CN110283860A - The Gracilaria tenuistipitata polysaccharide and its extracting method of ultrasonic wave assisted recombination enzymolysis and extraction - Google Patents
The Gracilaria tenuistipitata polysaccharide and its extracting method of ultrasonic wave assisted recombination enzymolysis and extraction Download PDFInfo
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Abstract
The present invention relates to the methods of ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide；This method takes the Gracilaria tenuistipitata powder by decolorization, and buffer solution is added, after stirring and evenly mixing, is ultrasonically treated；Complex enzyme is added and carries out enzymolysis processing；Complex enzyme is bromelain and the second component enzymes, and the second component enzymes are cellulase or pectase；After enzymatic hydrolysis, enzyme-deactivating is collected by being centrifuged and filtering extraction fluid by supernatant using boiling water bath；Extracting solution is subjected to alcohol precipitation, is centrifuged, is washed, it is dry, obtain Gracilaria tenuistipitata polyoses extract.Extracting method of the present invention has the advantages that mild condition, step enzymatic hydrolysis, easy to operate, extraction time is short, polysaccharide high income, and obtained Gracilaria tenuistipitata polysaccharide anti-oxidative ability is strong, and healthy nutritive value is high.
Description
Technical field
The present invention relates to Gracilaria tenuistipitata polysaccharide, more particularly to a kind of Gracilaria tenuistipitata of ultrasonic wave assisted recombination enzymolysis and extraction
Polysaccharide and its extracting method, the Gracilaria tenuistipitata polysaccharide that the method for the present invention obtains have good antioxidant activity；Belong to functionality
Food engineering development field.
Background technique
With the sharp increase of the mankind and size of animal on the earth, terrestrial resource is increasingly reduced；Seek the sustainable of human society
Property, it must just be engaged in the exploitation and research of marine resources.Since 1970, oneself from seaweed, sponge, coelenterate, mollusk,
Isolated natural products is many with antimycotic, antiviral, anticoagulant up to more than 3000 in echinoderm and microbial body
The research of the pharmacological activity such as blood, antitumor, marine natural products has become current heat subject.
In recent years, seaweed causes West Europe, North America, Northern Europe as functional food because of its peculiar flavour and nutritive value
The favor of equal developed countries.Existing research shows that seaweed can prevent the metabolic disease such as obesity, gall stone, constipation, enterogastritis
Disease and cancer-resisting, the formation for reducing blood pressure, preventing artery sclerosis and thrombus, reduction blood glucose and other effects.River hedge is economic valence
It is worth higher seaweed, is widely distributed in the global torrid zone, subtropical zone and temperate zone sea area, about 100 kinds named at present.I
State have hedge resource in river abundant, oneself identification clearly about more than 30 kinds, be China Hainan, Guangdong, Zhejiang, Taiwan along seaweeds support
One of kind is grown, mainly as fishery cultivating feed and production foodstuff glue raw material.There is in evolution for different types of river hedge
Difference, so that the property such as coagulation ability, setting temperature etc. to agar-agar brings Different Effects.Agar-agar or derivatives thereof can do biology
Medical material, health food, and there is antiviral activity, it is long-term to be used to make culture medium, in biochemical analysis and clinical assay
The technologies such as electrophoresis, chromatography commonly use agar or agarose, and agar is by promoting gastrointestinal peristalsis to prevent constipation, by preventing to rouge
The absorption of fat, cholesterol, chemical carcinogen is to prevent cardiovascular disease and colon cancer.
Gracilaria tenuistipitata (Gracilaria Tenuistipitata) is China's cultivation maximum river hedge of quantity, can be sent out
Bud breeding, seedling are easy to solve, and cultural method is also fairly simple, is the current Main Cultivation type in China.The biology of Gracilaria tenuistipitata
There has also been certain progress, Yeh et al. to have carried out grinding in vitro for anticancer effect to the methanolic extract of Gracilaria tenuistipitata for activity research
Study carefully, research shows that methanolic extract to Ca9-22 cancer cell of oral cavity there are cytotoxicity, can by inducing cell apoptosis,
Cause DNA damage and oxidative stress and makes cancer cell death (BMC Complement.Altern.Med., 2012,12,142-
151).Yangthong et al. studies the antioxidant activity of the boiling-water extract of four kinds of seaweed, studies have shown that thin base
The boiling-water extract of fragrant plant mentioned in ancient texts have due to there are polyphenolic substance certain oxidation resistance (Plant Foods Hum.Nutr.,
2009,64(3),218-223)；The water extract that Chen et al. then has studied Gracilaria tenuistipitata exempts from white shrimp under the influence of ammonia
Epidemic disease ability is replied situation and can be helped white studies have shown that Gracilaria tenuistipitata water extract has booster action to the improvement of its immune indexes
Prawn replys immunocompetence (Mar.Drugs, 2015,13 (6), 3606-3624) earlier.
