Intermittent enzyme catalytic reaction method

Abstract

The invention discloses a batch-type enzyme catalytic reaction method, which adopts a batch-type enzyme catalytic reaction device, the batch-type enzyme catalytic reaction device comprises a reaction kettle and a static mixer, and the reaction kettle and the static mixer are connected through an external circulation pipeline to form a closed loop; a reaction system circulates in a closed loop, and enzyme catalysis reaction is carried out at the same time; the reaction system comprises a reaction substrate and an enzyme catalysis system. By adopting the method, the enzyme catalytic reaction rate can be obviously improved, the production cost can be reduced, and the conversion rate can be improved.

Description

Technical Field

[0001] The present invention relates to an intermittent enzymatic reaction method. Background Art

[0002] In enzymatic reactions, there is generally a problem of low enzymatic reaction rate and efficiency due to the limited stability of enzymes and their relatively easy denaturation and inactivation. Further, in the process of scale-up production, there will be a problem of poor reproducibility. The conventional method is to adjust process conditions (such as increasing the amount of enzyme or reaction volume, etc.) or to modify the enzyme to improve its stability so as to increase the conversion rate. However, increasing the amount of enzyme or reaction volume will additionally increase the production cost; while evolving and modifying enzymes has a long cycle and high cost. Summary of the Invention

[0003] In order to solve the problems of low enzymatic reaction rate and efficiency in the prior art, the present invention provides an intermittent enzymatic reaction method. By using this method, the enzymatic reaction rate can be significantly increased, while the production cost can be reduced and the conversion rate can be improved.

[0004] The present invention solves the above technical problems through the following technical solutions.

[0005] The present invention provides an intermittent enzymatic reaction method, which uses an intermittent enzymatic reaction device. The intermittent enzymatic reaction device includes a reaction kettle and a static mixer, and the reaction kettle and the static mixer are connected into a closed loop through an external circulation pipeline; the reaction system circulates in the closed loop while the enzymatic reaction is carried out; wherein, the reaction system includes a reaction substrate and an enzymatic catalytic system.

[0006] In the present invention, preferably, a circulation pump is provided on the external circulation pipeline.

[0007] Among them, preferably, the circulation pump is preferably a peristaltic pump, a plunger pump, an injection pump, a diaphragm pump or a magnetic pump.

[0008] Among them, preferably, before the start of circulation, the reaction system is added to the reaction kettle; among them, preferably, the feeding sequence of the reaction system is: first add the enzymatic catalytic system, and then add the reaction substrate.

[0009] In the present invention, preferably, the ratio of the flow rate of the reaction system in the external circulation pipeline to the inner diameter of the external circulation pipeline is (20 - 400):1, wherein the unit of the flow rate is mL / min and the unit of the inner diameter is mm.

[0010] In the present invention, the intermittent enzymatic catalysis reaction method preferably further comprises the following steps: after the cycle ends, post-treat the product obtained from the enzymatic catalysis reaction; the post-treatment operation preferably includes: filtering the product to obtain a filter cake and a filtrate, rinsing the filter cake, extracting the filtrate, combining the rinsing liquid and the extraction liquid, and sequentially performing water removal, filtration and concentration to obtain the final product.

[0011] Among them, the filtration preferably uses diatomaceous earth.

[0012] Among them, the rinsing preferably uses ethyl acetate.

[0013] Among them, the extraction preferably uses ethyl acetate.

[0014] Among them, the water removal preferably uses anhydrous sodium sulfate.

[0015] Among them, the concentration is preferably vacuum concentration.

[0016] In the present invention, preferably, the flow rate of the reaction system in the outer circulation pipeline is 100 - 10000 mL / min, more preferably 150 - 3000 mL / min, and even more preferably 200 - 500 mL / min.

[0017] In the present invention, preferably, the inner diameter of the outer circulation pipeline is 0.5 - 200 mm, more preferably 1 - 10 mm, for example, 2 mm.

[0018] In the present invention, the reaction kettle can be a conventional reaction kettle in the art, for example, a three-neck glass jacket reaction flask.

[0019] In the present invention, preferably, a mechanical stirring device is provided on the reaction kettle.

[0020] Among them, the rotation speed of the mechanical stirring device is preferably 100 - 300 r / min.

[0021] In the present invention, the volume of the reaction kettle is preferably 0.25 - 1000 L, for example, 0.5 L.

