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Phanerochaete chrysosporium extract containing antioxidant enzyme and preparation method

A technology of Phanerochaete and antioxidant enzymes, applied in biochemical equipment and methods, oxidoreductases, enzymes, etc., can solve problems such as excessive production of free radicals, cell damage, disorder of free radical scavenging system, etc., and achieve small molecular weight , good solubility, stable antioxidant effect

Inactive Publication Date: 2016-08-17
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, under adversity, such as heavy metal stress, on the one hand, there are too many free radicals, and on the other hand, the free radical scavenging system will be disordered, which will damage the cells.

Method used

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  • Phanerochaete chrysosporium extract containing antioxidant enzyme and preparation method
  • Phanerochaete chrysosporium extract containing antioxidant enzyme and preparation method
  • Phanerochaete chrysosporium extract containing antioxidant enzyme and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] An extract of Phanerochaete chrysosporium containing antioxidant enzymes, using Phanerochaete chrysosporium as the extract and phosphate buffer (50 mM, pH 7.0) as the Phanerochaete thallus and phosphate buffer are mixed, ground, and centrifuged to obtain Phanerochaete chrysosporium extract containing antioxidant enzymes, Phanerochaete chrysosporium is Phanerochaete chrysosporium BKMF-1767, preserved The number is CCTCC AF96007, and it is deposited in the China Center for Type Culture Collection, and the deposit address is China. Wuhan. Wuhan University. The Phanerochaete chrysosporium extract containing antioxidant enzymes contains superoxide dismutase, glutathione peroxidase and catalase, and every 10 ml of Phanerochaete chrysosporium containing antioxidant enzymes In the bacterial extract, the activity of the superoxide dismutase was 33 U, the activity of the glutathione peroxidase was 61 U, and the activity of the catalase was 81 U.

[0024] The preparation method c...

Embodiment 2

[0031] An extract of Phanerochaete chrysosporium containing antioxidant enzymes, using Phanerochaete chrysosporium as the extract and phosphate buffer (50 mM, pH 7.0) as the Phanerochaete thallus and phosphate buffer are mixed, ground, and centrifuged to obtain Phanerochaete chrysosporium extract containing antioxidant enzymes, Phanerochaete chrysosporium is Phanerochaete chrysosporium BKMF-1767, preserved The number is CCTCC AF96007, and it is deposited in the China Center for Type Culture Collection, and the deposit address is China. Wuhan. Wuhan University. The Phanerochaete chrysosporium extract containing antioxidant enzymes contains superoxide dismutase, glutathione peroxidase and catalase, and every 10 mL of Phanerochaete chrysosporium containing antioxidant enzymes In the bacterial extract, the activity of the superoxide dismutase was 15 U, the activity of the glutathione peroxidase was 32 U, and the activity of the catalase was 20 U.

[0032] The preparation method c...

Embodiment 3

[0037] Superoxide dismutase assay

[0038] ①Determination of pyrogallol autoxidation rate: add 2.5 ml of buffer solution (0.1 M Tris-HCl, pH 8.0) at 25 °C, 0.1 ml of ultrapure water, add 0.15 ml of pyrogallol preheated at 25 °C , shake quickly, pour it into a cuvette with a light path of 1 cm, measure the absorbance value (A value) every 30 s at a wavelength of 320 nm, and use buffer (0.1 M Tris-HCl, pH 8.0) as a reference Likewise, the rate of pyrogallol autoxidation was determined.

[0039] ② Determination of SOD enzyme activity:

[0040] The amount of enzyme that inhibits the autooxidation rate of pyrogallol by 50% per minute is defined as a SOD activity unit, expressed in U.

[0041] The measurement method is as follows: add 2.5ml of buffer solution (0.1M Tris-HCl, pH 8.0) at 25°C, 0.1ml of Phanerochaete chrysosporium extract containing antioxidant enzymes, add 25°C preheated o-benzene Trisphenol 0.15ml, the total volume is 2.75ml, shake quickly, pour it into a cuvette ...

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Abstract

The invention relates to a panerochaete chrysosporium extracting solution with antioxidant enzyme. The panerochaete chrysosporium extracting solution with antioxidant enzyme is prepared by taking a panerochaete chrysosporium bacterium as an extracted substance, a phosphate buffer as an extracting agent, mixing up the panerochaete chrysosporium bacterium with the phosphate buffer, grinding and performing eccentric separation. The panerochaete chrysosporium extracting solution with antioxidant enzyme comprises superoxide dismutase, glutathione peroxidase and catalase. The panerochaete chrysosporium extracting solution is high in oxidation resistance. The invention further relates to a preparation method. The preparation method comprises the following steps: weighing the panerochaete chrysosporium bacterium, transferring into a glass homogenizer, adding the phosphate buffer, grinding, centrifuging to obtain the supernate, further adding the phosphate buffer into the glass homogenizer, grinding, centrifuging to obtain the supernate, and adding the phosphate buffer into the supernate until the constant volume is achieved. The panerochaete chrysosporium extracting solution is simple in process.

Description

technical field [0001] The invention relates to the technical field of antioxidant compositions, in particular to an extract of Phanerochaete chrysosporium containing antioxidant enzymes and a preparation method. Background technique [0002] Active oxygen is an inevitable product of microbial growth and metabolism, and the most common active oxygen in cells includes superoxide radicals (O 2 · - ), hydrogen peroxide (H 2 o 2 ) and hydroxyl radicals (·OH). Under normal circumstances, low levels of reactive oxygen species (ROS) are necessary for cells to perform biological functions, because ROS can participate in the regulation of signal transduction in many important pathways in cells. However, in some special cases, such as the redox metabolism disorder of the cell itself or external ion radiation, etc., a large amount of ROS will accumulate in the cell. Due to the high activity of free radicals and strong oxidation reaction ability, they can attack any molecule they ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12N9/08
CPCC12N9/0065C12N9/0089C12Y111/01006C12Y111/01009C12Y115/01001
Inventor 刘亮许飘陈明曾光明黄丹莲赖萃赵美花黄超李宁杰危臻
Owner HUNAN UNIV
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