CN1749281A - Process for extracting comb shell polysaccharide - Google Patents
Process for extracting comb shell polysaccharide Download PDFInfo
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- CN1749281A CN1749281A CN 200510047410 CN200510047410A CN1749281A CN 1749281 A CN1749281 A CN 1749281A CN 200510047410 CN200510047410 CN 200510047410 CN 200510047410 A CN200510047410 A CN 200510047410A CN 1749281 A CN1749281 A CN 1749281A
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- enzymolysis
- polysaccharide
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- slurries
- ultrasonication
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- 150000004676 glycans Chemical class 0.000 title claims abstract description 110
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 109
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 109
- 238000000034 method Methods 0.000 title claims description 58
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 63
- 238000001556 precipitation Methods 0.000 claims abstract description 45
- 230000007935 neutral effect Effects 0.000 claims abstract description 31
- 235000013372 meat Nutrition 0.000 claims abstract description 11
- 238000001035 drying Methods 0.000 claims abstract description 4
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 102
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 86
- 238000003756 stirring Methods 0.000 claims description 67
- 102000004190 Enzymes Human genes 0.000 claims description 62
- 108090000790 Enzymes Proteins 0.000 claims description 62
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 42
- 238000002525 ultrasonication Methods 0.000 claims description 38
- 239000000843 powder Substances 0.000 claims description 36
- 241000237509 Patinopecten sp. Species 0.000 claims description 33
- 235000020637 scallop Nutrition 0.000 claims description 33
- 239000002002 slurry Substances 0.000 claims description 31
- 238000006243 chemical reaction Methods 0.000 claims description 30
- 238000012546 transfer Methods 0.000 claims description 30
- 241000237516 Mizuhopecten yessoensis Species 0.000 claims description 27
- 238000005360 mashing Methods 0.000 claims description 22
- 241000238557 Decapoda Species 0.000 claims description 20
- 210000002784 stomach Anatomy 0.000 claims description 20
- 108090000145 Bacillolysin Proteins 0.000 claims description 19
- 239000000047 product Substances 0.000 claims description 12
- 239000002994 raw material Substances 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 11
- 238000005904 alkaline hydrolysis reaction Methods 0.000 claims description 10
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 8
- 239000012452 mother liquor Substances 0.000 claims description 8
- 238000004108 freeze drying Methods 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 230000002358 autolytic effect Effects 0.000 claims description 4
- QNEFNFIKZWUAEQ-UHFFFAOYSA-N carbonic acid;potassium Chemical compound [K].OC(O)=O QNEFNFIKZWUAEQ-UHFFFAOYSA-N 0.000 claims description 4
- 230000007062 hydrolysis Effects 0.000 claims description 4
- 238000006460 hydrolysis reaction Methods 0.000 claims description 4
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 4
- 235000017550 sodium carbonate Nutrition 0.000 claims description 4
- 238000012545 processing Methods 0.000 claims description 3
- 239000012141 concentrate Substances 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims 1
- 238000000605 extraction Methods 0.000 abstract description 6
- 239000000463 material Substances 0.000 abstract description 3
- 102000004142 Trypsin Human genes 0.000 abstract description 2
- 108090000631 Trypsin Proteins 0.000 abstract description 2
- 235000013376 functional food Nutrition 0.000 abstract description 2
- 239000012588 trypsin Substances 0.000 abstract description 2
- 102000057297 Pepsin A Human genes 0.000 abstract 1
- 108090000284 Pepsin A Proteins 0.000 abstract 1
- 108090000787 Subtilisin Proteins 0.000 abstract 1
- 229940111202 pepsin Drugs 0.000 abstract 1
- 238000009210 therapy by ultrasound Methods 0.000 abstract 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 18
- 238000005303 weighing Methods 0.000 description 11
- 239000000706 filtrate Substances 0.000 description 9
- 210000001835 viscera Anatomy 0.000 description 8
- 239000000284 extract Substances 0.000 description 6
- 238000011160 research Methods 0.000 description 5
- 230000004071 biological effect Effects 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 235000015170 shellfish Nutrition 0.000 description 3
- 241000233866 Fungi Species 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- -1 electuary Substances 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 210000004907 gland Anatomy 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000001603 reducing effect Effects 0.000 description 2
- 230000001568 sexual effect Effects 0.000 description 2
- 241001441955 Argopecten irradians Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 229920001491 Lentinan Polymers 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000237536 Mytilus edulis Species 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 229940115286 lentinan Drugs 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000020638 mussel Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000004223 radioprotective effect Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The extraction process of comb shell polysaccharide includes the technological steps of material treatment, homogenizing, ultrasonic treatment, enzymolysis, separating, concentration, alcohol precipitation and drying. In the material treatment, the comb shell meat tissue including leftover is taken and may be stoved or freeze dried; and in the enzymolysis, one or two of trypsin, pepsin and neutral subtilisin are used. The present invention provides the technological path of extracting comb shell polysaccharide and the comb shell polysaccharide product has high purity and high yield, and may be used in developing various functional foods.
Description
Technical field
What the present invention relates to is the extractive technique of polysaccharide, particularly is the technology that raw material extracts polysaccharide with the shellfish.
Background technology
In recent years, STUDY ON POLYSACHAROSE entered develop rapidly and the unprecedentedly active stage, people's such as Zhang Qian the research (progress of polysaccharide function " food research and development " 1998,9 (19)) prove that polysaccharide has many-sided biological activity, its energy immune cell activated improves body's immunological function, can form mixture with virus, disturb itself and the engaging of cell receptor, and normal cell is free from side effects.Physiological functions such as it is antitumor, raise immunity, hypoglycemic, reducing blood-fat, detoxifcation, antiviral, antibacterium, radioprotective are attracted attention by world the world of medicine.
