Method for preparing cefalonium by enzyme method

Abstract

The invention discloses a method for preparing cefalonium through catalysis of immobilized penicillin G acylase. According to the method, immobilized penicillin G acylase is used as a catalyst, 7-IACA and 2-thiophenemethyl acetate are catalyzed to react in a mixed system of water and an organic solvent, and cefalonium is prepared. The organic solvent is one or more of ethanol, methanol and acetone. The preparation method disclosed by the invention is simple to operate, mild in reaction condition, suitable for industrial production and environment-friendly; the used immobilized penicillin G acylase can be repeatedly used, so that the production cost can be reduced; the molar yield of the cefalonium is high, the purity is 99% or above, impurities are few, the quality is high, and compared with a chemical preparation method, the cefalonium preparation method has obvious advantages.

Description

Technical Field

[0001] The present invention relates to the technical field of drug preparation, and more specifically, to a method for preparing cefuroxime by catalysis of immobilized penicillin G acylase. Background Art

[0002] Cefalonium is a cephalosporin antibiotic for animals developed by Schering-Plough Pharmaceuticals in the United States. It was developed as an udder injection for dry period cows and launched in the UK in 1993. It is mainly used to treat latent mastitis in dry period cows and prevent new dry period infections caused by sensitive bacteria such as Staphylococcus, Streptococcus, and Escherichia coli. It achieves bactericidal effect by inhibiting the synthesis of bacterial cell walls, has broad-spectrum antibacterial activity, and is stable to penicillinase and β-lactamase. At present, cefuroxime udder injection has been marketed in many countries around the world, with products covering Europe, Oceania, South America, and Asia. Since 2019, many domestic companies have obtained Class II new veterinary drug certificates for cefuroxime udder injection (dry period).

[0003]

[0004] At present, the reported synthesis methods of cefuroxime are mainly chemical methods, which use 2-thiopheneacetyl chloride and cephalosporin substrates in aqueous solution or organic solvent system for synthesis. For example, patent CN 103242346 A records that 7-IACA is used to synthesize cefuroxime with 2-thiopheneacetyl chloride in 0°C aqueous solution or a mixed system of water and organic solvent, with a purity of 99% and a yield of 38.2-45%.

[0005]

[0006] This method has a simple process, but a low yield, uses 2-thiopheneacetyl chloride which is highly toxic, and produces a large amount of three waste liquids, which is not environmentally friendly.

[0007] Patent CN 103772415 B records that cefotaxime is synthesized in one step by using 7-aminocephalosporanic acid in a mixed system of water and an organic solvent at 0 to 10° C. with 2-thiopheneacetyl chloride and isonicotinamide, with a purity of 98% and a yield of 65%.

[0008]

[0009] This method does not require separation and purification of intermediates, and the yield is improved; however, the isonicotinamide reaction takes 60 to 150 hours, the reaction cycle is long, the energy consumption is high, and the efficiency is low.

[0010] Patent CN 107445978 B records that D-7ACA is used to synthesize a cefuroxime intermediate with 2-thiopheneacetyl chloride in dichloromethane at 0-5°C, and isonicotinamide is used to synthesize cefuroxime in aqueous solution at 55-65°C, with a purity of 98% and a total yield of about 32%.

[0011]

[0012] This method uses D-7ACA instead of 7-ACA to reduce production costs and has a smooth process; however, cephalosporin compounds are generally unstable in high-temperature aqueous solutions and may decompose to produce impurities, resulting in low product yield and low purity.

[0013] Since the existing chemical preparation route uses 2-thiopheneacetyl chloride, which is highly toxic and is a dangerous substance specified by the state, it reacts violently with water and releases toxic gases. Therefore, it is necessary to use an excess of 2-thiopheneacetyl chloride to react under low temperature conditions (2-thiopheneacetyl chloride reacts with water and consumes part of the material), resulting in high energy consumption and a lot of three wastes. In order to overcome these shortcomings, people pay more and more attention to the green and environmentally friendly enzymatic preparation of cephalosporin antibiotics. In view of the fact that enzymatic synthesis, compared with chemical synthesis, not only has mild conditions, simple synthesis routes, and high synthesis efficiency; it also has the significant advantages of less three wastes and environmental friendliness; therefore, the enzymatic preparation of β-lactam antibiotics is increasingly used; but there is currently no report on the use of enzymatic preparation of cephalosporin. Summary of the invention

[0014] The object of the present invention is to provide a method for preparing cefuroxime by catalysis of immobilized penicillin G acylase. The method uses a green and environmentally friendly enzymatic method to prepare cefuroxime, which can effectively solve the problems of poor safety, high energy consumption, more three wastes, poor quality and high cost in the preparation process of cefuroxime in the prior art.