The chemical composition of the extract of thin base river hedge is strongly depend on extraction system, and different extraction systems and condition imparting mention
Take the function that object is different.The resources development and utilization degree of thin base river hedge is lower, and the extraction process of Gracilaria tenuistipitata polysaccharide discloses at present
Be Zhao Mouming seminar ultrasound extraction process by water: 100W ultrasound 30min, solid-to-liquid ratio 1:30, extract 10h, highest recovery rate by 70 DEG C
6.25% (optimal procedure parameters), the best Antioxidant Indexes of resulting Gracilaria tenuistipitata polysaccharide are 1.25mgmL-1, hydroxyl free
Base clearance rate 51.54% (0.1wt% hydrogen peroxide) (food industry science and technology, 2005,2 (3), 67-69).Other presently disclosed rivers
Hedge Polyose extraction technology includes the method that the ultrasonic acid of 200810029075.7 A of Chinese invention patent mentions river hedge polysaccharide sulfate,
Solid-to-liquid ratio 1:40~1:60, concentration of hydrochloric acid 0.1~0.2%, 15~25min of 500W ultrasonic extraction (55~65 DEG C), polysaccharide highest
Recovery rate 10.93%.It is more that 200910001265.2 A of Chinese invention patent application discloses river hedge Aqueous extracts lyases, protease
The method for walking enzyme digestion reaction preparation river hedge polysaccharide, solid-to-liquid ratio 1:5~1:10 are extracted 8.5~12.5h (30~60 DEG C), and polysaccharide is most
High extraction 13.42%.201310694077.9 A of Chinese invention patent based on cellulase, compound protease stepwise discretization
The extracting method containing protein, the river hedge polysaccharide of river hedge polysaccharide multicomponent enzymolysis liquid of technique, solid-to-liquid ratio 1:8~1:14, enzymatic hydrolysis
It extracts 6~12h (40~65 DEG C), river hedge polysaccharide extract rate 8~10%.
On the whole, above-mentioned prior art river hedge polysaccharide yield is all also lower, and strong acid extracts the high requirements on the equipment, and
The problem of there are environmental pollutions, and extraction time is long, and energy consumption is high, and yield is low, low efficiency etc..
Summary of the invention
To overcome the extraction time of the prior art long, energy consumption is high, and yield is low, low efficiency, the anti-oxidant work of extract polysaccharide
Property it is relatively low, the invention proposes a kind of Phenol sulfuric acid procedure measurement recovery rate reach 25.01~38.60wt% ultrasonic wave auxiliary
The Gracilaria tenuistipitata polysaccharide and its extracting method that complex enzyme hydrolysis extracts；Resulting Gracilaria tenuistipitata polysaccharide reaches the clearance rate of hydroxy radical
To 50-66%.
The present invention is using ultrasound and multienzyme compound single step enzymatic hydrolysis to the more of the multiple dispersion of thin base river hedge powder and river hedge cell
Weight fragmentation, not only shortens extraction time, improves the recovery rate of Gracilaria tenuistipitata polysaccharide, and process conditions are mild, work
Skill is simple, and the recovery rate of Gracilaria tenuistipitata polysaccharide is significantly higher than the prior art, and resulting Gracilaria tenuistipitata polysaccharide is to hydroxy radical
Clearance rate reaches 50-66%, and the product of the more ultrasonic extraction process by water of the structure of the polysaccharide is improved, and antioxidation activity in vitro is obvious
Enhancing.
The object of the invention is achieved through the following technical solutions:
A kind of method of ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide, comprising the following steps:
1) the Gracilaria tenuistipitata powder by decolorization is taken, buffer solution is added, after stirring and evenly mixing, is carried out at ultrasound
Reason；
2) after ultrasound, complex enzyme is added and carries out enzymolysis processing；The complex enzyme is bromelain and second group
Divide enzyme, second component enzymes are cellulase or pectase, the mass ratio of the bromelain and the second component enzymes
For 1:1~1:2；Compound enzyme dosage is 1~5wt% of the quality of Gracilaria tenuistipitata powder；
3) after digesting, enzyme-deactivating is collected by being centrifuged and filtering extraction fluid by supernatant using boiling water bath；
4) extracting solution is subjected to alcohol precipitation, be centrifuged, washed, it is dry, obtain Gracilaria tenuistipitata polyoses extract.
To further realize the object of the invention, it is preferable that the buffer solution is that citrate-phosphate disodium hydrogen buffering is molten
Liquid, pH are 4.5~6.5, and the concentration of disodium hydrogen phosphate is 0.05 ± 0.005molL-1, the volume of buffer solution and the matter of powder
Amount is than being 40:1~100:1.
Preferably, bath temperature when being ultrasonically treated is 45~65 DEG C, and ultrasonic power is 90~150W, ultrasonic time
For 10~40min.
Preferably, the hydrolysis temperature of the enzymolysis processing is 45~65 DEG C, and enzymolysis time is 60~180min.
Preferably, the enzyme deactivation is using boiling water bath by enzyme-deactivating, and extracting solution 20min or more in boiling water bath loses enzyme
It is living.
Preferably, the revolving speed of centrifugation described in step 3) is 4500rpm or more, and the time of centrifugation is 8min or more；Step
4) revolving speed of the centrifugation described in is 3000rpm or more, and the time of centrifugation is 5min or more.
Preferably, the alcohol precipitation is the dehydrated alcohol precipitate polysaccharides that three times volume is added into supernatant, collects precipitating.
Preferably, the washing is to wash precipitating 2 times or more with dehydrated alcohol；The drying is at 50-70 DEG C
40-60h is dried in vacuum drying oven.
Preferably, it is measured by Phenol sulfuric acid procedure, the recovery rate of ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide
For 25.01~38.60wt%.
