CN106755190A - Based on the method that restructuring κ carrageenases prepare carrageenan oligosaccharide - Google Patents
Based on the method that restructuring κ carrageenases prepare carrageenan oligosaccharide Download PDFInfo
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- CN106755190A CN106755190A CN201611031738.XA CN201611031738A CN106755190A CN 106755190 A CN106755190 A CN 106755190A CN 201611031738 A CN201611031738 A CN 201611031738A CN 106755190 A CN106755190 A CN 106755190A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2468—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on beta-galactose-glycoside bonds, e.g. carrageenases (3.2.1.83; 3.2.1.157); beta-agarase (3.2.1.81)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01083—Kappa-carrageenase (3.2.1.83)
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Abstract
The invention discloses a kind of method that carrageenan oligosaccharide is prepared based on κ carrageenases.The characteristics of the method is that first culturing engineering bacterial strain makes it produce restructuring κ carrageenases, engineered strain is carried out into ultrasonic disruption Prepare restructuring enzyme liquid, by enzyme liquid according to 1：The ratio of 1.5 2.0 (V), is added in κ carragheen reaction solutions and is reacted, and obtains the crude extract containing carrageenan oligosaccharide.Carrageenan oligosaccharide is adsorbed using activated-charcoal column, using 20 40% ethanol desorption, concentration, freeze-drying, the carrageenan oligosaccharide that the degree of polymerization is 28 is obtained.It is an advantage of the invention that restructuring enzyme source is guaranteed, enzymolysis process is simple, stability is high, be adapted to industrialized production, κ carrageenan oligosaccharide of the degree of polymerization between 28 can quickly, be largely produced, has very big application prospect in terms of enzymatic isolation method prepares the production of κ carrageenan oligosaccharides.
Description
Technical field
The present invention relates to genetic engineering field, in particular to a kind of to prepare carrageenan oligosaccharide based on restructuring kappa-carrageenan enzyme
Method.
Background technology
It with 1,4- α-D- galactopyranoses and 1,3- β-D- galactopyranoses is to repeat that carragheen (Carrageenans) is
The sulfuric acid linear polysaccharide that units alternately is formed by connecting, is primarily present in the cell membrane of Rhodophyceae marine alga.The industrial card for using
Draw glue mainly include κ-, ι-and λ-three kinds.Carragheen not only has the characteristics such as gel, emulsification, thickening, film forming and stable dispersion, and
And can be blended with other foodstuff glues and with synergistic function, therefore in food industry, daily chemical industry and biochemistry, medical research
Deng in field, carragheen is widely used as gel, stabilizer, water binding agent suspending agent and thickener etc..Although OK a karaoke club
Glue possesses good application value, but self-molecules present amount is big, dissolubility is poor, absorbability is poor and has the shortcomings that some toxic,
It is strongly limit more to be widely applied.Carrageenan oligosaccharide (Carrageenan oligosaccharides) is used as carragheen
Catabolite, molecular weight is smaller, dissolubility is preferable, stability and security all make moderate progress, while because of the work on strand
Property group fully expose, its original activity is improved, or even generates new activity, further widened carragheen should
Use field.
The preparation of current carrageenan oligosaccharide has chemistry, three kinds of methods of physics and enzymolysis.Using chemically and physically grade tradition side
During method degraded carrageenan, because complex process, reaction condition are difficult to control, the carrageenan oligosaccharide degree of polymerization is uneven because obtained from
One, it is impossible to mass produced.Comparatively speaking, the method substrate using carrageenase hydrolysis production carrageenan oligosaccharide is special
First, product is homogeneous, and reaction condition is gentle, can avoid the shortcoming of above-mentioned conventional method, therefore carrageenase by more next
Utilization is researched and developed more.It is enzymatic isolation method to stablize, continuously, efficiently prepare high-quality carrageenase and optimization enzymolysis process
Prepare the precondition of carrageenan oligosaccharide, but the source that enzymatic isolation method prepares high-quality enzyme in the technique of OK a karaoke club plus oligosaccharides at present is one
Individual bottleneck problem.Japan Patent (patent No. JP2000116376) has used carrageenase to prepare kappa-carrageenin oligose, the method
The separation and Extraction carrageenase first from bacterial strain, then with this enzyme digest and obtain oligosaccharides mixture.Because this enzyme source is in sea
Foreign vibrios, strain enzyme-producing conditional instability, preparation process is cumbersome, the features such as the production cycle is long, thus it is expensive, cause cost
Height, is not suitable for industrialized production.
