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

A kind of enzyme for the preparation of antidepressant drug duloxetine intermediate, biocatalysis method and application thereof

A duloxetine and anti-depressant technology, applied in the biological field, can solve the problems of increasing operation difficulty and cost, and achieve the effects of high coenzyme cycle times, low discharge of three wastes, and high specific enzyme activity

Active Publication Date: 2021-12-31
NANJING LANGEN BIOLOGICAL SCI & TECH
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] U.S. Patent No. 8,426,178 discloses an engineered ketoreductase with improved properties, and also provides a polynucleotide encoding the engineered ketoreductase, a host cell capable of expressing the engineered ketoreductase, and a method for synthesizing various chiral compounds using the engineered ketoreductase; Engineered ketoreductase polypeptides optimized to catalyze the conversion of N,N-dimethyl-3-keto-3-(2-thienyl)-1-ketopropylamine to (S)-N,N-dimethyl-3 -Hydroxy-3-(2-thienyl)-1-propanamine; this scheme uses isopropanol as the coenzyme cycle method, but the reaction requires suction filtration to form a negative pressure to remove the by-product acetone of the reaction, so that the reaction can be carried out effectively , it will increase the operation difficulty and cost when the production is enlarged

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 enzyme for the preparation of antidepressant drug duloxetine intermediate, biocatalysis method and application thereof
  • A kind of enzyme for the preparation of antidepressant drug duloxetine intermediate, biocatalysis method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Through the method of whole gene synthesis, the secondary structure and codon bias of the gene are adjusted to achieve high expression in Escherichia coli.

[0032] Use Primer Premier (http: / / primer3.ut.ee / ) and OPTIMIZER (http: / / genomes.urv.es / OPTIMIZER / ) to design, and ensure that the Tm difference is controlled within 3°C, and the primer length is controlled within 60base , and the obtained primers were dissolved in double-distilled water, and added to the following reaction system, so that the final concentration of each primer was 30 nM, and the final concentration of the first and last primers was 0.6 μM.

[0033] 2mM dNTP mix (2mM each dNTP) 5μl 10×Pfu buffer 5μl Pfu DNA polymerase (10U / μl) 0.5μl ddH2O Bring the total volume of the reaction system to 50 μl

[0034] The prepared PCR reaction system was placed in the Bioer XP cycler gene amplification instrument, and the amplification was carried out according to the following p...

Embodiment 2

[0038] Through the method of whole gene synthesis, the secondary structure and codon bias of the gene are adjusted to achieve high expression in Escherichia coli.

[0039] Use Primer Premier (http: / / primer3.ut.ee / ) and OPTIMIZER (http: / / genomes.urv.es / OPTIMIZER / ) to design, and ensure that the Tm difference is controlled within 3°C, and the primer length is controlled within 60base , and the obtained primers were dissolved in double-distilled water, and added to the following reaction system, so that the final concentration of each primer was 30 nM, and the final concentration of the first and last primers was 0.6 μM.

[0040] 2mM dNTP mix (2mM each dNTP) 5μl 10×Pfu buffer 5μl Pfu DNA polymerase (10U / μl) 0.5μl ddH2O Bring the total volume of the reaction system to 50 μl

[0041] The prepared PCR reaction system was placed in the Bioer XP cycler gene amplification instrument, and the amplification was carried out according to the following p...

Embodiment 3

[0045] Through the method of whole gene synthesis, the secondary structure and codon bias of the gene are adjusted to achieve high expression in Escherichia coli.

[0046] Use Primer Premier (http: / / primer3.ut.ee / ) and OPTIMIZER (http: / / genomes.urv.es / OPTIMIZER / ) to design, and ensure that the Tm difference is controlled within 3°C, and the primer length is controlled within 60base , and the obtained primers were dissolved in double-distilled water, and added to the following reaction system, so that the final concentration of each primer was 30 nM, and the final concentration of the first and last primers was 0.6 μM.

[0047] 2mM dNTP mix (2mM each dNTP) 5μl 10×Pfu buffer 5μl Pfu DNA polymerase (10U / μl) 0.5μl wxya 2 o

Bring the total volume of the reaction system to 50 μl

[0048] The prepared PCR reaction system was placed in the Bioer XP cycler gene amplification instrument, and the amplification was carried out according to the foll...

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 enzyme for the preparation of an antidepressant drug duloxetine intermediate, a biocatalysis method and an application thereof, belonging to the field of biotechnology, which can convert 3-dimethylamino-1-(2-thienyl )-1-acetone hydrochloride is converted into S-3-dimethylamino-1-(2-thienyl)-1-propanol, which has higher alcohol dehydrogenase activity and has K49R, A68T, E101D, One or more mutations of F147E, T152A, S169V, A235S. The invention has obvious high specific enzyme activity, which is 2-10 times higher than that of wild-type ketoreductase, and the enzyme can biocatalyze 3-dimethylamino-1-(2-thienyl)-1-acetone hydrochloride Salt conversion generates S-3-dimethylamino-1-(2-thienyl)-1-propanol; the reaction conditions are mild, the requirements for equipment are low, the production process does not require high temperature or cooling, low energy consumption, convenient purification, and preparation The process is green and environmentally friendly.

Description

technical field [0001] The invention relates to an enzyme, a biocatalysis method and application thereof, in particular to an enzyme used for the preparation of an antidepressant drug duloxetine intermediate, a biocatalysis method and application thereof, and belongs to the field of biotechnology. Background technique [0002] Ketoreductases are versatile catalysts through the enantioselective reduction of aldehydes or ketones to the corresponding alcohols; (R)-specific ketoreductases have distinct properties from (S)-specific ketoreductases, and these catalysts are Optically active alcohols are used more and more frequently in the industrial synthesis; optical activity is a necessary condition for the selective action of many pharmaceutically and agrochemically active compounds, and in some cases one enantiomer has beneficial pharmaceutical However, enantiomers have genotoxic effects; therefore, in the synthesis of pharmaceutical and agricultural active compounds, it is nec...

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): C12N9/04C12N15/53C12N15/70C12N1/21C12P17/00C12R1/19
CPCC12N9/0006C12N15/70C12P17/00C12Y101/01
Inventor 丁雪峰李佳松
Owner NANJING LANGEN BIOLOGICAL SCI & 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