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Preparation method and application of polyvinyl alcohol immobilized acylase

A technology of polyvinyl alcohol and polyvinyl alcohol gel, which is applied in the direction of fixing on/in organic carriers, fermentation, etc., and can solve the problems of low activity of acylase, low stability of biological enzymes, and high price , to achieve stable acylase, good biocompatibility, and reduce application costs

Inactive Publication Date: 2014-01-22
JIANGSU HUITENG BIOMEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, a variety of acylases have been developed that can be used to catalyze the hydrolysis of the above substrates to obtain 7-ACA and 7-ADCA nuclei, but compared with chemical catalysts, biological enzymes still have low stability, relatively expensive prices, and The disadvantage of the acylase enzyme activity is still very low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Immobilized cephalosporin C acylase: prepare an aqueous solution containing 10 g of PVA (polyvinyl alcohol, degree of polymerization 2000), 6 g of PEG (polyethylene glycol, molecular weight 4000), and 80 g of deionized water, add 10 mL of cephalosporin C to acylate Enzyme (crude enzyme solution with a content of 25mg / mL), the concentration of acylase reached 2.5g / L, stirred evenly at room temperature, and then the sporin C acylase-PVA mixed aqueous solution prepared in this way was dropped onto a hard plate Dry it with air flow at room temperature, and dehydrate under reduced pressure at 35°C. Subsequently, the flaky gelled product was collected, transferred to 8% (w / w) sodium dihydrogen phosphate aqueous solution, soaked and hardened for 1 hour, and filtered for later use.

Embodiment 2

[0028] Immobilized cephalosporin C acylase: prepare an aqueous solution containing 5g PVA (polyvinyl alcohol, polymerization degree 2000), 5g HPMC (hydroxypropyl methylcellulose, viscosity 5), 100g deionized water, add 10mL cephalosporin C acylase (crude enzyme solution with a content of 25mg / mL), the concentration of acylase reached 2.4g / L, stirred at room temperature for 1 hour, and then the prepared cephalosporin C acylase-PVA mixed aqueous solution was added dropwise Into the constantly stirring 2% glutaraldehyde aqueous solution, after the dropwise addition is completed, filter and rinse with water until there is no residual glutaraldehyde, then transfer to 10% (w / w) potassium dihydrogen phosphate aqueous solution for immersion and hardening for 1 hour, Filter and set aside.

Embodiment 3

[0030] Immobilized cephalosporin C acylase: prepare PVA aqueous solution containing 15g PVA (polyvinyl alcohol, polymerization degree 2000), 100g deionized water, add 10mL cephalosporin C acylase (crude enzyme solution with a content of 25mg / mL ), to obtain the concentration of acylase is 5g / L, stir evenly at room temperature, then add the prepared cephalosporin C acylase-PVA mixed aqueous solution dropwise to the constantly stirring solution containing 5% (w / w) boric acid and 10 % (w / w) potassium dihydrogen phosphate 100g aqueous solution, immerse and harden for 1 hour, filter, wash with water for later use.

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Abstract

The invention discloses a preparation method of polyvinyl alcohol immobilized acylase. The preparation method comprises the steps: preparing a mixed aqueous solution from polyvinyl alcohol and a cross-linking agent, adding acylase into the mixed aqueous solution of the polyvinyl alcohol and the cross-linking agent, uniformly mixing to obtain an acylase polyvinyl alcohol gel aqueous solution, and carrying out formation and solidification process treatment on the acylase polyvinyl alcohol gel aqueous solution to obtain the polyvinyl alcohol immobilized acylase. The polyvinyl alcohol immobilized acylase is used for synthesizing a cephalosporin antibiotics midbody 7-ACA (Aminocephalosporanic Acid) or 7-ADCA (Aminodesacetoxycephalosporanic Acid). According to the preparation method disclosed by the invention, as PVA (Polyvinyl Alcohol) is used for immobilizing biological enzyme, the enzyme activity recovery rate can reach above 80 percent, immobilized acylase is also more stable and can be reused for above 50 times, so that the application cost of the acylase in the production of the 7-ACA and the 7-ADCA is greatly lowered, and the production efficiency and the product quality are improved.

Description

technical field [0001] The invention relates to a preparation method of immobilized acylase and its application in the synthesis of cephalosporin antibiotic intermediate 7-ACA or 7-ADCA. Background technique [0002] Cephalosporins are a class of antibiotics with broad antibacterial spectrum, strong antibacterial activity, high curative effect and low toxicity. They are also the most widely used antibiotics in clinical practice. Since they were developed in the 1970s, they have been developed and produced to the fourth generation of cephalosporins. Vegetarian varieties. [0003] Cephalosporins are mainly composed of 7-ACA (7-aminocephalosporanic acid, chemical name: 3-acetoxymethyl-5-sulfur-7-amino-8-oxo-1-azabicyclooct-2-ene- 2 carboxylic acid) and 7-ADCA (7-aminodesacetoxycephalosporanic acid) are semi-synthesized from the mother nucleus, and more than 70% of cephalosporin drugs are synthesized from 7-ACA and 7-ADCA through various routes . [0004] A variety of chemica...

Claims

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Application Information

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IPC IPC(8): C12N11/08C12P35/02
Inventor 胡惜朝葛忠平王小兵
Owner JIANGSU HUITENG BIOMEDICAL TECH
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