CN107474148A - A kind of method from sargassum extraction polysaccharide compound - Google Patents

Abstract

The present invention discloses a kind of method from sargassum extraction polysaccharide compound, belongs to technical field of polysaccharide extraction, the extracting method of polysaccharide：Including breaking-wall cell, concentration alcohol analysis, removing protein, freeze-drying are redissolved, breaking-wall cell step is：Take sargassum powder that algae solution is made, ultrasonic disruption, add bioactive peptide water-bath, centrifugation, except frond, the present invention significantly improves sargassum cell membrane percentage of damage by ultrasonic wave and bioactive peptide, improves the yield and purity of sargassan, the harmful effect of nutritive loss will not be caused to sargassum, the sargassan of extraction can be used for promoting in immunity, antitumor health products.

Description

A kind of method from sargassum extraction polysaccharide compound

Technical field

The invention belongs to technical field of polysaccharide extraction, and in particular to a kind of method from sargassum extraction polysaccharide compound.

Background technology

Sargassum (Sargassum pallidum) is a kind of marine alga in brown alga, category Phaeophyta (Phaeophyta), ink Angle Cutleriales (Fucales), Sargassaceae ((Sargassaceae), Sargassum ((Sargassum).Sargassum is warm temperate zone property Marine alga, perennial plant, therefore whole year can be seen.Abound with China's Huanghai Sea, Sea And Bohai Sea Coast various regions, be a kind of algae commonly seen Class, yield are very big.Foreign countries are distributed in the ground such as Sea of Okhotsk, Thousand Islands, Japan and Korea.Sargassum habitat requires sea Wave is unimpeded, and the wave is high greatly for wind, and water colour is clear, and transparency is high, sunny, is generally grown on the Shi Zhaozhong and subtidal zone water of intertidal zone On the rock of lower 4 meters of depths of 1-, for the dominant species in the waters, resource very abundant.

Recent 20 years come, and with going deep into for the secondary metabolite to brown alga-brown algae polyphenols compound research, it is unique Bioactivity be increasingly subject to the concern of people.Antioxidation activity report on marine alga reclaimed water, alcohol or fat-soluble solvent extract Road is a lot of.MaStSUk think marine alga water or alcohol extracting thing in oxidation-resistant active ingredient be the lipoxidase from Sargassum Inhibitor, or the brown algae polyphenols from Eisenia bicyclis.Antioxidation mechanism on brown algae polyphenols, it is considered that deposit In two kinds of approach:First, the dissociation by the light base of phenol.The reproducibility of the light base of phenol is one of general character of phenolic compound, with isophthalic three Phenol can easily be aoxidized for the light base of multiple phenol in the brown algae polyphenols molecule of monomer as the donor of hydrogen by the oxygen in air;Two It is by free radical approach.Biological anti-oxidant recognized by present is primarily referred to as that free radical can be removed, and suppresses lipid peroxy The active material of change, Yan and Li are purified into brown algae polyphenols from brown alga Sargassum kjellmanianum, and have studied it The inhibitory action become sour to fish oil, it is found that its anti-oxidant efficiency is higher than BHT 2.6 times.It is very strong that research shows that brown algae polyphenols have Radical scavenging activity, it is a kind of natural and freedom for having very much prospect of the application in fields such as pharmacy, food and daily use chemicals Base scavenger.

Prior art, such as Chinese invention granted patent document, the B of Authorization Notice No. CN 104829740, the invention is related to one The method that kind synchronously extracts blade of grass sargassan and blade of grass sargassum polyphenol from blade of grass sargassum, described method is using micro- Synchronously extraction obtains blade of grass sargassan and grass from blade of grass sargassum for ripple assisted extraction, aqueous two-phase extraction and membrane separation technique Leaf sargassum polyphenol.The invention additionally provides the blade of grass sargassan extract obtained by above method and blade of grass sargassum Polyphenol extract.It the advantage is that：Sargassan recovery rate is high, and bioactivity loss is small, and sargassan and sargassum are more The purity of phenol is all very high, while the method for the invention has that the time is short, treating capacity is big, operation can serialization, cost be cheap, environment The advantage of cleaning, therefore commercial Application is especially suitable for, but the sargassan yield in this method also has room for promotion.

The content of the invention

It is an object of the invention to provide a kind of method of the polysaccharide compound from sargassum extraction, shown by bioactive peptide Write and improve sargassum cell membrane percentage of damage, improve the yield and purity of sargassan, nutrition will not be caused to sargassan The harmful effect of loss, the sargassan compound of extraction can be used for having in anti-oxidant, antitumous effect health products.

