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Preparation and application method for 7-ACA immobilized enzyme LK118

A 7-ACA, immobilized enzyme technology, applied in the direction of immobilization on/in organic carrier, transferase, chemical industry, etc., can solve the problems of high CPC residue, short service life of enzyme, low conversion rate, etc. Good carrier strength, excellent product performance, and the effect of improving the conversion rate

Active Publication Date: 2015-11-25
AMICOGEN CHINA BIOPHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the problems in the existing one-step enzymatic 7-ACA production process, such as short service life of the enzyme, high residual CPC, low conversion rate, dark color, etc., the invention provides an immobilized 7-ACA acylase, and preparation of 7-ACA ACA's New Process

Method used

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  • Preparation and application method for 7-ACA immobilized enzyme LK118
  • Preparation and application method for 7-ACA immobilized enzyme LK118
  • Preparation and application method for 7-ACA immobilized enzyme LK118

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] In the enzyme cleavage reactor, add 5000u (enzyme activity 100u / g, 50g) of 7-ACA-immobilized enzyme LK118 to 500mL of CPC concentrate (titer 30000u / mL), and start stirring. Add ammonia water with a mass fraction of 5%, keep the reaction pH at 8.2-8.4, control the reaction temperature at 10° C., and react for 60 minutes. After the reaction, the obtained product passes through a sieve to separate the product and 7-ACA immobilized enzyme LK118. The temperature of the reaction product is lowered to 5-10°C, the pH value is adjusted to 5.2 with 5-6 mol / L hydrochloric acid solution, the crystal is grown, washed and dried to obtain the product 7-ACA.

Embodiment 2

[0049]In the enzyme cleavage reactor, add 3500u (enzyme activity 80u / g, 43.75g) of 7-ACA-immobilized enzyme LK118 to 500mL of CPC concentrate (titer 28000u / mL), and start stirring. Add ammonia water with a mass fraction of 2%, keep the reaction pH at 8.2-8.4, control the reaction temperature at 12-13° C., and react for 65 minutes. After the reaction, the obtained product passes through a sieve to separate the product and 7-ACA immobilized enzyme LK118. The temperature of the reaction product was lowered to 8°C, the pH value was adjusted to 5.2 with 5.5 mol / L hydrochloric acid solution, the crystal was grown, washed and dried to obtain the product 7-ACA.

Embodiment 3

[0051] In the enzyme cleavage reactor, add 5000u (enzyme activity 150u / g, 33.33g) of 7-ACA-immobilized enzyme LK118 to 500mL of CPC concentrate (titer 38000u / mL), and start stirring. Add ammonia water with a mass fraction of 6%, keep the reaction pH at 8.2-8.4, control the reaction temperature at 13-14° C., and react for 80 minutes. After the reaction, the obtained product passes through a sieve to separate the product and 7-ACA immobilized enzyme LK118. The temperature of the reaction product is lowered to 5-10°C, the pH value is adjusted to 5.2 with 5-6 mol / L hydrochloric acid solution, the crystal is grown, washed and dried to obtain the product 7-ACA.

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Abstract

The invention discloses preparation and an application process for a 7-ACA immobilized enzyme, and particularly relates to preparation for a durable 7-ACA immobilized enzyme LK118 and a process method for preparing 7-ACA by cracking CPC through the LK118 enzyme. According to the 7-ACA immobilized enzyme LK118, during production of 7-ACA, the problems that original 7-ACA acylase is short in service life, high in CPC residue during application, and low in conversion rate and the product 7-ACA is deep in color are solved. Preparation for the durable immobilized 7-ACA acylase and a new process for applying the durable immobilized 7-ACA acylase to 7-ACA are provided by the invention.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to the preparation of 7-ACA immobilized enzyme LK118 and a new process for preparing 7-ACA by one-step enzymatic method. Background technique [0002] 7-ACA (7-aminocephalosporanic acid) is a key intermediate of semi-synthetic cephalosporin antibiotics, which occupies an important position in the pharmaceutical industry. In the traditional 7-ACA production process, chemical methods are mainly used. With a large amount of toxic reagents, the pollution is extremely serious. In recent years, it has gradually turned to enzymatic production. [0003] In the enzymatic process, there are three-step enzymatic method and two-step enzymatic method. 7-ACA was prepared by a two-step enzymatic method, that is, in the first step, amino acid oxidase was used to convert CPC (cephalosporin C) to GL-7ACA, and in the second step, GL-deacylase was used to convert GL-7ACA to 7-ACA. [0004] Comp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08C12N9/10C12P35/02
CPCY02P20/52Y02P20/50
Inventor 林炳旺罗文军王玲赵新祥贾延宝尚新全
Owner AMICOGEN CHINA BIOPHARM CO LTD
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