CN105695537A - Preparation method of glycosylation enteromorpha oligosaccharide for reducing glucose level and regulating intestinal flora - Google Patents
Preparation method of glycosylation enteromorpha oligosaccharide for reducing glucose level and regulating intestinal flora Download PDFInfo
- Publication number
- CN105695537A CN105695537A CN201610110627.1A CN201610110627A CN105695537A CN 105695537 A CN105695537 A CN 105695537A CN 201610110627 A CN201610110627 A CN 201610110627A CN 105695537 A CN105695537 A CN 105695537A
- Authority
- CN
- China
- Prior art keywords
- oligosaccharide
- glycosylation
- enteromorpha
- volume
- entermorpha
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/18—Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Saccharide Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a preparation method of glycosylation enteromorpha oligosaccharide for reducing glucose level and regulating intestinal flora. The method includes the steps of smashing enteromorpha in a superfine mode after drying, conducting backflow extracting through ethyl alcohol, conducting concentrating, drying and smashing to obtain defatted enteromorpha powder, removing protein, polysaccharide and pigment, adding fucosyltransferase into a reaction system containing enteromorpha oligosaccharide, and conducting fucus glycosylation reaction to prepare fucus glycosylation enteromorpha oligosaccharide. It is proved that glycosylation enteromorpha oligosaccharide has strong restraining activity for alpha-glucosaccharase, remarkably increases the number of intestinal probiotics, and can be used for preparing medicines or health care products for preventing and treating diabetes and improving metabolic diseases of intestinal flora and the like.
Description
Technical field
The invention belongs to biological pharmacy technical field, relate to the glycosylation enteromorpha oligosaccharide preparation method of a kind of blood sugar lowering and regulating intestinal canal flora。
Background technology
Along with the raising of people's lives, the expanding day of hyperglycemia population, the hyperglycemia diabetes caused, blood circulation disease is increasing to human health risk。Data in recent years shows, intestinal microbial population is closely related with type 2 diabetes mellitus。Bacillus bifidus is the superiority bacteria spp in human body intestinal canal flora, is one of the important symbol of human health。The minimizing of bifidobacteria may result in whole alteration of intestinal flora and intestinal microecology is unbalance, thus causing the generation of numerous metabolic disease。The insulin secretion function that bacillus bifidus can be effectively improved carbohydrate tolerance and glucose brings out, reduces the diabetic mice endotoxemia of high fat diet induction and the proinflammatory inflammation factor level of blood plasma fatty tissue, thus improving the state that diabetic supersession is disorderly。Fucosido is transferred to by fucosyltransferase and is completed on oligosaccharide by the fucosylation of oligosaccharide。Fucosylated oligosaccharide plays a significant role in Eukaryotic multiple physiology and pathological process。Entermorpha is subordinate to Chlorophyta Ulvales Ulvaceae Enteromorpha plant, has convenience of drawing materials, cheap advantage, is the fabulous resource of exploitation marine drug。At present, functional enteromorpha oligosaccharide research relevant report is few, there is not yet glycosylation enteromorpha oligosaccharide blood sugar lowering and promotes relevant report or the patent of proliferation of intestinal probiotics functional study。á-glucosidase is had strong inhibitory activity by glycosylation enteromorpha oligosaccharide prepared by the present invention, can remarkably promote proliferation of intestinal probiotics, and the application treated and prevented in preparation on diabetes intestinal tract disease medicine and health product has important practical significance。
Summary of the invention
