Extraction and purification method of glutary-7-aminocefaphytanic acid acyltransferase

A technology of aminocephalosporanic acid acylase and purification method, applied in the direction of hydrolase, etc., can solve the problems of complex process, poor quality, serious pollution, etc., and achieve the effect of improving specific activity

Inactive Publication Date: 2003-07-09
HUNAN FLAG BIOTECHNOLOGY CO LTD
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And the key intermediate of semi-synthetic cephalosporin is 7-aminocephalosporanic acid (7-ACA), the method of traditional production in the past mainly is by chemical method, has four major steps, and process is complicated, and used reagent is expensive, and pollution is serious, And the yield is low, the quality is poor

Method used

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Effect test

Embodiment 1

[0012] The extraction and purification process of GL-7-ACA acylase is as follows: firstly adjust the pH of the fermentation broth produced by fermentation to 7.0 with dilute hydrochloric acid, then lower the pH to 6.6 with 20% calcium chloride, and then add 2% of the bacteria chitosan, and finally adjust the pH to 7.6 with dilute alkali. Centrifuge the bacterial liquid with a high-speed centrifuge, control the activity of the bacterial liquid to 5-8u / ml, and add surfactant cetyltrimethylammonium bromide equivalent to 2% (g / l) of the bacterial liquid. Stir and soak naturally for 3 hours. During the purification process, use 20% calcium chloride solution to lower the pH of the soaked bacterial solution to 5.8, collect the enzyme serum by plate and frame filtration, and use 10% sodium hydroxide solution to increase the pH of the enzyme serum to 7.6, and then collect the serum by plate and frame filtration. The specific activity of the serum was increased from 3.4u / mg to 7.8u / mg,...

Embodiment 2

[0014] The extraction and purification process of GL-7-ACA acylase is as follows: firstly adjust the pH of the fermentation broth produced by fermentation to 7.2 with dilute hydrochloric acid, then lower the pH to 6.8 with 20% calcium chloride, and then add 2% of the bacteria chitosan, and finally adjust the pH to 7.8 with dilute alkali. Centrifuge the bacterial liquid with a high-speed centrifuge, control the activity of the bacterial liquid to 5-8u / ml, and add cetyltrimethylammonium bromide equivalent to 2% (g / l) of the bacterial liquid. Stir and soak naturally for 5 hours. During the purification process, use 20% calcium chloride solution to lower the pH of the soaked bacterial solution to 6.2, collect the enzyme serum by plate and frame filtration, and use 10% sodium hydroxide solution to increase the pH of the enzyme serum to 8.0, and then collect the serum by plate and frame filtration. The specific activity of the serum was increased from 3.4u / mg to 7.8u / mg, and the pu...

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PUM

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Abstract

The method for extracting and purifying glutaryl-7-aminocephaphytanic acid acylase includes the following steps: firstly, using dilute hydrochloric acid to regulate pH of fermented liquor produced by fermentation, then adding calcium chloride solution, and adding chitosan, and using dilute alkali to regulate pH to 7.6-7.8, centrifuging to obtain centrifuged bacterial liquor, controlling activity of the bacterial liquor to 5-8 a/ml, adding surfactant, naturally stirring and soaking for 3-5 hr., and its purification process includes the following steps: using calcium chloride solution to downward regulate pH of soaked bacterial liquor, plate-frame filtering and collecting enzyme clear liquor, using sodium hydroxide solution to upward regulate pH of enzyme clear liquor to 7.6-80, and plate-frame filtering and collecting clear liquor.

Description

Technical field: [0001] The invention relates to a method for extracting and purifying glutaryl-7-aminocephalosporanic acid acylase, which is a key intermediate in the synthesis of semi-synthetic cephalosporins, 7-aminocephalosporanic acid. Background technique: [0002] Cephalosporins are antibiotics with important efficacy, while semi-synthetic cephalosporins have stronger antibacterial ability or can improve pharmacokinetic properties. And the key intermediate of semi-synthetic cephalosporin is 7-aminocephalosporanic acid (7-ACA), the method of traditional production in the past mainly is by chemical method, has four major steps, and process is complicated, and used reagent is expensive, and pollution is serious, And the yield is low and the quality is poor. The biotransformation method is a better method. Studies have shown that cephalosporin C can be oxidatively deaminated by D-amino acid oxidase, and further decarboxylated to generate GL-7-ACA, while the 7-side chain...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14C12N9/80
Inventor 许岗朱敏莫章华曾红宇柳杏辉唐卫红胡爱红
Owner HUNAN FLAG BIOTECHNOLOGY CO LTD
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