CN108299569A - The preparation method of oyster polysaccharide - Google Patents
The preparation method of oyster polysaccharide Download PDFInfo
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- CN108299569A CN108299569A CN201810129431.6A CN201810129431A CN108299569A CN 108299569 A CN108299569 A CN 108299569A CN 201810129431 A CN201810129431 A CN 201810129431A CN 108299569 A CN108299569 A CN 108299569A
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- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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Abstract
The invention discloses the preparation methods of oyster polysaccharide, including：Water carries, hot water carries, alkali carries, enzymolysis, takes off albumen, fresh oyster meat degreaser drying is obtained into skimmed milk first, by the progress of skimmed milk solution, normal-temperature water carries, hot water water carries, lye extracts and dialyses, alcohol precipitation, then it is digested through trypsase and pepsin, finally de- albumen is carried out to enzymolysis liquid handles up to oyster polysaccharide with the mixture of chloroform, n-butanol, acetic acid.It has the beneficial effect that：It is successively digested through alkali protease and acid protease, remaining protein in oyster Thick many candies is hydrolyzed to little albumen and polypeptide, and remove through subsequently de- albumen, the content and purity of the oyster polysaccharide in finished product can be greatly improved；Acetic acid and the L dimethyl tartrates of proper proportion can act on protein denaturation jelly with D dimethyl tartrates, reduce the amount that oyster polysaccharide is separated, improve the yield and content of oyster polysaccharide, improve purity of polysaccharide.
Description
Technical field
The present invention relates to the field of deep of oyster, more particularly, to the preparation method of oyster polysaccharide.
Technical background
Polysaccharide is a kind of natural products, is widely present in the organic matters such as plant, microorganism, animal.Polysaccharide clinically,
Food industry, fermentation industry and petroleum industry have a wide range of applications.Simultaneously with marine animal resource further exploitation,
Marine animal polysaccharide is had been to be concerned by more and more people due to its unique structure and multiple biological activities.Oyster
（ostreagigastnunb）And its entirety of nearly edge animal, it is seashell.In subtropical zone, coastal all the supporting suitable for oyster in the torrid zone
It grows, China's distribution is very wide, and North gets the Yalu River, south is coastal all to produce oyster to Hainan Island.Oyster, which is software, has shell, micropredation to move
Object, salt-fresh water boundary are produced particularly fertile.Oyster is mollusk, and there are two shells, and one small and flat, another is big and grand
It rises, the surface irregularity of shell.Meat is edible, and can obtain through refining oyster sauce.Meat, shell, oil can all be used as medicine, and also cry oyster or oyster.It is male
Oyster is commonly called as oyster, is four big cultivated shellfish of the big cultivated shellfish of the first in the world and China.It is peculiar containing marine organisms in oyster
Various active substance, wherein the polysaccharide contained, have adjust immunity of organism, inhibit tumour, anti-aging, hypoglycemic, blood fat,
Etc. bioactivity.
Invention content
The purpose of the present invention is to provide the preparation methods of oyster polysaccharide, and preparation method is simple for this, can drop significantly
Low-protein denaturation jelly wraps the probability of oyster polysaccharide, is separated so as to substantially reducing oyster polysaccharide
Amount improves the yield and content of oyster polysaccharide, reduces the content of protein in product, improves purity of polysaccharide.