The content of the invention
The present invention provides a kind of method for preparing carrageenan oligosaccharide based on restructuring kappa-carrageenan enzyme, the method restructuring enzyme source
Simply, enzyme activity is good, stability is high, enzymolysis process is simple, be adapted to industrialized production, can quickly, largely produce the degree of polymerization equal
One kappa-carrageenin oligose, has very big application prospect in terms of enzymatic isolation method prepares the production of kappa-carrageenin oligose.
In order to solve the above-mentioned technical problem, the technical solution adopted in the present invention is：
A kind of method for preparing carrageenan oligosaccharide based on restructuring kappa-carrageenan enzyme, comprises the following steps：
First, the preparation of kappa-carrageenan enzyme is recombinated
(1) the engineered strain BL21-Cly- κ-CAR (number of patent applications of restructuring kappa-carrageenan enzyme will be produced：
201610503827.3) it is seeded in the LB liquid medium that kanamycins concentration is 20pg/ml, 37 DEG C, 150-200rpm bars
Cultivated under part to bacterium solution OD600Value reaches 0.5-0.6, and LB culture medium prescriptions are：Peptone, 10g/L, yeast extract, 5g/L,
Sodium chloride, 10g/L, pH=7；
(2) induced expression is carried out to the IPTG for adding final concentration of 0.1mM in bacterium solution, inductive condition is 18 DEG C, 100-
120rpm, 24h, make it produce restructuring kappa-carrageenan enzyme；
(3) the BL21-Cly- κ-CAR bacterium solutions after induction are collected, 10000rpm, 10min is collected by centrifugation thalline, uses PBS phosphorus
After hydrochlorate cushioning liquid washing thalline 2-3 times, thalline is resuspended in cushioning liquid, ultrasonic disruption thalline, Ultrasonic Conditions are
Power 200W, each circulation includes ultrasound 5s, suspends 5s, and 20 circulations are obtained after crushing and contain restructuring kappa-carrageenan enzyme
Crude enzyme liquid；
2nd, the enzyme activity determination of kappa-carrageenan enzyme is recombinated
Different temperatures, pH, concentration of substrate, under conditions of the reaction time, determine restructuring kappa-carrageenan enzyme crude enzyme liquid enzyme
Vigor, so that it is determined that recombinating the optimum reaction conditionses of kappa-carrageenan enzyme, enzyme activity determination method is：Take 1mL crude enzyme liquids and add 1mL
Concentration is 0.5% carrageenan solutions, and control group is the enzyme liquid for boiling 5min inactivations, is incubated 30min at different conditions respectively
Afterwards, addition 2mLDNS solution, the absorbance that constant volume is surveyed at 540nm to 25mL, ultraviolet specrophotometer after boiling water bath reaction 5min,
Enzyme activity unit is defined as under above-mentioned condition, and the enzyme amount needed for 1 μm of ol reduced sugar of release per minute is an enzyme activity unit；
3rd, the preparation of carrageenan oligosaccharide mixed liquor
By volume 1：It is molten that the crude enzyme liquid obtained in step one (3) is added to aseptic kappa-carrageenan by the ratio of 1.5-2.0
In liquid, reacted in shaking table or fermentation tank under the optimum reaction conditionses for obtaining in step 2, acquisition contains carrageenan oligosaccharide
Mixed liquor；
4th, the purifying and identification of kappa-carrageenin oligose
The mixed liquor containing carrageenan oligosaccharide that step 3 is obtained processes 2-4h, absorption with the 2-3 times of activated carbon of volume
The less oligosaccharides of the degree of polymerization, it is the oligosaccharides and salt impurity for adsorbing to be eluted with pure water, recycles the ethanol of 20 (v) %-40 (v) %
Activated carbon after absorption is eluted, eluent vacuum concentration, freeze-drying obtains kappa-carrageenin oligose, using thin-layer chromatography
Technology, high-efficient liquid phase chromatogram technology and mass-spectrometric technique carry out the identification of carrageenan oligosaccharide structure.
A kind of method that carrageenan oligosaccharide is prepared based on restructuring kappa-carrageenan enzyme, the cushioning liquid in the step one (3)
It is PBS phosphate buffers or marine bacteria C.lytica M9 fermentation mediums, marine bacteria C.lytica M9 fermented and cultureds
The formula of base is：Contain carragheen 0.1%, peptone 0.5%, FeSO in per the old seawater of 100ml4.7H2O 0.002%.