The technical scheme taken to achieve the above object of the present invention is：A kind of side that polysaccharide compound is extracted from sargassum Method, including breaking-wall cell, concentration alcohol analysis, redissolve removing protein, are freeze-dried, concrete operation step is as follows：

1）Sargassum powder is taken to be made algae solution, the solid-to-liquid ratio of sargassum powder and water is 1:65-75, ultrasonic disruption, power 500w- 800w, time 10-20min, add algae solution weight 0.01%-0.03% bioactive peptide, and the amino acid sequence of bioactive peptide is QSGFCTSYYSKFCTLYGNLGICLHCSS, the water-bath 2-4h at 98 DEG C ~ 100 DEG C, 6500-9000r/min centrifugation 5-10min, Except frond；

2）Algae solution is put into the 1/4-1/3 that original volume is concentrated in vacuo in vacuum concentration pot, adds 90% ~ 95% ethanol solution, The volume ratio of ethanol and concentrate is 3-4：1, ethanol solution stands 2-4h in 0.1T-0.3T magnetic fields；

3）Redissolve, the trichloroacetic acid for adding 20% in the solution adjusts pH value to be stood overnight, 7000-8000r/min centrifugations to 3.5-4 10-18min, take supernatant；

4）The 1/10-3/10 of stoste volume is concentrated in vacuo to, is freeze-dried under the conditions of -30 DEG C to -25 DEG C, obtains sargassan Compound.

Compared with prior art, beneficial effects of the present invention are：

1）By carrying out cell membrane break process before sargassan extracts, made using ultrasonic wave, bioactive peptide and water-bath Cell wall rupture is more thorough, promotes intracellular organic matter dissolution, does not destroy wherein active ingredient, be obviously improved sargassan Yield；

2）The bioactive peptide used has hydrophily and lipophilicity, and hydrophily makes its soluble in water, and lipophilicity makes itself and sargassum Cell membrane combines, and makes to form aperture under cell membrane, causes intracellular content leaks, is advantageous to sargassan yield, utilizes Active peptide combination immersion method carries out clasmatosis, substantially reduces extraction time, extraction effect is good, also effectively prevent broken place The protein denaturation caused by heating and mechanical force is needed during reason；

3）The recovery rate of sargassan is more than 6.28%, and purity is up to 82.1%, and the sargassan compound tool extracted There are excellent anti-oxidant, antitumous effect to be used in health products；

4）The extraction process of the sargassan compound of the present invention is simple, and workable, production cost is low.

Embodiment

Technical scheme is described in further detail with reference to embodiments：

Embodiment 1：

A kind of method from sargassum extraction polysaccharide compound, including breaking-wall cell, concentration alcohol analysis, redissolve removing protein, and freezing is dry Dry, concrete operation step is as follows：

1）Sargassum powder is taken to be made algae solution, the solid-to-liquid ratio of sargassum powder and water is 1:68, ultrasonic disruption, power 700w, when Between be 14min, add the bioactive peptide of algae solution weight 0.02%, the amino acid sequence of bioactive peptide is QSGFCTSYYSKFCTLYGNLGICLHCSS, the water-bath 2h at 99 DEG C, 7500r/min centrifugation 8min, except frond；

2）Algae solution is put into the 1/4 of original volume is concentrated in vacuo in vacuum concentration pot, add 92% ethanol solution, ethanol with it is dense The volume ratio of contracting liquid is 3：1, ethanol solution stands 4h in 0.1T magnetic fields；

3）Redissolving, the trichloroacetic acid for adding 20% in the solution adjusts pH value to stand overnight to 3.5,7200r/min centrifugation 14min, Take supernatant；

4）The 1/5 of stoste volume is concentrated in vacuo to, is freeze-dried under the conditions of -30 DEG C, obtains sargassan compound.

