CN100336831C - Method for extracting abalone polysaccharide - Google Patents
Method for extracting abalone polysaccharide Download PDFInfo
- Publication number
- CN100336831C CN100336831C CNB200510047409XA CN200510047409A CN100336831C CN 100336831 C CN100336831 C CN 100336831C CN B200510047409X A CNB200510047409X A CN B200510047409XA CN 200510047409 A CN200510047409 A CN 200510047409A CN 100336831 C CN100336831 C CN 100336831C
- Authority
- CN
- China
- Prior art keywords
- abalone
- minutes
- supernatant liquor
- polysaccharide
- water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 150000004676 glycans Chemical class 0.000 title claims abstract description 99
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 98
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 98
- 238000000034 method Methods 0.000 title claims abstract description 56
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 98
- 102000004190 Enzymes Human genes 0.000 claims abstract description 97
- 108090000790 Enzymes Proteins 0.000 claims abstract description 97
- 238000005119 centrifugation Methods 0.000 claims abstract description 52
- 238000000605 extraction Methods 0.000 claims abstract description 30
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 20
- 230000003187 abdominal effect Effects 0.000 claims abstract description 18
- 238000001035 drying Methods 0.000 claims abstract description 16
- 210000001835 viscera Anatomy 0.000 claims abstract description 16
- 238000005507 spraying Methods 0.000 claims abstract description 10
- 239000000203 mixture Substances 0.000 claims abstract description 9
- 239000000047 product Substances 0.000 claims abstract description 8
- 235000013372 meat Nutrition 0.000 claims abstract description 7
- 239000006228 supernatant Substances 0.000 claims description 92
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 58
- 238000001556 precipitation Methods 0.000 claims description 57
- 238000010792 warming Methods 0.000 claims description 32
- 239000002994 raw material Substances 0.000 claims description 31
- 238000000703 high-speed centrifugation Methods 0.000 claims description 21
- 210000002784 stomach Anatomy 0.000 claims description 21
- 230000007062 hydrolysis Effects 0.000 claims description 19
- 238000006460 hydrolysis reaction Methods 0.000 claims description 19
- 239000002002 slurry Substances 0.000 claims description 17
- 238000012546 transfer Methods 0.000 claims description 17
- 230000007935 neutral effect Effects 0.000 claims description 15
- 239000000843 powder Substances 0.000 claims description 13
- 238000003756 stirring Methods 0.000 claims description 13
- 102000035092 Neutral proteases Human genes 0.000 claims description 12
- 108091005507 Neutral proteases Proteins 0.000 claims description 12
- 239000002253 acid Substances 0.000 claims description 12
- 238000001816 cooling Methods 0.000 claims description 12
- 108090000145 Bacillolysin Proteins 0.000 claims description 9
- 239000000284 extract Substances 0.000 claims description 9
- 238000002525 ultrasonication Methods 0.000 claims description 9
- 238000012545 processing Methods 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- 238000001291 vacuum drying Methods 0.000 claims description 6
- 238000005360 mashing Methods 0.000 claims description 5
- 230000002255 enzymatic effect Effects 0.000 claims description 4
- 239000012141 concentrate Substances 0.000 claims description 2
- 238000004108 freeze drying Methods 0.000 claims description 2
- 238000010298 pulverizing process Methods 0.000 claims description 2
- 239000011780 sodium chloride Substances 0.000 claims description 2
- 238000001694 spray drying Methods 0.000 claims description 2
- 238000011160 research Methods 0.000 abstract description 8
- 102000004142 Trypsin Human genes 0.000 abstract description 7
- 108090000631 Trypsin Proteins 0.000 abstract description 7
- 239000003513 alkali Substances 0.000 abstract description 5
- 238000002137 ultrasound extraction Methods 0.000 abstract description 3
- 230000002358 autolytic effect Effects 0.000 abstract description 2
- 239000012588 trypsin Substances 0.000 abstract description 2
- 229940088598 enzyme Drugs 0.000 abstract 7
- 239000000243 solution Substances 0.000 abstract 3
- 239000002131 composite material Substances 0.000 abstract 2
- 239000000126 substance Substances 0.000 abstract 2
- 102000057297 Pepsin A Human genes 0.000 abstract 1
- 108090000284 Pepsin A Proteins 0.000 abstract 1
- 229940111202 pepsin Drugs 0.000 abstract 1
- 150000004804 polysaccharides Polymers 0.000 abstract 1
- 239000010414 supernatant solution Substances 0.000 abstract 1
- 230000008569 process Effects 0.000 description 18
- 230000000694 effects Effects 0.000 description 17
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 16
- 241001465754 Metazoa Species 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229920000856 Amylose Polymers 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 208000002454 Nasopharyngeal Carcinoma Diseases 0.000 description 1
- 206010061306 Nasopharyngeal cancer Diseases 0.000 description 1
- 241000233805 Phoenix Species 0.000 description 1
- 241000206607 Porphyra umbilicalis Species 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 210000003746 feather Anatomy 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000010813 municipal solid waste Substances 0.000 description 1
- 201000011216 nasopharynx carcinoma Diseases 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000004223 radioprotective effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The present invention relates to a method for extracting abalone polysaccharide, which comprises the following steps: the shell of an abalone is removed, the meat (comprises the viscera) of the abalone is taken, and the wall is crashed by a tissue crashing machine; the crashed substances are put in water to be evenly mixed, the mixture is extracted for 2 to 6h at the temperature of 20 to 80 DEG C to be centrifugally separated, insoluble substances are put in water to be repeatedly extracted for 1 to 2 times, and then, supernatant solutions are mixed; the extracted solution is concentrated until the sugar content is from 1.5 to 3.0%, ethanol whose volume is 3 to 4 times of that of the extracted solution and whose concentration is 95% is added into the extracted solution, and then, the ethanol is deposited for 12 to 16h at the temperature of 0 to 4 DEG C; a product is obtained by a vacuum method after a centrifugation process is finished or a spraying method in a drying mode. An alkali adding extraction mode, an ultrasonic extraction mode and an enzyme extraction mode can also be used for extracting the abalone polysaccharide. A single enzyme, double enzymes, a composite enzyme, an autolytic enzyme and a composite method can be used by the enzyme extraction mode to extract the abalone polysaccharide; the enzymes comprise pepsin, trypsin, etc. The present invention develops the technological line for extracting abalone polysaccharide form an abalone (especially the abdominal feet and the viscera of the abalone), and the abalone polysaccharide can be effectively extracted. Because the research on the medicinal purposes of the abalone polysaccharide is deeply researched, the present invention provides the technological basis for further developing the abalone polysaccharide.
Description
Technical field
The present invention relates to a kind of extracting method of polysaccharide, belong to field of biological product.
Background technology
In recent years, along with development of molecular biology, people recognized one of polysaccharide, the same three class biomacromolecules that relate to vital movement essence with nucleic acid of protein gradually.Polysaccharide is that a class has extensive bioactive biomacromolecule material, claims saccharan again, and it is not only the essential composition of human life, and is present in all membrane structures, and participates in multiple vital functions activity.Over advancing year, find that polysaccharide has raise immunity to human body, therefore anticancer in addition, radioprotective, anti-inflammatory, physiologically active such as hypoglycemic are subjected to extensive concern and attention.
