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

Enzymatic preparation method of esomeprazole

A technology for esomeprazole and enzymatic preparation, which is applied in the field of enzymatic preparation of esomeprazole, can solve the problems of long reaction time, inability to further increase the yield, and large amount of enzyme input, and achieves efficient preparation and superiority. The effect of industrial application value and production cost reduction

Inactive Publication Date: 2018-07-06
ZHEJIANG JINGXIN PHARMA +1
View PDF3 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem solved by the present invention is to overcome the defects of the existing method for enzymatically synthesizing esomeprazole when the substrate concentration is large, the reaction time is too long, and the enzyme amount is large, and the yield cannot be further improved. , provide a kind of enzymatic preparation method of esomeprazole

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
  • Enzymatic preparation method of esomeprazole

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1: Construction of cyclohexanone monooxygenase genetically engineered bacteria

[0035] Commissioned Shanghai Jierui Bioengineering Co., Ltd. to custom synthesize cyclohexanone monooxygenase gene fragments SEQ ID NO.1, SEQ ID NO.3 and SEQ ID NO.5, and the corresponding encoded amino acid sequences are SEQ ID NO.2 and SEQ ID NO.2, respectively. ID NO.4 and SEQ ID NO.6. Then use the gene fragment as a template to amplify and expand by PCR (add Nde I and BamH I endonuclease fragments to both ends of the gene fragment), and insert the gene fragment into the pET28a plasmid using the Nde I and BamH I endonuclease sites At last, the vector obtained by ligation was transformed into Escherichia coli BL21 (DE3), and the recombinant Escherichia coli genetically engineered strains containing the cyclohexanone monooxygenase gene were constructed, respectively denoted as bacterial strain 1#, bacterial strain 2# and bacterial strain 3 #.

[0036] The primer sequences desi...

Embodiment 2

[0039] Embodiment 2: Construction of ketoreductase genetically engineered bacteria

[0040] Entrusted Shanghai Jierui Bioengineering Co., Ltd. to customize and synthesize the ketoreductase gene fragments SEQ ID NO.7, SEQ ID NO.9 and SEQ ID NO.11, and the corresponding encoded amino acid sequences are SEQ ID NO.8 and SEQ ID NO.10 respectively and SEQ ID NO.12. Then use the gene fragment as a template to amplify and expand by PCR (add Nde I and BamH I endonuclease fragments to both ends of the gene fragment), and insert the gene fragment into the pET28a plasmid using the Nde I and BamH I endonuclease sites , and finally the vector obtained by ligation was transformed into E. coli BL21(DE3), and recombinant E. coli genetically engineered strains containing the ketoreductase were constructed, which were respectively designated as strain 4#, strain 5# and strain 6#.

[0041] The primer sequences designed for strain 4# PCR amplification extension are as follows:

[0042] Forward p...

Embodiment 3

[0050] Embodiment 3: the construction of co-expression genetically engineered bacteria

[0051] Refer to "Molecular Cloning-A laboratory Manual" (third edition, 2001) to carry out the following enzyme digestion, ligation or competent cell preparation, transformation and other experiments.

[0052] Design following primers F5 and R5, take the gene fragment of SEQ ID NO.1 cyclohexanone monooxygenase as a template, expand the described cyclohexanone monooxygenase gene fragment by PCR amplification (add Nde I and BamH I endonuclease fragment); and using Nde I and BamH I endonuclease sites to insert the gene into the pET-21a plasmid, the vector after the ligation is transferred into Escherichia coli Trans-T1, and the recombinant plasmid is obtained by constructing, named pETC. Colony PCR verification was performed with primer T7 / R3, the recombinant plasmid was extracted and sequenced, and the recombinant plasmid pETC with correct results was obtained.

[0053] Forward primer F5: ...

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 an enzymatic preparation method of esomeprazole. The enzymatic preparation method comprises: carrying out a reaction on omeprazole thioether as a substrate and an oxidizing agent in a solvent under the actions of monooxygenase, an auxiliary component and a phase transfer catalyst to generate esomeprazole. According to the present invention, with the enzymatic preparation method, esomeprazole is efficiently prepared; and by using the phase transfer catalyst, the efficiency of the enzyme catalytic reaction is improved, the reaction time is substantially reduced, the concentration of the reaction substrate is increased, the conversion rate can be up to 99.8%, the production cost is reduced, the single-batch yield is substantially improved, and the excellent industrialapplication value is provided.

Description

technical field [0001] The invention relates to the technical fields of pharmacy and biochemical industry, in particular to an enzymatic preparation method of esomeprazole. Background technique [0002] Enzyme-catalyzed redox reactions in two-phase systems have been widely reported. Yu Yao (bromoperoxidase catalyzed cyclohexene epoxidation reaction, Yu Yao, Jin Yan, Wu Peichun, Zhang Wei, Acta Catalytica Sinica, 2007, Vol. 28, No. 10, pp. 915-918) studied bromine Peroxidase catalyzes the epoxidation of cyclohexene in a water-organic two-phase system, with a yield of 41.5% and a selectivity of 82.2%. Yang Zhonghua (Research on the Asymmetric Reduction of Acetophenone by Baker's Yeast to Synthesize Chiral Phenyl Alcohol in Water-Organic Solvent Two-phase System, Yang Zhonghua, Zeng Rong, Wu Gaoming, etc., Journal of Chemical Engineering of Universities, 2009, Volume 23, Issue 3, Pages 450-454) have reported that the water-organic solvent two-phase system promotes the asymmet...

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): C12P17/16
CPCC12P17/165
Inventor 黄悦张敏张敏洁
Owner ZHEJIANG JINGXIN PHARMA
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