[0022] In the present invention, the static mixer is preferably corrugated plate type, single spiral type, double spiral type or cross bar type.

[0023] In the present invention, preferably, the enzymatic catalysis reaction is an oxidation-reduction reaction, a hydrolysis reaction, an acetylation reaction or a transamination reaction.

[0024] Among them, the oxidation-reduction reaction is, for example, a ketone reduction reaction.

[0025] Among them, the hydrolysis reaction is, for example, an ester hydrolysis reaction.

[0026] In the present invention, preferably, the enzyme catalytic system includes an enzyme and a coenzyme recycling system.

[0027] Among them, the coenzyme recycling system includes, for example, glucose, glucose dehydrogenase (GDH), and NADP. NADP is a common coenzyme in the art, a substance formed by the ester bond binding of nicotinamide adenine dinucleotide and a phosphate molecule, and is widely present in the biological world.

[0028] In the present invention, the reaction system preferably further includes a buffer solution, and the buffer solution is preferably a phosphate buffer solution.

[0029] In the present invention, the reaction system preferably further includes a cosolvent, and the cosolvent is preferably n-heptane and / or dimethyl sulfoxide (DMSO).

[0030] In certain specific embodiments of the present invention, the enzyme-catalyzed reaction is a ketone reduction reaction, the reaction substrate includes a ketone, and the product formed by the ketone through the ketone reduction reaction includes an alcohol.

[0031] Among them, the type of the ketone is preferably a fatty ketone or an aromatic ketone.

[0032] Among them, the type of the alcohol is preferably a fatty alcohol or an aromatic alkyl alcohol.

[0033] Among them, in the enzyme catalytic system, the type of the enzyme is preferably ketoreductase (KRED) or alcohol dehydrogenase.

[0034] Among them, the reaction system preferably further includes a phosphate buffer solution, and the concentration of the phosphate buffer solution is, for example, 0.1 - 0.3 mol / L.

[0035] Among them, the pH value of the ketone reduction reaction is adjusted by a pH value regulator, and the pH value regulator is, for example, a NaOH solution.

[0036] Among them, the concentration of the NaOH solution is preferably 0.5 - 2 mol / L, for example, 1 mol / L.

[0037] Among them, the temperature of the ketone reduction reaction is preferably 10 - 60 °C, more preferably 25 - 35 °C.

[0038] In certain specific embodiments of the present invention, the ketone is an α-ester ketone, the product formed by the ketone through the ketone reduction reaction includes an α-ester alcohol, and the reaction formula of the ketone reduction reaction is as shown in Formula I:

[0039]

[0040] Among them, R1 is preferably selected from an aryl group, a fatty alkyl group, a cycloalkyl group, or a heterocyclic group.

[0041] Among them, the aryl group is, for example, a phenyl group.

[0042] Among them, R2 is preferably an alkyl group or an aryl group.

[0043] Among them, the alkyl group is, for example, a methyl group or an ethyl group.

[0044] Among them, the aryl group is, for example, a benzyl group.

[0045] Among them, the α-ester-based alcohol is preferably an R-type α-ester-based alcohol.

[0046] Among them, the mass ratio of the ketone to the enzyme is preferably 50:1.

[0047] Among them, the ketone reduction reaction is preferably carried out at a pH value of 7.5 - 8.5.

[0048] Among them, preferably, the enzyme catalytic system includes KRED, glucose, GDH, and NADP.

[0049] Among them, the mass ratio of KRED, GDH, and NADP is 1:0.5:1.25.

[0050] Among them, the mass ratio of glucose to the ketone is preferably (2 - 3):1.

[0051] Among them, preferably, the reaction system further includes a phosphate buffer solution and n-heptane.

[0052] Among them, the volume ratio of the phosphate buffer solution to the mass of the ketone is preferably (10 - 20):1, for example, 15:1. The unit of volume is mL, and the unit of mass is g.

[0053] In some specific embodiments of the present invention, the enzyme-catalyzed reaction is an ester hydrolysis reaction, the reaction substrate includes an ester, and the products formed by the enzyme-catalyzed reaction of the ester include an alcohol and a carboxylic acid.

[0054] Among them, the ester is, for example, a fatty ester, an aromatic ester, or a cyclic ester.