At present, study both at home and abroad more be eat, the medicinal fungus polysaccharide, along with to the going deep into of polysaccharide research, people also turn to the animal polysaccharide to sight gradually, marine animal STUDY ON POLYSACHAROSE especially also becomes the focus and the focus of researcher research day by day.Because abalone is long vegetative period, in this particular surroundings of high salt, high pressure, low temperature of ocean, live, in the health enrichment a large amount of physiologically active substances.In recent years, there is research from abalone, to separate and purifying can efficiently suppress the holosaccharide of nasopharyngeal carcinoma cell.A large amount of research paper reports show that also abalone polysaccharide has good antitumor activity and enhancing body immunologic function.Chinese Medicine information network (http://www.yy2000.com) published article claims, that abalone polysaccharide, laver amylose have proved is antitumor, prevent and treat the cardiovascular disorder effect, and this medicinal research continues, and will obtain more, better effect.Yet it is less to extract STUDY ON POLYSACHAROSE from animal, as for extract STUDY ON POLYSACHAROSE phoenix feathers and unicorn horns especially from marine animal.The research of effectively extracting abalone polysaccharide from abalone on one's body then yet there are no report.
Summary of the invention
The purpose of this invention is to provide a kind of processing method, is raw material with the abalone integral body of shelling, abalone abdominal foot, abalone internal organs, extracts abalone polysaccharide effectively.
Technical scheme of the present invention is: abalone through the processing of raw material, extraction, alcohol precipitation, drying and abalone polysaccharide.Concrete processing step is:
One, raw material is handled
The raw material abalone can be with fresh, and also available freezing product can also be used dry product.The abdominal foot and the internal organs that can divide abalone meat integral body (being regardless of abdominal foot and internal organs), abalone during processing.Abdominal foot of abalone and internal organs also can extract abalone polysaccharide separately.
Get abalone fresh or that thaw, cleaning shells obtains abalone meat integral body, also can separate obtaining abalone abdominal foot and internal organs, can distinguish or mixing to add 5～10 times of water stand-by with tissue mashing machine's broken walls, homogenate; Dry product is a band shell frozen abalone, and negative catalysis 5～10 minutes is shelled, and obtains abalone meat integral body, also can separate the abdominal foot and the internal organs that obtain abalone, and with it respectively or mixing freeze-drying or oven dry, pulverizing, its granularity is below 100 orders, and is stand-by.
Two, the extraction of abalone polysaccharide
The method of extracting abalone polysaccharide adopts water extract method, alkali liquor extraction method, ultrasonic extraction and enzyme extraction method, can also combine with first three methods by enzyme, to improve yield.
1, water extract method
The water that in slurry or dry powder, adds 10～50 times of weight, mixing, under 20～80 ℃ temperature, lixiviate is after 2～6 hours, and centrifugation obtains centrifuged supernatant and insolubles.Insolubles adds 10～40 times water again, and under 20～80 ℃ temperature, lixiviate is the recentrifuge separation after 2～6 hours, merges the supernatant liquor of twice gained, and is stand-by.
2, alkali liquor extraction method
The water that in slurry or dry powder, adds 10～50 times of weight, mixing, NaOH or the KOH solution of adding 0.5mol/L, transferring pH is 8～9, and under 20～60 ℃ temperature, lixiviate is after 2～6 hours, and centrifugation obtains supernatant liquor and insolubles.Insolubles adds 10～40 times water again, and keeping PH is 8～9, repeats aforesaid operations 1～2 time, merges the supernatant liquor of gained, and is stand-by with the acid neutralization.
3, ultrasonic extraction
The abalone slurry or the abalone dry powder that obtain during raw material handled add the water of 10～50 times of abalone amounts, and with 20～30KHZ ultrasonication 10～60 minutes, centrifugation obtained supernatant liquor and insolubles.Add 10～40 times water again in insolubles, used ultrasonication again 10～60 minutes, centrifugation obtains supernatant liquor and insolubles.The supernatant liquor that merges twice gained.
4, enzyme extraction method
Enzyme extraction method is that abalone slurry or abalone dry powder add water, under conditions such as suitable pH value of used enzyme and temperature, carry out enzymolysis with enzyme, used enzyme can be single enzyme, two enzyme, prozyme or autolytic enzyme, also can engage with the said extracted method, all can reach the extraction purpose.The kind of described single enzyme, two enzyme, prozyme is stomach en-, trypsinase, papoid, neutral protease etc.
(1) single, double, complex enzyme zymohydrolysis method
1) single enzyme enzymolysis process: the water that raw material after treatment adds 10～50 times of abalone amounts mixes, transferring pH with 6mol/L HCl is 1～5, the stomach en-that adds solution weight 0.05～3.00% stirred enzymolysis 2～5 hours at 35～50 ℃, kept above-mentioned pH value during hydrolysis; It is neutral transferring pH with the NaOH of 0.5mol/L or KOH solution again, is warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, is chilled to room temperature in 5 minutes, centrifugation, supernatant liquor and precipitation; Supernatant liquor is stand-by, and precipitation also can add water and repeat above-mentioned steps 1～2 time for 10～40 times, and the supernatant liquor of centrifugation merges stand-by.The enzymolysis operation of other kind enzyme (stomach en-, trypsinase, papoid, neutral protease etc.) is gone up the same, just is transferred to the suitable scope of used enzyme with alkali lye or acid solution.
2) two enzyme enzymolysis processs: the water that raw material after treatment adds 10～50 times of abalone amounts mixes, transferring pH with 6mol/L HCl is 1～5, the stomach en-that adds solution weight 0.05～3.00% stirred enzymolysis 1～6 hour at 20～60 ℃, kept above-mentioned pH value during hydrolysis; Transferring pH with the NaOH of 0.5mol/L or KOH solution again is 7～9, adds the trypsinase of solution weight 0.05～3.00%, 20～60 ℃ of stirred in water bath enzymolysis 1～6 hour, keeps above-mentioned pH value during hydrolysis; It is neutral transferring pH with acid behind the enzymolysis, is warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, is chilled to room temperature in 5 minutes, centrifugation, supernatant liquor and precipitation; Supernatant liquor is stand-by, and precipitation also can add 10～40 times of water and repeat above-mentioned steps 1～2 time, and the supernatant liquor of centrifugation merges stand-by.
3) complex enzyme zymohydrolysis method: with abalone slurry or the abalone dry powder that obtains in the raw material processing, the water that adds 10～50 times of abalone amounts, NaOH or KOH solution accent pH with 0.5mol/L are 7～9, add papoid, neutral protease and trypsinase, addition is 0.05～3.00% of a solution weight, the part by weight of three kinds of enzymes is 1.0: 0.1～1.0: 0.1～1.0, under 20～60 ℃ temperature, behind the enzymolysis 1～6 hour, transfer pH7 with acid, be warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, be chilled to room temperature in 5 minutes, centrifugation gets supernatant liquor and precipitation; Supernatant liquor is stand-by, and precipitation also can add 10～40 times of water and repeat above-mentioned steps 1～2 time, and the supernatant liquor of centrifugation merges stand-by.