A kind of Gracilaria tenuistipitata polysaccharide, is extracted to obtain by above-mentioned method；The Gracilaria tenuistipitata polysaccharide is to hydroxy radical
Clearance rate is 50-66%；WhereinTest method is as follows: taking
0.5mL concentration is 1.2mgmL-1For sample solution into 7mL centrifuge tube with cover, the phosphate-buffered that 1mL pH=7.4 is added is molten
Liquid, concentration 0.1M add 0.5mL 0.75mmolL-1Phen solution, 0.5mL is added after shaking up
0.75mmol·L-1L ferrous ammonium sulfate solution shakes up rapidly, and it is the molten of 0.01% hydrogen peroxide that 0.5mL mass concentration, which is then added,
Liquid, tube sealing shake up, and keep the temperature 60min at 37 DEG C, then cool to room temperature, and using distilled water as reference, measure and inhale at 510nm
Shading value, note absorbance value are Asample；Sample solution is substituted for distilled water, carries out identical processing, measurement, remembers absorbance
Value is Ablank；Sample solution and hydrogen peroxide are substituted for distilled water, carry out identical processing, measurement, note absorbance value is A0。
The type of Gracilaria tenuistipitata used herein is not particularly limited, and used Gracilaria tenuistipitata powder both can be purchased directly
It buys, can also voluntarily crush, decolourize, the powder for extracting sieves with 100 mesh sieve.
Bromelain and cellulase and pectase in the present invention are all the enzymes of plant origin, safe and reliable,
Wherein, bromelain is from the part such as pineapple fruits skin, stem, core through a kind of natural plant made from biotechnology
Enzyme, molecular weight 33000, isoelectric point 9.55 belong to thin base protease, and activity centre is to dredge base (- SH), can be carried out egg
The various biochemical reactions such as white matter hydrolysis, bromelain are a kind of natural, nontoxic, safe and healthy protease, it can
The industries such as food, medicine and biology are widely applied.Bromelain enzymatic structure belongs to glycoprotein, and (100 000U) optimal pH generally exists
7 or so, it is 55 DEG C of operating temperature most suitable.
Cellulase (beta-1,4-glucan -4- glucan hydrolase) is the glucogenic one group of enzyme of degraded cellulose
General name, its not instead of monomeric enzyme play the multicomponent enzyme system of synergistic effect, are a kind of complex enzymes, mainly by circumscribed beta glucan
The composition such as enzyme, Endo-β-glucanase and beta-glucosidase, there are also the zytases of very high vigor.Act on cellulose and
The product being derived from cellulose.Cellulase destroys at glucose and in juice thin in conversion insoluble fibrin
Cell wall has very important significance to improve fruit juice yield etc., is widely used in feed, alcohol, weaving and food
Deng.The optimal pH of cellulase is generally 4.5~6.5,40-60 DEG C of operating temperature.
Pectase is a multienzyme complex for decomposing pectin, generally includes protopectinase, pectinesterase hydrolase, pectin
Sour enzyme.Pectic substance is set to decompose completely by their synergy.Natural pectic substance is under protopectinase effect, conversion
At the pectin of water dissolvable；Pectin removes methyl esters group by pectinesterase hydrolysis enzymatic, generates pectic acid；Pectic acid is through pectin
Sour hydrolase and the degradation of pectate lyase class generate galacturonic acid.Pectase is mainly used for fruit-vegetable juice beverage and fruit wine
Juicing and clarification have good effect to pectin is decomposed.The optimal pH of cellulase is generally 3.0~6.0, operating temperature
15-55 DEG C, optimum temperature is 50 DEG C.
Importantly, it is a discovery of the invention that digesting multiple point to thin base river hedge powder using ultrasound and the compound single step of multienzyme
It dissipates and the multiple fragmentation of river hedge cell, especially bromelain and the second component enzymes (cellulase or pectase) exists
Coordinated, not only increases the recovery rate of Gracilaria tenuistipitata polysaccharide, and improves the composition of Gracilaria tenuistipitata polysaccharide, improves thin
The antioxidant activity of base Gracilaria gigas Harvey polysaccharides.Wherein the recovery rate of Gracilaria tenuistipitata polysaccharide reaches 25wt% or more, Gracilaria tenuistipitata polysaccharide pair
Scavenging action to hydroxyl free radical reaches 54% or more, this two indexs while the remote super prior art.It may be because of bromelain, fibre
It ties up plain enzyme and pectin enzymatic structure is close, while the PH of optimal enzymatic hydrolysis is close, multienzyme complex, which ties up to similar buffer system, to be had
Good dissolubility is easy to generate efficiently enzymatic hydrolysis synergistic effect in suitable PH point.And bromelain, cellulase and fruit
Glue enzyme is close with the cellulosic structure of river hedge powder, and by the effect of solvent in buffer system, these enzymes are readily adsorbed in river
In the surface of hedge powder and loose cavity.In addition the protein in river hedge powder is in the form of chemical bonding and cellulose mostly
And agarose is combined together in the form of glycoprotein, and in addition to glycoprotein in the hedge powder of river, most of macromolecule carbohydrate with
Cellulose and agarose exist, and the recovery rate of the river Yao Tigao hedge polysaccharide completes peptide bond, fiber it is necessary to carry out multidigit point digestion effect
The dissociation of plain glycosidic bond, agarose glycosidic bond significantly reduces the molecular weight of glycoprotein, cellulose and agarose, increases
The dissolubility of buffer system.The present invention further improves the speed of action of enzyme Yu river hedge powder by the effect of ultrasound, will be low
The good bromelain of the dissolution of concentration, cellulase and pectase are efficiently sent to reaction site, to greatly improve enzymatic hydrolysis
Efficiency and optimize molecular weight composition.
