Novel system for transferring biocatalytic reaction substrate based on high-molecular material

A polymer material and biocatalysis technology, which is applied in the field of new biocatalysis reaction substrate delivery system based on polymer materials, can solve the problem of no general and convenient selection method, negative enzyme activity and stability, and difficulty in obtaining the most suitable System and other issues, to achieve the effect of controllable release, increase the concentration of reaction substrate, and wide applicability

Inactive Publication Date: 2013-08-28
CHINA PHARM UNIV
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditionally, the most commonly used batch or continuous addition of substrates is complicated and usually needs to be used in combination with other methods; although the two-phase reaction can increase the amount of substrate input to a certain extent, organic reagents will affect enzyme activity and stability. Sexuality has a negative effect, and the two immiscible phases will face problems such as small contact area or difficulty in breaking after emulsification (see: Angew.Chem.Int.Ed.2012,51,1-5)
The application of ionic liquids and resins needs to be screened from a large number of candidate materials, which takes a long time and is not easy to obtain the most suitable system. At present, there is no general and convenient selection method (see: Biotechnol.Prog.2007,23:74-82 )

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
  • Novel system for transferring biocatalytic reaction substrate based on high-molecular material
  • Novel system for transferring biocatalytic reaction substrate based on high-molecular material
  • Novel system for transferring biocatalytic reaction substrate based on high-molecular material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Preparation of 2-oxo-4-phenylbutyric acid sodium salt. Weigh 20g of 2-oxo-4-phenylbutyric acid, stir and dissolve in 200ml ethanol; weigh 4.5g NaOH, stir and dissolve in 250ml ethanol to prepare NaOH ethanol solution. Pour the ethanol solution of NaOH into the ethanol solution of 2-oxo-4-phenylbutyric acid, stir for 1 hour to fully react, and stand at 4°C overnight. The precipitate was collected by suction filtration, washed with 1 L of ethanol at 4°C, and evaporated to dryness at room temperature. The precipitate obtained by HPLC analysis is indeed 2-oxo-4-phenylbutyric acid sodium salt with a purity greater than 97%.

[0049] The conversion rate and product purity of 2-oxo-4-phenylbutyric acid and its sodium salt and L-homophenylalanine were carried out by reverse phase C18 column (Waters Symmetry, 150mm×4.6mm, 5mm) for HPLC (Shimadzu2010A HT, Japan) analysis, the mobile phase is: A pump water (containing 0.1% trifluoroacetic acid), B pump acetonitrile (containing 0...

Embodiment 2

[0051] Calcium alginate substrate delivery system preparation. Weigh a certain amount of sodium alginate, stir and dissolve it in 6ml of water to prepare a sodium alginate solution with a concentration of 0.5-4% (w / w). Add the substrate 2-oxo-4-phenylbutyric acid or its sodium salt to the sodium alginate solution according to the polymer / substrate ratio of 1-2:1, and stir for 1 hour to dissolve or disperse evenly. The resulting solution was continuously added dropwise to 0.05M ~ 0.4M CaCl under stirring 2 In aqueous solution, sodium alginate solution and CaCl 2 The volume ratio of the solution was 1:1-1:10, and the cross-linking was promoted by continuous stirring for 0.5-10 hours. It was found that calcium alginate pellets could be formed after stirring for more than 1 hour. The obtained calcium alginate pellets were collected by suction filtration, washed three times with water, and dried at 60° C. for 12 to 24 hours to obtain substrate-loaded calcium alginate pellets.

...

Embodiment 3

[0054] Ethylcellulose substrate delivery system preparation. Weigh a certain amount of ethyl cellulose, prepare 5ml of 6% acetone solution, add the substrate 2-oxo-4-phenylbutyric acid or its sodium salt according to the ethyl cellulose / substrate ratio of 90:1~1:3 , Ultrasonic stirring for 2h to dissolve or disperse evenly, as the internal oil phase. Add the inner oil phase drop by drop into the outer oil phase composed of liquid paraffin containing 2-5% Span 80 under stirring, the ratio of the inner oil to the outer oil phase is 1:10, and the rotation speed is 100-600rpm. Continue stirring for 3h to volatilize acetone and form particles. When collecting particles, add n-hexane equal to the volume of the outer oil, let it stand, and pour off the upper liquid, filter to collect the particles, and wash with one of n-hexane, petroleum ether or n-heptane twice the volume of the outer oil phase, Evaporate the organic solvent at room temperature to obtain microparticles prepared f...

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 invention discloses a novel system for transferring a biocatalytic substrate based on high-molecular materials, wherein the system is composed of the following components by weight: 9-66.7% of the substrate and 33.7-91% of the high-molecular material. The system can effectively store high density substrate, can control the substrate to be slowly released in a water solution in a long time, and alleviates substrate inhibition. The invention further relates to several method for preparing the system for transferring the biocatalytic substrate, and an application of the transferring system for L-homophenylalanine synthesis reaction, wherein the substrate concentration reaches to 500mM, a 33h conversion rate reaches 79.1%. The system can effectively alleviate substrate inhibition, enables the substrate concentration raises 3 times, and has an important application value.

Description

technical field [0001] The invention belongs to the technical field of preparation of chiral pharmaceutical intermediates, and relates to a new method for preparing chiral pharmaceutical intermediates coupled with a novel substrate delivery system and a biocatalytic system, in particular to a substrate delivery system based on polymer materials Transfer the substrate 2-oxo-4-phenylbutyrate sodium, and use phenylalanine dehydrogenase as a biocatalyst to realize the efficient preparation of L-homophenylalanine with single optical purity. Background technique [0002] In recent decades, the advancement of science and technology has greatly promoted the development of biocatalysis technology as a practical and environmentally friendly technology that can be used as a substitute for traditional chemical catalysis in the process of chemical synthesis. It is used in pharmaceuticals, food, cosmetics and fine chemicals. The field shows broad application prospects. However, the appli...

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 Applications(China)
IPC IPC(8): C12P13/04
Inventor 陈依军张洁琳
Owner CHINA PHARM UNIV
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