(2) in conjunction with extraction method
1) self-dissolving and exogenous enzyme separate mutually legal: (1) self-dissolving: can carry out self-dissolving with following arbitrary method: 1. the abalone slurry adds 10～50 times of water of weight, with uviolizing 10～30 minutes, be to make it utilize self enzyme self-dissolving under 7.0～7.5,30～50 ℃ at pH with 0.06～0.08mol/L NaCl; 2. the abalone slurry is added 10～50 times of water of abalone amount and insert in the aseptic enzyme jar, self-dissolving is 6～8 hours under pH6.0～7.5, normal temperature; 3. the abalone slurry is added 10～50 times of water and insert in the aseptic enzyme jar, in pH6.0～7.5 time, 0～4 ℃ of self-dissolving 24～48 hours.(2) external source enzyme resolving: it is 1～5 that the solution of above-mentioned self-dissolving is transferred pH with the HCl of 6mol/L, the stomach en-that adds solution weight 0.3～0.5%, in 40～60 ℃ of enzymolysis 1～2 hour, it is neutral transferring pH behind the enzymolysis, be warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, be chilled to room temperature in 5 minutes, centrifugation gets supernatant liquor and precipitation; Supernatant liquor is stand-by, and precipitation also can add 10～40 times of water and repeat above-mentioned steps 1～2 time, and the supernatant liquor of centrifugation merges stand-by.
2) water is carried and two enzyme combined techniques: the abalone slurry or the abalone dry powder that obtain during raw material is handled add abalone and measure 10～50 times water, 20～80 ℃ of lixiviate 2～6 hours; Transferring pH with 6mol/L HCl is 1～5, adds the stomach en-of solution weight 0.05～3.00%, stirs enzymolysis 1～6 hour at 20～60 ℃, keeps above-mentioned pH value during hydrolysis; Transferring pH with the NaOH of 0.5mol/L or KOH solution again is 7～9, adds 0.05～3.00% trypsinase, 20～60 ℃ of stirred in water bath enzymolysis 1～6 hour, keeps above-mentioned pH value during hydrolysis; It is neutral transferring pH with acid behind the enzymolysis, is warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, is chilled to room temperature in 5 minutes, centrifugation, supernatant liquor and precipitation; Supernatant liquor is stand-by, and precipitation also can add 10～40 times of water and repeat above-mentioned steps 1～2 time, and the supernatant liquor of centrifugation merges stand-by.
Aforesaid operations is owing to need during two kinds of enzymic hydrolysiss pH value different, so divide front and back twice enzymolysis; Can add two kinds of enzymes simultaneously if two kinds of enzyme require pH values are identical, primary enzymolysis.Need the pH value identical as papoid, neutral protease during with trypsin hydrolyzing, be 7～9, just can under this pH value, add wherein two kinds of enzymes simultaneously.
3) ultrasonic wave and single enzyme combined techniques: the abalone slurry or the abalone dry powder that obtain during raw material handled, add the water of 10～50 times of abalone amounts, with 20～30KHZ ultrasonication 10～60 minutes, centrifugation obtained supernatant liquor and insolubles.In insolubles, add 10～40 times water again, NaOH or KOH solution accent pH with 0.5mol/L are 7～9, add papoid, addition is 0.05～3.00%, 20～60 ℃ of enzymolysis of solution weight after 1～6 hour, transfers pH7 with acid, be warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, be chilled to room temperature in 5 minutes, centrifugation gets supernatant liquor and precipitation; Supernatant liquor is stand-by, and precipitation also can add 10～40 times of water and repeat above-mentioned steps 1～2 time, and the supernatant liquor of centrifugation merges stand-by.
From aforesaid operations as can be seen, ultrasonic wave, alkali lye, water carry and single enzyme, two enzyme and prozyme combination are feasible all, only are to adjust the optimal pH value of enzyme and the addition of enzyme.
Above-mentioned all centrifugations are 3000～6000 rev/mins of high speed centrifugation operations about 10 minutes.
Three, abalone polysaccharide concentrates
With the centrifuged supernatant of above-mentioned gained, vacuum concentration to polysaccharide content is 1.5～3.0%.
Four, the alcohol precipitation of abalone polysaccharide
95% ethanol that adds 3～4 times of volumes in concentrated solution, under 0～4 ℃ temperature, alcohol precipitation is about 12～16 hours.
Five, abalone polysaccharide is centrifugal
Polysaccharide behind the alcohol precipitation, high speed centrifugation are about 10 minutes, and the precipitation that obtains is an abalone polysaccharide.
Six, the drying of abalone polysaccharide
The drying of abalone polysaccharide can all can achieve the goal with following arbitrary method:
1, the centrifugal insolubles that purifying is obtained, under 50～60 ℃ temperature, vacuum-drying obtains the exsiccant abalone polysaccharide.
2, the centrifugal insolubles that purifying is obtained, being diluted with water to proportion is about 1.05～1.10, carries out spraying drying, 160～180 ℃ of spray drying device temperature ins, 50～60 ℃ of temperature outs obtain the exsiccant abalone polysaccharide.
If desired, also the above-mentioned abalone polysaccharide that obtains can be crushed to below 160 orders with pulverizer.
The abalone polysaccharide that extracts with method of the present invention is a brown ceramic powder, and polysaccharide extract rate is 20%～40% (dry product ratio), and polysaccharide content is 6～18%, is soluble in hot water, is slightly soluble in cold water.Can be in the food service industry widespread use.If be further purified, deproteinated, polysaccharide content can reach 40%～60%, can be widely used in medicine trade again.
The characteristics that the present invention had are:
1. technology is reasonable, has started the operational path of (except that shell) extraction abalone polysaccharide in the abalone body, to the utmost abalone polysaccharide is extracted;
2. on extraction process, study widely and determined multiple effective extracting method;
3. because the research of the pharmaceutical usage of abalone polysaccharide deepens continuously, and its pharmaceutical use appreciates gradually, the present invention provides technical foundation for improving its economic benefit;
4. the abalone body except that shell all is used as and extracts raw material, particularly can utilize abdominal foot of abalone and internal organs to extract polysaccharide, not only makes full use of resource but also do not have trash discharge;
5. among the present invention, adopted halobiontic self-dissolving technology, can save the usage quantity of exogenous enzyme greatly, reduced cost.
Embodiment:
Example 1
Get whole 1000 grams of abalone, add water 5000 grams, use tissue mashing machine's homogenate, add 25000 gram water, stir, under 50 ℃ temperature, lixiviate is after 5 hours, and high speed centrifugation obtains supernatant liquor and precipitation.To precipitate adding 25000 gram water, and stir, under 80 ℃ temperature, lixiviate is after 3 hours, and high speed centrifugation obtains supernatant liquor and precipitation.The supernatant liquor that merges gained.With supernatant liquor vacuum concentration to polysaccharide content is 3% (detection of phenol sulfuric acid process); Add 21000 milliliters of 95% ethanol of 3 times of volumes in concentrated solution, under 4 ℃ temperature, alcohol precipitation 16 hours, high speed centrifugation obtained centrifugal insolubles about 10 minutes.Under 60 ℃ temperature, vacuum-drying obtains exsiccant abalone Crude polysaccharides 61.6 grams with this insolubles.Be crushed to below 160 orders with pulverizer.Detect with the phenol sulfuric acid process, its polysaccharide content is 8.1%.
