Lactamase and its application and method for enzymatic resolution and preparation of (1r, 4s)-venslide

A technology of lactamase and vince lactone, which is applied in the field of bioengineering, can solve the problems of low tolerance and activity, lower production efficiency, and low substrate concentration, so as to reduce production costs, improve production efficiency, The effect of high substrate tolerance

Active Publication Date: 2021-10-26
安徽红杉生物医药科技有限公司
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, optically pure (1R, 4S)-venslides are usually obtained by biocatalysis, but the tolerance and activity of enzymes currently used in biocatalysis are generally not high, resulting in a normal reaction system The substrate concentration is small, which reduces the production efficiency and the cost is high

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
  • Lactamase and its application and method for enzymatic resolution and preparation of (1r, 4s)-venslide
  • Lactamase and its application and method for enzymatic resolution and preparation of (1r, 4s)-venslide
  • Lactamase and its application and method for enzymatic resolution and preparation of (1r, 4s)-venslide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The method for the enzymatic resolution and preparation of (1R,4S)-venslide provided in this example specifically includes the following steps: dissolving 500 g of the substrate venslide with 1 L phosphate buffer solution with a concentration of 10 mM, and then adding lactam Enzyme 3g, start stirring, react at 25°C, take samples in the middle and control in the middle, after the reaction ≥ 99% (see figure 2), start post-processing, add 4% dichloromethane to the reaction system, stir for 10 minutes, then concentrate to 3 / 5 volume, use 2.5 times volume of dichloromethane to extract 3 times (1 times, 1 times, 0.5 times ), the concentrated dichloromethane phase after the extraction was completed, the solvent was recovered, and evaporated to dryness to obtain the target product (1R, 4S)-2-azabicyclo[2.2.1]hept-5-en-3-one 135.9g, yield 48.8%, chiral purity 100% (see image 3 ).

[0042] in, figure 2 and image 3 They are the detection result diagrams of the high perform...

Embodiment 2

[0054] The method for the enzymatic resolution and preparation of (1R,4S)-venslide provided in this example specifically includes the following steps: dissolving 500 g of the substrate venslide with 1 L phosphate buffer solution with a concentration of 10 mM, and then adding lactam Enzyme 3g, start stirring, react at 15°C, take samples in the middle and control in the middle, after the reaction is ≥99%, start post-processing, add 4% dichloromethane to the reaction system, stir for 10 minutes, then concentrate to 3 / 5 volume, use 2.5 times the volume of dichloromethane was extracted 3 times (1 times, 1 times, 0.5 times), after the extraction, the dichloromethane phase was concentrated, the solvent was recovered, and the target product (1R,4S)-2-azabicyclo was obtained after evaporation to dryness [2.2.1] 135.9 g of hept-5-en-3-one.

Embodiment 3

[0056] The method for the enzymatic resolution and preparation of (1R,4S)-venslide provided in this example specifically includes the following steps: dissolving 500 g of the substrate venslide with 1 L phosphate buffer solution with a concentration of 10 mM, and then adding lactam Enzyme 3g, start stirring, react at 20°C, take samples in the middle and control in the middle, after the reaction is ≥99%, start post-processing, add 4% dichloromethane to the reaction system, stir for 10 minutes, then concentrate to 3 / 5 volume, use 2.5 times the volume of dichloromethane was extracted 3 times (1 times, 1 times, 0.5 times), after the extraction, the dichloromethane phase was concentrated, the solvent was recovered, and the target product (1R,4S)-2-azabicyclo was obtained after evaporation to dryness [2.2.1] 135.9 g of hept-5-en-3-one.

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

PropertyMeasurementUnit
wavelengthaaaaaaaaaa
wavelengthaaaaaaaaaa
Login to view more

Abstract

The invention discloses a lactamase, its application and a method for enzymatic resolution and preparation of (1R, 4S)-vins lactone. The amino acid sequence of the lactamase is shown in SEQ ID NO.1. The nucleotide sequence of the nucleic acid molecule encoding the above-mentioned lactamase is shown in SEQ ID NO.2. The method for enzymatic resolution and preparation of (1R, 4S)-wens lactone comprises: in a buffer system, the substrate and the above-mentioned lactamase are mixed and reacted, and the substrate is 2-azabicyclo[2.2.1]heptane ‑5‑en‑3‑one. The lactamase has the characteristics of high substrate tolerance and high activity, and it is used in the method of enzymatic resolution and preparation of (1R, 4S)-Wens lactone, which can make a higher concentration of the substrate Fully reacting under the action of the lactamase, the (1R, 4S)-wens lactone with higher yield and chiral purity is obtained, which improves production efficiency and reduces production cost.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a lactamase and its application and a method for enzymatic resolution and preparation of (1R, 4S)-vins lactone. Background technique [0002] γ-lactam is the abbreviation of the compound 2-azabicyclo[2.2.1]hept-5-en-3-one (also commonly known as Wens lactone), and its molecular formula is C 6 h 7 NO. One of the isomers of γ-lactam is (-)-γ-lactam (1R, 4S)-wens lactone, which is mainly synthesized from optically pure (1R, 4S)-wens lactone There is the anti-HIV drug abacavir, and the anti-influenza drug peramivir is mainly synthesized from (1S,4R)-vinslide. Therefore, the separation and preparation of (1R, 4S)-venslide has important economic and social value. [0003] At present, optically pure (1R, 4S)-venslides are usually obtained by biocatalysis, but the tolerance and activity of enzymes currently used in biocatalysis are generally not high, resulting in a normal rea...

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): C12N9/86C12N15/55C12P41/00C12P17/10
CPCC12N9/86C12P17/10C12P41/001C12P41/006
Inventor 高仰哲吴法浩李钢王志航
Owner 安徽红杉生物医药科技有限公司
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