[0015] The above-mentioned object of the present invention is achieved through the following solutions:

[0016] A method for preparing cefuroxime by enzymatic method, using immobilized penicillin G acylase as a catalyst, catalyzing the reaction of 7-IACA and 2-thiopheneacetic acid methyl ester in a mixed system of water and an organic solvent, to prepare cefuroxime;

[0017] The organic solvent is one or more of ethanol, methanol and acetone.

[0018] Preferably, the organic solvent is ethanol and / or methanol.

[0019] The conventional synthesis method of the side chain at position 7 of cefuroxime is the chemical acyl chloride method, which uses 2-thiopheneacetyl chloride and cephalosporin substrate to react in an organic solvent or organic solvent-water system at 0-10°C. The 2-thiopheneacetyl chloride used is highly toxic and belongs to dangerous goods stipulated by the state. It reacts violently with water and releases toxic gases. Therefore, it is necessary to use an excessive amount of 2-thiopheneacetyl chloride to react under low temperature conditions, resulting in large energy consumption, many wastes, and poor safety. The present invention uses an enzyme method to replace the acyl chloride chemical method, and uses purified water and low-toxic organic solvents as the reaction solvent system at room temperature, which greatly reduces the three wastes, and the process is safer and more environmentally friendly; at the same time, the immobilized penicillin G acylase can be reused, effectively reducing costs, and is a green, environmentally friendly and economical method for synthesizing cefuroxime. And the reaction temperature is room temperature, which is a safe, environmentally friendly, mild and efficient method for synthesizing cefuroxime.

[0020] Preferably, the mass ratio of the total amount of water and organic solvent to 7-IACA is 10 to 20:1.

[0021] Preferably, the mass ratio of the total amount of water and organic solvent to 7-IACA is 10 to 15:1.

[0022] More preferably, the mass ratio of the total amount of water and organic solvent to 7-IACA is 15:1.

[0023] Preferably, the mass ratio of water to organic solvent is 11:4 to 14:1;

[0024] Preferably, the mass ratio of water to the organic solvent is 11:4, 13:2 or 14:1.

[0025] More preferably, when the organic solvent is ethanol, the mass ratio of water to ethanol is 11:4, 13:2 or 14:1.

[0026] More preferably, when the organic solvent is methanol, the mass ratio of water to methanol is 13:2 or 14:1.

[0027] More preferably, when the organic solvent is acetone, the mass ratio of water to acetone is 14:1.

[0028] Preferably, the mass ratio of the amount of immobilized penicillin G acylase to 7-IACA is 1.2-1.8:1.

[0029] Preferably, the mass ratio of the amount of immobilized penicillin G acylase to 7-IACA is 1.2-1.5:1.

[0030] More preferably, the mass ratio of the amount of immobilized penicillin G acylase to 7-IACA is 1.5:1.

[0031] Preferably, the reaction is carried out at 20-30°C.

[0032] More preferably, the reaction is carried out at 20-25°C.

[0033] Preferably, the reaction time is 4 to 7 hours.

[0034] More preferably, the reaction time is 4 to 5 hours.

[0035] Preferably, the molar ratio of 7-IACA to methyl 2-thiopheneacetate is 1:1.2 to 1.5.

[0036] Preferably, the pH value of the reaction system is 5.0-7.0.

[0037] More preferably, the pH value of the reaction system is 6.2±0.2.

[0038] Preferably, the pH value of the reaction system is adjusted using aqueous ammonia.

[0039] More preferably, the concentration of aqueous ammonia is 6M.

[0040] Preferably, the method for preparing cefuroxime by enzymatic method comprises: dissolving an appropriate amount of 7-IACA in water and an organic solvent, controlling the temperature, adjusting the pH value, then adding immobilized penicillin G acylase and 2-thiopheneacetic acid methyl ester, carrying out a catalytic reaction, separating and drying to obtain cefuroxime;

[0041] The organic solvent is one or more of ethanol, methanol and acetone.

[0042] Preferably, the organic solvent is ethanol and / or methanol.