Example 2
Get whole powder 1000 grams of freeze dried abalone, add water 50000 grams, stir, to wherein adding the 0.1N sodium hydroxide solution, transferring PH is 9, and under 20 ℃ temperature, lixiviate is after 6 hours, and being neutralized to pH with 6mol/L HCl is 7, and high speed centrifugation obtains supernatant liquor and precipitation.Add water 30000 grams to precipitation, stir, to the NaOH solution that wherein adds 0.5mol/L, transferring PH is 8, and under 80 ℃ temperature, lixiviate is after 3 hours, and being neutralized to pH with 6mol/L HCl is 7, and high speed centrifugation obtains supernatant liquor and precipitation.The supernatant liquor that merges gained.Concentrated, alcoholization, centrifugal as example 1.Under 50 ℃ temperature, vacuum-drying obtains exsiccant abalone Crude polysaccharides 298 grams with this insolubles.Be crushed to below 160 orders with pulverizer.Detect with the phenol sulfuric acid process, its polysaccharide content is 9.3%.
Example 3
Get abalone abdominal foot 1000 grams, handle as example 1, use 20KHZ ultrasonication 60 minutes, high speed centrifugation obtains centrifuged supernatant and precipitation.To precipitate adding 25000 gram water, and use 27KHZ ultrasonication 20 minutes, high speed centrifugation obtains centrifuged supernatant and precipitation.The supernatant liquor that merges gained.Concentrated, alcoholization, centrifugal as example 1.Under 50 ℃ temperature, vacuum-drying obtains thick 65.2 grams of exsiccant abalone polysaccharide with this insolubles.Be crushed to below 160 orders with pulverizer.Its polysaccharide content is 8.8%.
Example 4:
Get abalone abdominal foot 1000 grams, processing is as example 1, and to the NaOH solution that wherein adds 0.5mol/L, transferring pH is 8, add 900 gram trypsin enzyme activity 2500u/mg), under 37 ℃ temperature, enzymolysis is after 5 hours, and transferring pH with 6mol/L HCl is about 7, be warming up to 100 ℃ of enzymes 10 minutes of going out in 5 minutes, 5 minutes internal cooling are to room temperature, and high speed centrifugation obtains supernatant liquor and and precipitation.To precipitate and add 25000 gram water, to the KOH solution that wherein adds 0.5mol/L, transferring PH is 8, adds 150 gram trypsin enzyme activity 2500u/mg), under 50 ℃ temperature, behind the enzymolysis 2 hours, transferring pH with 6mol/L HCl is about 7, is warming up to 90 ℃ of enzymes 10 minutes of going out in 5 minutes, and internal cooling was to room temperature in 5 minutes, high speed centrifugation obtains supernatant liquor and and precipitation.The supernatant liquor that merges gained.Concentrated, alcoholization, centrifugal as example 1.With the centrifugal insolubles that obtains, being diluted with water to proportion is 1.07, carries out spraying drying, 160 ℃ of temperature ins, and 50 ℃ of temperature outs obtain exsiccant abalone polysaccharide 43.2 grams.Detect with the phenol sulfuric acid process, its polysaccharide content is 14.8%.
Example 5:
Get abalone abdominal foot dry powder 1000 gram, add 50000 gram water, stir, transferring pH with 6mol/L HCl is 3, adds the stomach en-s (enzyme 50u/mg alive) of 1500 grams, 50 ℃ of stirred in water bath enzymolysis 2 hours, keeps above-mentioned pH value during hydrolysis; It is neutral transferring pH with 0.5mol/L KOH liquid again, is warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, is chilled to room temperature in 5 minutes, centrifugation, supernatant liquor and precipitation; Precipitation adds water 30000 gram, adds the stomach en-s (enzyme live 50u/mg) of 500 grams, 40 ℃ of stirred in water bath enzymolysis 4 hours, keeps above-mentioned pH value during hydrolysis; It is neutral transferring pH with the KOH solution of 0.5mol/L again, is warming up to 98 ℃ of enzymes 10 minutes of going out in 5 minutes, is chilled to room temperature in 5 minutes, centrifugation, supernatant liquor and precipitation; The supernatant liquor of centrifugation merges stand-by.Concentrated, alcoholization, centrifugal as example 1.With the centrifugal insolubles that obtains, being diluted with water to proportion is 1.06, carries out spraying drying, 160 ℃ of temperature ins, and 50 ℃ of temperature outs obtain exsiccant abalone polysaccharide 236 grams.Detect with the phenol sulfuric acid process, its polysaccharide content is 13.9%.
Example 6:
Get abalone abdominal foot dry powder 1000 grams, add abalone amount 50000 gram water, stir, the neutral protease (enzyme activity is 100u/mg) that adds 1000 grams, 40 ℃ of stirred in water bath enzymolysis 5 hours, be warming up to 90 ℃ of enzymes 10 minutes of going out in 2 minutes, be chilled to room temperature in 5 minutes, centrifugation gets supernatant liquor and precipitation; Precipitation adds water 30000 gram, adds the neutral proteases (enzyme activity is 100u/mg) of 150 grams, 40 ℃ of stirred in water bath enzymolysis 3 hours, is warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, is chilled to room temperature in 5 minutes, centrifugation, supernatant liquor and precipitation.The supernatant liquor that merges gained.Concentrated, alcoholization, centrifugal as example 1.With the centrifugal insolubles that obtains, being diluted with water to proportion is 1.1, carries out spraying drying, 180 ℃ of temperature ins, and 60 ℃ of temperature outs obtain exsiccant abalone polysaccharide 225.5 grams.Detect with the phenol sulfuric acid process, its polysaccharide content is 14.2%.
Example 7:
Get abalone abdominal foot dry powder 1000 grams, add abalone amount 40000 gram water, stir, transferring pH with the KOH solution of 0.5mol/L is 8.5, add 600 gram papoids (enzyme activity is 1200u/mg),, keep above-mentioned pH value during hydrolysis 50 ℃ of stirred in water bath enzymolysis 4 hours; It is neutral transferring pH with 6mol/L HCl liquid again, is warming up to 98 ℃ of enzymes 10 minutes of going out in 5 minutes, is chilled to room temperature in 5 minutes, centrifugation, supernatant liquor and precipitation; Precipitation adds water 35000 gram, stirs, and transferring pH with the KOH solution of 0.5mol/L is 9.0, adds 175 and restrains papoids (enzyme activity is 1200u/mg), 35 ℃ of stirred in water bath enzymolysis 3 hours, keeps above-mentioned pH value during hydrolysis; It is neutral transferring pH with 6mol/L HCl liquid again, is warming up to 90 ℃ of enzymes 10 minutes of going out in 5 minutes, is chilled to room temperature in 5 minutes, centrifugation, supernatant liquor and precipitation; The supernatant liquor that merges gained.Concentrated, alcoholization, centrifugal as example 1.With the centrifugal insolubles that obtains, being diluted with water to proportion is 1.1, carries out spraying drying, 170 ℃ of temperature ins, and 55 ℃ of temperature outs obtain exsiccant abalone polysaccharide 234 grams.Detect with the phenol sulfuric acid process, its polysaccharide content is 15.2%.
