Unlock instant, AI-driven research and patent intelligence for your innovation.

A kind of preparation and application method of 7-aca immobilized enzyme lk118

A technology of 7-ACA and LK118, which is applied in the fields of immobilized on/in organic carrier, transferase, chemical industry, etc., can solve the problems of high CPC residue, short enzyme service life, low conversion rate, etc., and achieve carrier strength Good, excellent product performance, and the effect of improving conversion rate

Active Publication Date: 2018-09-28
AMICOGEN CHINA BIOPHARM CO LTD
View PDF3 Cites 0 Cited by
  • 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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of preparation and application method of 7-aca immobilized enzyme lk118
  • A kind of preparation and application method of 7-aca immobilized enzyme lk118
  • A kind of preparation and application method of 7-aca immobilized enzyme lk118

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0038] The preparation method of 7-ACA immobilized enzyme LK118, the specific steps are as follows:

[0039] (1) Preparation of epoxy carrier;

[0040] The monomer and cross-linking agent are used to prepare a macroporous adsorption resin epoxy carrier; and the epoxy value of the macroporous adsorption resin epoxy carrier is above 300 μmol / g; the turbidity is less than 200; the pore size is greater than 50nm.

[0041] (2) pretreatment of epoxy carrier;

[0042] Treat the epoxy-type carrier prepared in step (1) with a phosphate buffer solution;

[0043] (3) Immobilization of CPC-acylase;

[0044] Add the free-type CPC acylase from Pseudomonas sp. Shake in phosphate buffer for 15-40 hours at 20-30°C, filter and wash with phosphate buffer to obtain immobilized CPC acylase, namely 7-ACA immobilized enzyme LK118, and the resulting 7-ACA The enzyme activity of the immobilized enzyme LK118 is 80-150U / g.

[0045] The following examples were carried out using the 7-ACA-immobilized...

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.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
epoxy valueaaaaaaaaaa
pore sizeaaaaaaaaaa
Login to View More

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/08C12N9/10C12P35/02
CPCY02P20/50
Inventor 林炳旺罗文军王玲赵新祥贾延宝尚新全
Owner AMICOGEN CHINA BIOPHARM CO LTD