CN1752101A - Isolating albumen and application thereof from jellyfish with anti-oxidant activity - Google Patents

Isolating albumen and application thereof from jellyfish with anti-oxidant activity Download PDF

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CN1752101A
CN1752101A CNA2004100504813A CN200410050481A CN1752101A CN 1752101 A CN1752101 A CN 1752101A CN A2004100504813 A CNA2004100504813 A CN A2004100504813A CN 200410050481 A CN200410050481 A CN 200410050481A CN 1752101 A CN1752101 A CN 1752101A
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jellyfish
centrifugal
aequorin
phosphate buffer
buffer solution
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CN1324045C (en
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李鹏程
于华华
刘希光
邢荣娥
刘松
郭占勇
王丕波
李翠萍
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Institute of Oceanology of CAS
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Abstract

The present invention relates to marine organisms, specifically is the aequorin that two classes have anti-oxidant activity, and its separating step is after jellyfish is rubbed cytoclasis, to add in the phosphate buffer solution 2-6 ℃ of immersion 0.5-2h; Rotating speed 10000-20000rpm, centrifugal 10-30min gets supernatant liquor; The leaching of phosphate buffer solution secondary, rotating speed 10000-20000rpm, centrifugal 10-30min; Merge two times centrifugal supernatant; Use gel-filtration chromatography, the supernatant liquor that merges is separated, obtain two class aequorins; They can suppress ultra-oxygen anion free radical (O 2-), the generation of hydroxy radical qiao (OH), prevent that red corpuscle is because of hydrogen peroxide oxidation haemolysis and liver lipid peroxidation; Belong to natural antioxidants, the anti-oxidant activity height is to the people, animal toxicological harmless; Simultaneously, the present invention has utilized the jellyfish Living marine resources abundant, provides new way for its low value resource obtains high-valued development.

Description

Isolating albumen and application thereof from jellyfish with anti-oxidant activity
Technical field
The present invention relates to marine organisms, specifically is the aequorin that two classes have anti-oxidant activity, can be used as the medicine source of antiaging agent, and the while also provides new way for the development of natural additive for foodstuff.
Background technology
Active oxygen radical has huge damaging action to body, causes protein damage, enzyme deactivation, membranous peroxidation as meeting, causes the generation of numerous diseases such as aging, tumour, atherosclerosis.The butylated hydroxytoluene of synthetic (BHT), butylhydroxy methoxyl group (BHA), propylene glycol (PG) etc. have the effect of removing active oxygen radical preferably, are commonly used for foodstuff additive, but studies show that in recent years, and BHT, BHA, PG have carcinogenesis.Therefore screening is to human body safety, active oxygen scavenger has great significance efficiently.
Aequorin novel structure uniqueness has multiple biological activity, as: enzymic activity, influence ion running etc., but yet there are no report for anti-oxidant activity.China's jellyfish aboundresources, from Hainan Island, Guangdong, Shandong, until Liaoning is coastal distribution arranged all, when jellyfish exists in a large number, it will disturb the afloat nearly all activity of people, as swimming, support fishery, the more important thing is that it injures people's health, if stung by it, can produce multiple symptom, comprise pain, skin sting trace, oedema, neurally suppress, heartbeat stops, benumb even death.But to the utilization of jellyfish resource, present stage, just after simple processing, as food, added value was not high.So be necessary aequorin is studied, make this low value Biological resources of jellyfish obtain higher value application.
Summary of the invention
The objective of the invention is to utilize abundant jellyfish resource, from jellyfish, isolate albumen with anti-oxidant activity and the application of analyzing aequorin, make this low value Biological resources of jellyfish obtain higher value application.