Example 8:
Get abalone internal organs 1000 grams, add water 5000 grams, use tissue mashing machine's homogenate, add 25000 gram water, stir, transferring pH with 6mol/L HCl is 1, the stomach en-s (enzyme live 50u/mg) that add 900 grams 35 ℃ of stirred in water bath enzymolysis 2 hours, keep above-mentioned pH value during hydrolysis; Transferring pH with the KOH solution of 0.5mol/L again is 9, adds 300 gram trypsin enzyme activity 2500u/mg), 55 ℃ of stirred in water bath enzymolysis 4 hours, keep above-mentioned pH value during hydrolysis; It is neutral transferring pH with acid behind the enzymolysis, is warming up to 98 ℃ of enzymes 10 minutes of going out in 5 minutes, is chilled to room temperature in 5 minutes, centrifugation, supernatant liquor and precipitation; Precipitation adds water 10000 grams, repeats above-mentioned steps 1 time, and the supernatant liquor of centrifugation merges stand-by.Remaining operation obtains exsiccant abalone polysaccharide 50.2 grams as example 7.Detect with the phenol sulfuric acid process, its polysaccharide content is 8.4%.
Example 9:
Get whole 1000 grams of abalone, processing is as example 1, to the KOH solution that wherein adds 0.5mol/L, transferring pH is 7, and adding 300 gram papoids (enzyme activity is 1200u/mg), 100 gram neutral proteases (enzyme activity is 100u/mg) and 100 gram trypsin enzyme activities is 2500u/mg), under 37 ℃ temperature, behind the enzymolysis 6 hours, be warming up to 98 ℃ of enzymes 10 minutes of going out in 5 minutes, internal cooling was to room temperature (20 ℃) in 5 minutes, high speed centrifugation obtains supernatant liquor and precipitation.Precipitation adds water 20000 grams, to the KOH solution that wherein adds 0.5mol/L, transferring pH is 7, and adding 200 gram papoids (enzyme activity is 1200u/mg), 50 gram neutral proteases (enzyme activity is 100u/mg) and 20 gram trypsin enzyme activities is 2500u/mg), under 50 ℃ temperature, behind the enzymolysis 4 hours, transferring pH is about 7, is warming up to 98 ℃ of enzymes 10 minutes of going out in 5 minutes, and internal cooling was to room temperature (20 ℃) in 5 minutes, high speed centrifugation obtains supernatant liquor and precipitation.The supernatant liquor of centrifugation merges stand-by.Remaining operation obtains exsiccant abalone polysaccharide 50.9 grams as example 7.Detect with the phenol sulfuric acid process, its polysaccharide content is 14.6%.
Example 10:
Get whole 1000 grams of abalone, processing is as example 1, with the slurries that obtain, with uviolizing 30 minutes, the Nacl that with concentration is 0.06mol/l is in 7.5,50 ℃ of following self-dissolvings of PH, acquisition transfer pH to 5 from solution with 6mol/LHCl, add 150 gram stomach en-s (enzyme live 50u/mg), 40 ℃ of following enzymolysis 2 hours.Transfer PH to be warming up to 98 ℃ of enzymes 10 minutes of going out in 7,5 minutes, 5 minutes internal cooling are to room temperature, and high speed centrifugation obtains centrifuged supernatant and and precipitation.To precipitate and add 25000 gram water, transfer pH to 5 with 6mol/L HCl, add 80 gram stomach en-s (enzyme 50u/mg alive), under 60 ℃ temperature, enzymolysis is after 1 hour, and transferring pH with the KOH solution of 0.5mol/L is about 7, be warming up to 98 ℃ of enzymes 10 minutes of going out in 5 minutes, 5 minutes internal cooling are to room temperature, and high speed centrifugation obtains supernatant liquor and and precipitation.The supernatant liquor that merges gained.Remaining operation is with example 1.Obtain exsiccant abalone Crude polysaccharides 51.3 grams.Detect with the phenol sulfuric acid process, its polysaccharide content is 14.6%.
Example 11:
Get whole 1000 grams of abalone, handle as example 1, with the slurries that obtain, with uviolizing 10 minutes, with concentration be the Nacl of 0.08mol/l at PH7,30 ℃ of following self-dissolvings.Remaining operation is with example 10.Obtain exsiccant abalone Crude polysaccharides 50.9 grams.Detect with the phenol sulfuric acid process, its polysaccharide content is 14.4%.
Example 12:
Get abalone internal organs 1000 grams, handle as example 1, the slurries that obtain are put into aseptic enzymatic vessel, transferring pH is 6, normal temperature self-dissolving 8 hours; What will obtain then transferred pH to 3 from solution with 6mol/L HCl, adds 30 gram stomach en-s (enzyme live 50u/mg), 40 ℃ of following enzymolysis 2 hours.Transfer PH to be warming up to 90 ℃ of enzymes 10 minutes of going out in 7,3 minutes, 5 minutes internal cooling are to room temperature, and high speed centrifugation obtains centrifuged supernatant and and precipitation.Remaining operation is with example 10.Obtain exsiccant abalone Crude polysaccharides 66.2 grams.Detect with the phenol sulfuric acid process, its polysaccharide content is 6.5%.
Example 13:
Get abalone internal organs 1000 grams, handle as example 1, the slurries that obtain are put into aseptic enzymatic vessel, transferring pH is 7.5, normal temperature self-dissolving 6 hours.It is 5 that the solution of above-mentioned self-dissolving is transferred pH with 6mol/L HCl, the stomach en-(enzyme 50u/mg alive) that adds solution weight 100 grams, in 40 ℃ of enzymolysis 1～2 hour, transferring pH behind the enzymolysis be neutrality, is warming up to 98 ℃ of enzymes 10 minutes of going out in 5 minutes, is chilled to room temperature in 5 minutes, centrifugation, get supernatant liquor and precipitation, precipitation adds water and repeats above-mentioned steps 1 time, and the supernatant liquor of centrifugation merges stand-by.Remaining operation is with example 10.Obtain exsiccant abalone Crude polysaccharides 64.8 grams.Detect with the phenol sulfuric acid process, its polysaccharide content is 6.8%.
Example 14:
Get abalone abdominal foot 1000 gram, handle as 1,50 ℃ of lixiviate of example 3 hours, transferring pH with 6mol/L HCl is 3, adds the stomach en-s (enzyme 50u/mg alive) of 150 grams, 40 ℃ of stirred in water bath enzymolysis 5 hours, keeps above-mentioned pH value during hydrolysis; Transferring pH with the KOH solution of 0.5mol/L again is 7～9, adds the trypsinase of 100 grams .55 ℃ of stirred in water bath enzymolysis 2 hours, keeps above-mentioned pH value during hydrolysis; It is neutral transferring pH with acid behind the enzymolysis, is warming up to 98 ℃ of enzymes 10 minutes of going out in 5 minutes, is chilled to room temperature in 5 minutes, centrifugation, supernatant liquor and precipitation; Precipitation adds water and repeats above-mentioned steps 1 time, and the supernatant liquor of centrifugation merges stand-by.Concentrated, alcoholization, centrifugal as example 1.With the centrifugal insolubles that obtains, being diluted with water to proportion is 1.08, carries out spraying drying, 170 ℃ of temperature ins, and 55 ℃ of temperature outs obtain exsiccant abalone polysaccharide 50.3 grams.Detect with the phenol sulfuric acid process, its polysaccharide content is 16.1%.
