Extraction and purification method of haloacid dehalogenase

A technology of haloacid dehalogenase and purification method, which is applied in the field of designing haloacid dehalogenase, can solve problems such as differences in enzyme properties and activities, and achieve the effect of eliminating hazards

Active Publication Date: 2014-01-15
中科榆林能源技术运营有限责任公司
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ocean is rich in halogen ions, and the halogen organic compounds produced by marine organisms living in a high-halogen concentration environment are a

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
  • Extraction and purification method of haloacid dehalogenase
  • Extraction and purification method of haloacid dehalogenase
  • Extraction and purification method of haloacid dehalogenase

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0039] Hypersil GOLD C18 column (5μm, 250mm×4.6mm, Thermo); mobile phase, acetonitrile-water (15:85, pH 2.2), water containing 0.1% (v / v) concentrated H 3 PO 4 (85%, w / v); flow rate, 1mL / min; column temperature, 30℃; detection wavelength, 210nm. Example 1 Separation and purification of halogenated acid dehalogenase

[0040] Use 341mL buffer A (50mM K) with 16.9g of frozen bacteria Pseudomonas stutzeri 2 HPO 4 -K 2 HPO 4 +0.65mM DTT, pH 7.5) and resuspend the cells. After aliquoting about 25mL in each tube, the cells were disrupted by ultrasound (350w, ultrasound for 5s, interval of 5s, 60 times as a cycle. A total of 2 cycles). After centrifugation (12000rpm×30min, 4℃), take the supernatant for use. Solid ammonium sulfate was added to the supernatant to make 40% ammonium sulfate saturation. After stirring for 1 hour, the precipitate A was obtained after centrifugation (12000rpm×30min, 4℃). After centrifugation, the supernatant was continuously added with ammonium sulfate to 80% ...

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 separation and purification method of dehalogenase with stereoselectivity from a strain of marine bacteria and an application of the dehalogenase in biological resolution. According to the method, two types of 2-haloacid dehalogenase with stereoselectivity, namely L-2-haloacid dehalogenase and D-2-haloacid dehalogenase, are obtained from the strain of the marine bacteria, namely pseudomonas stutzeri by separation and purification. The apparent molecular weight of the enzyme of an enzyme subunit of the L-2-haloacid dehalogenase is about 26kDa, and the L-2-haloacid dehalogenase further has great reaction activity in the pH range of 8-11. D-hydroxy halogenated acid with configuration conversion is generated by selectively catalyzing the conversion of chiral L-2-haloacid through the enzyme. The D-2-haloacid dehalogenase can catalyze the conversion of D-2-haloacid and generate L-2-hydroxy acid, and further has good application prospects as a chiral resolution reagent.

Description

technical field [0001] The invention designs a haloacid dehalogenase, specifically a method for separating and purifying the haloacid dehalogenase in Pseudomonas stutzeri and its application. technical background [0002] Halogenated organic compounds are important raw materials and intermediates in fine chemicals and medicines. Dehalogenases are a class of enzymes that can release halogens from halogenated compounds. It not only plays an important role in the degradation of environmental organic halides, but also has good application prospects in chiral resolution and preparation of chiral compounds due to its stereoselective catalytic activity. [0003] At present, most of the bacteria that have been found to produce dehalogenase come from the terrestrial environment, and there are very few microorganisms from the marine environment. The ocean is rich in halogen ions, and marine organisms living in a high-halogen concentration environment produce very high halogen organi...

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): C12N9/88C12P7/42C12R1/38
CPCC12N9/14C12Y308/0101
Inventor 张锦友薛松曹旭鹏
Owner 中科榆林能源技术运营有限责任公司
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