[0055] Among them, the ester is, for example, a racemic ester; when the ester is a racemic ester, the products formed by the enzyme-catalyzed reaction of the ester further include a single configuration ester.

[0056] Among them, preferably, the products formed by the enzyme-catalyzed reaction of the ester include an S-ester, an R-alcohol, and a carboxylic acid.

[0057] Among them, preferably, the products formed by the enzyme-catalyzed reaction of the ester include an R-ester, an S-alcohol, and a carboxylic acid.

[0058] Among them, preferably, the products formed by the enzyme-catalyzed reaction of the ester include an S-ester, an alcohol, and an R-carboxylic acid.

[0059] Among them, preferably, the products formed by the enzyme-catalyzed reaction of the ester include R-ester, alcohol and S-carboxylic acid.

[0060] Among them, in the enzyme-catalyzed system, the type of the enzyme is preferably a hydrolase, and the hydrolase is, for example, one or more of lipase, esterase and protease.

[0061] Among them, the reaction system preferably further includes a phosphate buffer solution, and the concentration of the phosphate buffer solution is, for example, 0.1-0.3 mol / L.

[0062] Among them, the pH value of the ester hydrolysis reaction is adjusted by, for example, a pH value regulator, and the pH value regulator is, for example, a NaOH solution.

[0063] Among them, the concentration of the NaOH solution is preferably 0.5-2 mol / L, and is, for example, 1 mol / L.

[0064] Among them, the temperature of the ester hydrolysis reaction is preferably 10-60 °C, and more preferably 15-30 °C.

[0065] Among them, preferably, a mechanical stirring device is provided on the reaction kettle.

[0066] Among them, the rotation speed of the mechanical stirring device is preferably 50-1000 r / min, and more preferably 100-350 r / min.

[0067] In some specific embodiments of the present invention, the ester is a racemic ester, the products formed by the enzyme-catalyzed reaction of the ester include S-ester, R-alcohol and carboxylic acid, and the reaction formula of the ester hydrolysis reaction is as shown in Formula II:

[0068]

[0069] Among them, R3 is preferably selected from an aryl group, an aliphatic alkyl group, a cycloalkyl group, a heterocyclic group, an ester group or an alkoxy group.

[0070] Among them, the aryl group is, for example, a benzyl group.

[0071] Among them, the aliphatic alkyl group is, for example, a methyl group or an ethyl group.

[0072] Among them, R4 is preferably selected from an aryl group, an aliphatic alkyl group, a cycloalkyl group or a heterocyclic group.

[0073] Among them, the aliphatic alkyl group is, for example, a methyl group.

[0074] Among them, the mass ratio of the ester to the enzyme is preferably 5:1.

[0075] Among them, the type of the enzyme is, for example, lipase.

[0076] Among them, the hydrolysis reaction of the ester is preferably carried out at a pH value of 8.5 - 9.5.

[0077] Among them, preferably, the reaction system further includes a phosphate buffer solution and dimethyl sulfoxide.

[0078] Among them, the volume ratio of the phosphate buffer solution to the mass of the ester is preferably (15 - 30):1, for example, 20:1. The unit of volume is mL, and the unit of mass is g.

[0079] On the basis of conforming to the common knowledge in the art, the above preferred conditions can be combined arbitrarily to obtain various preferred examples of the present invention.

[0080] The reagents and raw materials used in the present invention are all commercially available.

[0081] The positive and progressive effects of the present invention are as follows:

[0082] The batch enzyme-catalyzed reaction method of the present invention has a high conversion rate, a high reaction rate, and a short reaction time. At the same time, it can also reduce the dosage of reactants such as enzymes, reduce production costs, and is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0083] Figure 1 It is a schematic diagram of the enzyme-catalyzed reaction device of the present invention.

[0084] Figure 2 It is a conversion rate diagram at different times for Example 2 and Comparative Example 4.

[0085] The reference numerals are as follows:

[0086] 1 - reaction kettle; 2 - static mixer, 3 - circulation pump. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0087] The present invention will be further described below by way of examples, but the present invention is not limited to the scope of the examples described herein. The experimental methods without specific conditions noted in the following examples are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0088] Example 1

[0089] Example 1 uses a batch enzyme-catalyzed reaction device. The batch enzyme-catalyzed reaction device includes a reaction kettle 1 and a static mixer 2. The static mixer 2 is single-spiral. The reaction kettle 1 and the static mixer 2 are connected into a closed loop through an external circulation pipeline. A circulation pump 3 is provided on the external circulation pipeline. The circulation pump 3 is a peristaltic pump, and the inner diameter of the external circulation pipeline is 2 mm; the batch enzyme-catalyzed reaction device is as Figure 1 shown.

[0090] Among them, the reaction kettle is a 250 mL three-necked glass jacketed reaction flask, equipped with a mechanical stirring device, and the rotation speed of the mechanical stirring device is 240 r / min.

[0091] The reaction system circulates in a closed loop while the enzymatic reaction is carried out; the reaction system includes a reaction substrate and an enzymatic catalytic system. The reaction formula of the ketone reduction reaction is as follows: Formula I:

[0092]

[0093] Among them, the ketone is an α-ester group ketone, R1 is a phenyl group, R2 is a methyl group, and the generated product includes the R-type α-ester group alcohol.

[0094] Among them, the enzymatic catalytic system includes 0.1 g of KRED, 12.8 g of glucose monohydrate, 0.125 g of NADP, and 0.05 g of GDH.

[0095] The reaction system also includes 75 mL of phosphate buffer solution with a concentration of 0.1 mol / L, and the enzymatic catalytic system is added to the phosphate buffer solution and stirred evenly. The temperature of the enzymatic reaction is controlled at 25 °C. Start the static mixer device to make the flow rate in the external circulation pipeline 200 mL / min. Then, a solution containing 5 g of the reaction substrate ketone and 2.5 mL of n-heptane is added dropwise to carry out the ketone reduction reaction.

[0096] During the reaction process, 1 mol / L NaOH solution is used to control the reaction pH to 8. The reaction solution is filtered with diatomaceous earth, the filter cake is rinsed with ethyl acetate, and the filtrate is extracted with ethyl acetate. Anhydrous sodium sulfate is added to remove water, and then filtered to obtain the organic phase, which is concentrated under reduced pressure to obtain the product alcohol.

[0097] Example 2

[0098] Example 2 uses the same batch enzymatic reaction device as Example 1.

[0099] The reaction formula of the ketone reduction reaction in Example 2 is as follows: Formula I:

[0100]

[0101] Among them, the ketone is an α-ester group ketone, R1 is a phenyl group, R2 is a methyl group, and the generated product includes the R-type α-ester group alcohol.

[0102] The addition amount of the phosphate buffer solution in Example 2 is 50 mL, and the other process parameters are the same as those in Example 1.

[0103] Comparative Example 1

[0104] The batch enzymatic reaction device in Comparative Example 1 only includes a reaction kettle and does not include an external circulation pipeline.

[0105] For Comparative Example 1, using the above reaction device, the addition amount of phosphate buffer solution was 50 mL, the enzyme catalytic system included 0.1 g of KRED, 12.8 g of glucose monohydrate, 0.125 g of NADP, and 0.25 g of GDH, and the remaining process parameters were the same as those in Example 1.

[0106] Comparative Example 2

[0107] Comparative Example 2 used the same batch enzyme catalytic reaction device as Comparative Example 1.

[0108] The addition amount of phosphate buffer solution in Comparative Example 2 was 50 mL, and the remaining process parameters were the same as those in Example 1.

[0109] Comparative Example 3

[0110] Comparative Example 3 used the same batch enzyme catalytic reaction device as Comparative Example 1.

[0111] The process parameters of Comparative Example 3 were the same as those in Example 1.

[0112] Comparative Example 4

[0113] Comparative Example 4 used the same batch enzyme catalytic reaction device as Comparative Example 1.

[0114] In the first 4 h of the reaction in Comparative Example 4, the process parameters were the same as those in Example 1. At 4 h of the reaction, 0.05 g of KRED was added and the reaction continued.

[0115] Example 3

[0116] Example 3 used a batch enzyme catalytic reaction device, which included a reaction kettle 1 and a static mixer 2. The reaction kettle 1 and the static mixer 2 were connected into a closed loop through an external circulation pipeline. A circulation pump 3 was provided on the external circulation pipeline, and the inner diameter of the external circulation pipeline was 2 mm; the batch enzyme catalytic reaction device was as Figure 1 shown.