Example 15:
Get abalone abdominal foot 1000 grams, handle as example 1, use 20KHZ ultrasonication 60 minutes, centrifugation obtains supernatant liquor and insolubles.In insolubles, add 25000 gram water again, transferring pH with the KOH solution of 0.5mol/L is 8.0, add 500 gram papoids (enzyme activity is 1200u/mg), behind the enzymolysis 4 hours, transfer pH to be warming up to 90～98 ℃ of enzymes 10 minutes of going out in 7,2～5 minutes with 6mol/L HCl, be chilled to room temperature in 5 minutes, centrifugation gets supernatant liquor and precipitation.Merge supernatant liquor.Concentrated, alcoholization, centrifugal as example 1.With the centrifugal insolubles that obtains, being diluted with water to proportion is 1.08, carries out spraying drying, 180 ℃ of temperature ins, and 50 ℃ of temperature outs obtain exsiccant abalone polysaccharide 49.2 grams.Detect with the phenol sulfuric acid process, its polysaccharide content is 15.8%.
Example 16:
Get abalone abdominal foot 1000 grams, handle as example 1, use 30KHZ ultrasonication 30 minutes, centrifugation obtains supernatant liquor and insolubles.Add 25000 gram water in insolubles again, transferring pH with 6mol/L HCl is 3, adds the stomach en-s (enzyme 50u/mg alive) of 100 grams, 40 ℃ of stirred in water bath enzymolysis 5 hours, keeps above-mentioned pH value during hydrolysis; Transferring pH with the KOH solution of 0.5mol/L again is 7～9, adds the trypsinase of 80 grams .55 ℃ of stirred in water bath enzymolysis 2 hours, keeps above-mentioned pH value during hydrolysis; It is neutral transferring pH with acid behind the enzymolysis, is warming up to 95 ℃ of enzymes 10 minutes of going out in 5 minutes, is chilled to room temperature in 5 minutes, centrifugation, supernatant liquor and precipitation.The supernatant liquor of centrifugation merges stand-by.Concentrated, alcoholization, centrifugal as example 1.With the centrifugal insolubles that obtains, being diluted with water to proportion is 1.10, carries out spraying drying, 180 ℃ of temperature ins, and 55 ℃ of temperature outs obtain exsiccant abalone polysaccharide 58.2 grams.Detect with the phenol sulfuric acid process, its polysaccharide content is 17.4%.
Claims (19)
1, the extracting method of abalone polysaccharide, comprise abalone through raw material handle, extract, concentrate, alcohol precipitation, exsiccant processing step and product;
It is the meat tissue mashing machine of the whole abalone broken wall that shells that wherein said raw material is handled, and adds 5～10 times of water homogenate;
Described extraction is treated good abalone raw material to be added 10～50 times of water mix, and 20～60 ℃ of following lixiviates 1～6 hour, centrifugation got insolubles and supernatant liquor, and insolubles adds 10～30 times of water, repeats 1～2 time aforesaid operations, and supernatant liquor merges;
Described concentrating is extraction gained clear liquid to be concentrated into contain sugar 1.5～3.0%;
Described alcohol precipitation is 95% ethanol with 3～4 times of volumes of concentrated solution, placed 12～16 hours down at 0～4 ℃, and centrifugation, getting insolubles is abalone polysaccharide;
Described drying is that 50～60 ℃ of vacuum-dryings of alcohol precipitation gained abalone polysaccharide are obtained product.
2, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that it is to separate from the whole abalone meat that shells to obtain abalone abdominal foot and internal organs that described raw material is handled, can distinguish or mixing to add 5～10 times of water stand-by with tissue mashing machine's broken walls, homogenate.
3, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that described raw material handle be shell whole abalone meat, separate the abdominal foot that obtains abalone and internal organs respectively or mixing freeze-drying or oven dry, pulverizing, the dry powder of its granularity for below 100 orders, forming.
4, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that described extraction is treated abalone raw material to be added 10～50 times of water mix, with 20～30KHZ ultrasonication 10～60 minutes, centrifugation gets insolubles and supernatant liquor adds 10～30 times of water, repeat 1～2 time aforesaid operations, supernatant liquor merges.
5, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that described extraction is treated abalone raw material to be added 10～50 times of water mix, adjust its pH value 8～9 with NaOH or KOH, extracted 1～6 hour at 20～60 ℃, centrifugation gets insolubles and supernatant liquor, adds 10～30 times of water, adjust its pH value 8～9 with NaOH or KOH once more, repeat 1～2 time aforesaid operations, supernatant liquor merges, and is neutralized to neutrality with HCl.
6, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that described extraction is treated abalone raw material to be added 10～50 times of water mix, adjust its pH value 2～5 with HCl, add the stomach en-of solution weight 0.05～3.00%, stir, handled 1～6 hour at 20～60 ℃, the speed enzyme 10 minutes of going out that heats up 90～98 ℃, speed is chilled to 20～30 ℃, and centrifugation gets insolubles and supernatant liquor, and it is neutral that supernatant liquor is adjusted its pH with NaOH or KOH.
7, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that described extraction is treated abalone raw material to be added 10～50 times of water mix, adjust its pH value 8～9 with NaOH or KOH, the trypsinase, papoid or the neutral protease that add solution weight 0.05～3.00%, stir, handled 1～6 hour at 20～60 ℃, the speed enzyme 10 minutes of going out that heats up 90～98 ℃, speed is chilled to 20～30 ℃, centrifugation gets insolubles and supernatant liquor, and it is neutral that supernatant liquor is adjusted its pH with NaOH or KOH.
8, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that described extraction is treated abalone raw material to be added 10～50 times of water mix, adjust its pH value 8～9 with NaOH or KOH, the papoid, the trypsinase that add solution weight 0.05～3.00%, the weight ratio of its two kinds of enzymes is 1: 0.01～1.0, stir, handled 1～6 hour at 20～60 ℃, the speed enzyme 10 minutes of going out that heats up 90～100 ℃, speed is chilled to 20～30 ℃, centrifugation gets insolubles and supernatant liquor, and supernatant liquor is neutralized to neutrality with Hcl.
9, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that described extraction is the abalone raw material that obtains after raw material is handled, add 10～50 times water, transfer pH1～5 with HCl, 0.05～3.00% stomach en-that adds solution weight, 20～60 ℃ of stirred in water bath enzymolysis 1～6 hour, keep pH simultaneously in above-mentioned scope; Transfer pH7～9 with NaOH or KOH again, add 0.05～3.00% trypsinase of solution weight,, keep pH simultaneously in above-mentioned scope 20～60 ℃ of stirred in water bath enzymolysis 1～6 hour; Transfer pH to be warming up to 90～100 ℃ of enzymes 10 minutes of going out in 7,2～5 minutes behind the enzymolysis, 5 minutes internal cooling are to room temperature, and high speed centrifugation gets insolubles and supernatant liquor.
10, extracting method according to the described abalone polysaccharide of claim 1, it is characterized in that described extraction is treated abalone raw material to be added 10～50 times of water mix, transferring pH with NaOH or KOH is 7～9, add papoid, neutral protease and trypsinase, adding total amount is 0.05～3.00% of solution weight, the part by weight of three kinds of enzymes is 1: 0.01～1: 0.01～1, under 20～60 ℃ temperature, behind the enzymolysis 1～6 hour, transferring pH is 7, be warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, 5 minutes internal cooling are to room temperature, and high speed centrifugation gets insolubles and supernatant liquor.
11, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that described extraction is the slurries with treated abalone raw material, with uviolizing 10～30 minutes, the NaCl that with concentration is 0.06～0.08mol/l is at PH 7～7.5, temperature is 30～50 ℃, makes it utilize the enzyme self-dissolving of self; Transfer pH to 1～5 with 6 mol HCl, add 0.05～3.00% stomach en-of solution weight, 20～60 ℃ of enzymolysis 1～6 hour; Transfer PH to be warming up to 90～98 ℃ of enzymes 10 minutes of going out in 7,2～5 minutes, 5 minutes internal cooling are to room temperature, high speed centrifugation about 10 minutes insolubles and supernatant liquor.