Polysaccharide extensively is present in plant, microorganism (bacterium and fungi) and the marine alga, and is wherein more, particularly clearer to the research of lentinan to edible fungi polysaccharide research.Along with going deep into that polysaccharide is studied, people turn to the animal body polysaccharide to sight gradually, particularly hydrocoles body polysaccharide.At present about the shellfish STUDY ON POLYSACHAROSE only relevant for fresh water mussel meat (" Yunnan University's journal (natural science edition) 1998,20 (3): 187～189 ") and the bay scallop shirt rim report of (" Chinese aquatic science " the 1st the 2nd phase of volume).
Patinopecten yessoensis is a kind of low temperature marine products bivalve shellfish, and the former Japan of originating from introduces China in the initial stage eighties.Patinopecten yessoensis delicious flavour, nutritious is rich in the amino acid of calcium, phosphorus, zinc, selenium and other trace elements and needed by human.Fresh Patinopecten yessoensis protein content is 14.5%, in the dried scallop up to 63.7%.Contain tens seed amino acids in the protein, the L-glutamic acid content that is delicate flavour is the highest in the fishery products, accounts for 7.15%.Modern science is discovered, except that containing rich in protein, also contains abundant polysaccharide in the Patinopecten yessoensis, but is not much accounted of always and studies.Research and report about Patinopecten yessoensis cultural technique aspect are a lot, but the research of relevant its nutritive value aspect and report are very few.Extracting method and technology for comb shell polysaccharide are not appeared in the newspapers as yet.
Summary of the invention
It is raw material with the Patinopecten yessoensis that purpose of the present invention aims to provide a kind of, utilizes enzyme process in conjunction with related process, avoids pyroprocessing, can keep the original biological activity of polysaccharide, can improve the novel process of the extraction active polysaccharide of polysaccharide extract rate again to greatest extent.
Technical scheme of the present invention is Patinopecten yessoensis to be carried out raw material handle, and obtains its polysaccharide product through extraction and purifying.In addition, the residue in the leaching process can prepare the Patinopecten yessoensis polypeptide products according to technological invention in the past.
One, raw material and processing
The raw material that the present invention uses can be whole corporality tissues (comprising scallop post, the cheek, sexual gland, internal organ, shirt rim etc.) that Patinopecten yessoensis shells, and also can be scallop post, " tankage " (cheek, sexual gland, internal organ, shirt rim etc. all or part of) or scallop post and partly " tankage ".
1. the smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out meat tissue with cutter, directly stand-by.
Or 2. will take out meat tissue by different sites, to containing moisture less than 4%, be crushed to 50～200 orders in 20～60 ℃ of oven dry or vacuum lyophilization again, obtain dry powder, stand-by.
Two, homogenate
1. add 0.5～5 times water by whole meat tissues of Patinopecten yessoensis (also can be the scallop post or/and part " tankage ", or all " tankage ") quality, use tissue mashing machine's homogenate, obtain uniform slurries.
Or the dry powder quality of 2. whole meat tissues by Patinopecten yessoensis (also can be the scallop post or/and part " tankage ", or all " tankage ") adds 10～50 times water, uses tissue mashing machine's homogenate, obtains uniform slurries.
Three, ultrasonication
Slurries are through ultrasonication 0.3～1 hour, 30～60 ℃ of operative temperatures, and power is 50～600W, thereby improves the effect of next step enzymolysis.
Four, enzymolysis
Extract the method that comb shell polysaccharide can adopt enzymolysis, carry out enzymolysis under conditions such as suitable pH value of used enzyme and temperature, used enzyme can be a single enzyme, also can be prozyme; Thereby can also before enzymolysis, carry out alkaline hydrolysis or use the autolytic enzyme technology to carry out pre-treatment reaching the purpose that improves extraction yield.
1. alkaline hydrolysis: in the slurries after ultrasonication, add 0.1～1.0% solid carbonic acid potassium or the yellow soda ash that grinds, stirred alkaline hydrolysis 1～5 hour in 30～60 ℃.
Or 2. autolytic enzyme technology: will be through the slurries after the ultrasonication through uviolizing after 10～60 minutes, in 40～60 ℃ of hydrolysis 2～6 hours.
3. single enzyme enzymolysis
1. trypsin digestion: is 7～9 with the slurries after ultrasonication with 0.5mol/L potassium hydroxide adjust pH, and keeps pH value in this scope in enzymolysis process, and adding the slurries massfraction is 0.1～2.0% enzyme work 5 * 10
4The trypsinase of u/g maintains the temperature at 40～60 ℃, stirs enzymolysis 1～8 hour.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 6mol/L hydrochloric acid.
Or 2. stomach en-enzymolysis: is 1～5 with the slurries after ultrasonication with 6mol/L hydrochloric acid adjust pH, and keeps pH value in this scope in enzymolysis process, and adding the slurries massfraction is 0.1～2.0% enzyme work 7.8 * 10
5The stomach en-of u/g maintains the temperature at 40～60 ℃, stirs enzymolysis 1～10 hour.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.
Or 3. bacillus subtilis neutral proteinase enzymolysis: is 6～7 with the slurries after ultrasonication with 6mol/L hydrochloric acid adjust pH, and keeps pH value in this scope in enzymolysis process, and adding the slurries massfraction is 0.1～2.0% enzyme work 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g maintains the temperature at 45～55 ℃, stirs enzymolysis 1～6 hour.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.