[0043] Preferably, the mass ratio of the total amount of water and organic solvent to 7-IACA is 10 to 20:1.

[0044] Preferably, the mass ratio of the total amount of water and organic solvent to 7-IACA is 10 to 15:1.

[0045] More preferably, the mass ratio of the total amount of water and organic solvent to 7-IACA is 15:1.

[0046] Preferably, the mass ratio of water to the organic solvent is 11:4 to 14:1.

[0047] Preferably, the mass ratio of water to the organic solvent is 11:4, 13:2 or 14:1.

[0048] More preferably, when the organic solvent is ethanol, the mass ratio of water to ethanol is 11:4, 13:2 or 14:1.

[0049] More preferably, when the organic solvent is methanol, the mass ratio of water to methanol is 13:2 or 14:1.

[0050] More preferably, when the organic solvent is acetone, the mass ratio of water to acetone is 13:2 or 14:1.

[0051] Preferably, the mass ratio of immobilized penicillin G acylase to 7-IACA is 1.2-1.8:1.

[0052] Preferably, the mass ratio of immobilized penicillin G acylase to 7-IACA is 1.2-1.5:1.

[0053] More preferably, the mass ratio of immobilized penicillin G acylase to 7-IACA is 1.5:1.

[0054] Preferably, the reaction is carried out at 20-30°C.

[0055] More preferably, the reaction is carried out at 20-25°C.

[0056] Preferably, the reaction time is 4 to 7 hours.

[0057] More preferably, the reaction time is 4 to 5 hours.

[0058] Preferably, the molar ratio of 7-IACA to methyl 2-thiopheneacetate is 1:1.2 to 1.5.

[0059] Preferably, the pH value of the reaction system is 5.0-7.0.

[0060] More preferably, the pH value of the reaction system is 6.2±0.2.

[0061] Preferably, the pH value of the reaction system is adjusted using aqueous ammonia.

[0062] More preferably, the concentration of aqueous ammonia is 6M.

[0063] Compared with the prior art, the present invention has the following beneficial effects:

[0064] (1) The preparation method of the present invention is simple to operate, has mild reaction conditions, and is suitable for industrial production;

[0065] (2) The preparation method of the present invention is environmentally friendly;

[0066] (3) The enzyme used in the preparation method of the present invention is immobilized penicillin G acylase, which can be reused, which is beneficial to reducing production costs;

[0067] (4) The preparation method of the present invention has a high molar yield of cefuroxime, a purity of more than 99%, few impurities and high quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0068] Figure 1 The HPLC spectrum of cefuroxime prepared in Example 2 is shown in FIG. DETAILED DESCRIPTION

[0069] The present invention is further described in detail below in conjunction with specific examples, which are only used to explain the present invention and are not used to limit the scope of the present invention. The test methods used in the following examples are conventional methods unless otherwise specified; the materials and reagents used are reagents and materials that can be obtained from commercial channels unless otherwise specified.

[0070] In the following examples, 7-IACA and 2-thiopheneacetic acid methyl ester were used as reaction materials, and cefuroxime was synthesized enzymatically under the catalysis of immobilized penicillin G acylase. The synthesis route is as follows:

[0071]

[0072] The activity of immobilized penicillin G acylase has a great influence on the catalytic reaction. Therefore, it is necessary to maintain a high activity of immobilized penicillin G acylase during the reaction to improve the yield of cefuroxime. Therefore, the effects of solvent type, dosage ratio, reaction temperature, reaction time and enzyme dosage during the reaction were investigated. The dosage of 7-IACA was calculated based on the mass of 7-IACA·HI·HCl.

[0073] Example 1 Influence of the type of reaction solvent

[0074] 1.1 Effect of water as reaction solvent

[0075] The cefuroxime intermediate 7-IACA (30 g) is added to purified water, the pH is adjusted to 6.2±0.2 with aqueous ammonia (6 M), and the mixture is stirred until it is clear. Immobilized penicillin G acylase is added, and then 2-thiopheneacetic acid methyl ester is added. The enzyme catalyzed reaction is carried out at a pH of 6.2±0.2 and 22° C. The reaction is carried out for 5 hours, and the immobilized penicillin G acylase and the product suspension are separated. Finally, the product suspension is filtered, washed, and dried to obtain cefuroxime.

[0076] The amount of immobilized penicillin G acylase used was 1.3 times that of 7-IACA (mass ratio); the amount of 2-thiopheneacetic acid methyl ester used was 1.5 times that of 7-IACA (molar ratio).

