Method for enzymatic synthesis of cefprozil in recyclable aqueous two-phase system by using immobilized penicillin acylase

A technology of penicillin acylase catalysis and penicillin acylase, which is applied in the direction of fermentation, can solve the problems of restricting large-scale production, inability to recycle, difficult to recycle, etc., and achieve increased yield, reduced by-products, and synthesis/hydrolysis ratio Improved effect

Inactive Publication Date: 2013-12-18
南通康鑫药业有限公司 +1
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0008] There are few studies on the catalytic synthesis of cefprozil by penicillin acylase, and the two-phase aqueous system used for enzyme-catalyzed synthesis is also a relatively expensive and difficult-to-recover polyethylene glycol and salt system
The traditional chemical synthesis method of cefprozil has disadvantages and needs to be improved urgently. Although the enzyme-catalyzed synthesis has certain advantages, it also has certain defects. The traditional two-phase aqueous system cannot be recycled, which limits large-scale production

Method used

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  • Method for enzymatic synthesis of cefprozil in recyclable aqueous two-phase system by using immobilized penicillin acylase
  • Method for enzymatic synthesis of cefprozil in recyclable aqueous two-phase system by using immobilized penicillin acylase
  • Method for enzymatic synthesis of cefprozil in recyclable aqueous two-phase system by using immobilized penicillin acylase

Examples

Experimental program
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Effect test

Embodiment 1

[0036] at P NB / P ADB Enzyme-catalyzed synthesis of cefprozil in a reusable two-phase aqueous system:

[0037]To P NB / P ADB A mixture with a total volume of 10 ml was formed in the two aqueous solution. Two polymers P NB with P ADB The concentration in the system is 2.5% (w / w). Then, add 3g (780u) of penicillin G acylase, add 60mmol / L ammonium sulfate to adjust the distribution coefficient of the product cefprozil in the two aqueous phases, adjust the pH of the system to 6.5, and shake the reaction mixture at 15°C After reacting for 80 hours, the yield of cefprozil as determined by liquid chromatography reached 78%, and the post-treatment was carried out by acid precipitation. The final yield was consistent with the yield determined by liquid chromatography, and the solid cefprozil product was obtained by filtration.

[0038] cefprozil 1 HNMR spectrum, mass spectrum and infrared spectrum (such as figure 1 shown). of which cefprozil 1 The HNMR data are shown in Tabl...

Embodiment 2

[0070] at P ADB / P ADBA Enzyme-catalyzed synthesis of cefprozil in a reusable two-phase aqueous system:

[0071] To P ADB / P ADBA A mixture with a total volume of 10 ml was formed in the two aqueous solution. Two polymers P ADB with P ADBA The concentration in the system is 2.5% (w / w). Then, add 3g (780u) of penicillin G acylase, add 60mmol / L ammonium sulfate to adjust the distribution coefficient of the product cefprozil in the two aqueous phases, adjust the pH of the system to 5.0, and shake the reaction mixture at 20°C After reacting for 80 hours, the yield of cefprozil determined by liquid chromatography reached 73%, and the post-treatment was carried out by acid precipitation method. The final yield was consistent with the yield determined by liquid chromatography, and the solid cefprozil product was obtained by filtration.

[0072] Cefprozil obtained through testing 1 HNMR data, mass spectral data and infrared spectrum data are the same as in Example 1.

Embodiment 3

[0074] at P ADB / Sodium citrate reusable enzyme-catalyzed synthesis of cefprozil in two-phase aqueous system:

[0075] To P ADB A mixture with a total volume of 10ml was formed in the aqueous sodium citrate two-phase solution, and the concentration of the two phase-forming substances in the system was 2.5% (w / w). Then, add 3g (780u) of penicillin G acylase, add 60mmol / L sodium dihydrogen phosphate to adjust the distribution coefficient of the product cefprozil in the two aqueous phases, adjust the pH of the system to 6.5, and place the reaction mixture at 20 The reaction was shaken at ℃ for 80 hours, and the yield of cefprozil as determined by liquid chromatography reached 75%. The acid precipitation method was used for post-treatment, and the final yield was consistent with the yield determined by liquid chromatography. The solid cefprozil product was obtained by filtration.

[0076] Cefprozil obtained through testing 1 HNMR data, mass spectral data and infrared spectrum d...

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Abstract

The invention discloses a method for the enzymatic synthesis of cefprozil in a recyclable aqueous two-phase system by using immobilized penicillin acylase. The method comprises the following steps: dissolving a cefprozil parent nucleus and an acylation reagent in an aqueous two-phase polymer solution, wherein the molar ratio of the parent nucleus to the acylation reagent is 1: (1-4); and adding immobilized penicillin G acylase, then adding a two-phase distribution conditioning agent, adjusting the pH value to 4.5-7, and reacting for 1 to 80 hours at the temperature of 5-30 DEG C, wherein the concentration of enzyme in a reaction system is 50-150U / ml. By adopting the method, the hydrolytic activity of the penicillin acylase can be effectively inhibited, thus the hydrolysis degrees of the acylation reagent and the cefprozil product can be reduced. Compared with a method using water as a medium, the method disclosed by the invention has the advantages that the product and the enzyme are distributed in two different water phases, a reactant is fully in contact with the enzyme, so that a synthesis / hydrolysis specific value is greatly increased, the yield of the cefprozil can be increased by 20%, and the highest yield can reach 78%; the adopted novel recyclable aqueous two-phase system can be recycled, so that the production cost is reduced.

Description

technical field [0001] The invention relates to a preparation method of cefprozil, in particular to a method for catalyzing the synthesis of cefprozil by using immobilized penicillin acylase in a reusable two-water phase system. Background technique [0002] Cefprozil (Cefprozil), is a broad-spectrum cephalosporin antibacterial drug. It has strong antibacterial activity against G+, G-bacteria and anaerobic bacteria, especially against G+ bacteria, and is stable against β-lactamase. It is mainly used for the treatment of otitis media, respiratory tract infections, skin and skin soft tissue infections, etc. Cefprozil has high bioavailability, complete gastrointestinal absorption, broad antibacterial spectrum and good stability. It is a semi-synthetic cephalosporin drug with good development prospects. [0003] The chemical synthesis of cefprozil mainly contains two kinds of methods, and one is to use the cephalosporin mother nucleus as starting material, introduce propenyl at...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P35/04
Inventor 曹学君祝超慧唐子安赵志炎李玉丹陈佳伶
Owner 南通康鑫药业有限公司
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