4. two enzyme enzymolysis
1. bacillus subtilis neutral proteinase and stomach en-carry out enzymolysis: is 6～7 with the slurries after ultrasonication with 6mol/L hydrochloric acid adjust pH, and adding the slurries massfraction is that 0.05～1.5% enzyme lives 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g keeps the pH value in this scope, stirs enzymolysis 1～3 hour in 45～55 ℃.Be 1～5 with 6mol/L hydrochloric acid adjust pH then, adding the slurries massfraction is that 0.05～1.5% enzyme lives 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 1～5 hour in 40～60 ℃.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.
Or 2. bacillus subtilis neutral proteinase and trypsinase carry out enzymolysis: is 7～8 with the slurries after ultrasonication with 0.5mol/L potassium hydroxide adjust pH, and adding the slurries massfraction is that 0.05～1.5% enzyme lives 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g keeps the pH value in this scope, stirs enzymolysis 1～3 hour in 45～55 ℃.Be 7～9 with 0.5mol/L potassium hydroxide adjust pH then, adding the slurries massfraction is that 0.05～1.5% enzyme lives 5 * 10
4The trypsinase of u/g keeps the pH value in this scope, in 40～60 ℃, stirs enzymolysis 1～4 hour.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 6mol/L hydrochloric acid.
Or 3. stomach en-and trypsinase carry out enzymolysis: is 1～5 with the slurries after ultrasonication with 6mol/L hydrochloric acid adjust pH, and adding the slurries massfraction is that 0.05～1.5% enzyme lives 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 1～5 hour in 40～60 ℃.Be 7～9 with 0.5mol/L potassium hydroxide adjust pH then, adding the slurries massfraction is that 0.05～1.5% enzyme lives 5 * 10
4The trypsinase of u/g keeps the pH value in this scope, in 40～60 ℃, stirs enzymolysis 1～4 hour.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 6mol/L hydrochloric acid.
Five, separate
1. centrifugal: with the Patinopecten yessoensis enzymolysis solution in 6000 rev/mins centrifugal 20 minutes, collect supernatant liquor, be the polysaccharide mother liquor.
Or 2. filter: the Patinopecten yessoensis enzymolysis solution is filtered through 200 eye mesh screens, obtain the polysaccharide mother liquor.
Six, concentrate
The polysaccharide mother liquor that obtains is placed vacuum concentration equipment, control vacuum tightness 85～98KPa, temperature is concentrated into 1/5～1/3 of original volume for 30～60 ℃.
Seven, alcohol precipitation
In the polysaccharide mother liquor, slowly add 2～5 times of 95% ethanol while stirring, in 2～6 ℃ of alcohol precipitations after 2～18 hours, centrifugal collection polysaccharide precipitation.
Eight, drying
With polysaccharide precipitation directly in 20～60 ℃ of oven dry, obtain product and be comb shell polysaccharide.
Or 2. place vacuum freeze, in vacuum tightness 70～150Pa, and 30～60 ℃ of plate temperature, lyophilize 6～15 hours obtains comb shell polysaccharide, can be used as a kind of product, also can further be processed into product forms such as capsule, electuary, tablet.
Advantage of the present invention is:
1. extraction process of the present invention is rationally effective, has started the operational path of (except the shell) extraction polysaccharide in the Patinopecten yessoensis body, to the utmost comb shell polysaccharide is extracted;
2. related process temperature of the present invention all is controlled at below 60 ℃, and the method that especially adopts low temperature press down to live substitutes in the past the high temperature enzyme that goes out, thereby guarantees to extract the biological activity of polysaccharide, makes the finished product holoside nutrient and functional having concurrently;
3. operational path of the present invention can not only be that raw material extracts polysaccharide with the higher scallop post of Patinopecten yessoensis price, and can be with the whole tissues of Patinopecten yessoensis except that shell, tankage are that raw material extracts polysaccharide even, and technical guarantee is provided for reducing cost, increasing economic efficiency;
4. the present invention has established the method that alkaline hydrolysis combines with enzymolysis and makes the raw material can decomposite polysaccharide more;
5. the present invention has established the method that the autolytic enzyme technology combines with enzymolysis and has not only made full use of Patinopecten yessoensis raw material self enzyme system, also will improve polysaccharide yield;
6. the residue in the leaching process of the present invention can prepare the Patinopecten yessoensis polypeptide products according to technological invention in the past;
7. the comb shell polysaccharide that extracts of the present invention, both can further be processed into product forms such as capsule, electuary, tablet separately as a kind of product, can also be as base-material, be aided with the multiple functional food of other food development, make the functional diversification day by day of protective foods.
Embodiment:
Example 1
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues with cutter.Take by weighing Patinopecten yessoensis and all organize 1 kilogram, add 2 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.3 hour, 60 ℃ of temperature, power is 600W.Be 6 with 6mol/L hydrochloric acid adjust pH then, add 1.5 gram (0.05%) enzymes and live 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g keeps the pH value in this scope, stirs enzymolysis 3 hours in 45 ℃.Be 1 with 6mol/L hydrochloric acid adjust pH then, add 45 gram (1.5%) enzymes and live 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 5 hours in 40 ℃.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.Centrifugal 20 minutes in 6000 rev/mins, collect supernatant liquor, place vacuum concentration equipment, control vacuum tightness 85KPa, temperature is concentrated into 1 liter for 30 ℃, slowly add 2 liter of 95% ethanol while stirring, in 2 ℃ of alcohol precipitations after 2 hours, centrifugal collection polysaccharide precipitation, directly in 20 ℃ of oven dry, obtain polysaccharide dry powder 24.2 grams, measure through the phenolsulfuric acid method, polysaccharide content is 17.1%.