[0077] The amount of water used is an investigation item, and the investigation range of the amount is 7 to 10 times (mass ratio) of 7-IACA.

[0078] The final yield of cefuroxime under different water dosages is shown in Table 1.

[0079] Table 1 Effect of water dosage on the yield of cefuroxime

[0080]

[0081] The results show that when the amount of solvent water is too little, as the reaction proceeds, the reaction system becomes viscous, which is not conducive to the contact between the reaction materials and the enzyme, thereby reducing the reaction rate; when the amount of solvent water is too much, the concentration of 7-IACA decreases, which also leads to a decrease in the reaction rate. From the above test results, it can be seen that when the solvent is water and its amount is 10 to 20 times that of 7-IACA (mass ratio), the yield of cefotaxime is relatively good, preferably 15 times the amount of 7-IACA (mass ratio).

[0082] However, in the above test results, the molar yield of cefotaxime still does not exceed 30%, which does not show an advantage in yield compared with the chemical preparation method. Therefore, the specific conditions of the reaction still need to be further optimized to improve the molar yield of the target product cefotaxime.

[0083] 1.2 Effect of the reaction solvent being a mixture of water and organic solvent

[0084] The test was performed with reference to the steps in 1.1, except that the solvent used was a mixture of water and an organic solvent, wherein the organic solvents were DMSO, ethanol, methanol and acetone, respectively, and the total amount of water and organic solvent was added at 15 times (mass ratio) of 7-IACA, and the influence of different usage ratios of water and organic solvent was investigated. The results are shown in Table 2.

[0085] Table 2 Effect of solvent type and ratio on the yield of cefuroxime

[0086]

[0087]

[0088] The results showed that when the reaction solvent was a mixed solvent of water and an organic solvent, the reaction speed was fast in a homogeneous reaction system, which increased the yield of the product; however, the type and dosage ratio of the organic solvent had different effects on the yield and quality of the product. When DMSO was used and the reaction system was 13 times water + 2 times DMSO, the yield of the product was low (38.3%) and the quality was poor (the residual amount of 7-IACA was 0.63%). The reason may be that DMSO had a destructive effect on the enzyme, resulting in a decrease in enzyme activity, which led to a low product yield; and when the organic solvent was ethanol, methanol or acetone, the yield of cefotaxime was significantly improved compared with the chemical preparation method and the enzymatic preparation method in 1.1 with only water as the reaction solvent; among them, the yields of experiments 6, 7, 8, 9, 10, 11, 12, 14, and 15 were better, all above 69.7%. When the organic solvent is ethanol, the yield of cefuroxime is good; when the organic solvent is methanol, the yield of cefuroxime is the best, which is 79.5%.

[0089] 1.3 Effect of reaction temperature

[0090] The test was performed with reference to the steps in 1.1, except that the solvent used was a mixture of water and methanol (mass ratio of 13:2), the total amount of water and methanol was added according to 15 times (mass ratio) of 7-IACA, the reaction temperature was changed, and its effect on the yield of cefuroxime was tested. The measured results are shown in Table 3.

[0091] Table 3 Effect of reaction temperature on the yield of cefuroxime

[0092]

[0093] The results showed that at 20-30°C, the enzyme-catalyzed synthesis of cefuroxime had little effect on the yield and quality. To facilitate the control of reaction conditions, the reaction temperature was preferably 20-25°C.

[0094] 1.4 Effect of reaction time

[0095] The test was performed with reference to the steps in 1.1, except that the solvent used was a mixture of water and methanol (mass ratio of 13:2), the total amount of water and methanol was added according to 15 times (mass ratio) of 7-IACA, the reaction time was changed, and its effect on the yield of cefuroxime was tested. The measured results are shown in Table 4.

[0096] Table 4 Effect of reaction time on the yield of cefuroxime

[0097]

[0098]

[0099] The results showed that when the reaction time was within 4 to 7 hours, there was no significant difference in the yield and quality of cefuroxime; considering the product preparation efficiency, the reaction time was preferably 5 hours.

[0100] 1.5 Effect of enzyme dosage

[0101] The test was performed with reference to the steps in 1.1, except that the solvent used was a mixture of water and methanol (mass ratio of 13:2), the total amount of water and methanol was added according to 15 times (mass ratio) of 7-IACA, the reaction time was 5h, and the amount of immobilized penicillin G acylase was changed to test its effect on the yield of cefuroxime. The measured results are shown in Table 5.