A kind of method for preparing carrageenan oligosaccharide based on restructuring kappa-carrageenan enzyme, reaction condition is in the step 3：κ-
Final concentration of 0.7 (wt) %-2.0 (wt) % of carragheen, 35-45 DEG C of temperature, pH=6-8, reaction time 2-4h, rotating speed is
100-150rpm。
The advantage of the invention is that the preparation condition of carrageenase is simple, enzyme activity stabilization and production of enzyme is high, enzymolysis process
Simply, yield is high, and the degree of polymerization of the kappa-carrageenin oligose of preparation is 2-8.The life of carrageenan oligosaccharide is prepared in scale enzymatic isolation method
Product aspect has very big prospect.
Brief description of the drawings
Fig. 1 is the zymologic property for recombinating kappa-carrageenan enzyme, and wherein A is the enzyme activity under condition of different temperatures；B is difference
Temperature and the enzyme activity under the conditions of the reaction time；C is the enzyme activity under condition of different pH；D is different pH and reaction time
Under the conditions of enzyme activity；E is the enzyme activity under the conditions of different concentration of substrate；F is degradation of substrates speed.
Fig. 2 is the thin-layer chromatogram of the kappa-carrageenin oligose for preparing, wherein, band M represents galactolipin, and band C represents card
Glue enzyme is drawn to represent the action time 10min of recombinase degraded carrageenan respectively from wild strain C.lytica M9, band 1-6,
The thin-layer chromatogram of the kappa-carrageenin oligose after 30min, 1h, 1.5h, 2.5h and 3h；The corresponding position of a, b, c be respectively eight sugar,
Six sugar and disaccharides).
Specific embodiment
With reference to specific embodiment and Figure of description, the present invention will be further described.
First, the preparation of kappa-carrageenan enzyme is recombinated
(1) the engineered strain BL21-Cly- κ-CAR for recombinating kappa-carrageenan enzyme are seeded to kanamycins concentration for 20pg/
In the LB liquid medium of ml, 37 DEG C, cultivate to OD under the conditions of 150rpm600Value reaches 0.5-0.6, and LB culture medium prescriptions are：Egg
White peptone, 10g/L, yeast extract, 5g/L, sodium chloride, 10g/L, pH=7；
(2) adding the IPTG of final concentration of 0.1mM carries out induced expression, and inductive condition is 18 DEG C, 100rpm 24h, makes it
Produce restructuring kappa-carrageenan enzyme；
(3) the BL21-Cly- κ-CAR bacterium solutions after induction are collected, 10000rpm, 10min is collected by centrifugation thalline, uses PBS phosphorus
After hydrochlorate cushioning liquid washing thalline 2-3 times, thalline is resuspended in marine bacteria C.lytica M9 fermentation mediums, ultrasound
Ripple crushes thalline, and Ultrasonic Conditions are power 200W, and each circulation includes ultrasound 5s, suspends 5s, and 20 circulations are obtained after crushing
The crude enzyme liquid of restructuring kappa-carrageenan enzyme must be contained；
2nd, the enzyme activity determination of kappa-carrageenan enzyme is recombinated
Different temperatures, pH, concentration of substrate, under conditions of the reaction time, determine restructuring kappa-carrageenan enzyme crude enzyme liquid enzyme
Vigor, so that it is determined that recombinating the optimum reaction conditionses of kappa-carrageenan enzyme, the assay method of enzyme activity is：Take the addition of 1mL crude enzyme liquids
1mL concentration is 0.5% carrageenan solutions, and control group is the enzyme liquid for boiling 5min inactivations, is incubated at different conditions respectively
After 30min, 2mLDNS solution, the suction that constant volume is surveyed at 540nm to 25mL, ultraviolet specrophotometer after boiling water bath reaction 5min are added
Luminosity, enzyme activity unit is defined as under above-mentioned condition, and the enzyme amount needed for 1 μm of ol reduced sugar of release per minute is an enzyme activity list
Position；
Wherein, enzyme reaction condition setting is as follows：
1st, the enzyme activity of restructuring kappa-carrageenan enzyme crude enzyme liquid is determined under the conditions of 25 DEG C, 30 DEG C, 35 DEG C, 40 DEG C, 45 DEG C, 50 DEG C
Power, is as a result shown in Figure 1A；
2nd, restructuring kappa-carrageenan enzyme crude enzyme liquid is respectively placed under the conditions of 35 DEG C, 40 DEG C, 45 DEG C, 50 DEG C and 55 DEG C, is incubated