For achieving the above object, the technical solution used in the present invention is: separate to have obtained the aequorin that two classes have anti-oxidant activity, the molecular weight of the first kind is greater than 800K, and the molecular weight of second class is less than 30K.(tentatively determining the molecular weight of aequorin according to the exclusion molecular weight of dextrane gel)
The separating technology step of this two classes aequorin is:
1) with after the even rubbing of the jellyfish of weighing, cytoclasis adds in the phosphate buffer solution of 0.5-5 by weight/volume doubly, puts under 2-6 ℃, soaks 0.5-2h;
2) under 4 ℃, rotating speed 10000-20000rpm (relative centrifugal force 9160-25480g), centrifugal 10-30min gets supernatant liquor; (5-20mM, PH4-8) secondary leaching under 4 ℃, are soaked 0.5-2h to residue with the phosphate buffer solution of 0.5-3 by weight/volume doubly; Again under 4 ℃, rotating speed 10000-20000rpm (relative centrifugal force 9160-25480g), centrifugal 10-30min; Merge two times centrifugal supernatant;
3) using the Xylene Brilliant Cyanine G method, is standard with the bovine serum albumin, measures the concentration of aequorin in the supernatant liquor that merges;
4) use gel-filtration chromatography, with 2) in the supernatant liquor of merging separate, obtain two class aequorins;
The proteic molecular weight of the first kind is greater than 800K, and iso-electric point PI precipitates in the ammoniumsulphate soln of percent saturation 30-60% at 3-9;
The molecular weight of second proteinoid is less than 30K, and iso-electric point PI precipitates in the ammoniumsulphate soln of percent saturation 0-30% at 4-8;
The amino acid masses percentage composition of two proteinoids that obtained is as follows:
The amino acid of table 1 liang proteinoid is formed (%)
Amino acid kind first kind albumen second proteinoid
Aspartic acid (Asp) 7.61 8.43
Threonine (THR) 4.12 3.89
Serine (SER) 2.52 2.62
Tryptophane (TRP) 3.23 3.43
L-glutamic acid (GLU) 13.29 13.15
Glycine (GLY) 13.09 12.18
L-Ala (ALA) 5.35 4.86
Gelucystine (CYS) 11.00 11.18
Xie Ansuan (VAL) 5.23 4.99
Methionine(Met) (MET) 3.85 3.89
Isoleucine (ILE) 3.06 3.11
Leucine (LEU) 3.85 3.82
Tyrosine (TYR) 2.06 2.07
Phenylalanine (PHE) 4.19 4.12
Methionin (LYS) 4.19 4.20
Histidine (HIS) 0.50 0.65
Arginine (ARG) 6.02 6.09
Proline(Pro) (PRO) 5.78 6.09
Total amino acid 98.94 98.77
Described jellyfish is fresh jellyfish or the fresh jellyfish that places-80～-20 ℃ the following preservation of low temperature; The concentration 5-20mM of phosphate buffer solution, PH4-8.
These two kinds of albumen can suppress ultra-oxygen anion free radical (O 2 -), the generation of hydroxy radical qiao (OH), prevent that red corpuscle is because of hydrogen peroxide oxidation haemolysis and liver lipid peroxidation.
The present invention has following advantage:
1) anti-oxidant activity height.Aequorin anti-oxidant activity height of the present invention can be used as the medicine source of antiaging agent, also provides new way for the development of natural additive for foodstuff simultaneously.
2) toxicological harmless.Aequorin of the present invention belongs to natural antioxidants, to the people, animal toxicological harmless.
3) production cost is low.Aequorin of the present invention has utilized this abundant Living marine resources of jellyfish, provides new approach for this low value resource of jellyfish obtains high-valued development.
Description of drawings
Figure 1A～C is that dextrane gel separates aequorin elution volume and absorbancy relation curve; Wherein, Figure 1A: dextrane gel SephadexG-100 separates aequorin; Figure 1B: dextrane gel SephadexG-200 separates first kind aequorin; Fig. 1 C: dextrane gel SephadexG-50 separates the second class aequorin;
Fig. 2 A～B is the infrared spectrogram of two proteinoids; Fig. 2 A: the proteic infrared spectrum of the first kind; Fig. 2 B: the infrared spectrum of second proteinoid;
The ammonium sulfate of Fig. 3 aequorin (classification) settling curve; First kind albumen is distributed in the ammonium sulfate precipitation of percent saturation 30%-60%; Second proteinoid is distributed in the ammonium sulfate precipitation of percent saturation 0-30%.
Embodiment
Protection scope of the present invention not only is confined in the following example.
