Protein separated from jellyfish and possessing antioxygenation activity and its application
An antioxidant activity, jellyfish technology, applied in the field of marine biology, can solve the problem of low added value, and achieve the effect of low production cost and high antioxidant activity
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Embodiment 1
[0047] (1) Break a certain amount of jellyfish with an ultrasonic breaker, add 0.5 times the weight / volume ratio of 5mM, pH4 phosphate buffer solution (PB), put it at 2°C, and soak for 0.5h;
[0048] (2) Centrifuge at 10,000 rpm for 30 minutes at 4°C, and take the supernatant; the residue is leached twice with 5mM phosphate buffer solution (PB) with a weight / volume ratio of 0.5 times, pH 4, and soaked at 4°C 0.5h; centrifuge at 10000rpm at 4°C for 30min; combine the supernatants from the two centrifugations;
[0049] (3) with Coomassie Brilliant Blue staining method (this method is selected from "Protein Molecular Basis", edited by Tao Weisun et al., Higher Education Press), taking bovine serum albumin as a standard, measuring the absorbance of jellyfish protein at 595nm (A 595 ), and then calculate the concentration (Cmg / ml) of aequorin in the supernatant according to Lambert Beer's law.
[0050] (4) with gel filtration chromatography (this method is selected from "Protein M...
Embodiment 2
[0052] (1) Break a certain amount of jellyfish with an ultrasonic breaker, add 2.0 times the weight / volume ratio of 10mM, pH6 phosphate buffer solution (PB), put it at 4°C, and soak for 1h;
[0053](2) Centrifuge at 15,000 rpm for 20 minutes at 4°C, and take the supernatant; the residue is leached twice with 10mM phosphate buffer solution (PB) with a weight / volume ratio of 1.5 times, pH 6, and soaked at 4°C 1h; centrifuge at 15000rpm at 4°C for 20min; combine the supernatants from the two centrifugations;
[0054] (3) with Coomassie Brilliant Blue staining method (this method is selected from "Protein Molecular Basis", edited by Tao Weisun et al., Higher Education Press), taking bovine serum albumin as a standard, measuring the absorbance of jellyfish protein at 595nm (A 595 ), according to Lambert Beer's law, and then calculate the concentration of aequorin in the supernatant (C mg / ml ).
[0055] (4) with gel filtration chromatography (this method is selected from "Protein ...
Embodiment 3
[0057] (1) Break a certain amount of jellyfish with an ultrasonic breaker, add 5.0 times the weight / volume ratio of 20mM, pH8 phosphate buffer solution (PB), put it at 4°C, and soak for 2h;
[0058] (2) Centrifuge at 20,000 rpm for 10 minutes at 4°C, and take the supernatant; the residue is leached twice with 20mM phosphate buffer solution (PB) with a weight / volume ratio of 1.5 times, pH8, and soaked at 4°C 1h; centrifuge at 20000rpm at 4°C for 10min; combine the supernatants from the two centrifugations;
[0059] (3) with Coomassie Brilliant Blue staining method (this method is selected from "Protein Molecular Basis", edited by Tao Weisun et al., Higher Education Press), taking bovine serum albumin as a standard, measuring the absorbance of jellyfish protein at 595nm (A 595 ), according to Lambert Beer's law, and then calculate the concentration of aequorin in the supernatant (C mg / ml ).
[0060] (4) with gel filtration chromatography (this method is selected from "Protein ...
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