Supercharge Your Innovation With Domain-Expert AI Agents!

Pond smelt anticoagulation protein and preparation method thereof

A pond male fish, anti-coagulation technology, applied in the direction of animal/human peptides, peptide sources, peptides, etc.

Inactive Publication Date: 2016-01-06
JILIN AGRICULTURAL UNIV
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Research on anticoagulant proteins at home and abroad mainly focuses on hirudin, ticks, snake venoms, hookworms, etc., while there are few reports on anticoagulant proteins from food sources, and even less reports on anticoagulant proteins from freshwater fish

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
  • Pond smelt anticoagulation protein and preparation method thereof
  • Pond smelt anticoagulation protein and preparation method thereof
  • Pond smelt anticoagulation protein and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Extraction of Pond Male Fish Anticoagulant Protein Coarse Powder

[0032] Male pond fish were purchased from Changchun Aquatic Products Wholesale Market in Jilin Province; DEAE-cellulose-52, SepharoseCL-6B and SephadexG-75 were purchased from Beijing Dingguo Biotechnology Co., Ltd.

[0033] 1) Pond male fish treatment

[0034] Remove the viscera, fins, skin and bones from fresh-frozen pond males. Mix the pond male fish in 20mM, pH7.60 Tris-HCl buffer solution according to 1:4 (w / v) and homogenize;

[0035] 2) Anticoagulant protein extraction

[0036] The homogenate was extracted at 4°C for 12 hours, during which the protease inhibitor PMSF was added every 30 minutes;

[0037] 3) Primary precipitation of anticoagulant protein

[0038] After centrifuging the anticoagulant protein extract at 4000rpm for 20min, discard the precipitate and take the supernatant. According to the ammonium sulfate saturation calculation table, add the ammonium sulfate saturation...

Embodiment 2

[0041] Example 2 Purification of Anticoagulant Protein from Pond Male Fish

[0042] 1) Anion chromatography

[0043] Weigh the anticoagulant protein powder and dissolve it in 20mM, pH7.60 Tris-HCl buffer solution, centrifuge at 10,000×g for 10min, take the supernatant and put it on a DEAE-cellulose-52 column, and use 0-1M NaCl gradient elution at 4℃ , the flow rate is 0.5ml / min, and one tube is collected every 10 minutes. Measured at 280nm with an ultraviolet detector, seven peaks of A, B, C, D, E, F, and G are obtained, such as figure 1 shown. The anticoagulant activity was measured by a coagulometer, such as figure 2 As shown, the activity of E is the highest;

[0044] 2) Agarose gel filtration chromatography

[0045]Dissolve the lyophilized powder obtained from peak E in 20mM, pH7.60 Tris-HCl buffer solution, centrifuge at 10,000×g for 10min, take the supernatant to pass through Sepharose CL-6B column, buffer with 20mM, pH7.60 Tris-HCl at 4°C The solution was elute...

Embodiment 3

[0053] Example 3 Determination of peak anticoagulant activity of anticoagulant protein E-Ⅱ-1 in pond male fish

[0054] Take 200 μL of anticoagulant protein solution and mix with 800 μL platelet-poor plasma (PPP), take 100 μL of the mixture, and use the APTT kit for detection. The results showed that: when the concentration of anticoagulant protein was 1mg / mL, the APTT value was 128.06s, and the normal control value was 24.56s.

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
Molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention discloses pond smelt anticoagulation protein and a preparation method thereof. The method includes the first step of pond smelt processing, the second step of anticoagulation protein extraction, the third step of anticoagulation protein primary sedimentation, the fourth step of dialysis desalination and concentration and the fifth step of anion chromatography, wherein the anticoagulation protein peak E is obtained through anion chromatography. A second peak, namely, an anticoagulation protein peak E-II, is collected after sepharose gel filtration chromatography is conducted on the anticoagulation protein peak E, and freeze-drying is conducted after dialysis. A first peak, namely, an anticoagulation protein peak E-II-1, is collected after sephadex gel filtration is conducted on the anticoagulation protein peak E-II. When the concentration of anticoagulation E-II-1 is 1 mg / mL, the APTT value is 128.06 s, and the normal contrast value is 24.56 s. The molecular weight of the anticoagulation protein is about 37 KDa, and the pond smelt anticoagulation protein has the in-vitro anticoagulation effect, comes from pond smelt, is high in safety and gentle in action, is used for anticoagulation functional food and anticoagulation medicine, has wide market prospects, and has great significance in prevention and control of thrombotic diseases.

Description

technical field [0001] The invention belongs to the field of health food and medicine, and relates to an anticoagulant protein of pond male fish and a preparation method thereof. Background technique [0002] Pond male fish, commonly known as cucumber fish, is a small freshwater economic fish with rich nutrition, delicious taste, and can be eaten as a whole. China's annual output is about 40 million tons, ranking among the top in the world. [0003] The blood coagulation system and the anticoagulation system in the human body are in a state of dynamic balance, once out of balance, it will lead to serious diseases such as myocardial infarction, cerebral thrombosis or hemorrhage. With the improvement of people's living standards and changes in diet structure, the number of people suffering from thrombotic diseases is increasing, and this disease has become the number one killer of human health in today's society. Therefore, the development of a food and anticoagulant drug wit...

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): C07K14/46C07K1/36C07K1/34C07K1/30C07K1/18C07K1/16
CPCC07K14/461
Inventor 刘学军王丽岩苟梦星赵冬兵王利嫒徐玉谭传欣王萌萌
Owner JILIN AGRICULTURAL UNIV
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com