[0102] The amount of immobilized penicillin G acylase used is expressed as a multiple (mass ratio) of 7-IACA.

[0103] Table 5 Effect of different dosages of immobilized penicillin G acylase on the yield of cefuroxime

[0104]

[0105] The results showed that when the amount of immobilized penicillin G acylase was 1.2 to 1.8 times that of 7-IACA, there was no significant difference in the yield of cefuroxime; from the perspective of reaction cost, 1.2 or 1.5 times was preferred.

[0106] The above test shows that when the reaction materials are 7-IACA and methyl 2-thiopheneacetate, the reaction solvent is water and an organic solvent (ethanol, methanol or acetone), the total amount is 15 times the amount of 7-IACA (mass ratio), the mass ratio of water to the organic solvent is 11:4 to 14:1, the reaction temperature is 20 to 25°C, the reaction time is 5h, and the amount of immobilized penicillin G acylase is 1.2 to 1.8 times that of 7-IACA (mass ratio), cefuroxime has a good yield.

[0107] Example 2

[0108] 30g of 7-IACA, an intermediate of cefuroxime, was added to an enzyme reactor, 390g of purified water and 60g of methanol were added, stirring was started, ammonia water was added dropwise at 20°C to adjust the pH to 6.2±0.2, 45g of immobilized penicillin G acylase and 12.6g (0.08mol) of methyl 2-thiopheneacetate were added, and the enzyme catalyzed reaction was carried out at 20°C for 4 hours, the reaction was stopped, the valve at the bottom of the enzyme reactor was opened, the feed liquid was filtered out, the enzyme was washed with 50mL of cold purified water, the feed liquid was combined, and the enzyme was retained in the enzyme reactor for repeated use. The feed liquid was filtered, the filter cake was washed with purified water, dried, and vacuum dried to obtain 25.10g of cefuroxime, with a molar yield of 78.2% and a purity of 99.3%.

[0109] The HPLC chart of the prepared cefuroxime is as follows: Figure 1 As shown below.

[0110] Example 3

[0111] 30g of 7-IACA, an intermediate of cefuroxime, was added to an enzyme reactor, 390g of purified water and 60g of ethanol were added, stirring was started, ammonia water was added dropwise at 22°C to adjust the pH to 6.2±0.2, 36g of immobilized penicillin G acylase and 11.4g of methyl 2-thiopheneacetate were added, and the enzyme catalytic reaction was carried out at 22°C for 5 hours, the valve at the bottom of the enzyme reactor was opened, the feed liquid was filtered out, the enzyme was washed with 50mL of cold purified water, the feed liquid was combined, and the enzyme was retained in the enzyme reactor for repeated use. The feed liquid was filtered, the filter cake was washed with purified water, dried, and vacuum dried to obtain 29.2g of cefuroxime, with a molar yield of 80.2% and a purity of 99.1%.

[0112] Example 4

[0113] 30g of 7-IACA, an intermediate of cefuroxime, was added to an enzyme reactor, 390g of purified water and 60g of acetone were added, stirring was started, 4mol / L ammonia water was added dropwise at 30°C to adjust the pH to 6.2±0.2, 54g of immobilized penicillin G acylase and 13.3g of methyl 2-thiopheneacetate were added, and the enzyme catalytic reaction was carried out at 30°C for 7 hours, the reaction was stopped, the valve at the bottom of the enzyme reactor was opened, the feed liquid was filtered out, the enzyme was washed with 50ml of cold purified water, the feed liquid was combined, and the enzyme was retained in the enzyme reactor for repeated use. The feed liquid was filtered, the filter cake was washed with purified water, dried, and vacuum dried to obtain 29.4g of cefuroxime, with a molar yield of 81.0% and a purity of 99.3%.

[0114] Example 5

[0115] 30g of 7-IACA, an intermediate of cefuroxime, was added to an enzyme reactor, 330g of purified water and 120g of methanol were added, stirring was started, ammonia water was added dropwise at 25°C to adjust the pH to 6.2±0.2, 45g of immobilized penicillin G acylase and 14.2g of methyl 2-thiopheneacetate were added, and the enzyme catalyzed reaction was carried out at 25°C for 5 hours, the reaction was stopped, the valve at the bottom of the enzyme reactor was opened, the feed liquid was filtered out, the enzyme was washed with 50mL of cold purified water, the feed liquid was combined, and the enzyme was retained in the enzyme reactor for repeated use. The feed liquid was filtered, the filter cake was washed with purified water, dried, and vacuum dried to obtain 28.5g of cefuroxime, with a molar yield of 78.3% and a purity of 99.0%.

