Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for preparing thymidine by immobilized Escherichia coli

A technology of Escherichia coli and thymidine, which is applied in the field of biocatalysis, can solve the problem of low yield of thymidine in bacteria, and achieve the effects of large-scale production under reaction conditions, improved tolerance, and high reusability

Active Publication Date: 2019-05-03
JIANGSU UNIV OF TECH
View PDF9 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problem that in the prior art chemical enzymatic method, the bacterium is easily inhibited by the reaction substrate and the yield of thymidine is low, the object of the present invention is to provide a method for preparing thymidine by immobilized Escherichia coli

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
  • Method for preparing thymidine by immobilized Escherichia coli
  • Method for preparing thymidine by immobilized Escherichia coli

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The method for preparing thymidine by immobilized Escherichia coli in this embodiment has the following steps:

[0025] (1) First, soak 3g of polyvinyl alcohol (PVA) in 50ml of distilled water for 24h, then add 0.3g of carrageenan, and keep it at 110℃ for 30min to sterilize and fully dissolve the PVA. Cool the mixed solution to 45℃ and keep it warm for use .

[0026] (2) Put a ring of mature E. coli seeds (about 1×10 6 A) Inoculate in 30mL liquid culture medium, 30℃, 100r / min shaking culture for 28h, and the culture solution is placed in a sterile centrifuge tube; centrifuged at 3000r / min for 10min, then washed with sterile saline, centrifuged again to obtain Escherichia coli cell.

[0027] (3) Add 50 mg of E. coli cells obtained in step ② and 10 mg of sterilized ultra-fine activated carbon to the mixed solution of step ①, and after mixing, use a syringe to drip sterile saturated boric acid and KCI solution (the quality of KCl The fraction is 7%) for 36 h to obtain immobiliz...

example 2～ example 7

[0031] The methods and procedures of Examples 2-7 are basically the same as those of Example 1, and the differences are shown in Table 1. The substrate 1 is 2-deoxy-α-D-ribose-1-phosphate dicyclohexylamine salt, and the substrate 2 is Thymine:

[0032] Table 1

[0033]

[0034] It can be seen from Table 1 that under the same other conditions, Example 6 has the best effect. The concentration of 2-deoxy-α-D-ribose-1-phosphate dicyclohexylamine salt is 150 mM, and the concentration of thymine is 150 mM. It is 600mM, the reaction temperature is 65°C, the thymidine yield reaches 90%, and the purity is 97.0%.

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 belongs to the field of biological catalysis, and in particular relates to a method for preparing thymidine by immobilized Escherichia coli. The method adopts PVA and carrageenan as theembedding substrates, takes activated carbon as an additive, and immobilizes the phosphorylase-producing Escherichia coli to obtain the immobilized Escherichia coli; in the preparation of thymidine, immobilized Escherichia coli is used as a catalyst, a phosphate buffer solution is used as a solvent, 2-deoxy-alpha-D-ribose-1-dicyclohexylamine phosphate and thymine are used as a reaction substrate,and thymidine is prepared. The method adopts the immobilized Escherichia coli to prepare thymidine, and can better improve the tolerance of the bacteria to the substrate, and the stability thereof isimproved, so that the yield of thymidine is improved; The results show that after purification, the purity of thymidine is more than 97%, and the yield is more than 90%. After 6 batches of reaction, no significant loss of viability in immobilized Escherichia coli is generated.

Description

Technical field [0001] The invention belongs to the field of biocatalysis, and specifically relates to a method for preparing thymidine by immobilized Escherichia coli. Background technique [0002] Thymidine is an important precursor substance of the anti-AIDS drug zidovudine and azidothymidine, which plays an irreplaceable role in anti-virus and anti-tumor. There are currently four main production methods for thymidine: chemical synthesis, fermentation, enzymatic and chemical enzymatic methods. [0003] There are many ways to synthesize thymidine by chemical synthesis, but there are many shortcomings in the synthesis of thymidine by chemical synthesis: the formed thymidine lacks stereospecificity in glycosidic bonds, raw materials are not easy to obtain, and the yield of thymidine is also very low. , And a large amount of waste water is produced during the production process, which causes great environmental pollution (Chinese invention patent, CN103450302B; CN102863493B). Alth...

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
IPC IPC(8): C12P19/38C12N11/08C12R1/19
Inventor 梁国斌汪斌贡洁翁居轼杨凤丽陈舟叶招莲刘维桥
Owner JIANGSU UNIV OF TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com