Residual enzyme activity is detected after 0.5h, 1h, 1.5h, 2h, Figure 1B is as a result seen；
3rd, Na is used2HPO4Respectively be adjusted to the pH value of carragheen substrate by-citrate buffer solution or Tris-HCl buffer solutions
4.0th, 5.0,6.0,7.0,8.0 and 9.0, the enzyme activity of restructuring kappa-carrageenan enzyme crude enzyme liquid is determined, as a result see Fig. 1 C；
4th, restructuring kappa-carrageenan enzyme crude enzyme liquid is pressed 1:1 mixes from different pH buffer solutions, under the conditions of putting 4 DEG C after 6h and 24h
With the carragheen substrate reactions of identical pH, remaining enzyme activity is determined according to above-mentioned enzyme activity determination method, as a result see Fig. 1 D；
5th, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% and 1% carragheen is respectively with concentration
As substrate, restructuring kappa-carrageenan enzyme activity is determined according to above-mentioned enzyme activity determination method, as a result see Fig. 1 E；
3rd, the preparation of carrageenan oligosaccharide mixed liquor
By volume 1：It is molten that the crude enzyme liquid obtained in step one (3) is added to aseptic kappa-carrageenan by the ratio of 1.5-2.0
In liquid, make final concentration of 0.7 (wt) %-2.0 (wt) % of kappa-carrageenan, reaction condition is 35-45 DEG C, pH=6-8, rotating speed are
100-150rpm, is placed in and 2-4h is reacted in shaking table or fermentation tank, obtains the mixed liquor containing carrageenan oligosaccharide；
4th, the purifying and identification of kappa-carrageenin oligose
The mixed liquor containing carrageenan oligosaccharide that step 3 is obtained processes 2-4h, absorption with the 2-3 times of activated carbon of volume
The less oligosaccharides of the degree of polymerization, it is the oligosaccharides and salt impurity for adsorbing to be eluted with pure water, recycles the ethanol of 20 (v) %-40 (v) %
Activated carbon after absorption is eluted, eluent vacuum concentration, freeze-drying obtains kappa-carrageenin oligose, using thin-layer chromatography
Technology, high-efficient liquid phase chromatogram technology and mass-spectrometric technique carry out the identification of carrageenan oligosaccharide structure, as a result see Fig. 2.
The above, the only present invention preferably specific embodiment, but protection scope of the present invention is not limited thereto,
Any one skilled in the art in the technical scope of present disclosure, according to the ordinary technical knowledge of this area
And universal method, technology according to the present invention scheme and its inventive concept are subject to equivalent or change, should all cover in this hair
Within bright protection domain.
Claims (3)
1. it is a kind of based on the restructuring kappa-carrageenan enzyme method for preparing carrageenan oligosaccharide, it is characterised in that to comprise the following steps：
First, the preparation of kappa-carrageenan enzyme is recombinated
(1) the engineered strain BL21-Cly- κ-CAR that will produce restructuring kappa-carrageenan enzyme are seeded to kanamycins concentration for 20pg/ml
LB liquid medium in, 37 DEG C, cultivate to bacterium solution OD under the conditions of 150-200rpm600Value reaches 0.5-0.6, and LB culture mediums are matched somebody with somebody
Fang Wei：Peptone, 10g/L, yeast extract, 5g/L, sodium chloride, 10g/L, pH=7；
(2) carry out induced expression to the IPTG for adding final concentration of 0.1mM in bacterium solution, inductive condition is 18 DEG C, 100-120rpm,
24h, makes it produce restructuring kappa-carrageenan enzyme；
(3) the BL21-Cly- κ-CAR bacterium solutions after induction are collected, 10000rpm, 10min is collected by centrifugation thalline, uses PBS phosphate
After cushioning liquid washing thalline 2-3 times, thalline is resuspended in cushioning liquid, ultrasonic disruption thalline, Ultrasonic Conditions are power
200W, each circulation includes ultrasound 5s, suspends 5s, and 20 circulations obtain the thick enzyme containing restructuring kappa-carrageenan enzyme after crushing
Liquid；
2nd, the enzyme activity determination of kappa-carrageenan enzyme is recombinated
Different temperatures, pH, concentration of substrate, under conditions of the reaction time, determine restructuring kappa-carrageenan enzyme crude enzyme liquid enzyme activity,