Example 2
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues with cutter.Take by weighing 1 kilogram on scallop post, add 5 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.5 hour, 55 ℃ of temperature, power is 300W.Be 7 with 0.5mol/L potassium hydroxide adjust pH then, and in enzymolysis process, keep the pH value, add 120 gram (2.0%) enzymes and live 5 * 10 in this scope
4The trypsinase of u/g maintains the temperature at 40 ℃, stirs enzymolysis 1 hour.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 6mol/L hydrochloric acid.Filter through 200 eye mesh screens, obtain filtrate and place vacuum concentration equipment, control vacuum tightness 92KPa, temperature is concentrated into 1.5 liters for 55 ℃, slowly adds 5.25 liter of 95% ethanol while stirring, in 3 ℃ of alcohol precipitations after 8 hours, centrifugal collection polysaccharide precipitation, place vacuum freeze, in vacuum tightness 150Pa, 30 ℃ of plate temperature, lyophilize 15 hours, obtain polysaccharide dry powder 33.5 grams, measure through the phenolsulfuric acid method, polysaccharide content is 18.4%.
Example 3
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues with cutter.Take by weighing 1 kilogram of tankage, add the water of 500 grams, after tissue mashing machine's homogenate, use ultrasonication 0.5 hour, 50 ℃ of temperature, power is 200W.Be 7 with 0.5mol/L potassium hydroxide adjust pH then, add 22.5 gram (1.5%) enzymes and live 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g keeps the pH value in this scope, stirs enzymolysis 1 hour in 45 ℃.Be 9 with 0.5mol/L potassium hydroxide adjust pH then, add 0.75 gram (0.05%) enzyme and live 5 * 10
4The trypsinase of u/g keeps the pH value in this scope, in 60 ℃, stirs enzymolysis 4 hours.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 6mol/L hydrochloric acid.In 6000 rev/mins centrifugal 20 minutes, collect supernatant liquor, place vacuum concentration equipment, control vacuum tightness 90KPa, temperature is concentrated into 0.3 liter for 50 ℃.Slowly add 0.9 liter of 95% ethanol while stirring, in 4 ℃ of alcohol precipitations after 10 hours, centrifugal collection polysaccharide precipitation, place vacuum freeze, in vacuum tightness 90Pa, 50 ℃ of plate temperature, lyophilize 10 hours, obtain polysaccharide dry powder 18.8 grams, measure through the phenolsulfuric acid method, polysaccharide content is 10.9%.
Example 4
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues with cutter.Take by weighing 1 kilogram in internal organ, add 4 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.5 hour, 50 ℃ of temperature, power is 200W.Be 1 with 6mol/L hydrochloric acid adjust pH then, add 2.5 gram (0.05%) enzymes and live 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 5 hours in 60 ℃.Be 9 with 0.5mol/L potassium hydroxide adjust pH then, add 75 gram (1.5%) enzymes and live 5 * 10
4The trypsinase of u/g keeps the pH value in this scope, in 60 ℃, stirs enzymolysis 1 hour.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 6mol/L hydrochloric acid.Filter through 200 eye mesh screens, obtain filtrate and place vacuum concentration equipment, control vacuum tightness 90KPa, temperature is concentrated into 1 liter for 50 ℃, slowly adds 4 liter of 95% ethanol while stirring, in 3 ℃ of alcohol precipitations after 10 hours, centrifugal collection polysaccharide precipitation directly in 45 ℃ of oven dry, obtains polysaccharide dry powder 12.6 grams, measure through the phenolsulfuric acid method, polysaccharide content is 11.7%.
Example 5
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues with cutter.Take by weighing 1 kilogram of shirt rim, add 3 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.5 hour, 40 ℃ of temperature, power is 100W.Be 1 with 6mol/L hydrochloric acid adjust pH then, and in enzymolysis process, keep the pH value, add 4 gram (0.1%) enzymes and live 7.8 * 10 in this scope
5The stomach en-of u/g maintains the temperature at 40 ℃, stirs enzymolysis 10 hours.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.Centrifugal 20 minutes in 6000 rev/mins, collect supernatant liquor, place vacuum concentration equipment, control vacuum tightness 90KPa, temperature is concentrated into 0.8 liter for 40 ℃, slowly add 3.2 liter of 95% ethanol while stirring, in 4 ℃ of alcohol precipitations after 14 hours, centrifugal collection polysaccharide precipitation, directly in 30 ℃ of oven dry, obtain polysaccharide dry powder 23.7 grams, measure through the phenolsulfuric acid method, polysaccharide content is 6.8%.
Example 6
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues with cutter.Take by weighing 1 kilogram in a organized way of institute after gilling, adds 4 kilograms water, after tissue mashing machine's homogenate, usefulness ultrasonication 0.5 hour, 40 ℃ of temperature, power is 200W.Be 6 with 6mol/L hydrochloric acid adjust pH then, and in enzymolysis process, keep the pH value, add 4 gram (0.1%) enzymes and live 5.0 * 10 in this scope
4The bacillus subtilis neutral proteinase of u/g maintains the temperature at 45 ℃, stirs enzymolysis 6 hours.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.Filter through 200 eye mesh screens, obtain filtrate and place vacuum concentration equipment, control vacuum tightness 90KPa, temperature is concentrated into 1 liter for 50 ℃, slowly adds 3 liter of 95% ethanol while stirring, in 5 ℃ of alcohol precipitations after 15 hours, centrifugal collection polysaccharide precipitation, place vacuum freeze, in vacuum tightness 90Pa, 50 ℃ of plate temperature, lyophilize 10 hours, obtain polysaccharide dry powder 26.4 grams, measure through the phenolsulfuric acid method, polysaccharide content is 10.2%.
