, purchased from general pharmaceutical industry, reagent number R308055) and 0.02g of NADP into the reaction solution, stirring, and adding 2g of 4-decanone; adjusting and controlling the temperature of the reaction solutionStirring was continued at 37 ℃ and pH 7.0 for 24 hours.
The reaction solution was extracted with 20mL of dichloromethane, and the organic phase was collected, dehydrated by addition of a small amount of anhydrous sodium sulfate, filtered, and concentrated by distillation at 30 ℃ under reduced pressure. The obtained light yellow liquid, namely the product (S) -4-decanol, has the crude yield of 90.1 percent, and the conversion rate of 96.2 percent and the enantiomeric excess value of 97.1 percent by analyzing through a high performance gas chromatography. The analysis conditions were the same as in example 1.
After the enzyme catalysis reaction is finished, the (S) -4-decanol product with high yield and high purity can be obtained through simple extraction, liquid separation and concentration operations.
Example 5
In a 50mL glass jacketed reaction flask, 20mL water, 0.212g KH was added2PO4And 0.557g K2HPO4Stirring until the solid is dissolved; adding potassium hydroxide to adjust the pH value to 6; to the reaction solution were added 4g of glucose, 1g of ketoreductase (commercially available reagent, available from general pharmaceutical industries, reagent No. R308055), 0.02g of NAD and 1g of GDH, and after stirring, 2g of 4-decanone was added; the reaction solution was stirred continuously for 24 hours while adjusting and controlling the temperature at 25 ℃ and pH 6.0.
The reaction solution was extracted with 22mL of methyl t-butyl ether, the organic phase was collected, a small amount of anhydrous sodium sulfate was added to remove water, and the mixture was filtered, and concentrated by distillation at 30 ℃ under reduced pressure. The obtained light yellow liquid, namely the product (S) -4-decanol, has the crude yield of 90.1 percent, and the conversion rate of 94.7 percent and the enantiomeric excess value of 97.1 percent by analyzing through a high performance gas chromatography. The analysis conditions were the same as in example 1.
After the enzyme catalysis reaction is finished, the (S) -4-decanol product with high yield and high purity can be obtained through simple extraction, liquid separation and concentration operations.
Example 6
In a 1L glass jacketed reaction flask, 500mL of water and 5.3g of KH were added2PO4And 13.9g K2HPO4Stirring until the solid is dissolved; adding potassium hydroxide to adjust the pH value to 7.0; adding 30 g of isopropanol, 25g of ketoreductase (commercial reagent, purchased from general pharmaceutical industry, reagent number R308055) and 500mg of NAD (nicotinamide adenine dinucleotide), stirring, and adding 50g of 4-decanone; the reaction solution was adjusted and controlled to have a temperature of 25 ℃ and a pH of 7.0, and stirred continuously for 24 hours.
After the reaction, 400mL and 300mL of dichloromethane and 50g of diatomite are respectively added into the reaction solution, stirred for 2h, extracted twice, filtered and separated, and then the lower organic phase is taken out. The celite cake was washed with another 100mL of dichloromethane and the organic phases were combined after repeating the above procedure. The organic phase was concentrated by vacuum distillation at 40 ℃ to obtain a pale yellow liquid, i.e., the product (S) -4-decanol, with a crude yield of 81.7%, a conversion of 99.9% and an enantiomeric excess of 97.5% as determined by high performance gas chromatography. Analysis conditions were as follows: shimadzu gas chromatograph, Agilent Cyclosil-B (30X 0.25mm, 0.25 μm) chiral chromatographic column, constant temperature 110 deg.C for 25min, carrier gas is nitrogen, carrier gas flow rate is 30mL/min, and diluent is ethanol. The product has a gas chromatography pattern shown in FIG. 3.
In summary, the above embodiments are merely preferred embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalents, improvements, etc. made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (15)
1. A method for preparing (S) -4-decanol by biological catalysis is characterized in that 4-decanone is used as a raw material, and the (S) -4-decanol is prepared by enzyme catalysis reaction in the presence of a hydrogen source and coenzyme; wherein,
the enzyme used for the enzyme-catalyzed reaction is at least one selected from ketoreductase, aldehyde ketoreductase, carbonyl reductase or alcohol dehydrogenase;
the pH of the enzyme catalysis reaction is 6-8, and the reaction temperature is 15-50 ℃;
the hydrogen source is selected from isopropanol, or glucose, or a mixture of isopropanol and glucose;
the coenzyme is selected from at least one of NAD, NADH, NADP or NADPH.
