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Psychrophilic glutathione reductase and its prepn process

A technology of glutathione and reductase, which is applied in the direction of oxidoreductase, etc., can solve the problems of low-temperature glutathione reductase separation and purification, properties and research, etc., so as to improve the enzymatic determination method and the adaptation mechanism , the effect of reducing the impact

Inactive Publication Date: 2007-08-22
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
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AI Technical Summary

Problems solved by technology

However, the isolation, purification, properties and research of low-temperature glutathione reductase have not yet been involved, especially for this enzyme from Antarctic microalgae

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0031] The invention is further illustrated by the following examples.

[0032] Initial extraction of enzyme solution:

[0033] The operation of glutathione reductase purification was carried out at 4°C. Weigh the freeze-dried Antarctic microalgae, 20g in total, add quartz sand and liquid nitrogen to grind several times, add 4 times of about 80ml of enzyme extract (containing 0.1mol / L potassium phosphate buffer solution of pH7.8, 5mmol / L glutathione, 1mmol / L EDTA, 0.1mmol / L DDT, 1% PVP polyvinylpyrrolidone), repeated freezing and thawing for 3 times, and filtered through four layers of gauze; Then, centrifuge at a speed of 14000 rpm for 15 minutes to remove insoluble starch, then centrifuge the supernatant at a speed of 11000 rpm for 15 minutes, discard the precipitate and keep the supernatant. The supernatant was dialyzed against the enzyme extract for 2 days with several buffer changes during the dialysis.

[0034] Ammonium Sulfate Precipitation:

[0035] Measure the vo...

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Abstract

The present invention is psychrophilic glutathione reductase and its preparation process. The psychrophilic glutathione reductase is separated and purified in a four-step process including ammonium sulfate precipitation, ion exchanging, affinity chromatography and gel filtering. It consists of two identical subunits of apparent molecular weight 54.6 kDa each, and has optimal temperature of 25 deg.c, optimal pH value of 7.5, optimal Mg2+ concentration of 7.5 mmol / L, and promotion effect of Ca2+ and Mg2+. The psychrophilic glutathione reductase may be well used in low temperature enzyme catalysis mechanism research, in resistant variety improvement, etc.

Description

Technical field: [0001] The invention belongs to the fields of microbiology, biochemistry, enzymology and polar biology, and in particular relates to a psychrophilic glutathione reductase and a preparation method thereof. Background technique: [0002] Due to the unique geographical and climate characteristics of the polar region, a natural environment of dryness, severe cold, strong radiation, high salinity and low light is formed, and the organisms that can survive in it must have corresponding unique physiological, biochemical and molecular mechanisms to adapt to extreme conditions. environment of. The unique physiological and biochemical characteristics of Antarctic microalgae, its adaptability to extreme environmental stress and its mechanism have broad prospects in basic research, development and application, and it has become a good experimental material for studying extreme environmental biology. [0003] The glutathione system plays a very important role in maintai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02
Inventor 丁燏简纪常吴灶和
Owner GUANGDONG OCEAN UNIVERSITY
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