One, the isolating embodiment of aequorin:
Embodiment 1
(1) a certain amount of jellyfish is used the fragmentation of ultrasonic disruption instrument, added the 5mM of 0.5 times by weight/volume, in the phosphate buffer solution of pH4 (PB), put under 2 ℃, soak 0.5h;
(2) under 4 ℃, rotating speed 10000rpm, centrifugal 30min gets supernatant liquor; Residue leaches with the 5mM of 0.5 times by weight/volume, the phosphate buffer solution of pH4 (PB) secondary, under 4 ℃, soaks 0.5h; Again under 4 ℃, rotating speed 10000rpm, centrifugal 30min; Merge two times centrifugal supernatant;
(3) with Coomassie brilliant blue staining (this method is selected from " protein molecule basis ", volumes such as Tao Weisun, Higher Education Publishing House), be standard, survey the absorbancy (A of aequorin at the 595nm place with the bovine serum albumin 595), according to langbobier law, and then calculate the concentration (Cmg/ml) of aequorin in the supernatant liquor.
(4) with gel-filtration chromatography (this method is selected from " protein molecule basis ", volumes such as Tao Weisun, Higher Education Publishing House), select gel SephadexG-100 for use, the supernatant liquor separation with the merging in (3) obtains two class aequorins, elutriant is 50mM, the phosphate buffer solution of PH6.The proteic molecular weight of the first kind (separates it with SephadexG-200 greater than 800K, elution volume and void volume are suitable, the exclusion molecular weight of SephadexG-200 is 800K), the molecular weight of second proteinoid (separates it with SephadexG-50 less than 30K, elution volume is greater than void volume, and the exclusion molecular weight of SephadexG-50 is 30K).
Embodiment 2
(1) a certain amount of jellyfish is used the fragmentation of ultrasonic disruption instrument, added the 10mM of 2.0 times by weight/volume, in the phosphate buffer solution of pH6 (PB), put under 4 ℃, soak 1h;
(2) under 4 ℃, rotating speed 15000rpm, centrifugal 20min gets supernatant liquor; Residue leaches with the 10mM of 1.5 times by weight/volume, the phosphate buffer solution of pH6 (PB) secondary, under 4 ℃, soaks 1h; Again under 4 ℃, rotating speed 15000rpm, centrifugal 20min; Merge two times centrifugal supernatant;
(3) with Coomassie brilliant blue staining (this method is selected from " protein molecule basis ", volumes such as Tao Weisun, Higher Education Publishing House), be standard, survey the absorbancy (A of aequorin at the 595nm place with the bovine serum albumin 595), according to langbobier law, and then calculate the concentration (C of aequorin in the supernatant liquor Mg/ml).
(4) with gel-filtration chromatography (this method is selected from " protein molecule basis ", volumes such as Tao Weisun, Higher Education Publishing House), select gel SephadexG-100 for use, the supernatant liquor separation with the merging in (3) obtains two class aequorins, elutriant is 50mM, the phosphate buffer solution of PH6.The proteic molecular weight of the first kind (separates it with SephadexG-200 greater than 800K, elution volume and void volume are suitable, the exclusion molecular weight of SephadexG-200 is 800K), the molecular weight of second proteinoid (separates it with SephadexG-50 less than 30K, elution volume is greater than void volume, and the exclusion molecular weight of SephadexG-50 is 30K).
Embodiment 3
(1) a certain amount of jellyfish is used the fragmentation of ultrasonic disruption instrument, added the 20mM of 5.0 times by weight/volume, in the phosphate buffer solution of pH8 (PB), put under 4 ℃, soak 2h;
(2) under 4 ℃, rotating speed 20000rpm, centrifugal 10min gets supernatant liquor; Residue leaches with the 20mM of 1.5 times by weight/volume, the phosphate buffer solution of pH8 (PB) secondary, under 4 ℃, soaks 1h; Again under 4 ℃, rotating speed 20000rpm, centrifugal 10min; Merge two times centrifugal supernatant;
(3) with Coomassie brilliant blue staining (this method is selected from " protein molecule basis ", volumes such as Tao Weisun, Higher Education Publishing House), be standard, survey the absorbancy (A of aequorin at the 595nm place with the bovine serum albumin 595), according to langbobier law, and then calculate the concentration (C of aequorin in the supernatant liquor Mg/ml).
(4) with gel-filtration chromatography (this method is selected from " protein molecule basis ", volumes such as Tao Weisun, Higher Education Publishing House), select gel SephadexG-100 for use, the supernatant liquor separation with the merging in (3) obtains two class aequorins, elutriant is 50mM, the phosphate buffer solution of PH6.The proteic molecular weight of the first kind (separates it with SephadexG-200 greater than 800K, elution volume and void volume are suitable, the exclusion molecular weight of SephadexG-200 is 800K), the molecular weight of second proteinoid (separates it with SephadexG-50 less than 30K, elution volume is greater than void volume, and the exclusion molecular weight of SephadexG-50 is 30K).
Table 2: dextrane gel SephadexG-100 separates aequorin
Column diameter (mm) Effective column length (mm) Void volume Vo (ml) Sample size (ml) Flow velocity (ml/h) Elution volume Ve1 (ml) Elution volume Ve2 (ml) Resolving power ρ *
16 170 14 2 9 13.2 35.8 1.56
16 170 14 2 19 12.4 35.4 1.53
16 170 14 2 30 13.4 37.8 1
16 370 28 2 22 35.0 73.2 0.71
16 370 28 2 28 37.1 71 0.61
16 370 28 2 31 44.2 89.6 0.81
10 370 14 1 10 14.6 35.2 1.6
10 370 14 1 14 16.7 35.5 1.2
10 370 14 1 21 16.8 35.8 1.16
10 750 29 1 9 First peak disappears 66.2 -
10 750 29 1 15 28.2 64.4 1.80
10 750 29 1 20 29.3 64.2 1.04
Shown in Figure 1A～C, dextrane gel separates aequorin elution volume and absorbancy relation curve.Wherein, Figure 1A separates aequorin for dextrane gel SephadexG-100; Figure 1B separates first kind aequorin for dextrane gel SephadexG-200; Fig. 1 C separates the second class aequorin for dextrane gel SephadexG-50.
Two, the embodiment of aequorin anti-oxidant activity:
1. aequorin is to the scavenging(action) of ultra-oxygen anion free radical
Adopt phenazine methosulfate-NADH system to produce free radical.Reaction system is the Tris-HCl damping fluid (16mmol/L of 3.0ml, pH8.0), wherein contain 78 μ mol/L DPNHs (NADH), 50 μ mol/L nitro blue tetrazoliums (NBT), 10 μ mol/L phenazine methosulfates (PMS), and the aequorin solution of different concns.The color reaction of ultra-oxygen anion free radical and NBT adopts the absorbancy of spectrophotometry assaying reaction liquid under the 560nm wavelength.In the blank experiment, replace NADH with the Tris-HCl damping fluid.
Experimental result is represented with clearance rate: clearance rate %=(A-A 1)/(A-A 0) * 100%;
Experimental result is with reference to shown in the table 3, wherein, and A: blank value, A 1: the light absorption value behind the adding sample, A 0: reference;
Table 3: aequorin is to the scavenging(action) of superoxide radical
First kind albumen Second proteinoid
Concentration (μ g/mL) Absorbance A 560 Clearance rate % Concentration (μ g/mL) Absorbance A 560 Clearance rate %
3.13 0.748±0.004 27.86±0.41 0.47 0.564±0.016 36.22±1.80
6.27 0.616±0.031 40.60±3.00 1.87 0.347±0.008 65.60±0.77
9.40 0.234±0.019 72.97±2.21 2.81 0.113±0.003 86.92±0.33
12.54 0.214±0.025 79.41±2.39 3.75 0.079±0.001 92.22±0.07
18.81 0.105±0.003 89.87±0.27 7.49 0.019±0.001 98.17±0.07
25.07 0.054±0.014 94.79±1.36 14.99 0.012±0.001 98.86±0.07
31.34 0.040±0.023 96.19±2.25 18.73 0.012±0.002 98.86±0.14
Blank: 1.037 (first kind albumen), 1.009 (second proteinoids)
Interpretation of result: two proteinoids have very strong scavenging(action) to superoxide anion, and scavenging(action) has dependency to concentration, and promptly along with the raising of concentration, scavenging(action) strengthens, EC 50Be respectively 6.12 μ g/ml, 0.88 μ g/ml, far below BHA, BHT, vitamin-E, their EC 50Be respectively 31 μ g/ml, 61 μ g/ml, 88 μ g/ml (M ü nirOktay, G ü lin and .
Figure A20041005048100092
Figure A20041005048100093
Determination of in vitro antioxidantactivity of fennel (Foeniculum vulgare) seed extracts, Lebensmittel-Wissenschaftund-Technologie, Volume 36, and Issue 2, March 2003, Pages 263-271).
2. aequorin is to the scavenging(action) of hydroxy radical qiao
OH is by EDTANa 2-Fe (II)-H 2O 2System produces, because OH can make the Stigma Croci redness fade specifically, weighs the content of OH with colorimetry according to fading extent.The phosphoric acid buffer 1.5ml that contains pH7.4 in the reaction system, Stigma Croci (260 μ g/ml) 0.2ml, 3% H 2O 20.8ml, EDTANa 2-Fe (II) 0.7ml, the aequorin solution of a certain amount of different concns mixes the back in 37 ℃ of water bath heat preservation 30min, surveys the absorbance A value then in the 520nm place.Blank group replaces sample liquid with distilled water, and control group replaces sample liquid and EDTANa with distilled water 2-Fe (II), the reaction cumulative volume is 4.0ml.
Experimental result is represented with clearance rate: clearance rate %=(A Sample-A Blank)/(A Contrast-A Blank) * 100%; Experimental result is with reference to shown in the table 4;
Table 4: aequorin is to the scavenging(action) of hydroxy radical qiao
First kind albumen Second proteinoid
Concentration (μ g/mL) Absorbance A 560 Clearance rate % Concentration (μ g/mL) Absorbance A 560 Clearance rate %
1.12 0.022±0.001 3.46±0.17 0.62 0.109±0.014 17.17±2.23
2.25 0.106±0.004 16.69+0.67 1.25 0.294±0.011 46.22±1.67
4.49 0.130±0.006 20.55±1.00 2.50 0.469±0.011 73.78±1.67
9.98 0.166±0.001 26.14±0.17 4.99 0.564±0.004 88.74±0.56
17.96 0.230±0.006 36.14±1.00 6.24 0.575±0.001 90.47±0.11
22.46 0.289±0.004 45.51±0.67 9.99 0.600±0.003 94.49±0.44
44.91 0.436±0.004 68.58±0.56
Blank: 0.635
Interpretation of result: two proteinoids have scavenging(action) to hydroxy radical qiao, and the scavenging(action) of second proteinoid is very strong, and when concentration was 9.99 μ g/ml, clearance rate reached 94.49%.Scavenging(action) has dependency to concentration, and promptly along with the raising of concentration, scavenging(action) strengthens.Two proteinoids are to the EC of hydroxy radical qiao 50Be respectively 45.42 μ g/ml, 1.52 μ g/ml, far below N.F,USP MANNITOL and vitamin-E, their EC 50Be respectively 4536 μ g/ml, 1907 μ g/ml.