Example 7
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues, to containing moisture, be crushed to 200 orders again, obtain dry powder less than 4% in 60 ℃ of oven dry with cutter.Get Patinopecten yessoensis and all organize dry powder 100 grams, add 5 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.6 hour, 35 ℃ of temperature, power is 80W.Be 9 with 0.5mol/L potassium hydroxide adjust pH then, and in enzymolysis process, keep the pH value, add 5.1 gram (0.1%) enzymes and live 5 * 10 in this scope
4The trypsinase of u/g maintains the temperature at 60 ℃, stirs enzymolysis 8 hours.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 6mol/L hydrochloric acid.Filter through 200 eye mesh screens, obtain filtrate and place vacuum concentration equipment, control vacuum tightness 92KPa, temperature is concentrated into 1.5 liters for 45 ℃, slowly adds 3.75 liter of 95% ethanol while stirring, in 4 ℃ of alcohol precipitations after 14 hours, centrifugal collection polysaccharide precipitation, place vacuum freeze, in vacuum tightness 70Pa, 60 ℃ of plate temperature, lyophilize 6 hours, obtain polysaccharide dry powder 12.1 grams, measure through the phenolsulfuric acid method, polysaccharide content is 17.6%.
Example 8
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues, to containing moisture, be crushed to 50 orders again, obtain dry powder less than 4% in 20 ℃ of oven dry with cutter.Get scallop post dry powder 100 grams, add 1 kilogram water, after tissue mashing machine's homogenate, use ultrasonication 1 hour, 30 ℃ of temperature, power is 50W.Be 7 with 6mol/L hydrochloric acid adjust pH then, add 16.5 gram (1.5%) enzymes and live 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g keeps the pH value in this scope, stirs enzymolysis 1 hour in 55 ℃.Be 5 with 6mol/L hydrochloric acid adjust pH then, add 0.55 gram (0.05%) enzyme and live 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 1 hour in 60 ℃.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.Centrifugal 20 minutes in 6000 rev/mins, collect supernatant liquor, place vacuum concentration equipment, control vacuum tightness 98KPa, temperature is concentrated into 0.3 liter for 60 ℃, slowly add 1.5 liter of 95% ethanol while stirring, in 6 ℃ of alcohol precipitations after 18 hours, centrifugal collection polysaccharide precipitation, directly in 60 ℃ of oven dry, obtain polysaccharide dry powder 13.4 grams, measure through the phenolsulfuric acid method, polysaccharide content is 18.1%.
Example 9
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, with cutter it is cut open, shell, press different sites and take out all tissues, vacuum lyophilization is crushed to 150 orders again to containing moisture less than 4%, obtains dry powder.Get tankage dry powder 100 grams, add 4 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.5 hour, 40 ℃ of temperature, power is 100W.Be 5 with 6mol/L hydrochloric acid adjust pH then, and in enzymolysis process, keep the pH value, add 82 gram (2.0%) enzymes and live 7.8 * 10 in this scope
5The stomach en-of u/g maintains the temperature at 60 ℃, stirs enzymolysis 1 hour.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.Centrifugal 20 minutes in 6000 rev/mins, collect supernatant liquor, place vacuum concentration equipment, control vacuum tightness 90KPa, temperature is concentrated into 1 liter for 40 ℃, slowly add 4 liter of 95% ethanol while stirring, in 4 ℃ of alcohol precipitations after 14 hours, centrifugal collection polysaccharide precipitation, directly in 30 ℃ of oven dry, obtain polysaccharide dry powder 9.8 grams, measure through the phenolsulfuric acid method, polysaccharide content is 11.3%.
Example 10
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues, to containing moisture, be crushed to 100 orders again, obtain dry powder less than 4% in 30 ℃ of oven dry with cutter.Get internal organ dry powder 100 grams, add 2 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.5 hour, 40 ℃ of temperature, power is 200W.Be 7 with 6mol/L hydrochloric acid adjust pH then, and in enzymolysis process, keep the pH value, add 42 gram (2.0%) enzymes and live 5.0 * 10 in this scope
4The bacillus subtilis neutral proteinase of u/g maintains the temperature at 55 ℃, stirs enzymolysis 1 hour.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.Filter through 200 eye mesh screens, obtain filtrate and place vacuum concentration equipment, control vacuum tightness 90KPa, temperature is concentrated into 0.5 liter for 50 ℃, slowly adds 1.5 liter of 95% ethanol while stirring, in 5 ℃ of alcohol precipitations after 15 hours, centrifugal collection polysaccharide precipitation, place vacuum freeze, in vacuum tightness 90Pa, 50 ℃ of plate temperature, lyophilize 10 hours, obtain polysaccharide dry powder 7.4 grams, measure through the phenolsulfuric acid method, polysaccharide content is 12.8%.
Example 11
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, with cutter it is cut open, shell, press different sites and take out all tissues, vacuum lyophilization is crushed to 120 orders again to containing moisture less than 4%, obtains dry powder.Get shirt rim dry powder 100 grams, add 3 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.6 hour, 50 ℃ of temperature, power is 200W.Be 8 with 0.5mol/L potassium hydroxide adjust pH then, add 1.55 gram (0.05%) enzymes and live 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g keeps the pH value in this scope, stirs enzymolysis 3 hours in 55 ℃.Be 7 with 0.5mol/L potassium hydroxide adjust pH then, add 46.5 gram (1.5%) enzymes and live 5 * 10
4The trypsinase of u/g keeps the pH value in this scope, in 40 ℃, stirs enzymolysis 1 hour.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 6mol/L hydrochloric acid.In 6000 rev/mins centrifugal 20 minutes, collect supernatant liquor, place vacuum concentration equipment, control vacuum tightness 90KPa, temperature is concentrated into 0.8 liter for 50 ℃, slowly adds 2.4 liter of 95% ethanol while stirring, after 10 hours, centrifugal collection polysaccharide precipitation places vacuum freeze in 4 ℃ of alcohol precipitations, in vacuum tightness 90Pa, 50 ℃ of plate temperature, lyophilize 10 hours obtains polysaccharide dry powder 10.8 grams, measure through the phenolsulfuric acid method, polysaccharide content is 6.4%.