Compared with prior art, the present invention has the following advantages and beneficial effects:
(1) present invention prepares Gracilaria tenuistipitata polysaccharide using ultrasonic wave added complex enzyme hydrolysis technology, which utilizes ultrasound and multienzyme
Compound single step enzymatic hydrolysis is to the multiple dispersion of thin base river hedge powder and the multiple fragmentation of river hedge cell, when not only shortening extraction
Between, the recovery rate (25.01~38.60wt% of recovery rate) of Gracilaria tenuistipitata polysaccharide is substantially increased, and process conditions are mild, work
Skill is simple.
(2) the method for the present invention Gracilaria tenuistipitata polysaccharide anti-oxidative ability obtained is strong, has preferable remove to free radical
Effect, (the polysaccharide concentration 1.2mgmL of Scavenging action to hydroxyl free radical 54.94~65.54%-1), it can be used as a kind of natural anti-oxidation
Agent uses, and healthy nutritive value is high.
Specific embodiment
For a better understanding of the invention, explanation, but reality of the invention are further explained to the present invention below with reference to example
It is without being limited thereto to apply mode.
Experimental method used in following embodiments or condition are conventional method, normal condition unless otherwise specified
Or method proposed by instrument manufacturing manufacturer and condition.
Recovery rate is measured by Phenol sulfuric acid procedure in following embodiments.Method of the Phenol sulfuric acid procedure referring to propositions such as Cuesta
It is prepared and measures (J.Microbiol.Methods., 2003,52 (1), 69-73).Concrete operations are as follows: it is appropriate to learn from else's experience
The phenol solution of 1mL 5wt% is added in test tube in diluted extracting solution 2mL, and 98% sulfuric acid of 5mL is added after shaking up and shakes up,
After placing 30min at room temperature, in measuring absorbance on ultraviolet-visible spectrophotometer, and in the glucose of 105 DEG C of dry 2h
Standard curve is drawn as reference substance and calculates recovery rate, and recovery rate is calculated with the content of glucose.
It is indicated in following embodiments by measuring the clearance rate to hydroxy radical, the method for bibliography is measured
It is 1.2mgmL that (food industry science and technology, 2005,26 (3), 67-69), which take 0.5mL concentration,-1Sample solution is to 7mL centrifuge tube with cover
In, the phosphate buffer solution (0.1M) of 1mL pH=7.4 is added, adds 0.5mL 0.75mmolL-1Phen it is molten
0.5mL 0.75mmolL is added in liquid after shaking up-1L ferrous ammonium sulfate solution shakes up rapidly, and 0.5mL 0.01% is then added
(w/w) solution of hydrogen peroxide, tube sealing shake up, and keep the temperature 60min at 37 DEG C, then cool to room temperature, and are ginseng with distilled water
Than measuring absorbance value at 510nm, note absorbance value is Asample；Sample solution is substituted for distilled water, is carried out identical
Processing, measurement, note absorbance value are Ablank；Sample solution and hydrogen peroxide are substituted for distilled water, carry out identical processing, survey
Fixed, note absorbance value is A0.1. Scavenging action to hydroxyl free radical is calculated by formula:
Embodiment 1
Gracilaria tenuistipitata powder 1.0g is taken, disodium hydrogen phosphate-citric acid solution of prepared pH=5.5 is added
40mL, stirring makes powder sufficiently absorb water and mix on magnetic stirring apparatus, is then placed into and has been heated to 55 DEG C of ultrasonic wave
In cleaning device, setting ultrasonic time is 40min, and ultrasonic power 150W is ultrasonically treated.
After ultrasonic treatment, 1mL bromelain enzyme solutions (15mgmL is respectively added according to the compound enzyme dosage of 3wt%-1)
With 1mL cellulase solution (15mgmL-1), it is transferred in 55 DEG C of waters bath with thermostatic control excessively and is digested immediately after, the time is
120min.Bromelain Bromelain herein, nine front yard chemical products Co., Ltd (pineapple used in following example of Zhengzhou
Albumen enzyme source is identical)；Cellulase CELLUCLAST 1.5L, Novi believe that (China) Bioisystech Co., Ltd (implements below
Cellulase source used in example is identical).
After enzymatic hydrolysis, it is transferred to boiling water bath 20min in boiling water bath, is centrifuged immediately after, condition is
5000rpm, be centrifuged 10min, collect supernatant, into supernatant be added 3 times of volumes dehydrated alcohol precipitate polysaccharides, then from
The heart, condition 3000rpm are centrifuged 5min, after centrifugation, collect ethanol pellet with washes of absolute alcohol 3 times, then 50
48h is dried in~60 DEG C of vacuum drying ovens, obtained solid object is Gracilaria tenuistipitata polysaccharide.
Use Phenol sulfuric acid procedure measurement recovery rate for 28.36 ± 3.84%.
Clearance rate to hydroxy radical is 64.00% (polysaccharide concentration 1.2mgmL-1)。
The best Antioxidant Indexes 1.25mgmL of document Gracilaria tenuistipitata polysaccharide-1, hydroxyl radical free radical clearance rate 51.54%
(food industry science and technology, 2005,26 (3), 67-69).