The present invention is directed to the problem of being mentioned in background technology, and the technical solution taken is：The preparation method of oyster polysaccharide, packet
It includes：Water carries, hot water carries, alkali carries, enzymolysis, takes off albumen, specifically includes following steps：
Water carries：Oyster fresh meat is rubbed, homogenate, obtains skimmed milk with acetone degreasing and low temperature drying, extracting degreasing powder adds 20-25 times of matter
The normal-temperature distilled water for measuring volume, is stirred at room temperature 3-5 hours, centrifuges, and residue is repeated as stated above to extract, merges extracting solution,
Concentration；The 95-98% ethyl alcohol of 4-5 times of quality, 1-4 DEG C of precipitates overnight is added, next day centrifugation collects precipitation, uses absolute ethyl alcohol respectively
It is dehydrated with acetone, is dried under reduced pressure at 45-48 DEG C and carries Thick many candies up to oyster water；After being carried by degreasing and water, it can remove water-soluble
Property protein, and provide advantage for further deproteination；
Hot water carries：15-18 times of quality volume distilled water is added in residue after above-mentioned room temperature is extracted, magnetic force at a temperature of 60-66 DEG C
Stirring 2-3 hours centrifuges, concentration, and the 95-98% ethyl alcohol of 4-5 times of quality volume, 1-4 DEG C of precipitates overnight is added, and next day centrifugation is received
Collection precipitation uses absolute ethyl alcohol and acetone to be dehydrated, is dried under reduced pressure at 45-48 DEG C and carries Thick many candies up to oyster hot water respectively；Pass through heat
Water purifies, and can improve the solubility of water soluble protein, further separation and concentration goes out the protein in Thick many candies；
Alkali carries：The sodium hydroxide solution of 6-8 times of volume 2.5-3.0%, 60-65 DEG C of magnetic is added in residue after above-mentioned hot water is extracted
Power stirs 2-3 hours, and neutrality is neutralized to dilute hydrochloric acid, centrifuges, and concentration is dialysed 48-72 hours with 3.5kDa bag filters, in bag taking
The 95-98% ethyl alcohol of 4-5 times of quality volume, 1-4 DEG C of precipitates overnight are added after liquid concentration, next day centrifugation is collected precipitation, used respectively
Absolute ethyl alcohol and acetone are dehydrated, and are dried under reduced pressure at 45-48 DEG C up to oyster alkali carries Thick many candies；It, can be further by alkali carries
Extra Separation of Proteins is gone out on ground, is ready for further enzymolysis；
Enzymolysis：Oyster Thick many candies are merged, the solution that mass fraction is 8-10% is configured to pure water, adjusts pH to 8.2-8.4,
Addition trypsase end mass fraction is 0.3-0.5%, insulation reaction 4-5 hours at a temperature of 52-55 DEG C；It uses after completion of the reaction
It in 95-98 DEG C of enzyme deactivation 12-18 minutes after dilute hydrochloric acid tune neutrality, is cooled to room temperature, adjusts pH to 2.3-2.4, pepsin is added
It is 0.5-0.7% to whole mass fraction, insulation reaction 3-4 hours at a temperature of 37-40 DEG C, after completion of the reaction through dilute sodium hydroxide
Solution neutralizes, and low temperature is concentrated into 10-15% up to enzymolysis liquid；Successively digested through alkali protease and acid protease, oyster is thick
Remaining protein is hydrolyzed to little albumen and polypeptide in polysaccharide, and is removed through subsequently de- albumen, can greatly improve in finished product
The content and purity of oyster polysaccharide；Oyster polysaccharide can have an impact bacterial cell membrane, change cell membrane fluidity and penetrating
Property, it destroys the supramolecular structure of cell film and cellular content is caused to be lost in, bacteria cell wall influence can also be acted on
Bacterial metabolism, to play fungistatic effect；
De- albumen：By chloroform, n-butanol, acetic acid according to volume ratio 45-48:10-15:1 ratio is uniformly mixed, in mixed solution
Dimethyl tartrate also containing 1.5-1.8 ‰, L-TARTARIC ACID dimethyl ester and D- dimethyl tartrates in dimethyl tartrate
Ratio is 15.6-15.7:1；Under constant stirring, according to volume ratio 1:The ratio of 7-8 pours into enzymolysis liquid in mixed solution, close
It is honored as a queen and uses magnetic stirrer 45-60 minutes, pour into separatory funnel, upper solution is collected after static layering, and centrifuge and divide
A small amount of denatured protein is separated out, by above-mentioned steps repetitive operation 4-5 times, keeps the removal of protein in extracting solution complete, in merging
Clear liquid is concentrated through low temperature up to oyster polysaccharide；Chloroform, n-butanol mixed solution floating preteins and polypeptide can be changed into not
Soluble substance is removed through centrifugation, can achieve the purpose that remove protein in oyster polysaccharide, the L- winestones of acetic acid and proper proportion
Dimethyl phthalate can act on protein denaturation jelly with D- dimethyl tartrates, reduce protein denaturation jelly and wrap
The probability of oyster polysaccharide improves the yield of oyster polysaccharide and contains so as to substantially reduce the amount that oyster polysaccharide is separated
Amount reduces the content of protein in product, improves purity of polysaccharide.