[0117] Among them, the reaction kettle was a 500 mL three-necked glass jacketed reaction flask, and a mechanical stirring device was provided. The rotation speed of the mechanical stirring device was 300 r / min.

[0118] The reaction system circulated in the closed loop while the ester hydrolysis reaction was carried out; the reaction system included a reaction substrate and an enzyme catalytic system. The reaction formula of the ester hydrolysis reaction was as follows: Formula II

[0119]

[0120] Among them, the reaction substrate was a racemic ester, R3 was a benzyl group, R4 was a methyl group, and the products formed included S-ester, R-alcohol, and carboxylic acid.

[0121] Among them, the enzyme catalytic system includes 2 g of lipase.

[0122] The reaction system also includes 200 mL of phosphate buffer solution with a concentration of 0.1 mol / L. The enzyme catalytic system is added to the phosphate buffer solution and stirred evenly. The temperature of the enzyme catalytic reaction is controlled at 15°C. Start the static mixer device to make the flow rate in the external circulation pipeline 200 mL / min. Then, a solution containing 10 g of reaction substrate ketone and 80 mL of DMSO is added dropwise to carry out the ester hydrolysis reaction.

[0123] During the reaction process, 1 mol / L NaOH solution is used to control the reaction pH to 9. The reaction solution is filtered with diatomaceous earth, the filter cake is rinsed with ethyl acetate, and the filtrate is extracted with ethyl acetate. Anhydrous sodium sulfate is added to remove water, and then the organic phase is obtained by filtration and concentrated under reduced pressure to obtain the target product S-ester.

[0124] Comparative Example 5

[0125] The batch enzyme catalytic reaction device of Comparative Example 5 only includes a reaction kettle and does not include an external circulation pipeline, which is the same as the reaction device of Comparative Example 1.

[0126] The process parameters of Comparative Example 5 are the same as those of Example 3.

[0127] Effect Example 1

[0128] The molar amounts of each substance in Examples 1-2 and Comparative Examples 1-4 were determined using a liquid chromatography-mass spectrometry instrument.

[0129] Among them, the calculation formula for the conversion rate of Examples 1-2 and Comparative Examples 1-4 is: conversion rate = [(molar amount of initial reaction substrate - molar amount of current reaction substrate) / molar amount of initial reaction substrate] * 100%.

[0130] The contents of each substance in Example 3 and Comparative Example 5 were determined using a supercritical fluid chromatograph.

[0131] Among them, the calculation formula for the ee value of Example 3 and Comparative Example 5 is: ee value = (content of S-ester - content of racemic ester) / (content of S-ester + content of racemic ester).

[0132] The ee value, i.e., the enantiomeric excess value, is used to represent the optical purity of a chiral compound. The higher the ee value, the higher the optical purity.

[0133] The test results of Examples 1-2 and Comparative Examples 1-4 are listed in Table 1. The conversion rate diagrams of Example 2 and Comparative Example 4 at different times are as Figure 2 shown:

[0134] Table 1

[0135]

[0136] As can be seen from Table 1, the conversion rates of Example 1 and Example 2 can reach over 70% in 2 h, over 95% in 4 h, and after a long reaction time, the final conversion rate can reach over 98%.

[0137] Compared with the examples, the conversion rates of Comparative Example 2 and Comparative Example 3 decreased significantly.

[0138] Although the final conversion rates of Comparative Example 1 and Comparative Example 4 can reach over 95% after a long reaction time, the dosages of ketoreductase KRED or coenzyme GDH are high, the reaction time is long, and the production cost is high, which is not conducive to industrial production.

[0139] Compared with Comparative Example 3, Example 2 can not only reduce the dosage of buffer solution, but also increase the conversion rate.

[0140] The test results of Example 3 and Comparative Example 5 are listed in Table 2:

[0141] Table 2

[0142]

[0143] Note: / indicates not monitored.

[0144] As can be seen from Table 2, Example 3 can reach the reaction end point in 20 h, which can greatly shorten the reaction time. Compared with Comparative Example 5, the reaction time is shortened to 1 / 3 of that of Comparative Example 5, which can accelerate the R & D cycle and is suitable for industrial production.