Embodiment 2
Gracilaria tenuistipitata powder 1.0g is taken, disodium hydrogen phosphate-citric acid solution of prepared pH=5.5 is added
100mL, stirring makes powder sufficiently absorb water and mix on magnetic stirring apparatus, is then placed into and has been heated to 55 DEG C of ultrasonic wave
In cleaning device, setting ultrasonic time is 40min, and ultrasonic power 150W is ultrasonically treated.
After ultrasonic treatment, 1mL bromelain enzyme solutions (10mgmL is respectively added according to the compound enzyme dosage of 3wt%-1)
With 1mL cellulase solution (20mgmL-1), it is transferred in 55 DEG C of waters bath with thermostatic control excessively and is digested immediately after, the time is
120min。
After enzymatic hydrolysis, it is transferred to boiling water bath 20min in boiling water bath, is centrifuged immediately after, condition is
5000rpm, be centrifuged 10min, collect supernatant, into supernatant be added 3 times of volumes dehydrated alcohol precipitate polysaccharides, then from
The heart, condition 3000rpm are centrifuged 5min, after centrifugation, collect ethanol pellet with washes of absolute alcohol 3 times, then 50
48h is dried in~60 DEG C of vacuum drying ovens, obtained solid object is Gracilaria tenuistipitata polysaccharide.
Use Phenol sulfuric acid procedure measurement recovery rate for 38.52 ± 3.93%.
Clearance rate to hydroxy radical is 63.59% (polysaccharide concentration 1.2mgmL-1)。
Embodiment 3
Gracilaria tenuistipitata powder 1.0g is taken, disodium hydrogen phosphate-citric acid solution of prepared pH=5.5 is added
80mL, stirring makes powder sufficiently absorb water and mix on magnetic stirring apparatus, is then placed into and has been heated to 55 DEG C of ultrasonic wave
In cleaning device, setting ultrasonic time is 20min, and ultrasonic power 150W is ultrasonically treated.
After ultrasonic treatment, 1mL bromelain enzyme solutions (12.5mgmL is respectively added according to the compound enzyme dosage of 3wt%-1) and 1mL cellulase solution (17.5mgmL-1), it is transferred in 55 DEG C of waters bath with thermostatic control excessively and is digested immediately after, the time
For 120min.
After enzymatic hydrolysis, it is transferred to boiling water bath 20min in boiling water bath, is centrifuged immediately after, condition is
5000rpm, be centrifuged 10min, collect supernatant, into supernatant be added 3 times of volumes dehydrated alcohol precipitate polysaccharides, then from
The heart, condition 3000rpm are centrifuged 5min, after centrifugation, collect ethanol pellet with washes of absolute alcohol 3 times, then 50
48h is dried in~60 DEG C of vacuum drying ovens, obtained solid object is Gracilaria tenuistipitata polysaccharide.
Use Phenol sulfuric acid procedure measurement recovery rate for 33.16 ± 1.78%.
Clearance rate to hydroxy radical is 64.96% (polysaccharide concentration 1.2mgmL-1)。
Embodiment 4
Gracilaria tenuistipitata powder 1.0g is taken, disodium hydrogen phosphate-citric acid solution of prepared pH=5.5 is added
60mL, stirring makes powder sufficiently absorb water and mix on magnetic stirring apparatus, is then placed into and has been heated to 55 DEG C of ultrasonic wave
In cleaning device, setting ultrasonic time is 40min, and ultrasonic power 90W is ultrasonically treated.
After ultrasonic treatment, 1mL bromelain enzyme solutions (10mgmL is respectively added according to the compound enzyme dosage of 3wt%-1)
With 1mL cellulase solution (20mgmL-1), it is transferred in 55 DEG C of waters bath with thermostatic control excessively and is digested immediately after, the time is
120min。
After enzymatic hydrolysis, it is transferred to boiling water bath 20min in boiling water bath, is centrifuged immediately after, condition is
5000rpm, be centrifuged 10min, collect supernatant, into supernatant be added 3 times of volumes dehydrated alcohol precipitate polysaccharides, then from
The heart, condition 3000rpm are centrifuged 5min, after centrifugation, collect ethanol pellet with washes of absolute alcohol 3 times, then 50
48h is dried in~60 DEG C of vacuum drying ovens, obtained solid object is Gracilaria tenuistipitata polysaccharide.
Use Phenol sulfuric acid procedure measurement recovery rate for 25.01 ± 3.82%.
Clearance rate to hydroxy radical is 54.94% (polysaccharide concentration 1.2mgmL-1)。
Embodiment 5
Gracilaria tenuistipitata powder 1.0g is taken, disodium hydrogen phosphate-citric acid solution of prepared pH=5.5 is added
60mL, stirring makes powder sufficiently absorb water and mix on magnetic stirring apparatus, is then placed into and has been heated to 55 DEG C of ultrasonic wave
In cleaning device, setting ultrasonic time is 40min, and ultrasonic power 120W is ultrasonically treated.
After ultrasonic treatment, 1mL bromelain enzyme solutions (10mgmL is respectively added according to the compound enzyme dosage of 3wt%-1)
With 1mL cellulase solution (20mgmL-1), it is transferred in 55 DEG C of waters bath with thermostatic control excessively and is digested immediately after, the time is
120min。
After enzymatic hydrolysis, it is transferred to boiling water bath 20min in boiling water bath, is centrifuged immediately after, condition is
5000rpm, be centrifuged 10min, collect supernatant, into supernatant be added 3 times of volumes dehydrated alcohol precipitate polysaccharides, then from
The heart, condition 3000rpm are centrifuged 5min, after centrifugation, collect ethanol pellet with washes of absolute alcohol 3 times, then 50
48h is dried in~60 DEG C of vacuum drying ovens, obtained solid object is Gracilaria tenuistipitata polysaccharide.