Example 12
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues, to containing moisture, be crushed to 150 orders again, obtain dry powder less than 4% in 40 ℃ of oven dry with cutter.Get institute's dry powder 100 gram in a organized way except that internal organ, adds 2 kilograms water, after tissue mashing machine's homogenate, with ultrasonication 0.5 hour, 50 ℃ of temperature, power is 200W.Be 5 with 6mol/L hydrochloric acid adjust pH then, add 31.5 gram (1.5%) enzymes and live 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 1 hour in 40 ℃.Be 7 with 0.5mol/L potassium hydroxide adjust pH then, add 1.05 gram (0.05%) enzymes and live 5 * 10
4The trypsinase of u/g keeps the pH value in this scope, in 40 ℃, stirs enzymolysis 4 hours.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 6mol/L hydrochloric acid.Filter through 200 eye mesh screens, obtain filtrate and place vacuum concentration equipment, control vacuum tightness 90KPa, temperature is concentrated into 0.5 liter for 50 ℃, slowly adds 2 liter of 95% ethanol while stirring, in 3 ℃ of alcohol precipitations after 10 hours, centrifugal collection polysaccharide precipitation directly in 45 ℃ of oven dry, obtains polysaccharide dry powder 12.8 grams, measure through the phenolsulfuric acid method, polysaccharide content is 10.7%.
Example 13
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues with cutter.Take by weighing 1 kilogram of tankage, add 4 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.5 hour, 30 ℃ of temperature, power is 100W.Add the solid carbonic acid potassium that 50 grams (1.0%) grind then, stirred alkaline hydrolysis 1 hour in 30 ℃.With 6mol/L hydrochloric acid adjust pH is 6.5, adds 50 gram (1.0%) enzymes and lives 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g keeps the pH value in this scope, stirs enzymolysis 2 hours in 50 ℃.Be 4 with 6mol/L hydrochloric acid adjust pH then, add 50 gram (1.0%) enzymes and live 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 3 hours in 50 ℃.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.Centrifugal 20 minutes in 6000 rev/mins, collect supernatant liquor, place vacuum concentration equipment, control vacuum tightness 90KPa, temperature is concentrated into 1.25 liters for 50 ℃, slowly add 3.75 liter of 95% ethanol while stirring, in 3 ℃ of alcohol precipitations after 10 hours, centrifugal collection polysaccharide precipitation, directly in 50 ℃ of oven dry, obtain polysaccharide dry powder 19.7 grams, measure through the phenolsulfuric acid method, polysaccharide content is 10.4%.
Example 14
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, with cutter it is cut open, shell, press different sites and take out all tissues, vacuum lyophilization is crushed to 180 orders again to containing moisture less than 4%, obtains dry powder.Get internal organ dry powder 100 grams, add 3 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.8 hour, 40 ℃ of temperature, power is 200W.Add the solid carbonic acid potassium that 3.1 grams (0.1%) grind then, stirred alkaline hydrolysis 5 hours in 60 ℃.With 6mol/L hydrochloric acid adjust pH is 6.5, and keeps the pH value in this scope in enzymolysis process, adds 46.5 gram (1.5%) enzymes and lives 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g maintains the temperature at 50 ℃, stirs enzymolysis 5 hours.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.Filter through 200 eye mesh screens, obtain filtrate and place vacuum concentration equipment, control vacuum tightness 95KPa, temperature is concentrated into 1 liter for 40 ℃, slowly adds 4 liter of 95% ethanol while stirring, in 5 ℃ of alcohol precipitations after 15 hours, centrifugal collection polysaccharide precipitation, place vacuum freeze, in vacuum tightness 85Pa, 45 ℃ of plate temperature, lyophilize 8 hours, obtain polysaccharide dry powder 7.9 grams, measure through the phenolsulfuric acid method, polysaccharide content is 12.3%.
Example 15
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues with cutter.Take by weighing 1 kilogram in internal organ, add 3 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.6 hour, 45 ℃ of temperature, power is 400W.Add the solid sodium carbonate that 4 grams (0.1%) grind then, stirred alkaline hydrolysis 5 hours in 30 ℃.With 6mol/L hydrochloric acid adjust pH is 4, adds 48 gram (1.2%) enzymes and lives 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 2 hours in 45 ℃.Be 8 with 0.5mol/L potassium hydroxide adjust pH then, add 20 gram (0.5%) enzymes and live 5 * 10
4The trypsinase of u/g keeps the pH value in this scope, in 50 ℃, stirs enzymolysis 2 hours.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 6mol/L hydrochloric acid.Filter through 200 eye mesh screens, obtain filtrate and place vacuum concentration equipment, control vacuum tightness 95KPa, temperature is concentrated into 1 liter for 45 ℃, slowly adds 3 liter of 95% ethanol while stirring, in 5 ℃ of alcohol precipitations after 16 hours, centrifugal collection polysaccharide precipitation directly in 40 ℃ of oven dry, obtains polysaccharide dry powder 26.3 grams, measure through the phenolsulfuric acid method, polysaccharide content is 11.0%.
