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
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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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