Use Phenol sulfuric acid procedure measurement recovery rate for 30.62 ± 0.99%.
Clearance rate to hydroxy radical is 61.72% (polysaccharide concentration 1.2mgmL-1)。
Embodiment 6
Gracilaria tenuistipitata powder 1.0g is taken, disodium hydrogen phosphate-citric acid solution of prepared pH=5.5 is added
60mL, stirring makes powder sufficiently absorb water and mix on magnetic stirring apparatus, is then placed into and has been heated to 55 DEG C of ultrasonic wave
In cleaning device, setting ultrasonic time is 40min, and ultrasonic power 90W is ultrasonically treated.
After ultrasonic treatment, 1mL bromelain enzyme solutions (10mgmL is respectively added according to the compound enzyme dosage of 3wt%-1)
With 1mL cellulase solution (20mgmL-1), it is transferred in 55 DEG C of waters bath with thermostatic control excessively and is digested immediately after, the time is
180min。
After enzymatic hydrolysis, it is transferred to boiling water bath 20min in boiling water bath, is centrifuged immediately after, condition is
5000rpm, be centrifuged 10min, collect supernatant, into supernatant be added 3 times of volumes dehydrated alcohol precipitate polysaccharides, then from
The heart, condition 3000rpm are centrifuged 5min, after centrifugation, collect ethanol pellet with washes of absolute alcohol 3 times, then 50
48h is dried in~60 DEG C of vacuum drying ovens, obtained solid object is Gracilaria tenuistipitata polysaccharide.
Use Phenol sulfuric acid procedure measurement recovery rate for 37.17 ± 4.51%.
Clearance rate to hydroxy radical is 55.85%, (polysaccharide concentration 1.2mgmL-1)。
Embodiment 7
Gracilaria tenuistipitata powder 1.0g is taken, disodium hydrogen phosphate-citric acid solution of prepared pH=4.5 is added
60mL, stirring makes powder sufficiently absorb water and mix on magnetic stirring apparatus, is then placed into and has been heated to 45 DEG C of ultrasonic wave
In cleaning device, setting ultrasonic time is 40min, and ultrasonic power 150W is ultrasonically treated.
After ultrasonic treatment, 1mL bromelain enzyme solutions (3.33mgmL is respectively added according to the compound enzyme dosage of 1wt%-1) and 1mL cellulase solution (6.67mgmL-1), it is transferred in 45 DEG C of waters bath with thermostatic control excessively and is digested immediately after, the time
For 60min.
After enzymatic hydrolysis, it is transferred to boiling water bath 20min in boiling water bath, is centrifuged immediately after, condition is
5000rpm, be centrifuged 10min, collect supernatant, into supernatant be added 3 times of volumes dehydrated alcohol precipitate polysaccharides, then from
The heart, condition 3000rpm are centrifuged 5min, after centrifugation, collect ethanol pellet with washes of absolute alcohol 3 times, then 50
48h is dried in~60 DEG C of vacuum drying ovens, obtained solid object is Gracilaria tenuistipitata polysaccharide.
Use Phenol sulfuric acid procedure measurement recovery rate for 34.51 ± 1.14%.
Clearance rate to hydroxy radical is 65.54%, (polysaccharide concentration 1.2mgmL-1)。
Embodiment 8
Gracilaria tenuistipitata powder 1.0g is taken, disodium hydrogen phosphate-citric acid solution of prepared pH=4.5 is added
60mL, stirring makes powder sufficiently absorb water and mix on magnetic stirring apparatus, is then placed into and has been heated to 65 DEG C of ultrasonic wave
In cleaning device, setting ultrasonic time is 40min, and ultrasonic power 150W is ultrasonically treated.
After ultrasonic treatment, 1mL bromelain enzyme solutions (10mgmL is respectively added according to the compound enzyme dosage of 3wt%-1)
With 1mL pectinase solution (20mgmL-1, it is pectase Pectinase C80max, Dutch DSM group), it shifts immediately after
It is digested into 65 DEG C of waters bath with thermostatic control excessively, time 60min.
After enzymatic hydrolysis, it is transferred to boiling water bath 20min in boiling water bath, is centrifuged immediately after, condition is
5000rpm, be centrifuged 10min, collect supernatant, into supernatant be added 3 times of volumes dehydrated alcohol precipitate polysaccharides, then from
The heart, condition 3000rpm are centrifuged 5min, after centrifugation, collect ethanol pellet with washes of absolute alcohol 3 times, then 50
48h is dried in~60 DEG C of vacuum drying ovens, obtained solid object is Gracilaria tenuistipitata polysaccharide.
Use Phenol sulfuric acid procedure measurement recovery rate for 37.83 ± 0.40%.
Clearance rate to hydroxy radical is 65.49%, (polysaccharide concentration 1.2mgmL-1)。
Embodiment 9
Gracilaria tenuistipitata powder 1.0g is taken, disodium hydrogen phosphate-citric acid solution of prepared pH=5.5 is added
60mL, stirring makes powder sufficiently absorb water and mix on magnetic stirring apparatus, is then placed into and has been heated to 65 DEG C of ultrasonic wave
In cleaning device, setting ultrasonic time is 40min, and ultrasonic power 150W is ultrasonically treated.