Example 16
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues with cutter.Take by weighing Patinopecten yessoensis and all organize 1 kilogram, add 4 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.8 hour, 40 ℃ of temperature, power is 500W.Add the solid sodium carbonate that 50 grams (1.0%) grind then, stirred alkaline hydrolysis 1 hour in 60 ℃.With 6mol/L hydrochloric acid adjust pH is 6.7, adds 10 gram (0.2%) enzymes and lives 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g keeps the pH value in this scope, stirs enzymolysis 2 hours in 50 ℃.Be 4 with 6mol/L hydrochloric acid adjust pH then, add 15 gram (0.3%) enzymes and live 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 3 hours in 50 ℃.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.Centrifugal 20 minutes in 6000 rev/mins, collect supernatant liquor, place vacuum concentration equipment, control vacuum tightness 90KPa, temperature is concentrated into 1 liter for 40 ℃, slowly add 3 liter of 95% ethanol while stirring, in 4 ℃ of alcohol precipitations after 12 hours, centrifugal collection polysaccharide precipitation, directly in 30 ℃ of oven dry, obtain polysaccharide dry powder 24.1 grams, measure through the phenolsulfuric acid method, polysaccharide content is 15.7%.
Example 17
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues with cutter.Take by weighing Patinopecten yessoensis and all organize 1 kilogram, add 2 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.3 hour, 60 ℃ of temperature, power is 600W.Through uviolizing after 60 minutes, in 60 ℃ of hydrolysis 2 hours.Be 6 with 6mol/L hydrochloric acid adjust pH then, add 1.5 gram (0.05%) enzymes and live 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g keeps the pH value in this scope, stirs enzymolysis 3 hours in 45 ℃.Be 1 with 6mol/L hydrochloric acid adjust pH then, add 30 gram (1.0%) enzymes and live 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 4 hours in 40 ℃.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 0.5mol/L potassium hydroxide.Centrifugal 20 minutes in 6000 rev/mins, collect supernatant liquor, place vacuum concentration equipment, control vacuum tightness 85KPa, temperature is concentrated into 1 liter for 30 ℃, slowly add 2 liter of 95% ethanol while stirring, in 2 ℃ of alcohol precipitations after 2 hours, centrifugal collection polysaccharide precipitation, directly in 20 ℃ of oven dry, obtain polysaccharide dry powder 25.6 grams, measure through the phenolsulfuric acid method, polysaccharide content is 17.4%.
Example 18
The smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, press different sites and take out all tissues with cutter.Take by weighing 1 kilogram in internal organ, add 3 kilograms water, after tissue mashing machine's homogenate, use ultrasonication 0.6 hour, 45 ℃ of temperature, power is 400W.Through uviolizing after 10 minutes, in 40 ℃ of hydrolysis 6 hours.With 6mol/L hydrochloric acid adjust pH is 4, adds 48 gram (1.2%) enzymes and lives 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 2 hours in 45 ℃.Be 8 with 0.5mol/L potassium hydroxide adjust pH then, add 20 gram (0.5%) enzymes and live 5 * 10
4The trypsinase of u/g keeps the pH value in this scope, in 50 ℃, stirs enzymolysis 2 hours.Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction.Transfer pH to neutral with 6mol/L hydrochloric acid.Filter through 200 eye mesh screens, obtain filtrate and place vacuum concentration equipment, control vacuum tightness 95KPa, temperature is concentrated into 1 liter for 45 ℃, slowly adds 3 liter of 95% ethanol while stirring, in 5 ℃ of alcohol precipitations after 16 hours, centrifugal collection polysaccharide precipitation directly in 40 ℃ of oven dry, obtains polysaccharide dry powder 26.3 grams, measure through the phenolsulfuric acid method, polysaccharide content is 11.0%.
Claims (12)
1, a kind of comb shell polysaccharide extracting method is characterized in that:
(1) raw material and processing: the smooth scallop water of fresh shelled shrimp is rinsed well, after draining, it is cut open, shell, take out whole meat tissues by different sites with cutter,
(2) homogenate: press the water that Patinopecten yessoensis meat tissue quality adds 0.5～5 times, use tissue mashing machine's homogenate, obtain uniform slurries,
(3) ultrasonication: slurries are through ultrasonication 0.3～1 hour, 30～60 ℃ of operative temperatures, and power is 50～600W,
(4) enzymolysis: is 7～9 with the slurries after ultrasonication with 0.5mol/L potassium hydroxide adjust pH, and keeps the pH value in this scope in enzymolysis process, and adding massfraction is that 0.1～2.0% enzyme lives 5 * 10
4The trypsinase of u/g maintains the temperature at 40～60 ℃, stirs enzymolysis 1～8 hour, afterwards, is cooled to rapidly below 30 ℃, keeps 30 minutes with termination reaction, transfers pH to neutral with 6mol/L hydrochloric acid,
(5) separate: with the Patinopecten yessoensis enzymolysis solution in 6000 rev/mins centrifugal 20 minutes, collect supernatant liquor, be the polysaccharide mother liquor,
(6) concentrate: the polysaccharide mother liquor that obtains is placed vacuum concentration equipment, control vacuum tightness 85～98KPa, temperature is concentrated into 1/5～1/3 of original volume for 30～60 ℃,
(7) alcohol precipitation: in the polysaccharide mother liquor, slowly add 2～5 times of 95% ethanol while stirring, in 2～6 ℃ of alcohol precipitations after 2～18 hours, centrifugal collection polysaccharide precipitation,
(8) drying: polysaccharide precipitation directly in 20～60 ℃ of oven dry, obtains product and is comb shell polysaccharide.