2. The method of claim 1, wherein the enzyme catalyzing the reaction is a ketoreductase.
3. The method of claim 2, wherein the ketoreductase is a reductase available from the syndet pharmaceutical industry as reagent No. R308055, R130709, R113020, R103014, or R103022.
4. The method of claim 1, wherein the amount of enzyme used in the enzyme-catalyzed reaction is 0.002 to 10 times the mass of the raw material.
5. The method according to claim 4, wherein the amount of the enzyme is 0.02 to 1 times the mass of the raw material.
6. The method of claim 1, wherein the hydrogen source is used in an amount of 0.5 to 10 times the mass of the feedstock.
7. The method of claim 6, wherein glucose dehydrogenase may be further added to the hydrogen source; the dosage of the glucose dehydrogenase is 0.01-1 time of the mass of the raw materials.
8. The method of claim 1, wherein the coenzyme is a combination of NAD and NADP.
9. The method of claim 8, wherein the amount of NAD is 0.001 to 0.1 times and the amount of NADP is 0.001 to 0.1 times the mass of the raw material.
10. The method of claim 1, wherein the pH of the enzymatic reaction is 6.2 to 7.5 and the reaction temperature is 20 to 40 ℃.
11. The method of claim 1, wherein the solvent of the enzyme-catalyzed reaction is water, or a mixture of water and an organic solvent.
12. The method of claim 11, wherein the water is pure water or an aqueous phosphate buffered solution; the phosphate buffer aqueous solution is KH containing phosphate with a concentration of 7.0-9.0 in the range of 0.1-2 mmol/L, pH2PO4-K2HPO4Or NaH2PO4-Na2HPO4And (4) a buffer solution.
13. The process of claim 1 further comprising isolating the product (S) -4-decanol by work-up after the reaction is complete.
14. The method of claim 13, wherein the post-treatment is extraction or chromatography with an organic solvent.
15. The method of claim 14, wherein the post-treatment is extraction using dichloromethane or methyl tert-butyl ether, and after layering, an organic layer is separated, i.e. a crude product solution; and (3) directly concentrating the crude product solution under reduced pressure at the temperature of not higher than 45 ℃ until the crude product solution is dried, and finally obtaining colorless to light yellow liquid, namely the product (S) -4-decanol.
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| CN114958927A (en) * | 2021-02-22 | 2022-08-30 | 尚科生物医药（上海）有限公司 | Method for preparing (S) -1- (3-chlorphenyl) -1, 3-propylene glycol |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009042984A1 (en) * | 2007-09-28 | 2009-04-02 | Codexis, Inc. | Ketoreductase polypeptides and uses thereof |
| CN102876733A (en) * | 2012-10-23 | 2013-01-16 | 凯莱英医药集团（天津）股份有限公司 | Method for preparing alcohol compound with high optical purity |
| CN103131734A (en) * | 2013-02-21 | 2013-06-05 | 凯莱英医药集团（天津）股份有限公司 | Method for reducing ketone compound by using two alcohol dehydrogenases |
| CN107574194A (en) * | 2017-09-27 | 2018-01-12 | 上海合全药物研发有限公司 | The method that living things catalysis prepares the hydroxy piperidine of (R) 1 N benzene methoxycarbonyl group 3 |
-
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009042984A1 (en) * | 2007-09-28 | 2009-04-02 | Codexis, Inc. | Ketoreductase polypeptides and uses thereof |
| CN102876733A (en) * | 2012-10-23 | 2013-01-16 | 凯莱英医药集团（天津）股份有限公司 | Method for preparing alcohol compound with high optical purity |
| CN103131734A (en) * | 2013-02-21 | 2013-06-05 | 凯莱英医药集团（天津）股份有限公司 | Method for reducing ketone compound by using two alcohol dehydrogenases |
| CN107574194A (en) * | 2017-09-27 | 2018-01-12 | 上海合全药物研发有限公司 | The method that living things catalysis prepares the hydroxy piperidine of (R) 1 N benzene methoxycarbonyl group 3 |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114958927A (en) * | 2021-02-22 | 2022-08-30 | 尚科生物医药（上海）有限公司 | Method for preparing (S) -1- (3-chlorphenyl) -1, 3-propylene glycol |
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