Compared with the prior art, the advantages of the present invention are as follows：
1）Acetic acid and the L-TARTARIC ACID dimethyl ester of proper proportion can act on protein denaturation glue with D- dimethyl tartrates
Object reduces the probability that protein denaturation jelly wraps oyster polysaccharide, is separated so as to substantially reduce oyster polysaccharide
The amount gone improves the yield and content of oyster polysaccharide, reduces the content of protein in product, improves purity of polysaccharide；
2）It is successively digested through alkali protease and acid protease, remaining protein in oyster Thick many candies is hydrolyzed to little albumen
And polypeptide, and removed through subsequently de- albumen, the content and purity of the oyster polysaccharide in finished product can be greatly improved；Oyster polysaccharide energy
It is enough that bacterial cell membrane is had an impact, change cell membrane fluidity and permeability, destroy the supramolecular structure of cell film and
Cellular content is caused to be lost in, can also act on bacteria cell wall influences bacterial metabolism, plays fungistatic effect.
Specific implementation mode
The present invention program is described further below by embodiment：
Embodiment 1：
The preparation method of oyster polysaccharide, includes the following steps：
1）Oyster fresh meat is rubbed, homogenate, obtains skimmed milk with acetone degreasing and low temperature drying, extracting degreasing powder adds 20 times of quality volumes
Normal-temperature distilled water, be stirred at room temperature 3 hours, centrifuge, by residue as stated above repeat extract, merge extracting solution, concentration；It is added
95% ethyl alcohol of 4 times of quality, 1 DEG C of precipitates overnight, next day centrifugation collect precipitation, absolute ethyl alcohol and acetone are used to be dehydrated respectively, 45 DEG C
Under be dried under reduced pressure and carry Thick many candies up to oyster water；2）15 times of quality volume distilled water are added in residue after above-mentioned room temperature is extracted,
Magnetic agitation 2 hours at a temperature of 60 DEG C, centrifuge, concentration, be added 4 times of quality volumes 95% ethyl alcohol, 1 DEG C of precipitates overnight, next day from
The heart collects precipitation, uses absolute ethyl alcohol and acetone to be dehydrated respectively, is dried under reduced pressure at 45 DEG C and carries Thick many candies up to oyster hot water；
3）Residue after above-mentioned hot water is extracted is added the sodium hydroxide solution of 6 times of volumes 2.5%, 60 DEG C of magnetic agitations 2 hours,
It is neutralized to neutrality with dilute hydrochloric acid, is centrifuged, concentration is dialysed 48 hours with 3.5kDa bag filters, and 4 times of matter are added after liquid concentration in bag taking
95% ethyl alcohol of volume, 1 DEG C of precipitates overnight are measured, next day centrifugation collects precipitation, uses absolute ethyl alcohol and acetone to be dehydrated respectively, at 45 DEG C
It is dried under reduced pressure up to oyster alkali carries Thick many candies；4）Oyster Thick many candies are merged, the solution that mass fraction is 8% is configured to pure water,
PH to 8.2 is adjusted, it is 0.3% that trypsase end mass fraction, which is added, the insulation reaction 4 hours at a temperature of 52 DEG C；After completion of the reaction
With, in 95 DEG C of enzyme deactivations 12 minutes, being cooled to room temperature after dilute hydrochloric acid tune neutrality, pH to 2.3 is adjusted, pepsin is added to whole quality
Score is 0.5%, and insulation reaction 3 hours, neutralizes through diluted sodium hydroxide solution after completion of the reaction at a temperature of 37 DEG C, low temperature concentration
To 10% up to enzymolysis liquid；5）By chloroform, n-butanol, acetic acid according to volume ratio 45:10:1 ratio is uniformly mixed, mixed solution
In also containing 1.5 ‰ dimethyl tartrate, the ratio of L-TARTARIC ACID dimethyl ester and D- dimethyl tartrates in dimethyl tartrate
Example is 15.6:1；Under constant stirring, according to volume ratio 1:7 ratio pours into enzymolysis liquid in mixed solution, and magnetic is used after sealing
Power blender stirs 45 minutes, pours into separatory funnel, upper solution is collected after static layering, and centrifuge out a small amount of denaturation
Protein keeps the removal of protein in extracting solution complete by above-mentioned steps repetitive operation 4 times, merges supernatant and is concentrated through low temperature
Up to oyster polysaccharide.