So that it is determined that recombinating the optimum reaction conditionses of kappa-carrageenan enzyme, enzyme activity determination method is：Take 1mL crude enzyme liquids and add 1mL concentration
It is 0.5% carrageenan solutions, control group is the enzyme liquid for boiling 5min inactivations, respectively at different conditions after insulation 30min, plus
Enter 2mLDNS solution, the absorbance that constant volume is surveyed at 540nm to 25mL, ultraviolet specrophotometer after boiling water bath reaction 5min, enzyme activity
Unit of force is defined as under above-mentioned condition, and the enzyme amount needed for 1 μm of ol reduced sugar of release per minute is an enzyme activity unit；
3rd, the preparation of carrageenan oligosaccharide mixed liquor
By volume 1：Be added to the crude enzyme liquid obtained in step one (3) in aseptic kappa-carrageenan solution by the ratio of 1.5-2.0,
Reacted in shaking table or fermentation tank under the optimum reaction conditionses for obtaining in step 2, obtain the mixing containing carrageenan oligosaccharide
Liquid；
4th, the purifying and identification of kappa-carrageenin oligose
The mixed liquor containing carrageenan oligosaccharide that step 3 is obtained processes 2-4h, adsorpting polymerization with the 2-3 times of activated carbon of volume
Less oligosaccharides is spent, it is the oligosaccharides and salt impurity for adsorbing to be eluted with pure water, recycle the ethanol of 20 (v) %-40 (v) % to inhaling
Attached activated carbon is eluted, eluent vacuum concentration, and freeze-drying obtains kappa-carrageenin oligose, using thin-layer chromatography technology,
High-efficient liquid phase chromatogram technology and mass-spectrometric technique carry out the identification of carrageenan oligosaccharide structure.
2. according to claim 1 a kind of based on the method that kappa-carrageenan enzyme prepares carrageenan oligosaccharide is recombinated, its feature exists
In, cushioning liquid in the step one (3) be PBS phosphate buffers or marine bacteria C.lytica M9 fermentation mediums,
The formula of marine bacteria C.lytica M9 fermentation mediums is：Contain carragheen 0.1%, peptone in per the old seawater of 100ml
0.5%th, FeSO4.7H2O 0.002%.
3. according to claim 1 a kind of based on the method that kappa-carrageenan enzyme prepares carrageenan oligosaccharide is recombinated, its feature exists
In optimum reaction conditionses are in the step 3：Final concentration of 0.7 (wt) %-2.0 (wt) % of kappa-carrageenan, temperature 35-45
DEG C, pH=6-8, reaction time 2-4h, rotating speed is 100-150rpm.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107997081A (en) * | 2017-11-24 | 2018-05-08 | 安徽省林锦记食品工业有限公司 | The processing method that a kind of xylitol preserved fruit freezes |
| CN111303310A (en) * | 2020-03-12 | 2020-06-19 | 大连大学 | Preparation method of marine sulfuric acid galactohexaose and application of marine sulfuric acid galactohexaose in preparation of medicines for resisting Alzheimer disease |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103290079A (en) * | 2012-02-29 | 2013-09-11 | 宁波大学 | Preparation method of lambda-carrageenan oligosaccharide |
| CN105274162A (en) * | 2015-09-18 | 2016-01-27 | 集美大学 | Preparation method of different polymerization degrees of kappa-carrageenan oligosaccharides |
| CN105950640A (en) * | 2016-06-30 | 2016-09-21 | 大连大学 | Preparation method of marine bacteria-derived kappa-carrageenase gene and recombinase |
-
2016
- 2016-11-22 CN CN201611031738.XA patent/CN106755190A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103290079A (en) * | 2012-02-29 | 2013-09-11 | 宁波大学 | Preparation method of lambda-carrageenan oligosaccharide |
| CN105274162A (en) * | 2015-09-18 | 2016-01-27 | 集美大学 | Preparation method of different polymerization degrees of kappa-carrageenan oligosaccharides |
| CN105950640A (en) * | 2016-06-30 | 2016-09-21 | 大连大学 | Preparation method of marine bacteria-derived kappa-carrageenase gene and recombinase |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107997081A (en) * | 2017-11-24 | 2018-05-08 | 安徽省林锦记食品工业有限公司 | The processing method that a kind of xylitol preserved fruit freezes |
| CN111303310A (en) * | 2020-03-12 | 2020-06-19 | 大连大学 | Preparation method of marine sulfuric acid galactohexaose and application of marine sulfuric acid galactohexaose in preparation of medicines for resisting Alzheimer disease |
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