It is an object of the invention to provide the glycosylation enteromorpha oligosaccharide preparation method of a kind of blood sugar lowering and regulating intestinal canal flora。Preparation technology route is simply effective, enhances the activity of functional oligosaccharide, be a kind of green high-efficient effective ways of preparing glycosylation functional oligosaccharide, the significant increase economic value added of chlorella Entermorpha resource。á-glucosidase is had strong inhibitory activity by the glycosylation enteromorpha oligosaccharide of preparation, can remarkably promote proliferation of intestinal probiotics, and have the effect improving intestinal microbial population, and beneficial bacteria of intestinal tract bacillus bifidus propagation is obviously promoted effect。Glycosylation enteromorpha oligosaccharide of the present invention can be applicable to the preparation of blood sugar lowering probiotics preparation medicine or health product, and technical problem to be solved is to provide a kind of good effect and the high diabetes of preventing and treating of availability improve the glycosylation oligosaccharide of intestinal microbial population。
For achieving the above object, the present invention adopts the following technical scheme that
(1) Entermorpha powder dry, micronizing, excessively 80-100 mesh sieve is taken, volume ratio (g/mL) 1:20-30 adds mass concentration by weight is the alcoholic solution of 60%-80%, 200-400W supersound extraction 1-1.5 hour at 60-90 DEG C, after extraction terminates, it is filtered, filtering residue repeat the above steps is carried out 2-3 time and extracts, united extraction liquid, carry out being evaporated to the 3-8% of original volume after the filtrate of acquisition being mixed, pulverize after vacuum lyophilization, cross 50-80 mesh sieve；
(2) adding distilled water in step (1) gained powder, wherein powder quality and distilled water volume ratio are (g/mL) 1:10-20, are stirred redissolving, with one or more removing protein 3-5 time in Sevag method, trifluorotrichloroethane method, trichloroacetic acid method；
(3) by solution obtained for step (2), by HPD-826 macroporous adsorbent resin packed column, eluent is further crosses non-polar adsorbent activated carbon, activated carbon makes consumption remove pigment with eluent volume ratio by weight (g/mL) 1:30-50,60-80r/min vibration 1-2h in shaking table, 4000-6000rpm is centrifuged 15-20min, collects supernatant；
(4) supernatant concentrated by rotary evaporation step (3) obtained is to the 3-8% of original volume, then 1:3-4 adds mass concentration by volume is the ethanol of 95%, standing 24h under 4-10 DEG C of condition is put into after stirring, 6000-8000rpm is centrifuged 15-20min, take supernatant concentrating under reduced pressure, lyophilization obtains the solid of Entermorpha functional oligosaccharide, crosses 50-80 mesh sieve after powder；
(5) enteromorpha oligosaccharide crude powder in step (4) adds distilled water dissolve, powder quality and water volume ratio (g/mL) are 1:20-50, solution molecular cut off 2000D bag filter processes, collect dialysis solution, the rotary evaporation when 37-40 DEG C and rotating speed are 60-80r/min, is concentrated into the 1/10-1/5 of original volume；Concentrated solution adopts molecular cut off 500D bag filter to process, and by trapped fluid lyophilization, prepares non-glycosylation enteromorpha oligosaccharide highly finished product；
(6) the enteromorpha oligosaccharide highly finished product that step (5) obtains are dissolved in distilled water by quality with volume ratio (mg/mL) 1:1-2, commercialization fucosyltransferase is joined in reaction system and carry out biological respinse, fucosyltransferase consumption presses quality and volume ratio (mg/mL) 1:3-5 with reaction system, enteromorpha oligosaccharide and guanosine diphosphate (GDP)-fucose concentration ratio (mM) 1:1.0-1.5, MgCl in reaction system2Final concentration of 15-20mM, pH6.5-8.0, be placed in 35-40 DEG C of temperature, reacts 24-36h when 80-100r/min shaking table。
(7) step (6) reaction system 1:1 by volume adding 95% ethanol, terminate reaction protein precipitation, the centrifugal 15-20min of 8000-10000r/min removes precipitation, collects supernatant concentration, obtains glycosylation Entermorpha activated oligosaccharide crude product after vacuum lyophilization。
(8) Bio-GelP-2 polyacrylamide gel will be crossed after step (7) gained dissolving crude product, distilled water eluting, measure through 3,5-dinitrosalicylic acid systems and collect oligosaccharide compositions, concentration final vacuum lyophilization obtains glycosylation Entermorpha activated oligosaccharide。