Embodiment 2：
The preparation method of oyster polysaccharide, includes the following steps：
1）Water carries：Oyster fresh meat is rubbed, homogenate, obtains skimmed milk with acetone degreasing and low temperature drying, extracting degreasing powder adds 25 times of matter
The normal-temperature distilled water for measuring volume, is stirred at room temperature 5 hours, centrifuges, and residue is repeated as stated above to extract, merges extracting solution, dense
Contracting；98% ethyl alcohol of 5 times of quality, 4 DEG C of precipitates overnights is added, next day centrifugation collects precipitation, uses absolute ethyl alcohol and acetone de- respectively
Water is dried under reduced pressure at 48 DEG C and carries Thick many candies up to oyster water；After being carried by degreasing and water, water soluble protein can be removed, and
And provide advantage for further deproteination；
2）Hot water carries：18 times of quality volume distilled water are added in residue after above-mentioned room temperature is extracted, magnetic agitation 3 at a temperature of 66 DEG C
Hour, it centrifuges, concentration, 98% ethyl alcohol of 5 times of quality volumes, 4 DEG C of precipitates overnights is added, next day centrifugation is collected precipitation, used respectively
Absolute ethyl alcohol and acetone are dehydrated, and are dried under reduced pressure at 48 DEG C and are carried Thick many candies up to oyster hot water；It is purified by hot water, water can be improved
The solubility of soluble proteins, further separation and concentration go out the protein in Thick many candies；
3）Alkali carries：The sodium hydroxide solution of 8 times of volumes 3.0%, 65 DEG C of magnetic agitations 3 are added in residue after above-mentioned hot water is extracted
Hour, it is neutralized to neutrality with dilute hydrochloric acid, is centrifuged, concentration is dialysed 72 hours with 3.5kDa bag filters, is added after liquid concentration in bag taking
98% ethyl alcohol of 5 times of quality volumes, 4 DEG C of precipitates overnights, next day centrifugation collect precipitation, absolute ethyl alcohol and acetone are used to be dehydrated respectively,
It is dried under reduced pressure at 48 DEG C up to oyster alkali carries Thick many candies；By alkali carries, further extra Separation of Proteins can be gone out
It goes, is ready for further enzymolysis；
4）Enzymolysis：Oyster Thick many candies are merged, the solution that mass fraction is 10% is configured to pure water, adjusts pH to 8.4, be added
Trypsase end mass fraction is 0.5%, the insulation reaction 5 hours at a temperature of 55 DEG C；After completion of the reaction with after dilute hydrochloric acid tune neutrality
It in 98 DEG C of enzyme deactivations 18 minutes, is cooled to room temperature, adjusts pH to 2.4, it is 0.7% that pepsin to whole mass fraction, which is added, at 40 DEG C
At a temperature of insulation reaction 4 hours, neutralized after completion of the reaction through diluted sodium hydroxide solution, low temperature is concentrated into 15% up to enzymolysis liquid；First
It is digested by alkali protease and acid protease, remaining protein in oyster Thick many candies is hydrolyzed to little albumen and polypeptide,
And removed through subsequently de- albumen, the content and purity of the oyster polysaccharide in finished product can be greatly improved；Oyster polysaccharide can be to thin
Bacterium cell membrane has an impact, and changes cell membrane fluidity and permeability, destroys the supramolecular structure of cell film and causes thin
Intracellular ingredients from lossing, can also act on bacteria cell wall influences bacterial metabolism, to play fungistatic effect；
5）De- albumen：By chloroform, n-butanol, acetic acid according to volume ratio 48:15:1 ratio is uniformly mixed, and is also contained in mixed solution
There is a dimethyl tartrate of 1.5-1.8 ‰, the ratio of L-TARTARIC ACID dimethyl ester and D- dimethyl tartrates in dimethyl tartrate
It is 15.7:1；Under constant stirring, according to volume ratio 1:8 ratio pours into enzymolysis liquid in mixed solution, and magnetic force is used after sealing
Blender stirs 60 minutes, pours into separatory funnel, upper solution is collected after static layering, and centrifuges out a small amount of denaturation egg
White matter keeps the removal of protein in extracting solution complete by above-mentioned steps repetitive operation 5 times, merges supernatant and is through low temperature concentration
Obtain oyster polysaccharide；Chloroform, n-butanol mixed solution floating preteins and polypeptide can be changed into insoluble substance, through centrifugation go
It removes, can achieve the purpose that remove protein in oyster polysaccharide, the L-TARTARIC ACID dimethyl ester and D- winestones of acetic acid and proper proportion
Dimethyl phthalate can act on protein denaturation jelly, reduce the probability that protein denaturation jelly wraps oyster polysaccharide,
So as to substantially reduce the amount that oyster polysaccharide is separated, the yield and content of oyster polysaccharide are improved, egg in product is reduced
The content of white matter improves purity of polysaccharide.
Embodiment 3：
The preparation method of oyster polysaccharide, including：Water carries, hot water carries, alkali carries, enzymolysis, takes off albumen, specifically includes following steps：
Water carries：Oyster fresh meat is rubbed, homogenate, obtains skimmed milk with acetone degreasing and low temperature drying, extracting degreasing powder adds 24 times of quality
The normal-temperature distilled water of volume is stirred at room temperature 4 hours, centrifugation, and residue is repeated as stated above to extract, merges extracting solution, concentration；
96% ethyl alcohol of 4 times of quality, 2 DEG C of precipitates overnights is added, next day centrifugation collects precipitation, absolute ethyl alcohol and acetone is used to be dehydrated respectively,
It is dried under reduced pressure at 46 DEG C and carries Thick many candies up to oyster water；After being carried by degreasing and water, water soluble protein can be removed, and be
Further deproteination provides advantage；
Hot water carries：16 times of quality volume distilled water are added in residue after above-mentioned room temperature is extracted, magnetic agitation 2.5 at a temperature of 65 DEG C
Hour, it centrifuges, concentration, 96% ethyl alcohol of 4 times of quality volumes, 1-4 DEG C of precipitates overnight is added, next day centrifugation collects precipitation, respectively
It is dehydrated with absolute ethyl alcohol and acetone, is dried under reduced pressure at 46 DEG C and carries Thick many candies up to oyster hot water；It is purified, can be improved by hot water
The solubility of water soluble protein, further separation and concentration go out the protein in Thick many candies；
Alkali carries：The sodium hydroxide solution of 7 times of volumes 2.8%, 64 DEG C of magnetic agitations 2.5 are added in residue after above-mentioned hot water is extracted
Hour, it is neutralized to neutrality with dilute hydrochloric acid, is centrifuged, concentration is dialysed 60 hours with 3.5kDa bag filters, is added after liquid concentration in bag taking
96% ethyl alcohol of 4 times of quality volumes, 2 DEG C of precipitates overnights, next day centrifugation collect precipitation, absolute ethyl alcohol and acetone are used to be dehydrated respectively,
It is dried under reduced pressure at 46 DEG C up to oyster alkali carries Thick many candies；By alkali carries, further extra Separation of Proteins can be gone out
It goes, is ready for further enzymolysis；
Enzymolysis：Oyster Thick many candies are merged, the solution that mass fraction is 9% is configured to pure water, adjusts pH to 8.3, pancreas egg is added
White enzyme end mass fraction is 0.4%, the insulation reaction 4 hours at a temperature of 54 DEG C；After completion of the reaction with after dilute hydrochloric acid tune neutrality 96