12,, it is characterized in that described extraction is that the slurries of abalone internal organ are put into aseptic enzymatic vessel, in pH6～7.5, normal temperature self-dissolving 6～8 hours according to the extracting method of the described abalone polysaccharide of claim 1; Transfer pH to 1～5 with HCl, add 0.05～3.00% stomach en-of solution weight, 20～60 ℃ of enzymolysis 1～6 hour.Transfer PH to be warming up to 90～98 ℃ of enzymes 10 minutes of going out in 7,2～5 minutes, 5 minutes internal cooling are to room temperature, and high speed centrifugation 10 minutes obtains insolubles and supernatant liquor.
13, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that described extraction is that the slurries of abalone internal organ are put into aseptic enzymatic vessel, self-dissolving is 24～48 hours in pH6～7.5, under 0～4 ℃ of temperature; Transfer pH to 1～5 with HCl, add 0.05～3.00% stomach en-of solution weight, 20～60 ℃ of enzymolysis 1～6 hour; Transfer PH to be warming up to 90～98 ℃ of enzymes 10 minutes of going out in 7,2～5 minutes, 5 minutes internal cooling are to room temperature, high speed centrifugation 10 minutes, insolubles and supernatant liquor.
14,, it is characterized in that described extraction is the abalone raw material after raw material is handled, and adds the water of 10～50 times of abalone amounts, 20～80 ℃ of lixiviates 3～6 hours according to the extracting method of the described abalone polysaccharide of claim 1; Transferring pH with HCl is 1～5, adds the stomach en-of solution weight 0.05～3.00%, 20～60 ℃ of enzymolysis 1～6 hour, keeps above-mentioned pH value during hydrolysis; Transferring pH with 0.5mol/LNaOH or KOH liquid again is 7～9, adds 0.05～3.00% trypsinase, 20～60 ℃ of enzymolysis 1～6 hour, keeps above-mentioned pH value during hydrolysis; It is neutral transferring pH with acid behind the enzymolysis, is warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, is chilled to room temperature in 5 minutes, and centrifugation gets insolubles and supernatant liquor.
15,, it is characterized in that described extraction is the abalone raw material after raw material is handled, and adds the water of 10～50 times of abalone amounts, 20～80 ℃ of lixiviates 3～6 hours according to the extracting method of the described abalone polysaccharide of claim 1; Transferring pH with NaOH or KOH liquid is 7～9, adds papoid, neutral protease, the tryptic combination in twos of solution weight 0.05～3.00%, 20～60 ℃ of stirred in water bath enzymolysis 1～6 hour, keeps above-mentioned pH value during hydrolysis; It is neutral transferring pH with acid behind the enzymolysis, is warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, is chilled to room temperature in 5 minutes, and centrifugation gets insolubles and supernatant liquor.
16, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that described extraction is the abalone raw material after raw material is handled, the water that adds 10～50 times of abalone amounts, with 20～30KHZ ultrasonication 10～60 minutes, centrifugation obtained supernatant liquor and insolubles; In insolubles, add 15～25 times water again, transferring pH with 0.5mol/LNaOH or KOH liquid is 7～9, add papoid, trypsinase, neutral protease or it makes up in twos, addition is 0.05～3.00% of a solution weight, at 20～60 ℃ of enzymolysis after 1～6 hour, transfer pH7 with acid, be warming up to 90～98 ℃ of enzymes 10 minutes of going out in 2～5 minutes, be chilled to room temperature in 5 minutes, centrifugation gets insolubles and supernatant liquor; Get supernatant liquor and precipitation; Precipitation adds water and repeats above-mentioned steps 1～2 time, and centrifugation gets supernatant liquor.
17, according to the extracting method of the described abalone polysaccharide of each claim in the claim 6～15, it is characterized in that the insolubles of centrifugation gained in the described extraction adds 10～40 times of water, repeat to extract 1～2 time, whole supernatant liquors of centrifugation gained merge.
18,, it is characterized in that described drying is 50～60 ℃ of following vacuum-dryings with alcohol precipitation gained abalone polysaccharide according to the extracting method of the described abalone polysaccharide of claim 1.
19, according to the extracting method of the described abalone polysaccharide of claim 1, it is characterized in that described drying is that purifying alcohol precipitation gained abalone polysaccharide is diluted with water to its proportion is 1.05～1.10, carry out spraying drying, spray drying unit entry and exit temperature is respectively 50～60 ℃ and 160～180 ℃.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200510047409XA CN100336831C (en) | 2005-10-11 | 2005-10-11 | Method for extracting abalone polysaccharide |
| JP2008533850A JP4420470B2 (en) | 2005-10-11 | 2006-10-10 | Abalone polysaccharide extraction method |
| PCT/CN2006/002650 WO2007041951A1 (en) | 2005-10-11 | 2006-10-10 | Extraction method of abalone polysaccharide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200510047409XA CN100336831C (en) | 2005-10-11 | 2005-10-11 | Method for extracting abalone polysaccharide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1749280A CN1749280A (en) | 2006-03-22 |
| CN100336831C true CN100336831C (en) | 2007-09-12 |
Family
ID=36604952
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB200510047409XA Active CN100336831C (en) | 2005-10-11 | 2005-10-11 | Method for extracting abalone polysaccharide |
Country Status (3)
| Country | Link |
|---|---|
| JP (1) | JP4420470B2 (en) |
| CN (1) | CN100336831C (en) |
| WO (1) | WO2007041951A1 (en) |
Families Citing this family (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101503721B (en) * | 2008-11-27 | 2012-01-11 | 浙江珍世堂生物科技有限公司 | Method for extracting swan-mussel polysaccharide |
| KR101257494B1 (en) * | 2010-05-27 | 2013-04-26 | 조선대학교산학협력단 | Composition for the treatment and the prevention of inflammatory diseases in immune system containing Abalone gastrointestinal digests |
| CN101955548B (en) * | 2010-09-26 | 2012-05-23 | 陕西农产品加工技术研究院 | Method for enzyme-assisted extraction of fructus tribuli polysaccharide |
| CN102140141B (en) * | 2011-05-13 | 2012-10-03 | 陈锦权 | Process for preparing polysaccharide of abalones |
| CN102839149B (en) * | 2012-09-21 | 2013-11-13 | 中国海洋大学 | Application of abalone viscera polysaccharide-alternative serum in cell culture medium |
| CN103819575B (en) * | 2014-03-07 | 2016-03-02 | 潍坊医学院 | The purposes of a kind of extracting method of Ground Beetle polysaccharide, the polysaccharide of extraction and this polysaccharide |
| CN104381791B (en) * | 2014-11-25 | 2015-08-26 | 烟台新海水产食品有限公司 | A kind of instant abalone raspberry particle electuary and preparation method thereof |
| CN104356249B (en) * | 2014-11-27 | 2016-07-13 | 厦门蓝湾科技有限公司 | The preparation technology of a kind of high-purity Carnis Haliotidis polysaccharide and application thereof |
