Preparation method and application of 7-aminocephalosporanic acid

A technology of aminocephalosporanic acid and cephalosporins, which is applied in the direction of fermentation, etc., can solve the problems of reduced reaction speed of immobilized enzymes, and achieve the effects of improving catalytic stability, shortening catalytic time, and easy operation

Active Publication Date: 2019-11-08
UNIV OF SCI & TECH BEIJING
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is because for the catalysis of immobilized CPC acylase, similar to the catalysis of most immobilized enzymes, the initial stage of the reaction is due to the higher substrate concentration and lower product concentration (the product is often an inhibitor of the enzyme) , the initial catalytic rate is the fastest; in the later stage of the reaction, due to the decrease of the substrate concentration and the increase of the product concentration (the corresponding product inhibition will be enhanced), the reaction rate of the immobilized enzyme is greatly reduced

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
  • Preparation method and application of 7-aminocephalosporanic acid
  • Preparation method and application of 7-aminocephalosporanic acid
  • Preparation method and application of 7-aminocephalosporanic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0032] The present invention is described in more detail below by way of examples. However, the scope of the present invention is not limited to the examples.

[0033] Analytical method in the embodiment:

[0034] (1) Establish the standard curve of product 7-ACA :

[0035] 1) Prepare 20mg / mL CPC solution and 3mg / mL 7-ACA solution with 0.1mol / L, pH8.5 sodium phosphate buffer solution, and adjust the pH to 8.5 with 1mol / L NaOH solution.

[0036] 2) Add 0, 1, 2, 4, 8, 12, 16, and 20 μL of 7-ACA solution into centrifuge tubes, and make up to 20 μL with 0.1 mol / L sodium phosphate buffer solution at pH 8.5 one by one.

[0037] 3) Add 20 μL of CPC solution (preheated at 37°C for 3 minutes) to each tube and mix well, let stand at 37°C for 5 minutes, then add 200 μL of stop solution (50 mmol / L NaOH solution and 20% glacial acetic acid solution at a ratio of 1:2 volume ratio), and shake well to mix.

[0038] 4) Centrifuge the above mixed solution at 12000rpm for 3min, then take 200...

Embodiment 1

[0060] Preparation of immobilized CPC acylase: Add CPC acylase and 1.25M sodium phosphate buffer (pH8.0) twice the volume of enzyme solution to about 20 g of EPC carrier, so that the final concentration of sodium phosphate buffer is 0.83M. Under the condition of 25°C and 160rpm, let it stand for 24h. After 24 hours, the carrier was washed with deionized water and 0.1M PBS, recovered by suction filtration and stored at 4°C. And use the aforementioned method to measure the enzyme activity.

[0061] Catalytic reaction: The immobilized enzyme that obtains in the step (1) adopts the enzyme dosage of 6U / ml, drops in the cephalosporin C aqueous solution of 25mg / ml, and regulates reaction system pH8.5 with 2M ammonia water, is 10 with circulating water control reaction temperature. °C was maintained for 30 minutes, and then the reaction temperature was controlled at 37 °C until the end of the reaction.

[0062] During the whole reaction process, regular samples were taken to mea...

Embodiment 2-6

[0065] Examples 2-6 were carried out as described in Example 1, except for the temperature profile as follows.

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

No PUM Login to view more

Abstract

The present invention discloses a 7-aminocephalosporanic acid preparation method, and in particular relates to a one-step enzymatic method 7-aminocephalosporanic acid preparation method by use of immobilized cephalosporin C acylase to catalyze cephalosporin C. The method is as follows: the immobilized cephalosporin C acylase and the cephalosporin C are mixed in a liquid, and in a reaction cycle reaction system temperature is continuously or discontinuously raised from initial reaction temperature to 16-37 DEG C. The method has the advantages of stable catalytic rate, shorter reaction time and multiple batches of use of the immobilized cephalosporin C acylase.

Description

technical field [0001] The present invention relates to a preparation method and application of 7-aminocephalosporanic acid, in particular to the method and application of using immobilized cephalosporin C acylase to catalyze cephalosporin C to prepare 7-aminocephalosporanic acid by one-step enzymatic method . Background technique [0002] Cephalosporin C (CPC) acylase is an acylase for the one-step enzymatic production of 7-aminocephalosporanic acid (7-ACA), which can directly catalyze the substrate cephalosporin C , take off the side chain of D-α-amino adipic acid, and produce 7-ACA in one step. Compared with the traditional two-step enzymatic method, the one-step enzymatic method is more simple, convenient, and low-cost, and has been paid more and more attention. [0003] Immobilized enzyme technology was produced in the 1960s and has been widely used in industrial production in the field of biocatalysis. Compared with free enzymes, immobilized enzymes have the advanta...

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): C12P35/02
Inventor 罗晖刘秀红常雁红彭钦成李玺何华于慧敏沈忠耀
Owner UNIV OF SCI & TECH BEIJING
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap