CN105648007A - Preparation method of mustelus griseus chondroprotein antioxidative peptide - Google Patents
Preparation method of mustelus griseus chondroprotein antioxidative peptide Download PDFInfo
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Abstract
The invention discloses a preparation method of mustelus griseus chondroprotein antioxidative peptide. In the method, mustelus griseus cartilage is taken as a raw material; an enzymatic hydrolysate is obtained through total protein extraction, acetone fractional precipitation, and trypsin enzymatic hydrolysis; and the enzymatic hydrolysate is separated and purified through ultrafiltration, ion exchange resin chromatography, gel column chromatography, and reversed-phase high performance liquid chromatography to obtain the antioxidative peptide Gly-Glu-Arg-Glu-Ala-Asn-Val-Met. The molecular weight of the antioxidative peptide is measured through ESI-MS and is 905.00 Da. The prepared high-activity antioxidative peptide has high free-radical scavenging activity and an excellent lipid-peroxidation inhibiting effect, and can be used for developing a medicine, a health-care food, and a food additive.
Description
Technical field
The preparation method that the present invention relates to a kind of animal cartilage bioactive peptide, the preparation method being specifically related to a kind of Mustelus griseus (pietschmann) chondroprotein anti-oxidation peptide.
Background technology
Mustelus griseus (pietschmann) (Mustelusgriseus) popular name sand bar, also known as ash ermine shark, for the one of Chondrichthyes Carcharhiniforme Triakidae Mustelus. It is distributed in northwest Pacific district, the marine site such as including South China Sea, the East China Sea and the Yellow Sea, Taiwan, and Busan, Korea and the sea area such as Mokpo and South Japan.
Mustelus griseus (pietschmann) regulating liver-QI, meat, skin, fin etc. all can be used as medicine. Mustelus griseus (pietschmann) has the effects such as solution fish and shrimp poison and promoting digestion and removing stagnation. Mustelus griseus (pietschmann) meat has the effects such as invigorating the spleen and benefiting QI, removing blood stasis to reduce swelling, strengthening by means of tonics, it is possible to treatment body is empty, the diseases such as wound healing is slow, insufficiency of the spleen edema. Mustelus griseus (pietschmann) has the effects such as strengthening by means of tonics, QI invigorating, qi-restoratives. Mustelus griseus (pietschmann) has removing food stagnancy, solves the effects such as all ichthyotoxin. Mustelus griseus (pietschmann) has heat-clearing and toxic substances removing, detumescence, the effect such as invigorate blood circulation. Mustelus griseus (pietschmann) bone has the effects such as expelling wind and removing dampness, invigorating the spleen to arrest diarrhea, promoting blood circulation and stopping pain. The Mustelus griseus (pietschmann) heart has invigorating spleen and reinforcing stomach, and expectorant dissipating fluid-retention, QI invigorating opens the effects such as diaphragm. The interior retention of phlegm drink because weakness of the spleen and stomach causes, indigestion, breast wrist distension can be treated, vomit diseases such as having loose bowels.
Summary of the invention
The technical problem to be solved be to provide a kind of can the preparation method of Mustelus griseus (pietschmann) chondroprotein anti-oxidation peptide of scavenging free radicals and anti-lipid peroxidation effect.
The present invention solves that the technical scheme that above-mentioned technical problem is taked is: the preparation method of Mustelus griseus (pietschmann) chondroprotein anti-oxidation peptide, it comprises the following steps:
1) preparation of Mustelus griseus (pietschmann) cartilage total protein crude extract:Weigh Mustelus griseus (pietschmann) cartilage and be cut into fine grained chippings, add appropriate distilled water, in high-speed tissue mashing machine, homogenate is to pasty state, the Mustelus griseus (pietschmann) cartilage of pasty state is dipped in the 1.0mol/L guanidine hydrochloride solution of 4��6 times of volumes (containing 0.02mol/LMES and 0.02mol/LEDTA, pH7��8), at 4 DEG C, stir extracting 36��48h. Extracting solution with the centrifugal 15��25min of 4000��6000r/min, is abandoned residue and is collected supernatant at 4 DEG C; Supernatant continues, at 4 DEG C, the centrifugal 15��25min of 10000��12000r/min, to take supernatant and remove insoluble impurities with 0.22 ��m of filtering with microporous membrane. Filtrate being loaded in the bag filter that molecular retention amount is 8000, with Tris-HCl(pH7.6) buffer dialyses 24��48h at 4 DEG C, and in bag filter, solution is Mustelus griseus (pietschmann) cartilage total protein crude extract.
2) preparation of Mustelus griseus (pietschmann) cartilage acetone fractional precipitation albumen:Taking Mustelus griseus (pietschmann) cartilage total protein crude extract, to be slowly added to pre-cold acetone to acetone concentration under ice bath be 30%, after standing 4��6h at-20 DEG C, in 4 DEG C, 10000��12000r/min high speed centrifugation 20min, take supernatant, adding pre-cold acetone to the final acetone concentration of solution is 60%, obtain Mustelus griseus (pietschmann) cartilage 60% acetone precipitation albumen, lyophilizing, be 60% acetone fractional precipitation albumen.
3) enzymolysis of Mustelus griseus (pietschmann) cartilage acetone fractional precipitation albumen:Take Mustelus griseus (pietschmann) cartilage 60% acetone fractional precipitation albumen and be dissolved in Tris-HCl buffer (0.05mol/L, pH7.5��8.5) according to solid-liquid ratio 1g:3��5mL, add trypsin 1.9 �� 10 according to the 2.0��3.0% of 60% acetone fractional precipitation protein by weight4U/g), in 40 DEG C of enzymolysis 3��5h, then in 90 DEG C, the work of 10min enzyme denaturing, enzymolysis solution, in 4 DEG C, the centrifugal 15��25min of 10000��12000r/min, is abandoned residue and is collected supernatant, obtain chondroprotein enzymolysis solution.
4) preparation of Mustelus griseus (pietschmann) chondroprotein anti-oxidation peptide:Adopt 3kDa ultrafilter membrane to carry out hyperfiltration treatment above-mentioned enzymolysis solution, collect molecular weight less than 3kDa part, lyophilizing, obtain ultrafiltration zymolyte; By ultrafiltration zymolyte time through resinbed analysis, gel filtration chromatography and reversed-phase high-performance liquid chromatography (RP-HPLC) purification, obtain anti-oxidation peptide.
As preferably, the described Mustelus griseus (pietschmann) in step 1) be Mustelus griseus (pietschmann) (Mustelusgriseus).
As preferably, the detailed process of the resinbed analysis of described step 4), gel filtration chromatography and RP-HPLC purification is:
Resinbed is analysed: above-mentioned ultrafiltration zymolyte is dissolved in distilled water and is made into the solution that concentration is 45��55mg/mL, post is analysed through DEAE-52 resinbed, eluting is carried out with water, 0.1mol/L, 0.5mol/L and 1.0mol/LNaCl solution, flow velocity is 0.6��0.8mL/min, the every 3min of wash solution collects a pipe and detects in 280nm, solution in test tube is merged by peak, compare the DPPH free radical at each peak and the scavenging capacity of hydroxyl radical free radical, select the strongest component lyophilizing of activity, be ion-exchange chromatography zymolyte;
Gel filtration chromatography: above-mentioned ion-exchange chromatography zymolyte is dissolved in distilled water and is made into the solution that concentration is 45��55mg/mL, through polydextran gel SephadexG-15 column chromatography for separation, eluting is carried out with distilled water, flow velocity is 0.6��0.8mL/min, the every 3min of wash solution collects a pipe and detects in 280nm, merges solution in test tube by peak, compares the DPPH free radical at each peak and the scavenging capacity of hydroxyl radical free radical, select the strongest component lyophilizing of activity, be gel chromatography zymolyte;
RP-HPLC purification: above-mentioned gel chromatography zymolyte distilled water is made into the solution of 80��100 �� g/mL, RP-HPLC is utilized to be purified, obtaining 1 high activity anti-oxidation peptide Gly-Glu-Arg-Glu-Ala-Asn-Val-Met (GEREANVM) according to the scavenging capacity of DPPH free radical and hydroxyl radical free radical, it is 905.00Da that ESI-MS measures molecular weight.
Further preferably, described RP-HPLC condition is: sample size 10��15 �� L; Chromatographic column ZorbaxSB-C18(250mm �� 4.6mm, 5 ��m); Mobile phase: water-acetonihile gradient elution (0��32min acetonitrile concentration is at the uniform velocity risen to 50% by 0); Elution speed 0.6 ~ 1.0mL/min; Ultraviolet detection wavelength 280nm.
Compared with prior art, DPPH free radical, hydroxyl radical free radical, ABTS free radical and ultra-oxygen anion free radical are had good scavenging action by Mustelus griseus (pietschmann) chondroprotein anti-oxidation peptide provided by the present invention;Meanwhile, Gly-Glu-Arg-Glu-Ala-Asn-Val-Met (GEREANVM) also demonstrates good Lipid peroxidation; Gly-Glu-Arg-Glu-Ala-Asn-Val-Met (GEREANVM) has safe without toxic side effect, antioxidant activity is strong and is prone to advantages such as digesting and assimilating, it is possible to as the additive of medicine, health food and food.
Accompanying drawing explanation
Fig. 1 is the DEAE-52 resinbed analysis figure of the present invention.
Fig. 2 is the polydextran gel SephadexG-15 tomographic map of the present invention.
Fig. 3 polydextran gel SephadexG-15 prepares the RP-HPLC of zymolyte and analyzes.
The mass spectrum of Fig. 4 Gly-Glu-Arg-Glu-Ala-Asn-Val-Met (GEREANVM).
The lipid peroxidation inhibition ability of Fig. 5 Gly-Glu-Arg-Glu-Ala-Asn-Val-Met (GEREANVM).
Detailed description of the invention
Below in conjunction with accompanying drawing embodiment, the present invention is described in further detail.
Embodiment:
The preparation method of a kind of Mustelus griseus (pietschmann) chondroprotein anti-oxidation peptide, preparation technology flow process is as follows: hammerhead cartilage " cartilage total protein " cartilage 60% acetone precipitation albumen " trypsin digestion " zymolyte " ultrafiltration " ion-exchange chromatography " gel permeation chromatography " high performance liquid chromatography prepares " anti-oxidation peptide.
1) preparation of Mustelus griseus (pietschmann) cartilage total protein crude extract:Weigh Mustelus griseus (pietschmann) (Mustelusgriseus) cartilage is cut into fine grained chippings, adding appropriate distilled water, in high-speed tissue mashing machine, homogenate is to pasty state, is dipped in the 1.0mol/L guanidine hydrochloride solution of 4 times of volumes the Mustelus griseus (pietschmann) cartilage of pasty state (containing 0.02mol/LMES and 0.02mol/LEDTA, pH7.0), at 4 DEG C, extracting 48h is stirred. Extracting solution with the centrifugal 20min of 5000r/min, is abandoned residue and is collected supernatant at 4 DEG C; Supernatant continues, at 4 DEG C, the centrifugal 20min of 12000r/min, to take supernatant and remove insoluble impurities with 0.22 ��m of filtering with microporous membrane. Filtrate being loaded in the bag filter that molecular retention amount is 8000, with Tris-HCl(pH7.6) buffer dialyses 48h at 4 DEG C, and in bag filter, solution is Mustelus griseus (pietschmann) cartilage total protein crude extract.
2) preparation of Mustelus griseus (pietschmann) cartilage acetone fractional precipitation albumen:Take Mustelus griseus (pietschmann) cartilage total protein crude extract, being slowly added to pre-cold acetone to acetone concentration under ice bath is 30%, after standing 5h at-20 DEG C, in 4 DEG C, 10000r/min high speed centrifugation 20min, taking supernatant, adding pre-cold acetone to the final acetone concentration of solution is 60%, it is thus achieved that Mustelus griseus (pietschmann) cartilage 60% acetone precipitation albumen, lyophilizing, obtains 60% acetone fractional precipitation albumen.
3) enzymolysis of Mustelus griseus (pietschmann) cartilage acetone fractional precipitation albumen:Take Mustelus griseus (pietschmann) cartilage 60% acetone fractional precipitation albumen and be dissolved in Tris-hydrochloride buffer (0.05mol/L, pH8.0) according to solid-liquid ratio 1g:4mL, add trypsin 1.9 �� 10 according to the 2.5% of 60% acetone fractional precipitation protein by weight4U/g), in 40 DEG C of enzymolysis 4h, then in 90 DEG C, the work of 10min enzyme denaturing, enzymolysis solution, in 4 DEG C, the centrifugal 20min of 12000r/min, is abandoned residue and is collected supernatant, obtain chondroprotein enzymolysis solution.
4) preparation of Mustelus griseus (pietschmann) chondroprotein anti-oxidation peptide:Adopt 3kDa ultrafilter membrane to carry out hyperfiltration treatment above-mentioned enzymolysis solution, collect molecular weight less than 3kDa part (MP60-I), obtain ultrafiltration enzymolysis solution, by ultrafiltration enzymolysis solution successively through resinbed analysis, gel filtration chromatography and reversed-phase high-performance liquid chromatography (RP-HPLC) purification, obtain anti-oxidation peptide.
1. resinbed analysis: MP60-I is dissolved in distilled water and is made into the solution that concentration is 50mg/mL, post is analysed through DEAE-52 resinbed, eluting is carried out with water, 0.1mol/L, 0.5mol/L and 1.0mol/LNaCl solution, flow velocity is 0.6mL/min, the every 3min of wash solution collects a pipe and detects in 280nm, solution in test tube is merged by peak, compare DPPH free radical and the Hydroxyl radical-scavenging ability at each peak, select the strongest component lyophilizing of oxidation resistance, be ion-exchange chromatography zymolyte MP60-3(Fig. 1);
2. gel filtration chromatography: MP60-3 is dissolved in distilled water and is made into the solution that concentration is 50mg/mL, through polydextran gel SephadexG-15 column chromatography for separation, eluting is carried out with distilled water, flow velocity is 0.6��0.8mL/min, the every 3min of wash solution collects a pipe and detects in 280nm, merges solution in test tube by peak, compares DPPH free radical and the Hydroxyl radical-scavenging ability at each peak, select the strongest component lyophilizing of oxidation resistance, be gel chromatography zymolyte MP60-3-1(Fig. 2);
3. RP-HPLC purification: MP60-3-1 distilled water is made into the solution of 80��100 �� g/mL, (RP-HPLC condition is: sample size 15 �� L to utilize RP-HPLC to be purified; Chromatographic column ZorbaxSB-C18(250mm �� 4.6mm, 5 ��m); Mobile phase: water-acetonihile gradient elution (0��32min acetonitrile concentration is at the uniform velocity risen to 50% by 0); Elution speed 0.8mL/min; Ultraviolet detection wavelength 280nm), obtain 1 high activity anti-oxidation peptide DCPE-B(Fig. 3 according to the scavenging capacity of DPPH free radical and hydroxy radical).
4. structure detection: collect DPPH free radical and the highest anti-oxidation peptide DCPE-B of Scavenging activity on hydroxyl free radical, it is simple spike through RP-HPLC detection, utilizing protein/polypeptide sequenator to measure aminoacid sequence is Gly-Glu-Arg-Glu-Ala-Asn-Val-Met (GEREANVM), and it is 905.00Da([M+H that ESI-MS measures molecular weight]+906.68Da) (Fig. 4).
Prepared Mustelus griseus (pietschmann) chondroprotein anti-oxidation peptide Gly-Glu-Arg-Glu-Ala-Asn-Val-Met (GEREANVM) is carried out free radical scavenging experiment and lipid peroxidation Inhibition test, test result indicate that: Gly-Glu-Arg-Glu-Ala-Asn-Val-Met (GEREANVM) is to DPPH free radical (EC502.76mg/mL), hydroxyl radical free radical (EC500.22mg/mL), ABTS free radical (EC500.08mg/mL) with ultra-oxygen anion free radical (EC500.13mg/mL) there is good scavenging action; Meanwhile, Gly-Glu-Arg-Glu-Ala-Asn-Val-Met (GEREANVM) also demonstrates good Lipid peroxidation (Fig. 5).
Finally, in addition it is also necessary to be only the specific embodiment of the present invention it is noted that listed above. It is clear that the invention is not restricted to above example, it is also possible to there are many deformation. All deformation that those of ordinary skill in the art can directly derive from present disclosure or associate, are all considered as protection scope of the present invention.
SEQUENCELISTING
<110>Oceanography Institute Of Zhejiang
<120>preparation method of Mustelus griseus (pietschmann) chondroprotein anti-oxidation peptide
<130>zjou-wb-201511-402
<160>1
<170>PatentInversion3.5
<210>1
<211>8
<212>PRT
<213>synthetic
<400>1
GlyGluArgGluAlaAsnValMet
15
Claims (5)
1. the preparation method of Mustelus griseus (pietschmann) chondroprotein anti-oxidation peptide, it is characterised in that comprise the following steps:
1) preparation of Mustelus griseus (pietschmann) cartilage total protein crude extract:Weighing Mustelus griseus (pietschmann) cartilage and be cut into fine grained chippings, add appropriate distilled water, in high-speed tissue mashing machine, homogenate is to pasty state, is dipped in the 1.0mol/L guanidine hydrochloride solution of 4��6 times of volumes by the Mustelus griseus (pietschmann) cartilage of pasty state, stirs extracting 36��48h at 4 DEG C; Extracting solution with the centrifugal 15��25min of 4000��6000r/min, is abandoned residue and is collected supernatant at 4 DEG C; Supernatant continues, at 4 DEG C, the centrifugal 15��25min of 10000��12000r/min, to take supernatant and remove insoluble impurities with 0.22 ��m of filtering with microporous membrane; Filtrate being loaded in the bag filter that molecular retention amount is 8000, dialyse 24��48h with the Tris-HCl buffer that pH value is 7.6 at 4 DEG C, in bag filter, solution is Mustelus griseus (pietschmann) cartilage total protein crude extract;
2) preparation of Mustelus griseus (pietschmann) cartilage acetone fractional precipitation albumen:Taking Mustelus griseus (pietschmann) cartilage total protein crude extract, to be slowly added to pre-cold acetone to acetone concentration under ice bath be 30%, after standing 4��6h at-20 DEG C, in 4 DEG C, 10000��12000r/min high speed centrifugation 20min, take supernatant, adding pre-cold acetone to the final acetone concentration of solution is 60%, obtain Mustelus griseus (pietschmann) cartilage 60% acetone precipitation albumen, lyophilizing, be 60% acetone fractional precipitation albumen;
3) enzymolysis of Mustelus griseus (pietschmann) cartilage acetone fractional precipitation albumen:Take Mustelus griseus (pietschmann) cartilage 60% acetone fractional precipitation albumen and be dissolved in Tris-HCl buffer (0.05mol/L, pH7.5��8.5) according to solid-liquid ratio 1g:3��5mL, add trypsin 1.9 �� 10 according to the 2.0��3.0% of 60% acetone fractional precipitation protein by weight4U/g), in 40 DEG C of enzymolysis 3��5h, then in 90 DEG C, the work of 10min enzyme denaturing, enzymolysis solution, in 4 DEG C, the centrifugal 15��25min of 10000��12000r/min, is abandoned residue and is collected supernatant, obtain chondroprotein enzymolysis solution;
4) preparation of Mustelus griseus (pietschmann) chondroprotein anti-oxidation peptide:Adopt 3kDa ultrafilter membrane to carry out hyperfiltration treatment above-mentioned enzymolysis solution, collect molecular weight less than 3kDa part, lyophilizing, obtain ultrafiltration zymolyte; By ultrafiltration zymolyte time through resinbed analysis, gel filtration chromatography and reversed-phase high-performance liquid chromatography purification, obtain anti-oxidation peptide;
Wherein, the guanidine hydrochloride solution in described step 1) is containing 0.02mol/LMES and 0.02mol/LEDTA, and pH value is 7��8; Tris-HCl buffer in described step 3) is 0.05mol/L, and pH value is 7.5��8.5; Trypsin is 1.9 �� 104U/g��
2. preparation method according to claim 1, it is characterised in that the Mustelus griseus (pietschmann) in described step 1) is Mustelus griseus (pietschmann) isMustelusgriseus��
3. preparation method according to claim 1, it is characterised in that the detailed process of the resinbed analysis of described step 4), gel filtration chromatography and reversed-phase high-performance liquid chromatography purification is:
Resinbed is analysed: above-mentioned ultrafiltration zymolyte is dissolved in distilled water and is made into the solution that concentration is 45��55mg/mL, post is analysed through DEAE-52 resinbed, eluting is carried out with water, 0.1mol/L, 0.5mol/L and 1.0mol/LNaCl solution, flow velocity is 0.6��0.8mL/min, the every 3min of wash solution collects a pipe and detects in 280nm, solution in test tube is merged by peak, compare the DPPH free radical at each peak and the scavenging capacity of hydroxyl radical free radical, select the strongest component lyophilizing of activity, be ion-exchange chromatography zymolyte;
Gel filtration chromatography: above-mentioned ion-exchange chromatography zymolyte is dissolved in distilled water and is made into the solution that concentration is 45��55mg/mL, through polydextran gel SephadexG-15 column chromatography for separation, eluting is carried out with distilled water, flow velocity is 0.6��0.8mL/min, the every 3min of wash solution collects a pipe and detects in 280nm, merges solution in test tube by peak, compares the DPPH free radical at each peak and the scavenging capacity of hydroxyl radical free radical, select the strongest component lyophilizing of activity, be gel chromatography zymolyte;
Reversed-phase high-performance liquid chromatography purification: above-mentioned gel chromatography zymolyte distilled water is made into the solution of 80��100 �� g/mL, reversed-phase high-performance liquid chromatography purification is utilized to be purified, according to the scavenging capacity of DPPH free radical and hydroxyl radical free radical obtains 1 high activity anti-oxidation peptide Gly-Glu-Arg-Glu-Ala-Asn-Val-Met, ESI-MS, to measure molecular weight be 905.00Da.
4. preparation method according to claim 1, it is characterised in that described reversed-phase high-performance liquid chromatography condition is: sample size 10��15 �� L;Chromatographic column is specification is 250mm �� 4.6mm, the ZorbaxSB-C18 of 5 ��m; Mobile phase is water-acetonihile gradient elution, and 0��32min acetonitrile concentration is at the uniform velocity risen to 50% by 0; Elution speed 0.6 ~ 1.0mL/min; Ultraviolet detection wavelength 280nm.
5. preparation method according to claim 1, it is characterised in that prepared anti-oxidation peptide Gly-Glu-Arg-Glu-Ala-Asn-Val-Met is octapeptide compounds, it is 905.00Da that ESI-MS measures molecular weight.
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| CN110108808A (en) * | 2019-04-12 | 2019-08-09 | 广西壮族自治区兽医研究所 | Method that is a kind of while evaluating oxidation-resistant active ingredient and content in downy rosemyrtle root |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103275180A (en) * | 2013-05-23 | 2013-09-04 | 浙江海洋学院 | Navodon septentrionalos fish head protein antioxidant peptide as well as preparation method and application thereof |
-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103275180A (en) * | 2013-05-23 | 2013-09-04 | 浙江海洋学院 | Navodon septentrionalos fish head protein antioxidant peptide as well as preparation method and application thereof |
| CN103275180B (en) * | 2013-05-23 | 2015-03-11 | 浙江海洋学院 | Navodon septentrionalos fish head protein antioxidant peptide as well as preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 胡发远: "孔鳐软骨多肽及泥蚶多肽的制备及活性研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110108808A (en) * | 2019-04-12 | 2019-08-09 | 广西壮族自治区兽医研究所 | Method that is a kind of while evaluating oxidation-resistant active ingredient and content in downy rosemyrtle root |
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