DEG C enzyme deactivation 15 minutes, is cooled to room temperature, and adjusts pH to 2.3, and it is 0.6% that pepsin to whole mass fraction, which is added, in 38 DEG C of temperature
Lower insulation reaction 3.5 hours, neutralizes through diluted sodium hydroxide solution after completion of the reaction, and low temperature is concentrated into 12% up to enzymolysis liquid；Successively
It is digested through alkali protease and acid protease, remaining protein in oyster Thick many candies is hydrolyzed to little albumen and polypeptide, and
It is removed through subsequently de- albumen, the content and purity of the oyster polysaccharide in finished product can be greatly improved；Oyster polysaccharide can be to bacterium
Cell membrane has an impact, and changes cell membrane fluidity and permeability, destroys the supramolecular structure of cell film and causes cell
Interior ingredients from lossing, can also act on bacteria cell wall influences bacterial metabolism, to play fungistatic effect；
De- albumen：By chloroform, n-butanol, acetic acid according to volume ratio 45:14:1 ratio is uniformly mixed, and is also contained in mixed solution
1.6 ‰ dimethyl tartrate, the ratio of L-TARTARIC ACID dimethyl ester and D- dimethyl tartrates is 15.6 in dimethyl tartrate:
1；Under constant stirring, according to volume ratio 1:7 ratio pours into enzymolysis liquid in mixed solution, is stirred with magnetic stirring apparatus after sealing
It mixes 45 minutes, pours into separatory funnel, upper solution is collected after static layering, and centrifuge out a small amount of denatured protein, press
Above-mentioned steps repetitive operation 4 times, keeps the removal of protein in extracting solution complete, and merging supernatant concentrates more up to oyster through low temperature
Sugar；Chloroform, n-butanol mixed solution floating preteins and polypeptide can be changed into insoluble substance, removed through centrifugation, can be with
Achieve the purpose that remove protein in oyster polysaccharide, the L-TARTARIC ACID dimethyl ester and D- dimethyl tartrates of acetic acid and proper proportion
Protein denaturation jelly can be acted on, the probability that protein denaturation jelly wraps oyster polysaccharide is reduced, so as to
The amount that oyster polysaccharide is separated is substantially reduced, improves the yield and content of oyster polysaccharide, reduces containing for protein in product
Amount improves purity of polysaccharide.
Routine operation in operating procedure of the present invention is well known to those skilled in the art, herein without repeating.
Technical scheme of the present invention is described in detail in embodiment described above, it should be understood that the above is only
For specific embodiments of the present invention, it is not intended to restrict the invention, all any modifications made in the spirit of the present invention,
Supplement or similar fashion replacement etc., should all be included in the protection scope of the present invention.
Claims (9)
1. the preparation method of oyster polysaccharide, including：Water carries, hot water carries, alkali carries, enzymolysis, takes off albumen, it is characterised in that：It is described de-
Albumen step is：Chloroform, n-butanol, acetic acid are uniformly mixed, the also tartaric acid diformazan containing 1.5-1.8 ‰ in mixed solution
Ester；Under constant stirring, according to volume ratio 1:The ratio of 7-8 pours into enzymolysis liquid in mixed solution, and magnetic agitation is used after sealing
Device stirs 45-60 minutes, pours into separatory funnel, upper solution is collected after static layering, and centrifuges out a small amount of denaturation egg
White matter keeps the removal of protein in extracting solution complete by above-mentioned steps repetitive operation 4-5 times, merges supernatant and is concentrated through low temperature
Up to oyster polysaccharide.
2. the preparation method of oyster polysaccharide according to claim 1, it is characterised in that：In the de- albumen step, chloroform,
N-butanol, acetic acid ratio be 45-48:10-15:1.