After ultrasonic treatment, 1mL bromelain enzyme solutions (16.7mgmL is respectively added according to the compound enzyme dosage of 5wt%-1) and 1mL cellulase solution (33.3mgmL-1), it is transferred in 65 DEG C of waters bath with thermostatic control excessively and is digested immediately after, the time
For 60min.
After enzymatic hydrolysis, it is transferred to boiling water bath 20min in boiling water bath, is centrifuged immediately after, condition is
5000rpm, be centrifuged 10min, collect supernatant, into supernatant be added 3 times of volumes dehydrated alcohol precipitate polysaccharides, then from
The heart, condition 3000rpm are centrifuged 5min, after centrifugation, collect ethanol pellet with washes of absolute alcohol 3 times, then 50
48h is dried in~60 DEG C of vacuum drying ovens, obtained solid object is Gracilaria tenuistipitata polysaccharide.
Use Phenol sulfuric acid procedure measurement recovery rate for 38.60 ± 1.13%.
Clearance rate to hydroxy radical is 63.67%, (polysaccharide concentration 1.2mgmL-1)。
Embodiment 10
Gracilaria tenuistipitata powder 1.0g is taken, disodium hydrogen phosphate-citric acid solution of prepared pH=5.5 is added
60mL, stirring makes powder sufficiently absorb water and mix on magnetic stirring apparatus, is then placed into and has been heated to 65 DEG C of ultrasonic wave
In cleaning device, setting ultrasonic time is 30min, and ultrasonic power 150W is ultrasonically treated.
After ultrasonic treatment, 1mL bromelain enzyme solutions (16.7mgmL is respectively added according to the compound enzyme dosage of 5wt%-1) and 1mL cellulase solution (33.3mgmL-1), it is transferred in 65 DEG C of waters bath with thermostatic control excessively and is digested immediately after, the time
For 60min.
After enzymatic hydrolysis, it is transferred to boiling water bath 20min in boiling water bath, is centrifuged immediately after, condition is
5000rpm, be centrifuged 10min, collect supernatant, into supernatant be added 3 times of volumes dehydrated alcohol precipitate polysaccharides, then from
The heart, condition 3000rpm are centrifuged 5min, after centrifugation, collect ethanol pellet with washes of absolute alcohol 3 times, then 50
48h is dried in~60 DEG C of vacuum drying ovens, obtained solid object is Gracilaria tenuistipitata polysaccharide.
Use Phenol sulfuric acid procedure measurement recovery rate for 38.20 ± 1.08%.
Clearance rate to hydroxy radical is 64.90%, (polysaccharide concentration 1.2mgmL-1)。
Embodiment 11
Gracilaria tenuistipitata powder 1.0g is taken, disodium hydrogen phosphate-citric acid solution of prepared pH=5.5 is added
60mL, stirring makes powder sufficiently absorb water and mix on magnetic stirring apparatus, is then placed into and has been heated to 65 DEG C of ultrasonic wave
In cleaning device, setting ultrasonic time is 10min, and ultrasonic power 150W is ultrasonically treated.
After ultrasonic treatment, 1mL bromelain enzyme solutions (16.7mgmL is respectively added according to the compound enzyme dosage of 5wt%-1) and 1mL cellulase solution (33.3mgmL-1), it is transferred in 65 DEG C of waters bath with thermostatic control excessively and is digested immediately after, the time
For 60min.
After enzymatic hydrolysis, it is transferred to boiling water bath 20min in boiling water bath, is centrifuged immediately after, condition is
5000rpm, be centrifuged 10min, collect supernatant, into supernatant be added 3 times of volumes dehydrated alcohol precipitate polysaccharides, then from
The heart, condition 3000rpm are centrifuged 5min, after centrifugation, collect ethanol pellet with washes of absolute alcohol 3 times, then 50
48h is dried in~60 DEG C of vacuum drying ovens, obtained solid object is Gracilaria tenuistipitata polysaccharide.
Use Phenol sulfuric acid procedure measurement recovery rate for 33.19 ± 1.90%.
Clearance rate to hydroxy radical is 59.63% (polysaccharide concentration 1.2mgmL-1)。
It should be appreciated that the above examples are only used to illustrate the technical scheme of the present invention, rather than the guarantor that limitation is of the invention
Protect range.Those skilled in the art can make various modifications or changes to the present invention, the shape of equal value of these various changes or modification
Formula is equally fallen in the claim of this application limited range.
Claims (10)
1. the method for ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide, it is characterised in that the following steps are included:
1) the Gracilaria tenuistipitata powder by decolorization is taken, buffer solution is added, after stirring and evenly mixing, is ultrasonically treated；
2) after ultrasound, complex enzyme is added and carries out enzymolysis processing；The complex enzyme be bromelain and the second component enzymes,
Second component enzymes are cellulase or pectase, and the mass ratio of the bromelain and the second component enzymes is 1:1
~1:2；Compound enzyme dosage is 1~5wt% of the quality of Gracilaria tenuistipitata powder；
3) after digesting, enzyme-deactivating is collected by being centrifuged and filtering extraction fluid by supernatant using boiling water bath；
4) extracting solution is subjected to alcohol precipitation, be centrifuged, washed, it is dry, obtain Gracilaria tenuistipitata polyoses extract.