2, comb shell polysaccharide extracting method according to claim 1 is characterized in that the Patinopecten yessoensis meat tissue in the described raw material processing is scallop post, the tankage except that the scallop post, part tankage, scallop post and part tankage or the whole meat tissues that comprise scallop post and tankage except that shell.
3, comb shell polysaccharide extracting method according to claim 1 and 2, it is characterized in that it is whole tissues that Patinopecten yessoensis is taken out by different sites that described raw material is handled, in 20～60 ℃ the oven dry or vacuum lyophilization to containing moisture less than 4%, be crushed to 50～200 orders again, obtain dry powder; Described homogenate is by the water of 10～50 times of Patinopecten yessoensis meat tissue dry powder quality addings, uses tissue mashing machine's homogenate, obtains uniform slurries.
4, comb shell polysaccharide extracting method according to claim 1, it is characterized in that described enzymolysis is is 1～5 with the slurries after ultrasonication with 6mol/L hydrochloric acid adjust pH, and in enzymolysis process, keep the pH value in this scope, adding massfraction is that 0.1～2.0% enzyme lives 7.8 * 10
5The stomach en-of u/g maintains the temperature at 40～60 ℃, stirs enzymolysis 1～10 hour; Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction; Transfer pH to neutral with 0.5mol/L potassium hydroxide.
5, comb shell polysaccharide extracting method according to claim 1, it is characterized in that described enzymolysis is is 6～7 with the slurries after ultrasonication with 6mol/L hydrochloric acid adjust pH, and in enzymolysis process, keep the pH value in this scope, adding massfraction is that 0.1～2.0% enzyme lives 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g maintains the temperature at 45～55 ℃, stirs enzymolysis 1～6 hour; Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction; Transfer pH to neutral with 0.5mol/L potassium hydroxide.
6, process for extracting comb shell polysaccharide according to claim 1, it is characterized in that described enzymolysis is two enzyme enzymolysis: is 6～7 with the slurries after ultrasonication with 6mol/L hydrochloric acid adjust pH, adding massfraction is that 0.05～1.5% enzyme lives 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g keeps the pH value in this scope, stirs enzymolysis 1～3 hour in 45～55 ℃; Be 1～5 with 6mol/L hydrochloric acid adjust pH then, adding massfraction is that 0.05～1.5% enzyme lives 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 1～5 hour in 40～60 ℃; Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction; Transfer pH to neutral with 0.5mol/L potassium hydroxide.
7, process for extracting comb shell polysaccharide according to claim 1, it is characterized in that described enzymolysis is two enzyme enzymolysis: is 7～8 with the slurries after ultrasonication with 0.5mol/L potassium hydroxide adjust pH, and adding massfraction is that 0.05～1.5% enzyme lives 5.0 * 10
4The bacillus subtilis neutral proteinase of u/g keeps the pH value in this scope, stirs enzymolysis 1～3 hour in 45～55 ℃; Be 7～9 with 0.5mol/L potassium hydroxide adjust pH then, adding massfraction is that 0.05～1.5% enzyme lives 5 * 10
4The trypsinase of u/g keeps the pH value in this scope, in 40～60 ℃, stirs enzymolysis 1～4 hour; Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction; Transfer pH to neutral with 6mol/L hydrochloric acid.
8, process for extracting comb shell polysaccharide according to claim 1, it is characterized in that described enzymolysis is two enzyme enzymolysis: is 1～5 with the slurries after ultrasonication with 6mol/L hydrochloric acid adjust pH, adding massfraction is that 0.05～1.5% enzyme lives 7.8 * 10
5The stomach en-of u/g keeps the pH value in this scope, stirs enzymolysis 1～5 hour in 40～60 ℃; Be 7～9 with 0.5mol/L potassium hydroxide adjust pH then, adding massfraction is that 0.05～1.5% enzyme lives 5 * 10
4The trypsinase of u/g keeps the pH value in this scope, in 40～60 ℃, stirs enzymolysis 1～4 hour; Afterwards, be cooled to rapidly below 30 ℃, keep 30 minutes with termination reaction; Transfer pH to neutral with 6mol/L hydrochloric acid.
9, according to the described process for extracting comb shell polysaccharide of each claim in the claim 1,4～8, it is characterized in that described enzymolysis, be to carry out alkaline hydrolysis before the enzymolysis earlier: in the slurries after ultrasonication, add 0.1～1.0% solid carbonic acid potassium or the yellow soda ash that grinds, stirred alkaline hydrolysis 1～5 hour in 30～60 ℃.
10, according to the described process for extracting comb shell polysaccharide of each claim claim in the claim 1,4～8, it is characterized in that described enzymolysis, be to utilize the autolytic enzyme technology to carry out self-dissolving before the enzymolysis earlier: will be through the slurries after the ultrasonication through uviolizing after 10～60 minutes, in 40～60 ℃ of hydrolysis 2～6 hours.
11, process for extracting comb shell polysaccharide according to claim 1 is characterized in that described separation is to adopt 200 eye mesh screens to filter, and obtains the polysaccharide mother liquor.
12, process for extracting comb shell polysaccharide according to claim 1 is characterized in that described drying is to adopt vacuum lyophilization: control vacuum tightness 70～150Pa, 30～60 ℃ of plate temperature, lyophilize 6～15 hours.
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| CN109053922A (en) * | 2018-07-20 | 2018-12-21 | 大连工业大学 | Scallop Viscus Polysaccharide, extracting method and its application and pharmaceutical composition |
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