3. the preparation method of oyster polysaccharide according to claim 1, it is characterised in that：In the de- albumen step, winestone
The ratio of L-TARTARIC ACID dimethyl ester and D- dimethyl tartrates is 15.6-15.7 in dimethyl phthalate:1.
4. the preparation method of oyster polysaccharide according to claim 1, it is characterised in that：The aqueous extraction step is：By oyster
Fresh meat rubs, homogenate, and skimmed milk is obtained with acetone degreasing and low temperature drying, and water, which carries, to be operated extracting solution and to concentrate；4-5 times of matter is added
The 95-98% ethyl alcohol of amount, 1-4 DEG C of precipitates overnight, next day centrifugation collect precipitation, use absolute ethyl alcohol and acetone to be dehydrated respectively, 45-48
It is dried under reduced pressure at DEG C and carries Thick many candies up to oyster water.
5. the preparation method of oyster polysaccharide according to claim 4, it is characterised in that：In the aqueous extraction step, water carries behaviour
As：Extracting degreasing powder adds the normal-temperature distilled water of 20-25 times of quality volume, is stirred at room temperature 3-5 hours, centrifugation, by residue by above-mentioned
Method repeats to extract, and merges extracting solution.
6. the preparation method of oyster polysaccharide according to claim 1, it is characterised in that：The hot aqueous extraction step is：It will be upper
It states the residue after room temperature extraction and 15-18 times of quality volume distilled water is added, magnetic agitation 2-3 hours at a temperature of 60-66 DEG C, from
The 95-98% ethyl alcohol of 4-5 times of quality volume, 1-4 DEG C of precipitates overnight is added in the heart, concentration, and next day centrifugation is collected precipitation, used respectively
Absolute ethyl alcohol and acetone are dehydrated, and are dried under reduced pressure at 45-48 DEG C and are carried Thick many candies up to oyster hot water.
7. the preparation method of oyster polysaccharide according to claim 1, it is characterised in that：The alkali carries step is：It will be above-mentioned
The sodium hydroxide solution of 6-8 times of volume 2.5-3.0%, 60-65 DEG C of magnetic agitation 2-3 hour, use is added in residue after hot water extraction
Dilute hydrochloric acid is neutralized to neutrality, centrifuges, and concentration is dialysed 48-72 hours with 3.5kDa bag filters, is collected after liquid concentration alcohol precipitation in bag taking
Precipitation is used absolute ethyl alcohol and acetone to be dehydrated, is dried under reduced pressure at 45-48 DEG C up to oyster alkali carries Thick many candies respectively.
8. the preparation method of oyster polysaccharide according to claim 1, it is characterised in that：The enzymolysis step is：By oyster
Thick many candies merge, and are configured to the solution that mass fraction is 8-10% with pure water, trypsin digestion are added 4-5 hours, reaction finishes
Afterwards with, in 95-98 DEG C of enzyme deactivation 12-18 minutes, being cooled to room temperature after dilute hydrochloric acid tune neutrality, pepsin is added and digests 3-4 hours,
It is neutralized after completion of the reaction through diluted sodium hydroxide solution, low temperature is concentrated into 10-15% up to enzymolysis liquid.
9. the preparation method of oyster polysaccharide according to claim 8, it is characterised in that：In the enzymolysis step, tryptose
Enzyme enzymolysis, which operates, is：PH to 8.2-8.4 is adjusted, it is 0.3-0.5%, hydrolysis temperature 52-55 that trypsase end mass fraction, which is added,
℃；Pepsin enzymolysis, which operates, is：PH to 2.3-2.4 is adjusted, it is 0.5-0.7%, enzymolysis that pepsin to whole mass fraction, which is added,
Temperature is 37-40 DEG C.
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| CN114853916A (en) * | 2022-04-22 | 2022-08-05 | 东北农业大学 | Method for extracting oyster polysaccharide and application thereof |
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| CN114853916A (en) * | 2022-04-22 | 2022-08-05 | 东北农业大学 | Method for extracting oyster polysaccharide and application thereof |
| CN114853916B (en) * | 2022-04-22 | 2023-03-03 | 东北农业大学 | Method for extracting oyster polysaccharide and application thereof |
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