Embodiment 2：

A kind of method from sargassum extraction polysaccharide compound, including breaking-wall cell, concentration alcohol analysis, redissolve removing protein, and freezing is dry Dry, preferable concrete operation step is as follows：

1）Sargassum powder is taken to be made algae solution, the solid-to-liquid ratio of sargassum powder and water is 1:70, ultrasonic disruption, power 750w, when Between be 14min, add the bioactive peptide of algae solution weight 0.02%, the amino acid sequence of bioactive peptide is QSGFCTSYYSKFCTLYGNLGICLHCSS, the water-bath 4h at 100 DEG C, 8000r/min centrifugation 7min, except frond；

2）Algae solution is put into the 1/4 of original volume is concentrated in vacuo in vacuum concentration pot, add 92% ethanol solution, ethanol with it is dense The volume ratio of contracting liquid is 4：1, ethanol solution stands 4h in 0.2T magnetic fields；

3）Redissolve, the trichloroacetic acid for adding 20% in the solution adjusts pH value to stand overnight, 7000r/min centrifugation 10min, take to 4 Supernatant；

4）The 3/10 of stoste volume is concentrated in vacuo to, is freeze-dried under the conditions of extremely -28 DEG C, obtains sargassan compound.

Embodiment 3：

A kind of method from sargassum extraction polysaccharide compound, including breaking-wall cell, concentration alcohol analysis, redissolve removing protein, and freezing is dry Dry, concrete operation step is as follows：

1）Sargassum powder is taken to be made algae solution, the solid-to-liquid ratio of sargassum powder and water is 1:72, ultrasonic disruption, power 580w, when Between be 12min, add the bioactive peptide of algae solution weight 0.018%, the amino acid sequence of bioactive peptide is QSGFCTSYYSKFCTLYGNLGICLHCSS, the water-bath 3h at 100 DEG C, 7500r/min centrifugation 8min, except frond；

2）Algae solution is put into the 1/4 of original volume is concentrated in vacuo in vacuum concentration pot, add 90% ethanol solution, ethanol with it is dense The volume ratio of contracting liquid is 3：1, ethanol solution stands 2h in 0.1T magnetic fields；

3）Redissolving, the trichloroacetic acid for adding 20% in the solution adjusts pH value to stand overnight to 3.7,7800r/min centrifugation 18min, Take supernatant；

4）The 1/10 of stoste volume is concentrated in vacuo to, is freeze-dried under the conditions of -27 DEG C, obtains sargassan compound.

Routine operation in the operating procedure of the present invention is well known to those skilled in the art, herein without repeating.

Technical scheme is described in detail embodiment described above, it should be understood that it is described above only For the specific embodiment of the present invention, it is not intended to limit the invention, all any modifications made in the spirit of the present invention, Supplement or similar fashion replacement etc., should be included in the scope of the protection.

SEQUENCE LISTING

<110>Pujiang County Mei Ze bio tech ltd

<120>A kind of method from sargassum extraction polysaccharide compound

<130> 1

<160> 1

<170> PatentIn version 3.5

<210> 1

<211> 27

<212> PRT

<213>It is artificial synthesized

<400> 1

Gln Ser Gly Phe Cys Thr Ser Tyr Tyr Ser Lys Phe Cys Thr Leu Tyr

1 5 10 15

Gly Asn Leu Gly Ile Cys Leu His Cys Ser Ser

20 25

Claims (6)

1. a kind of method from sargassum extraction polysaccharide compound, including breaking-wall cell, concentration alcohol analysis, redissolve removing protein, freezing Dry, it is characterised in that：The breaking-wall cell step is：Take sargassum powder that algae solution is made, ultrasonic disruption, add bioactive peptide Water-bath, centrifugation, except frond.

A kind of 2. method from sargassum extraction polysaccharide compound according to claim 1, it is characterised in that：The cell The solid-to-liquid ratio of algae powder and water in broken wall step in algae solution is 1:65-75, ultrasonic disruption power are 500w-800w, and the time is 10-20min。

A kind of 3. method from sargassum extraction polysaccharide compound according to claim 1, it is characterised in that：The cell Algae solution weight 0.01%-0.03% bioactive peptide is added in broken wall step, the amino acid sequence of the bioactive peptide is QSGFCTSYYSKFCTLYGNLGICLHCSS。

A kind of 4. method from sargassum extraction polysaccharide compound according to claim 1, it is characterised in that：The cell Bath temperature is 98 DEG C ~ 100 DEG C in broken wall step, time 2-4h.

A kind of 5. method from sargassum extraction polysaccharide compound according to claim 1, it is characterised in that：The concentration Alcohol analysis step is that algae solution is concentrated into the 1/4-1/3 of original volume, adds the body of 90% ~ 95% ethanol solution, ethanol and concentrate Product ratio is 3-4：1, the ethanol solution stands 2-4h in 0.1T-0.3T magnetic fields.

A kind of 6. method from sargassum extraction polysaccharide compound according to claim 1, it is characterised in that：It is described to remove egg White step uses trichloroacetic acid method removing protein.