| CN104513843B (en) * | 2014-12-30 | 2018-01-19 | 集美大学 | A kind of combined preparation process of polysaccharide and protein peptides |
| CN104497164A (en) * | 2015-01-26 | 2015-04-08 | 吉林大学 | High-voltage pulsed electric field assisted enzymolysis based clam polysaccharide extracting process |
| CN105087713A (en) * | 2015-09-01 | 2015-11-25 | 集美大学 | Preparation method of abalone viscera polysaccharide |
| CN105272887B (en) * | 2015-10-20 | 2017-06-23 | 集美大学 | A kind of method for extracting taurine and polysaccharide in the internal organ from abalone simultaneously |
| CN111513269B (en) * | 2020-04-16 | 2022-07-19 | 自然资源部第三海洋研究所 | Abalone viscera extract and application thereof |
| CN113150178B (en) * | 2021-04-14 | 2023-05-02 | 大连工业大学 | Sulfated abalone polysaccharide and application thereof in inhibiting new coronavirus |
| CN114133459A (en) * | 2021-11-23 | 2022-03-04 | 福建师范大学 | Preparation method and application of abalone polysaccharide |
| CN114805623A (en) * | 2022-04-22 | 2022-07-29 | 福建大众健康生物科技有限公司 | Ultralow-temperature freeze-thawing pressurization extraction process for abalone polysaccharide |
| CN115025515B (en) * | 2022-06-10 | 2023-11-14 | 陈建 | Pleurotus tuber-regium polysaccharide draws splitter |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1202749A (en) * | 1997-04-11 | 1998-12-23 | 莫列斯公司 | Input/output connector with resilient connecting means |
| CN1057776C (en) * | 1997-06-27 | 2000-10-25 | 吉林大学 | Combined extraction technology of ginseng and American ginseng polysaccharide saponin |
| CN1473508A (en) * | 2003-08-07 | 2004-02-11 | 大连太平洋海龙科技有限公司 | Nutrient solution of abalone, sea cucumber and sea chestnut and its producing method |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1202749C (en) * | 2002-08-30 | 2005-05-25 | 大连轻工业学院 | Abalone food and its preparation method |
-
2005
- 2005-10-11 CN CNB200510047409XA patent/CN100336831C/en active Active
-
2006
- 2006-10-10 JP JP2008533850A patent/JP4420470B2/en active Active
- 2006-10-10 WO PCT/CN2006/002650 patent/WO2007041951A1/en active Application Filing
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1202749A (en) * | 1997-04-11 | 1998-12-23 | 莫列斯公司 | Input/output connector with resilient connecting means |
| CN1057776C (en) * | 1997-06-27 | 2000-10-25 | 吉林大学 | Combined extraction technology of ginseng and American ginseng polysaccharide saponin |
| CN1473508A (en) * | 2003-08-07 | 2004-02-11 | 大连太平洋海龙科技有限公司 | Nutrient solution of abalone, sea cucumber and sea chestnut and its producing method |
Non-Patent Citations (2)
| Title |
|---|
| 鲍鱼多糖Hal-A的甲醇解研究 佘志刚 等，有机化学，第22卷第5期 2002 * |
| 鲍鱼多糖的提取及抗肿瘤试验 陈倩超，中国现代应用药学，第15卷第1期 1998 * |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2007041951A1 (en) | 2007-04-19 |
| CN1749280A (en) | 2006-03-22 |
| JP4420470B2 (en) | 2010-02-24 |
| JP2009510234A (en) | 2009-03-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100336831C (en) | Method for extracting abalone polysaccharide | |
| CN100336832C (en) | Process for extracting comb shell polysaccharide | |
| CN1281628C (en) | Method for extracting edible tree fungi polysaccharide | |
| WO2017076112A1 (en) | Oyster peptide capable of enhancing sexual function, and preparation method and application thereof | |
| CN101451157B (en) | Method for preparing low molecular weight sea cucumber polysaccharide | |
| CN1202751C (en) | Seacucumbus whole powder food and its preparation method | |
| CN101724677B (en) | Method for extracting collagen polypeptide and hydroxyapatite in fish scales by cooking hot extrusion | |
| CN102086464B (en) | Method for preparing chitin | |
| CN111685286B (en) | Oyster peptide with blood lipid reducing function and preparation method and application thereof | |
| CN106213523A (en) | A kind of extracting method of Salicornia Bigelovii Torr. dietary fiber | |
| CN1344565A (en) | Fucoidin ester as antiviral immunoregulator and its prepn | |
| CN1211019C (en) | Compsns. contg. peptide and electrolyte excretion promoter and foods contg. same | |
| CN102499967B (en) | Ultrasonic-assisted method for preparing garlic blood-pressure-lowering functional factors | |
| CN114395603A (en) | Method for preparing small molecular peptide by hydrolyzing oyster protein with complex enzyme | |
| WO2014194707A1 (en) | Calculus enzyme and preparation method therefor | |
| CN113584108A (en) | Preparation method of spleen aminopeptide | |
| CN101372514B (en) | Scaphium scaphigerum polysaccharide, preparation and use thereof | |
| CN102524797A (en) | Preparation method of pearl oyster peptide calcium | |
| CN1749282A (en) | Belly bottom snail polysaccharide and its preparing method | |
| CN108866135B (en) | Method for preparing horseshoe crab blood protein hydrolysate peptide with alpha-glucosidase inhibitory activity | |
| CN1552892A (en) | Physiological active buck peptide by compound proteinase catalysis and use of products thereby | |
| CN108175771A (en) | The purposes of mushroom polysaccharide | |
| CN1181750C (en) | Prawn feed immunopolysaccharide | |
| CN111675769A (en) | Sea cucumber extract and preparation process thereof | |
| RU2623738C1 (en) | Biologically active additive from marine hydrobionts - source of hondroitinsulfate and method of its production |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: DALIAN POLYTECHNIC COLLEGE; APPLICANT Free format text: FORMER OWNER: DALIAN POLYTECHNIC COLLEGE Effective date: 20060616 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20060616 Address after: 116034 No. 1 light industry garden, Ganjingzi District, Liaoning, Dalian Applicant after: Dalian Instutute of Light Industry Co-applicant after: Dalian Zhangzidao Fishery Group Co., Ltd. Address before: 116034 No. 1 light industry garden, Ganjingzi District, Liaoning, Dalian Applicant before: Dalian Instutute of Light Industry |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee |
Owner name: DALIAN POLYTECHNIC UNIVERSITY Free format text: FORMER NAME: DALIAN LIGHT INDUSTRY COLLEGE |
|
| CP01 | Change in the name or title of a patent holder |
Address after: 116034 Ganjingzi Light Industry Zone, Liaoning City, No. 1 Co-patentee after: Dalian Zhangzidao Fishery Group Co., Ltd. Patentee after: Dalian Polytechnic University Address before: 116034 Ganjingzi Light Industry Zone, Liaoning City, No. 1 Co-patentee before: Dalian Zhangzidao Fishery Group Co., Ltd. Patentee before: Dalian Instutute of Light Industry |
|
| C56 | Change in the name or address of the patentee | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 116034 Ganjingzi Light Industry Zone, Liaoning City, No. 1 Patentee after: Dalian Polytechnic University Patentee after: Zhangzidao Island Group Co., Ltd. Address before: 116034 Ganjingzi Light Industry Zone, Liaoning City, No. 1 Patentee before: Dalian Polytechnic University Patentee before: Dalian Zhangzidao Fishery Group Co., Ltd. |