Claims

1. An intermittent enzymatic reaction method, characterized in that, It adopts a batch enzyme-catalyzed reaction device, and the batch enzyme-catalyzed reaction device includes a reaction kettle and a static mixer. The reaction kettle and the static mixer are connected into a closed loop through an external circulation pipeline; the reaction system circulates in the closed loop while undergoing an enzyme-catalyzed reaction; wherein, the reaction system includes a reaction substrate and an enzyme-catalyzed system.

2. The intermittent enzyme-catalyzed reaction method according to claim 1, characterized in that, A circulation pump is provided on the external circulation pipeline; wherein, the circulation pump is preferably a peristaltic pump, a plunger pump, an injection pump, a diaphragm pump or a magnetic pump; And / or, before the start of circulation, the reaction system is added to the reaction kettle; wherein, preferably, the feeding sequence of the reaction system is: first add the enzyme-catalyzed system, and then add the reaction substrate; And / or, the ratio of the flow rate of the reaction system in the external circulation pipeline to the inner diameter of the external circulation pipeline is (20 - 400):1, wherein the unit of the flow rate is mL / min and the unit of the inner diameter is mm; And / or, the batch enzyme-catalyzed reaction method further includes the following steps: after the circulation ends, post-treat the product obtained by the enzyme-catalyzed reaction; the post-treatment operation preferably includes: filtering the product to obtain a filter cake and a filtrate, washing the filter cake, extracting the filtrate, combining the washing liquid and the extract, and successively performing water removal, filtration and concentration to obtain a final product.

3. The intermittent enzymatic reaction method according to claim 1, characterized in that, The flow rate of the reaction system in the external circulation pipeline is 100 - 10000 mL / min, preferably 150 - 3000 mL / min, more preferably 200 - 500 mL / min; And / or, the inner diameter of the external circulation pipeline is 0.5 - 200 mm, preferably 1 - 10 mm, for example 2 mm; And / or, a mechanical stirring device is provided on the reaction kettle; wherein, the rotation speed of the mechanical stirring device is preferably 100 - 300 r / min; And / or, the volume of the reaction kettle is 0.25 - 1000 L, for example 0.5 L; And / or, the static mixer is in a corrugated plate shape, a single spiral shape, a double spiral shape or a horizontal strip shape.

4. The intermittent enzymatic catalysis reaction method according to claim 1, characterized in that, The enzyme-catalyzed reaction is an oxidation-reduction reaction, a hydrolysis reaction, an acetylation reaction or a transamination reaction; wherein, the oxidation-reduction reaction is, for example, a ketone reduction reaction; wherein, the hydrolysis reaction is, for example, an ester hydrolysis reaction; And / or, the enzyme-catalyzed system includes an enzyme and a coenzyme recycling system; wherein, the coenzyme recycling system includes, for example, glucose, glucose dehydrogenase and NADP; And / or, the reaction system further includes a buffer solution, and the buffer solution is preferably a phosphate buffer solution; And / or, the reaction system further includes a co-solvent, and the co-solvent is preferably n-heptane and / or dimethyl sulfoxide.

5. The intermittent enzymatic reaction method according to any one of claims 1-4, characterized in that, The enzyme-catalyzed reaction is a ketone reduction reaction, the reaction substrate includes a ketone, and the product formed by the ketone through the ketone reduction reaction includes an alcohol.

6. The intermittent enzyme-catalyzed reaction method according to claim 5, characterized in that, The type of the ketone is a fatty ketone or an aromatic ketone; And / or, the type of the alcohol is a fatty alcohol or an aromatic alkyl alcohol; And / or, in the enzyme-catalyzed system, the type of the enzyme is a ketoreductase or an alcohol dehydrogenase; And / or, the reaction system further includes a phosphate buffer solution, and the concentration of the phosphate buffer solution is, for example, 0.1 - 0.3 mol / L; And / or, the pH value of the ketone reduction reaction is adjusted with a pH value regulator, such as a NaOH solution; wherein, the concentration of the NaOH solution is preferably 0.5 - 2 mol / L, such as 1 mol / L; And / or, the temperature of the ketone reduction reaction is 10 - 60 °C, preferably 25 - 35 °C.

