A method of extracting and purifying thioredoxin reductase from chicken livers
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A liver thioredoxin, separation and purification technology, applied in the field of separation and purification of thioredoxin reductase, can solve the problems of difference in activity, high requirements, difficulty in obtaining materials from cattle liver, etc. Get difficult effects
Active Publication Date: 2013-09-25
UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
View PDF0 Cites 3 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
Currently, thioredoxin reductases from Escherichia coli, spinach, Arabidopsis thaliana, cattle, and rats have been purified. Generally, the liver of cattle or rats is used as raw material to extract TrxR f
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
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0028] Example 1. Extraction and purification of chicken liver thioredoxin reductase
[0029] Material
[0030] Fresh chicken liver, anion exchange chromatography filler (DEAE-Cellulose DE52, Whatman), ADP affinity chromatography filler (2,5-ADP Sepharose TM 4B, Amersham), and other reagents were of analytical grade.
[0031] main instrument
[0032] High-speed tissue masher (DS200, Jiangsu Jiangyin Zhouzhuang Scientific Research Equipment Factory), super constant temperature water tank (DKB-501A, Shanghai Precision Experimental Equipment Co., Ltd.), spectrophotometer (UV-VIS8500, KEDA), low-temperature chromatography cabinet (DW -1. Shengxian Refrigerator Factory in Zhejiang Province), fully automatic high-speed refrigerated centrifuge (GL-20, Xiangxi Instrument Factory Mechanical Instrument Factory), constant temperature magnetic stirrer (JB-3, Xinjing Branch of Shanghai Leici Instrument Factory) factory), circulating water multi-purpose vacuum pump (SHB-III, Zhengzhou G...
Embodiment 2
[0046] Embodiment 2: the concentration, activity, purity and sequence determination of chicken thioredoxin reductase
[0047] main instrument
[0048] Stabilized voltage and current electrophoresis instrument (DYY-6B, Beijing Liuyi Instrument Factory), decolorization shaker (TS-1, Jiangsu Haimen Qilin Medical Instrument Factory), pH meter (PHS-3D, SANXIN)
[0049] method:
[0050] The enzyme solution finally extracted in Example 1 was used to measure the protein concentration by Coomassie Brilliant Blue staining. Take 20 μl of 2-fold diluted enzyme solution and 20 μl of bovine serum albumin at standard concentrations of 0, 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, and 0.50 mg / ml, respectively, and add them to a 96-well plate Add 180 μl of Coomassie Brilliant Blue staining solution, and monitor the light absorption value of each well at 595 nm after 5 to 10 minutes (because the complex formed by Coomassie Brilliant Blue and protein has specific absorption at 595 ...
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
Login to view more
Abstract
The invention relates to an enzyme extraction and purification technology, specifically to a method of extracting and purifying thioredoxin reductase from chicken livers. The method comprises steps of obtaining a crude extract from fresh chicken livers; salting out; and extracting and purifying thioredoxin reductase from chicken livers by employing chromatography that uses two chromatography media comprising a DEAE anion exchange chromatography column and an ADP affinity chromatography column. The method provided by the invention is advantageous in that the raw material chicken livers have wide source and low cost; process of the extraction and purification technology is simple, and can be operated conveniently; finished products have high purity and the same capability of catalyzing human Trx reduction as cattle or mouse thioredoxin reductase, and therefore the finished products can be applied to scientific researches and practices, in place of cattle or mouse thioredoxin reductase, for human-related disease researches; and problems, of extraction and purification processes for cattle or mouse TrxR, comprising difficulty in obtaining raw materials, large individual differences, etc. are solved. The method provided by the invention fills in the blank of biological reagent preparations in our country and has high economic value.
Description
technical field [0001] The invention relates to an enzyme extraction and purification process, in particular to a method for separating and purifying thioredoxin reductase from fresh chicken liver. Background technique [0002] Thioredoxin system (thioredoxin system) is an important redox regulation system in organisms, involved in cell signal transduction, defense against oxidative stress, regulation of growth and other processes. The thioredoxin system is composed of thioredoxin (Trx), thioredoxin reductase (TrxR) and nicotinamide adenine dinucleotide phosphate (NADPH). Thioredoxin reductase, which catalyzes the electron transfer from NADPH to Trx, plays a central role in this system. Thioredoxin reductase is a member of the pyridine nucleotide disulfide oxidoreductase family. Its structure varies greatly in different organisms, but they are all homodimers. Each subunit has a NADPH binding site point, FAD coenzyme and redox active site, while mammalian thioredoxin reduct...
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
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to view more 
Login to view more