Method for separating and purifying various components from cefaclor enzymatic reaction solution

A cefaclorase, separation and purification technology, applied in the biological field, can solve the problems of polluting the environment, increasing separation and purification, and threats to the health of operators, and achieves the effects of pollution-free health, reduction of production costs, and avoidance of waste.

Inactive Publication Date: 2013-07-10
CHENGDU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The resulting aqueous mixture contains cefaclor in an amount >10 (w/w), 7-ACCA in an amount <2 (w/w), and D- Phenylglycine, this patent also discloses the separation and purification method of cefaclor, the disadvantage of this patent is that the separation of unreacted 7-ACCA and by-product D-phenylglycine has not been studied
Patent CN 200810038896.7 discloses a method for purifying cefaclor from cefaclor and naphthol complex, the method is mainly to use organic solvent to selectively extract the naphthol in the compound of cefaclor and naphthol, and then collect and obtain The aqueous phase is adsorbed by a macroporous adsorption resin for the separation and purification of cefaclor. The disadvantage of this patent is that the complexing agent naphthol is added to the enzymatic reaction solution, and then the compound of cefaclor and naphthol is obtained. , and then used for furt

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Embodiment 1

[0035] A method for separating and purifying each component from cefaclor enzymatic reaction liquid, comprising the following steps:

[0036] (1) Recovery of 7-ACCA: first measure 200ml of the reaction solution of cefaclor synthesized by enzymatic method, remove the immobilized enzyme by filtration, wash the immobilized enzyme repeatedly with deionized water, combine the filtrate and water washing solution to obtain 780ml of cefaclor Enzymatic reaction solution, which contains 1.47g 7-ACCA, 7.3g cefaclor, 4.13g D-phenylglycine, adjust the pH value to 1.6 with 6mol / L hydrochloric acid, maintain the temperature of the process at 20°C, and stir at 200rpm for 30 minutes , filter the above cefaclor reaction solution through a G4 glass sand funnel, wash with 5ml of water with a pH value of 1.6, and repeat three to four times. Vacuum filtration and drying yielded 1.25 g of 7-ACCA with a yield of 85% and a purity of 99.62% by HPLC. After the combined filtrate and washing water were m...

Embodiment 2

[0040] A method for separating and purifying each component from cefaclor enzymatic reaction liquid, comprising the following steps:

[0041] (1) Recovery of 7-ACCA: First, weigh 200ml of the reaction solution of cefaclor synthesized by enzymatic method, remove the immobilized enzyme by filtration, wash the immobilized enzyme repeatedly with deionized water, combine the filtrate and water washing solution to obtain 780ml of cefaclor Enzymatic reaction solution, which contains 1.47g 7-ACCA, 7.3g cefaclor, 4.13g D-phenylglycine, adjust the pH value to 2.0 with 6mol / L hydrochloric acid, maintain the temperature of the process at 20°C, and stir at 200rpm for 30 minutes , filter the above cefaclor reaction solution through a G4 glass sand funnel, wash with 5 ml of water with a pH value of 2.0, and repeat three to four times. Vacuum filtration and drying yielded 1.22 g of 7-ACCA with a yield of 83% and a purity of 99.82% by HPLC. After the combined filtrate and washing water were m...

Embodiment 3

[0045] A method for separating and purifying each component from cefaclor enzymatic reaction liquid, comprising the following steps:

[0046] (1) Recovery of 7-ACCA: First, weigh 200ml of the reaction solution of cefaclor synthesized by enzymatic method, remove the immobilized enzyme by filtration, wash the immobilized enzyme repeatedly with deionized water, combine the filtrate and water washing solution to obtain 780ml of cefaclor Enzymatic reaction solution, which contains 1.47 g 7-ACCA, 7.3 g cefaclor, 4.13 g D-phenylglycine, adjust the pH value to 1.34 with 6 mol / L hydrochloric acid, maintain the temperature of the process at 20 ° C, 200 rpm magnetic stirring for 30 Minutes later, the cefaclor reaction solution was filtered through a G4 glass sand funnel, washed with 5 ml of water with a pH value of 1.34, and repeated three to four times. Vacuum filtration and drying gave 1.17 g of 7-ACCA with a yield of 80% and a purity of 99.82% by HPLC. After the combined filtrate and...

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Abstract

The invention relates to a method for separating and purifying various components from a cefaclor enzymatic reaction solution, and the method comprises the following steps of: (1) recovering an unreacted mother nucleus 7-ACCA from the cefaclor enzymatic reaction solution by adopting an acid precipitation method; (2) separating and purifying cefaclor by adopting macroporous adsorption resin; and (3) separating and purifying D-phenylglycine by adopting cation exchange resin. The method disclosed by the invention can be used for separating and purifying the unreacted mother nucleus 7-ACCA, the product cefaclor and the by-product D-phenylglycine from the reaction solution used for enzymatically synthesizing the cefaclor, sufficiently utilizes the raw materials and reduces the production cost; and products obtained by adopting the method disclosed by the invention are high in yield purity and suitable for popularization and usage.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for separating and purifying components from cefaclor enzymatic reaction liquid. Background technique [0002] Cefaclor (Cefaclor) is a semi-synthetic second-generation cephalosporin antibiotic, which has good antibacterial activity against Gram-positive bacteria, high efficiency, broad spectrum, and good chemical stability. One of the main drugs for bacterial infections. Since its launch in the 1970s, cefaclor has been highly praised as an antibiotic with high efficiency, broad spectrum and good clinical safety. Its chemical structural formula is as follows: [0003] ; [0004] In industry, cefaclor is mainly prepared by condensation of 7-amino-3-chloro-deacetoxy-cephalosporanic acid (7-ACCA) and activated D-phenylglycine. There are two processes of chemical condensation and enzymatic condensation. Patent WO2006 / 069984 discloses a method for enzymatic condensation of ce...

Claims

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Application Information

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IPC IPC(8): C07D501/59C07D501/12C07C229/36C07C227/40
Inventor 王佳珉李端华秦川萍王辂褚以文潘月李进军
Owner CHENGDU UNIV
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