7. The intermittent enzymatic reaction method according to claim 6, wherein, The ketone is an α-ester group ketone, and the product formed by the ketone through the ketone reduction reaction includes α-ester group alcohol. The reaction formula of the ketone reduction reaction is as shown in Formula I: Wherein, R1 is preferably selected from an aromatic group, an aliphatic alkyl group, a cycloalkyl group or a heterocyclic group; wherein, the aromatic group is, for example, a phenyl group; Wherein, R2 is preferably an alkyl group or an aromatic group; wherein, the alkyl group is, for example, a methyl group or an ethyl group; wherein, the aromatic group is, for example, a benzyl group; Wherein, the α-ester group alcohol is preferably an R-type α-ester group alcohol; Wherein, the mass ratio of the ketone to the enzyme is preferably 50:1; Wherein, the ketone reduction reaction is preferably carried out at a pH value of 7.5 - 8.5; Wherein, preferably, the enzyme catalytic system includes KRED, glucose, GDH and NADP; wherein, the mass ratio of KRED, GDH and NADP is preferably 1:0.5:1.25; wherein, the mass ratio of glucose to the ketone is preferably (2 - 3):1; Wherein, preferably, the reaction system further includes a phosphate buffer solution and n-heptane; wherein, the volume ratio of the phosphate buffer solution to the mass of the ketone is preferably (10 - 20):1, such as 15:1, where the unit of volume is mL and the unit of mass is g.

8. The intermittent enzymatic catalysis reaction method according to any one of claims 1-4, characterized in that The enzyme-catalyzed reaction is an ester hydrolysis reaction. The reaction substrate includes an ester, and the product formed by the ester through the enzyme-catalyzed reaction includes an alcohol and a carboxylic acid.

9. The intermittent enzymatic reaction method according to claim 8, wherein The ester is a fatty ester, an aromatic ester or a cyclic ester; And / or, the ester is a racemic ester; when the ester is a racemic ester, the product formed by the ester through the enzyme-catalyzed reaction further includes a single configuration ester; wherein, preferably, the product formed by the ester through the enzyme-catalyzed reaction includes an S-ester, an R-alcohol and a carboxylic acid; wherein, preferably, the product formed by the ester through the enzyme-catalyzed reaction includes an R-ester, an S-alcohol and a carboxylic acid; wherein, preferably, the product formed by the ester through the enzyme-catalyzed reaction includes an S-ester, an alcohol and an R-carboxylic acid; wherein, preferably, the product formed by the ester through the enzyme-catalyzed reaction includes an R-ester, an alcohol and an S-carboxylic acid; And / or, in the enzyme catalytic system, the type of the enzyme is a hydrolase, such as one or more of a lipase, an esterase and a protease; And / or, the reaction system further includes a phosphate buffer solution, and the concentration of the phosphate buffer solution is, for example, 0.1 - 0.3 mol / L; And / or, the pH value of the ester hydrolysis reaction is adjusted with a pH value regulator, such as a NaOH solution; wherein, the concentration of the NaOH solution is preferably 0.5 - 2 mol / L, such as 1 mol / L; And / or, the temperature of the ester hydrolysis reaction is 10 - 60 °C, preferably 15 - 30 °C; And / or, a mechanical stirring device is provided on the reactor; wherein, the rotation speed of the mechanical stirring device is preferably 50 - 1000 r / min, more preferably 100 - 350 r / min.

10. The intermittent enzymatic reaction method according to claim 9, characterized in that, The ester is a racemic ester, and the products formed by the enzymatic reaction of the ester include S-ester, R-alcohol and carboxylic acid. The reaction formula of the ester hydrolysis reaction is as shown in Formula II: Wherein, R3 is preferably selected from an aryl group, an aliphatic alkyl group, a cycloalkyl group, a heterocyclic group, an ester group or an alkoxy group; the aryl group is, for example, a benzyl group; the aliphatic alkyl group is, for example, a methyl group or an ethyl group; Wherein, R4 is preferably selected from an aryl group, an aliphatic alkyl group, a cycloalkyl group or a heterocyclic group; the aliphatic alkyl group is, for example, a methyl group; Wherein, the mass ratio of the ester to the enzyme is preferably 5:1; Wherein, the type of the enzyme is, for example, lipase; Wherein, the ester hydrolysis reaction is preferably carried out at a pH value of 8.5 - 9.5; Wherein, preferably, the reaction system further includes a phosphate buffer solution and dimethyl sulfoxide; wherein, the volume ratio of the phosphate buffer solution to the mass of the ester is preferably (15 - 30):1, for example, 20:1, wherein the unit of volume is mL and the unit of mass is g.

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