It is an advantage of the current invention that: the present invention utilizes fucosyltransferase biological respinse to prepare to have preventing and treating type 2 diabetes mellitus and improve the glycosylation enteromorpha oligosaccharide of intestinal microbial population, there is not yet fucosylation enteromorpha oligosaccharide and promote the report of proliferation of intestinal probiotics。The present invention is simple to operation, improves original activity of functional oligosaccharide, decreases production cost prepared by fucosylated oligosaccharide。Meanwhile, the comprehensive utilization Entermorpha resource that the present invention can be effective and reasonable, improves its commodity value and economic value added。
Accompanying drawing explanation
Fig. 1 enteromorpha oligosaccharide inhibitory action to á-glucosidase activity。
Fig. 2 glycosylation oligosaccharide is to bifidobacteria infantisBifidobacteriuminfantisProliferative effect。
Detailed description of the invention
Embodiment 1
(1) Entermorpha powder dry, micronizing, excessively 80 mesh sieves is taken, volume ratio (g/mL) 1:20 adds mass concentration by weight is the alcoholic solution of 60%, 200W supersound extraction 1 hour at 60 DEG C, after extraction terminates, it is filtered, filtering residue repeat the above steps is carried out 2 times and extracts, united extraction liquid, carry out being evaporated to the 3% of original volume after the filtrate of acquisition being mixed, pulverize after vacuum lyophilization, cross 50 mesh sieves；
(2) adding distilled water in step (1) gained powder, wherein powder quality and distilled water volume ratio be (g/mL) 1:10, are stirred redissolution, with trifluorotrichloroethane method removing protein 3 times；
(3) by solution obtained for step (2), by HPD-826 macroporous adsorbent resin packed column, eluent is further crosses non-polar adsorbent activated carbon, activated carbon makes consumption remove pigment with eluent volume ratio by weight (g/mL) 1:30, speed is 60r/min vibration 1h, 4000rpm is centrifuged 15min, collects supernatant；
(4) supernatant concentrated by rotary evaporation step (3) obtained is to the 3% of original volume, then 1:3 adds mass concentration by volume is the ethanol of 95%, standing 24h under 4 DEG C of conditions is put into after stirring, 6000rpm is centrifuged 15min, take supernatant concentrating under reduced pressure, lyophilization obtains the solid of Entermorpha functional oligosaccharide, crosses 50 mesh sieves after powder；
(5) enteromorpha oligosaccharide crude powder in step (4) adds distilled water dissolve, powder quality and water volume ratio (g/mL) are 1:20, and solution molecular cut off 2000D bag filter processes, and collects dialysis solution, the rotary evaporation when 37 DEG C and rotating speed are 60r/min, is concentrated into the 1/10 of original volume；Concentrated solution adopts molecular cut off 500D bag filter to process, and by trapped fluid lyophilization, prepares non-glycosylation enteromorpha oligosaccharide highly finished product；
(6) the enteromorpha oligosaccharide highly finished product that step (5) obtains are dissolved in distilled water by quality with volume ratio (mg/mL) 1:1, commercialization fucosyltransferase is joined in reaction system and carry out biological respinse, fucosyltransferase consumption presses quality and volume ratio (mg/mL) 1:3 with reaction system, enteromorpha oligosaccharide and guanosine diphosphate (GDP)-fucose concentration ratio (mM) 1:1.0, MgCl in reaction system2Final concentration of 15mM, pH6.5, be placed in 35 DEG C of temperature, reacts 24h when 80r/min shaking table。
(7) step (6) reaction system 1:1 by volume adding 95% ethanol, terminate reaction protein precipitation, the centrifugal 20min of 10000r/min removes precipitation, collects supernatant concentration, obtains glycosylation Entermorpha activated oligosaccharide crude product after vacuum lyophilization。
(8) Bio-GelP-2 polyacrylamide gel will be crossed after step (7) gained dissolving crude product, distilled water eluting, measure through 3,5-dinitrosalicylic acid systems and collect oligosaccharide compositions, concentration final vacuum lyophilization obtains glycosylation Entermorpha activated oligosaccharide。
Embodiment 2
(1) Entermorpha powder dry, micronizing, excessively 100 mesh sieves is taken, volume ratio (g/mL) 1:30 adds mass concentration by weight is the alcoholic solution of 80%, 400W supersound extraction 1.5 hours at 90 DEG C, after extraction terminates, it is filtered, filtering residue repeat the above steps is carried out 3 times and extracts, united extraction liquid, carry out being evaporated to the 8% of original volume after the filtrate of acquisition being mixed, pulverize after vacuum lyophilization, cross 80 mesh sieves；
