Preparation method of snail respiratory protein

A snail and protein technology, applied in the fields of medicine extraction and biology, can solve the problems of slow gel filtration purification, many impurities in hemocyanin, and difficulty in extracting hemolymph.

Inactive Publication Date: 2012-07-25
GANNAN NORMAL UNIV
View PDF0 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Oncomelania snails are small, and the heart is even smaller, so it is difficult to extract hemolymph directly with a syringe; the hemocyanin obtained by single ammonium sulfate precipitation and low-speed centrifugation als

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
  • Preparation method of snail respiratory protein
  • Preparation method of snail respiratory protein
  • Preparation method of snail respiratory protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Take the snails in batches on a glass plate, mechanically crush and remove the shells, intercept the required tissues, and mechanically squeeze the Oncomelania hemolymph to a centrifuge tube; add Tris-HCl pH7.0 buffer solution, mechanically squeeze and stir to fully dissolve, and centrifuge at 8000g After 20 minutes, discard the precipitate; add ammonium sulfate to the supernatant to a saturation of 33%, let it stand at 4°C for 2 hours, and centrifuge to remove the supernatant to obtain a crude hemocyanin blue precipitate.

[0022] The hemocyanin precipitate was redissolved with the aforementioned Tris-HCl buffer, dialyzed overnight at 4°C; transferred to a centrifuge tube at 180,000g, centrifuged at 4°C for 3.5h, and the supernatant was discarded to obtain a blue protein precipitate again, which was dissolved in 0.05M Tris -HCl (pH7.0, containing 5mM CaCl 2 , 5 mM MgCl 2 ) in the buffer solution. The protein purity is above 95%.

Embodiment 2

[0024] Take snails in batches on a glass plate, mechanically crush and remove the shells, intercept the required tissues, mechanically squeeze and collect Oncomelania hemolymph into a centrifuge tube; mechanically squeeze and collect hemolymph, and centrifuge at 10,000g for 50 minutes to remove impurities such as remaining tissue fragments; add Tris-HCl pH 7.0 buffer was used to dissolve hemolymph, and ammonium sulfate was added to reach a saturation of 43%, and the protein was precipitated at 8°C for 3 hours, and the supernatant was removed by centrifugation to obtain a crude blue precipitate of hemocyanin.

[0025] Hemocyanin precipitate was redissolved with the aforementioned Tris-HCl buffer, dialyzed overnight at 8°C; transferred to a centrifuge tube at 180,000g, centrifuged at 6°C for 4.5h, discarded the supernatant, and obtained blue protein precipitate again, and dissolved the precipitate in 0.08M Tris -HCl (pH7.0, containing 5mM CaCl 2 , 5 mM MgCl 2 ) in the buffer so...

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 present invention relates to a preparation method of a snail respiratory protein. A respiratory protein containing copper ions is extracted from a snail vivo used as an intermediate host of Schistosoma japonicum by virtue of mechanical extrusion, tissue dissection, hemolymph extraction, ammonium sulfate precipitation, super high-speed centrifugal separation and other means. The respiratory protein contains 0.2-0.3% of copper, the UV characteristic absorption peak is at 275nm and 340nm, and a cylindrical agreegate structure is observed under a transmission electron microscope.

Description

technical field [0001] The invention belongs to biological and medical extraction technology, and relates to a method for extracting and separating Oncomelania hemocyanin from Oncomelania, specifically, a three-step method of tissue stripping, mechanical extrusion, ammonium sulfate precipitation and ultra-high-speed centrifugation from Oncomelania hemocyanin. A preparation method for extracting high-purity Oncomelania hemocyanin from Background technique [0002] The respiratory proteins that perform the function of oxygen transport in the long evolutionary process generally include hemoglobin, hemerythrin and hemocyanin. Hemocyanin, a respiratory protein that turns blue when oxygenated, exists in the hemolymph of two major invertebrates - arthropods and molluscs. Hemocyanin is a huge complex molecule with a multi-subunit quaternary structure. A pair of copper ions is bound to the oxygenated amino acid residue site. The deoxygenated state is monovalent, and the protein is c...

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): C07K14/795C07K1/30C07K1/14
Inventor 范小林李永东郭道义李勋张云曾丹
Owner GANNAN NORMAL UNIV
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