3. aequorin is to H 2O 2The experiment of inductive oxidative hemolysis of erythrocyte
Healthy Wistar rat eye socket is got blood, make anticoagulation, the centrifugal 10min of 1000 * g, move and abandon blood plasma and white corpuscle, in sedimentary red corpuscle, add isoosmotic physiological saline, mixing, the centrifugal 10min of 1000 * g, abandon supernatant liquor, 2 Washed Red Blood Cells are so repeatedly made red corpuscle 0.5% suspension.Get red blood cell suspension 1ml, add the aequorin solution of different concns, add the H of 100mmol/l at last 2O 2, mixing, 37 ℃ of temperature are bathed 60min, and with 5 times of physiological saline dilutions, the centrifugal 10min of 1000 * g, supernatant liquor measure absorbance in the 415nm place.Experimental result is with reference to shown in the table 5;
Table 5: aequorin is to H 2O 2The effect of inductive oxidative hemolysis of erythrocyte
First kind albumen Second proteinoid
Concentration (μ g/mL) Absorbance A 560 Inhibiting rate % Concentration (μ g/mL) Absorbance A 560 Inhibiting rate %
1.68 0.337±0.002 68.88±0.17 0.47 0.170±0.017 84.35±1.63
3.37 0.242±0.004 77.65±0.68 0.94 0.158±0.016 85.36±1.50
6.74 0.204±0.004 81.21±0.68 1.87 0.153±0.008 85.92±0.72
13.47 0.177±0.001 83.66±0.09 2.81 0.147±0.007 86.43±0.65
33.68 0.169±0.004 84.40±0.68 3.75 0.134±0.003 87.62±0.26
67.37 0.159±0.004 85.36±0.68 5.62 0.133±0.001 87.77±0.07
Interpretation of result: two proteinoids are to H 2O 2The inductive oxidative hemolysis of erythrocyte has very strong restraining effect, when the proteic concentration of the first kind is 6.74 μ g/ml, inhibiting rate reaches 81.21%, when the concentration of second proteinoid is 0.47 μ g/ml, inhibiting rate reaches 84.35%, inhibiting rate is not very strong to the dependency of concentration, and along with the raising of concentration, it is not clearly that inhibiting rate changes.
4. aequorin is to the effect of rat liver homogenate lipid peroxidation
Healthy Wistar rat, the cervical vertebra dislocation causes death, and separates hepatic tissue rapidly, makes 20% homogenate with ice-cold Tris-HCl damping fluid (20mmol/l), the centrifugal 20min of 9810 * g, precipitation is washed once also centrifugal again, the merging supernatant liquor.(pH7.4) adds and contains liver homogenate liquid 0.2ml, FeSO in 0.2mol/l Tris-HCl damping fluid 410 μ mol/l, the aequorin solution of xitix 0.12mmol/l and different concns is bathed 60min 37 ℃ of temperature, and insulation finishes the back and adds 20% trichoroacetic acid(TCA) (TCA) 1.0ml termination reaction.Mixing adds 0.67% thiobarbituricacid (TBA) 1.5ml again, boiling water bath heating 15min.Behind the centrifugal removal protein precipitation, measure absorbance in 532nm.Experimental result is with reference to shown in the table 6.
Table 6: aequorin is to the rat liver homogenate lipid peroxidation
First kind albumen Second proteinoid
Concentration (μ g/mL) Absorbance A 560 Clearance rate % Concentration (μ g/mL) Absorbance A 560 Clearance rate %
6.74 0.288±0.001 21.88±0.19 1.87 0.270±0.003 26.63±0.77
13.47 0.212±0.002 42.53±0.58 3.75 0.194±0.002 47.42±0.58
26.95 0.134±0.003 63.59±0.77 7.50 0.164±0.001 55.43±0.38
40.42 0.109±0.002 70.52±0.58 11.24 0.143±0.004 61.28±0.96
53.89 0.097±0.001 73.78±0.19 14.98 0.130±0.003 64.7±0.76
67.37 0.070±0.002 81.11±0.58 18.73 0.104±0.001 71.88±0.19
Interpretation of result: two proteinoids have obvious suppression effect, EC to the liver homogenate lipid peroxidation 50Be respectively 18.74 μ g/ml, 5.74 μ g/ml, two proteinoids all show concentration dependent to suppressing lipid peroxidation.