(2) adding distilled water in step (1) gained powder, wherein powder quality and distilled water volume ratio be (g/mL) 1:20, are stirred redissolution, with Sevag method removing protein 5 times；
(3) by solution obtained for step (2), by HPD-826 macroporous adsorbent resin packed column, eluent is further crosses non-polar adsorbent activated carbon, activated carbon makes consumption remove pigment with eluent volume ratio by weight (g/mL) 1:50,80r/min vibration 2h in shaking table, 5000rpm is centrifuged 17min, collects supernatant；
(4) supernatant concentrated by rotary evaporation step (3) obtained is to the 8% of original volume, then 1:4 adds mass concentration by volume is the ethanol of 95%, standing 24h under 10 DEG C of conditions is put into after stirring, 8000rpm is centrifuged 20min, take supernatant concentrating under reduced pressure, lyophilization obtains the solid of Entermorpha functional oligosaccharide, crosses 100 mesh sieves after powder；
(5) enteromorpha oligosaccharide crude powder in step (4) adds distilled water dissolve, powder quality and water volume ratio (g/mL) are 1:50, and solution molecular cut off 2000D bag filter processes, and collects dialysis solution, the rotary evaporation when 40 DEG C and rotating speed are 80r/min, is concentrated into the 1/5 of original volume；Concentrated solution adopts molecular cut off 500D bag filter to process, and by trapped fluid lyophilization, prepares non-glycosylation enteromorpha oligosaccharide highly finished product；
(6) the enteromorpha oligosaccharide highly finished product that step (5) obtains are dissolved in distilled water by quality with volume ratio (mg/mL) 1:2, commercialization fucosyltransferase is joined in reaction system and carry out biological respinse, fucosyltransferase consumption presses quality and volume ratio (mg/mL) 1:5 with reaction system, enteromorpha oligosaccharide and guanosine diphosphate (GDP)-fucose concentration ratio (mM) 1:1.5, MgCl in reaction system2Final concentration of 20mM, pH8.0, be placed in 40 DEG C of temperature, reacts 36h when 100r/min shaking table。
(7) step (6) reaction system 1:1 by volume adding 95% ethanol, terminate reaction protein precipitation, the centrifugal 20min of 10000r/min removes precipitation, collects supernatant concentration, obtains glycosylation Entermorpha activated oligosaccharide crude product after vacuum lyophilization。
(8) Bio-GelP-2 polyacrylamide gel will be crossed after step (7) gained dissolving crude product, distilled water eluting, measure through 3,5-dinitrosalicylic acid systems and collect oligosaccharide compositions, concentration final vacuum lyophilization obtains glycosylation Entermorpha activated oligosaccharide。
Embodiment 3
(1) Entermorpha powder dry, micronizing, excessively 90 mesh sieves is taken, volume ratio (g/mL) 1:26 adds mass concentration by weight is the alcoholic solution of 75%, 300W supersound extraction 1.3 hours at 75 DEG C, after extraction terminates, it is filtered, filtering residue repeat the above steps is carried out 3 times and extracts, united extraction liquid, carry out being evaporated to the 5% of original volume after the filtrate of acquisition being mixed, pulverize after vacuum lyophilization, cross 70 mesh sieves；
(2) adding distilled water in step (1) gained powder, wherein powder quality and distilled water volume ratio be (g/mL) 1:16, are stirred redissolution, with trichloroacetic acid method removing protein 4 times；
(3) by solution obtained for step (2), by HPD-826 macroporous adsorbent resin packed column, eluent is further crosses non-polar adsorbent activated carbon, activated carbon makes consumption remove pigment with eluent volume ratio by weight (g/mL) 1:40,70r/min vibration 1.5h in shaking table, 6000rpm is centrifuged 20min, collects supernatant。
(4) supernatant concentrated by rotary evaporation step (3) obtained is to the 6% of original volume, then 1:3 adds mass concentration by volume is the ethanol of 95%, standing 24h under 5 DEG C of conditions is put into after stirring, 7000rpm is centrifuged 17min, take supernatant concentrating under reduced pressure, lyophilization obtains the solid of Entermorpha functional oligosaccharide, crosses 70 mesh sieves after powder；
(5) enteromorpha oligosaccharide crude powder in step (4) adds distilled water dissolve, powder quality and water volume ratio (g/mL) are 1:40, and solution molecular cut off 2000D bag filter processes, and collects dialysis solution, the rotary evaporation when 38 DEG C and rotating speed are 70r/min, is concentrated into the 1/10 of original volume；Concentrated solution adopts molecular cut off 500D bag filter to process, and by trapped fluid lyophilization, prepares non-glycosylation enteromorpha oligosaccharide highly finished product；