2. the method for ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide according to claim 1, which is characterized in that
The buffer solution is citrate-phosphate disodium hydrogen buffer solution, and pH is 4.5~6.5, and the concentration of disodium hydrogen phosphate is 0.05
±0.005mol·L-1, the volume of buffer solution and the mass ratio of powder are 40:1~100:1.
3. the method for ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide according to claim 1, which is characterized in that
Bath temperature when being ultrasonically treated is 45~65 DEG C, and ultrasonic power is 90~150W, and ultrasonic time is 10~40min.
4. the method for ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide according to claim 1, which is characterized in that
The hydrolysis temperature of the enzymolysis processing is 45~65 DEG C, and enzymolysis time is 60~180min.
5. the method for ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide according to claim 1, which is characterized in that
The enzyme deactivation is using boiling water bath by enzyme-deactivating, and extracting solution 20min or more in boiling water bath inactivates enzyme.
6. the method for ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide according to claim 1, which is characterized in that
The revolving speed of centrifugation described in step 3) is 4500rpm or more, and the time of centrifugation is 8min or more；Centrifugation described in step 4) turns
Speed is 3000rpm or more, and the time of centrifugation is 5min or more.
7. the method for ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide according to claim 1, which is characterized in that
The alcohol precipitation is the dehydrated alcohol precipitate polysaccharides that three times volume is added into supernatant, collects precipitating.
8. the method for ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide according to claim 1, which is characterized in that
The washing is to wash precipitating 2 times or more with dehydrated alcohol；The drying is to dry 40- in 50-70 DEG C of vacuum drying oven
60h。
9. the method for ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide according to claim 1, which is characterized in that
It being measured by Phenol sulfuric acid procedure, the recovery rate of ultrasonic wave assisted recombination enzymolysis and extraction Gracilaria tenuistipitata polysaccharide is 25.01~
38.60wt%.
10. a kind of Gracilaria tenuistipitata polysaccharide, which is characterized in that it is extracted by method described in any one of claim 1 to 9
It arrives, the Gracilaria tenuistipitata polysaccharide is 50-66% to the clearance rate of hydroxy radical；WhereinTest method is as follows: taking 0.5mL concentration is 1.2mg
mL-1Sample solution is into 7mL centrifuge tube with cover, the phosphate buffer solution of addition 1mL pH=7.4, concentration 0.1M, then plus
Enter 0.5mL 0.75mmolL-1Phen solution, after shaking up be added 0.5mL 0.75mmolL-1Iron ammonium sulfate is molten
Liquid shakes up rapidly, and the solution that 0.5mL mass concentration is 0.01% hydrogen peroxide is then added, and tube sealing shakes up, protects at 37 DEG C
Warm 60min, then cools to room temperature, and using distilled water as reference, absorbance value is measured at 510nm, note absorbance value is
Asample；Sample solution is substituted for distilled water, carries out identical processing, measurement, note absorbance value is Ablank；By sample solution
It is substituted for distilled water with hydrogen peroxide, carries out identical processing, measurement, note absorbance value is A0。
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116178585A (en) * | 2023-03-20 | 2023-05-30 | 江苏沿江地区农业科学研究所 | Combined method for extracting okra polysaccharide |
| CN116444692A (en) * | 2023-05-19 | 2023-07-18 | 四川农业大学 | Enzymolysis extraction method of pollen polysaccharide |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101475651A (en) * | 2009-01-16 | 2009-07-08 | 伍曾利 | Preparation of Gracilaria gigas Harvey polysaccharides |
| CN103074321A (en) * | 2013-02-01 | 2013-05-01 | 山东省巨野晨农天然产物有限公司 | Immobilized compound enzyme and method for treating alliin waste liquid by using same |
| CN105542026A (en) * | 2016-01-19 | 2016-05-04 | 济南大学 | Method for efficiently extracting okra polysaccharide |
| CN108148147A (en) * | 2018-01-25 | 2018-06-12 | 安徽诚亚生物科技有限公司 | A kind of extracting method of tremella polysaccharides |
-
2019
- 2019-05-31 CN CN201910466353.3A patent/CN110283860B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101475651A (en) * | 2009-01-16 | 2009-07-08 | 伍曾利 | Preparation of Gracilaria gigas Harvey polysaccharides |
| CN103074321A (en) * | 2013-02-01 | 2013-05-01 | 山东省巨野晨农天然产物有限公司 | Immobilized compound enzyme and method for treating alliin waste liquid by using same |
| CN105542026A (en) * | 2016-01-19 | 2016-05-04 | 济南大学 | Method for efficiently extracting okra polysaccharide |
| CN108148147A (en) * | 2018-01-25 | 2018-06-12 | 安徽诚亚生物科技有限公司 | A kind of extracting method of tremella polysaccharides |
Non-Patent Citations (2)
| Title |
|---|
| 刘慧燕，等: "江蓠低分子量多糖的提取以及抗氧化活性研究", 《食品工业科技》 * |
| 郝晓敏，等: "超声酸提江蓠硫酸酯多糖工艺及其性质研究", 《食品科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116178585A (en) * | 2023-03-20 | 2023-05-30 | 江苏沿江地区农业科学研究所 | Combined method for extracting okra polysaccharide |
| CN116444692A (en) * | 2023-05-19 | 2023-07-18 | 四川农业大学 | Enzymolysis extraction method of pollen polysaccharide |
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