Claims (4)

1. isolating albumen with anti-oxidant activity from jellyfish is characterized in that:
Its separating step is,
1) with after the even rubbing of the jellyfish of weighing, cytoclasis adds in the phosphate buffer solution of 0.5-5 by weight/volume doubly, puts under 2-6 ℃, soaks 0.5-2h;
2) under 4 ℃, rotating speed 10000-20000rpm, centrifugal 10-30min gets supernatant liquor; Residue under 4 ℃, soaks 0.5-2h with the phosphate buffer solution secondary leaching of 0.5-3 by weight/volume doubly; Again under 4 ℃, rotating speed 10000-20000rpm, centrifugal 10-30min; Merge two times centrifugal supernatant;
3) use gel-filtration chromatography, with 2) in the supernatant liquor of merging separate, obtain two class aequorins;
The proteic molecular weight of the first kind is greater than 800K, and iso-electric point PI precipitates in the ammoniumsulphate soln of percent saturation 30-60% at 3-9; Protein mass percentage composition: aspartic acid 7.61%, Threonine 4.12%, Serine 2.52%, tryptophane 3.23%, L-glutamic acid 13.29%, glycine 13.09%, L-Ala 5.35%, Gelucystine 11.00%, Xie Ansuan 5.23%, methionine(Met) 3.85%, Isoleucine 3.06%, leucine 3.85%, tyrosine 2.06, phenylalanine 4.19%, Methionin 4.19%, Histidine 0.50%, arginine 6.02%, proline(Pro) 5.78%, total amino acid 98.94%;
The molecular weight of second proteinoid is less than 30K, and iso-electric point PI precipitates in the ammoniumsulphate soln of percent saturation 0-30% at 4-8; Protein mass percentage composition: aspartic acid 8.43%, Threonine 3.89%, Serine 2.62%, tryptophane 3.43%, L-glutamic acid 13.15%, glycine 12.18%, L-Ala 4.86%, Gelucystine 11.18%, Xie Ansuan 4.99%, methionine(Met) 3.89%, Isoleucine 3.11%, leucine 3.82%, tyrosine 2.07%, phenylalanine 4.12%, Methionin 4.20%, Histidine 0.65%, arginine 6.09%, proline(Pro) 6.09%, total amino acid 98.77%.
According to claim 1 described from jellyfish isolating albumen with anti-oxidant activity, it is characterized in that: described jellyfish is fresh jellyfish or the fresh jellyfish that places-80～-20 ℃ low temperature to preserve down.
According to claim 1 described from jellyfish isolating albumen with anti-oxidant activity, it is characterized in that: the concentration 5-20mM of phosphate buffer solution, PH4-8.
4. described proteic application of claim 1, it is characterized in that: these two kinds of albumen can suppress ultra-oxygen anion free radical (O 2 -), the generation of hydroxy radical qiao (OH), prevent that red corpuscle is because of hydrogen peroxide oxidation haemolysis and liver lipid peroxidation.
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CN101130572B (en) * 2007-08-08 2010-11-24 中国科学院海洋研究所 Method for extracting toxin from jellyfish tentacle
CN102516379A (en) * 2011-12-13 2012-06-27 中国科学院海洋研究所 Protein having superoxide anion-clearing activity and separated from Stomolophus meleagris L. Agassiz nematocyst venom
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JP2003321497A (en) * 2002-04-24 2003-11-11 Ryoyo Sangyo Kk Jellyfish collagen
JP2004099513A (en) * 2002-09-09 2004-04-02 Toshiba Corp Method and system for recovering collagen from jellyfish

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CN101130572B (en) * 2007-08-08 2010-11-24 中国科学院海洋研究所 Method for extracting toxin from jellyfish tentacle
WO2009114597A1 (en) * 2008-03-11 2009-09-17 Quincy Bioscience, Llc Apoaequorin-containing compositions and methods of using same
US8796213B2 (en) 2008-03-11 2014-08-05 Qunicy Bioscience, LLC Apoaequorin-containing compositions and methods of using same
CN102516379A (en) * 2011-12-13 2012-06-27 中国科学院海洋研究所 Protein having superoxide anion-clearing activity and separated from Stomolophus meleagris L. Agassiz nematocyst venom
CN102516379B (en) * 2011-12-13 2013-10-16 中国科学院海洋研究所 Separating from Stomolophus meleagris L. Agassiz nematocyst venom of protein having superoxide anion-clearing activity
WO2018058050A1 (en) * 2016-09-23 2018-03-29 Quincy Bioscience, Llc Apoaequorin and vitamin d-containing compositions and methods of using same

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