(6) the enteromorpha oligosaccharide highly finished product that step (5) obtains are dissolved in distilled water by quality with volume ratio (mg/mL) 1:1.5, commercialization fucosyltransferase is joined in reaction system and carry out biological respinse, fucosyltransferase consumption presses quality and volume ratio (mg/mL) 1:4 with reaction system, enteromorpha oligosaccharide and guanosine diphosphate (GDP)-fucose concentration ratio (mM) 1:1.2, MgCl in reaction system2Final concentration of 18mM, pH7, be placed in 37 DEG C of temperature, reacts 30h when 90r/min shaking table。
(7) step (6) reaction system 1:1 by volume adding 95% ethanol, terminate reaction protein precipitation, the centrifugal 18min of 9000r/min removes precipitation, collects supernatant concentration, obtains glycosylation Entermorpha activated oligosaccharide crude product after vacuum lyophilization。
(8) Bio-GelP-2 polyacrylamide gel will be crossed after step (7) gained dissolving crude product, distilled water eluting, measure through 3,5-dinitrosalicylic acid systems and collect oligosaccharide compositions, concentration final vacuum lyophilization obtains glycosylation Entermorpha activated oligosaccharide。
á-glucosidase is had strong inhibitory activity by Entermorpha activated oligosaccharide prepared by the present invention, and its IC50 value is 1.36mg/mL, and presents dose dependent (Fig. 1)。
By bifidobacteria infantisBifidobacteriuminfantisIt is seeded in commercialization BBL culture medium (BBL culture medium prescription: peptone 15g/L, yeast extract powder 2g/L, glucose 20g/L, soluble starch 0.5g/L, sodium chloride 5g/L, cysteine 0.5g/L, tween 80 1mL/L, hepar siccatum 0.3g/L, agar 15g/L, pH6.8 ± 0.2), glycosylation oligosaccharide replaces glucose as given the test agent group simultaneously, it is placed in anaerobic culture box 37 DEG C and cultivates 84h, measure culture fluid OD600 value to monitor bacillus bifidus proliferative conditions every 12h。From Figure 2 it can be seen that glycosylation oligosaccharide can significantly improve beneficial bacteria of intestinal tract bifidobacteria。
The foregoing is only presently preferred embodiments of the present invention, all equalizations done according to the present patent application the scope of the claims change and modify, and all should belong to the covering scope of the present invention。
Claims (3)
1. the glycosylation enteromorpha oligosaccharide preparation method of a blood sugar lowering and regulating intestinal canal flora, it is characterised in that: described method includes as follows:
(1) Entermorpha powder dry, micronizing, excessively 80-100 mesh sieve is taken, the ratio 1:20-30 of g and volume mL adds mass concentration by weight is the alcoholic solution of 60%-80%, 200-400W supersound extraction 1-1.5 hour at 60-90 DEG C, after extraction terminates, it is filtered, filtering residue repeat the above steps is carried out 2-3 time and extracts, united extraction liquid, carry out being evaporated to the 3-8% of original volume after the filtrate of acquisition being mixed, pulverize after vacuum lyophilization, cross 50-80 mesh sieve；
(2) adding distilled water in step (1) gained powder, wherein powder quality g and distilled water mL volume ratio are 1:10-20, are stirred redissolving, with one or more removing protein 3-5 time in Sevag method, trifluorotrichloroethane method, trichloroacetic acid method；
(3) by solution obtained for step (2), by HPD-826 macroporous adsorbent resin packed column, eluent is further crosses non-polar adsorbent activated carbon, activated carbon makes consumption remove pigment with eluent volume ratio 1:30-50 by weight, 60-80r/min vibration 1-2h in shaking table, 4000-6000rpm is centrifuged 15-20min, collects supernatant；
(4) supernatant concentrated by rotary evaporation step (3) obtained is to the 3-8% of original volume, then 1:3-4 adds mass concentration by volume is the ethanol of 95%, standing 24h under 4-10 DEG C of condition is put into after stirring, 6000-8000rpm is centrifuged 15-20min, take supernatant concentrating under reduced pressure, lyophilization obtains the solid of Entermorpha functional oligosaccharide, crosses 50-80 mesh sieve after powder；