[0116] Example 6

[0117] 30g of 7-IACA, an intermediate of cefuroxime, was added to an enzyme reactor, 330g of purified water and 120g of ethanol were added, stirring was started, ammonia water was added dropwise at 25°C to adjust Ph to 6.2±0.2, 45g of immobilized penicillin G acylase and 12.6g of methyl 2-thiopheneacetate were added, and the enzyme catalytic reaction was carried out at 25°C for 5 hours, the reaction was stopped, the valve at the bottom of the enzyme reactor was opened, the feed liquid was filtered out, the enzyme was washed with 50mL of cold purified water, the feed liquid was combined, and the enzyme was retained in the enzyme reactor for repeated use. The feed liquid was filtered, the filter cake was washed with purified water, dried, and vacuum dried to obtain 29.3g of cefuroxime, with a molar yield of 80.4% and a purity of 99.1%.

[0118] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. For ordinary technicians in this field, other different forms of changes or modifications can be made based on the above descriptions and ideas. It is not necessary and impossible to list all the implementation methods here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A method for preparing cefuroxime by enzymatic method, characterized in that: Using immobilized penicillin G acylase as a catalyst, 7-IACA and 2-thiopheneacetic acid methyl ester are catalyzed to prepare cefuroxime in a mixed system of water and organic solvent. The organic solvent is one or more of ethanol, methanol and acetone.

2. The method for preparing cefuroxime by enzymatic method according to claim 1, characterized in that: The organic solvent is ethanol and / or methanol.

3. The method for preparing cefuroxime by enzymatic method according to claim 1, characterized in that: The mass ratio of the total amount of water and organic solvent to 7-IACA is 10 to 20:1; Preferably, the mass ratio of the total amount of water and organic solvent to 7-IACA is 10 to 15:1; More preferably, the mass ratio of the total amount of water and organic solvent to 7-IACA is 15:

1.

4. The method for preparing cefuroxime by enzymatic method according to claim 3, characterized in that: The mass ratio of water to organic solvent is 11:4 to 14:1; Preferably, the mass ratio of water to organic solvent is 11:4, 13:2 or 14:

1.

5. The method for preparing cefuroxime by enzymatic method according to claim 4, characterized in that: The mass ratio of the immobilized penicillin G acylase to 7-IACA is 1.2 to 1.8:1; Preferably, the mass ratio of the immobilized penicillin G acylase to 7-IACA is 1.2-1.5:

1.

6. The method for preparing cefuroxime by enzymatic method according to claim 5, characterized in that: The reaction is carried out at 20-30°C and the reaction time is 4-7h; Preferably, the reaction is carried out at 20 to 25°C; Preferably, the reaction time is 4 to 5 hours.

7. The method for preparing cefuroxime by enzymatic method according to claim 6, characterized in that: The molar ratio of 7-IACA to 2-thiopheneacetic acid methyl ester is 1:1.2-1.5; the pH value of the reaction system is 5.0-7.0; Preferably, the pH value of the reaction system is 6.2±0.

2.

8. The method for preparing cefuroxime by enzymatic method according to any one of claims 1 to 7, characterized in that: Dissolve an appropriate amount of 7-IACA in water and an organic solvent, control the temperature, adjust the pH value, then add immobilized penicillin G acylase and 2-thiopheneacetic acid methyl ester to carry out a catalytic reaction, and after the reaction is completed, separate and dry to obtain cefuroxime; The organic solvent is one or more of ethanol, methanol and acetone.

9. The method for preparing cefuroxime by enzymatic method according to claim 8, characterized in that: The mass ratio of the total amount of water and organic solvent to 7-IACA is 10 to 20:1; Preferably, the mass ratio of the total amount of water and organic solvent to 7-IACA is 10 to 15:1; More preferably, the mass ratio of the total amount of water and organic solvent to 7-IACA is 15:

1.

10. The method for preparing cefuroxime by enzymatic method according to claim 8, characterized in that: The mass ratio of water to organic solvent is 11:4 to 14:1; Preferably, the mass ratio of water to organic solvent is 11:4, 13:2 or 14:1.

Images