(5) enteromorpha oligosaccharide crude powder in step (4) adds distilled water dissolve, powder quality g and water mL volume ratio are 1:20-50, solution molecular cut off 2000D bag filter processes, collect dialysis solution, the rotary evaporation when 37-40 DEG C and rotating speed are 60-80r/min, is concentrated into the 1/10-1/5 of original volume；Concentrated solution adopts molecular cut off 500D bag filter to process, and by trapped fluid lyophilization, prepares non-glycosylation enteromorpha oligosaccharide highly finished product；
(6) the enteromorpha oligosaccharide highly finished product that step (5) obtains are dissolved in distilled water by the ratio 1:1-2 of quality mg and volume mL, fucosyltransferase is joined reaction system carries out biological respinse, fucosyltransferase consumption and reaction system press quality mg and volume mL than 1:3-5, and in reaction system, enteromorpha oligosaccharide and guanosine diphosphate (GDP)-fucose concentration are than 1:1.0-1.5, MgCl2Final concentration of 15-20mM, pH6.5-8.0, be placed in 35-40 DEG C of temperature, reacts 24-36h when 80-100r/min shaking table；
(7) step (6) reaction system 1:1 by volume adding 95% ethanol, terminate reaction protein precipitation, the centrifugal 15-20min of 8000-10000r/min removes precipitation, collects supernatant concentration, obtains glycosylation Entermorpha activated oligosaccharide crude product after vacuum lyophilization；
(8) Bio-GelP-2 polyacrylamide gel will be crossed after step (7) gained dissolving crude product, distilled water eluting, measure through 3,5-dinitrosalicylic acid systems and collect oligosaccharide compositions, concentration final vacuum lyophilization obtains glycosylation Entermorpha activated oligosaccharide。
2. the glycosylation Entermorpha activated oligosaccharide that a preparation method as claimed in claim 1 prepares。
3. glycosylation Entermorpha activated oligosaccharide as claimed in claim 2 is prevented and treated diabetes in preparation and is improved the application in the medicine of intestinal flora metabolism disease or health product。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610110627.1A CN105695537B (en) | 2016-02-29 | 2016-02-29 | Glycosylation enteromorpha oligosaccharide preparation method that is a kind of hypoglycemic and adjusting intestinal flora |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610110627.1A CN105695537B (en) | 2016-02-29 | 2016-02-29 | Glycosylation enteromorpha oligosaccharide preparation method that is a kind of hypoglycemic and adjusting intestinal flora |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105695537A true CN105695537A (en) | 2016-06-22 |
| CN105695537B CN105695537B (en) | 2019-11-19 |
Family
ID=56222638
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610110627.1A Active CN105695537B (en) | 2016-02-29 | 2016-02-29 | Glycosylation enteromorpha oligosaccharide preparation method that is a kind of hypoglycemic and adjusting intestinal flora |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105695537B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106632712A (en) * | 2016-11-01 | 2017-05-10 | 福建海兴保健食品有限公司 | Process for extracting oligosaccharide by utilizing polysaccharide extracted green algae waste liquor |
| CN107319540A (en) * | 2017-08-18 | 2017-11-07 | 青岛瑞思德生物科技有限公司 | A kind of health food for improving diabetes containing Enteromorpha and preparation method thereof |
| CN109734761A (en) * | 2019-02-28 | 2019-05-10 | 福建农林大学 | A kind of preparation method and applications of Enteromorpha fucose complex compound |
| CN111629732A (en) * | 2017-11-20 | 2020-09-04 | 高丽大学校产学协力团 | Process for preparing various fucosyl oligose and its use |
| CN112715964A (en) * | 2020-12-16 | 2021-04-30 | 中国科学院南海海洋研究所 | Spirulina oligosaccharide and application thereof in preparation of preparation for regulating intestinal health function |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102898536A (en) * | 2012-10-19 | 2013-01-30 | 中国科学院海洋研究所 | Method for degrading enteromorpha polysaccharide |
| CN103201380A (en) * | 2010-10-11 | 2013-07-10 | 詹尼温生物技术有限责任公司 | Novel fucosyltransferases and their applications |
| CN103328630A (en) * | 2011-01-20 | 2013-09-25 | 詹尼温生物技术有限责任公司 | Novel fucosyltransferases and their applications |
| CN103951761A (en) * | 2014-05-13 | 2014-07-30 | 浙江工商大学 | Method for degrading enteromorpha prolifera polysaccharides by enzymic method |
-
2016
- 2016-02-29 CN CN201610110627.1A patent/CN105695537B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103201380A (en) * | 2010-10-11 | 2013-07-10 | 詹尼温生物技术有限责任公司 | Novel fucosyltransferases and their applications |
| CN103328630A (en) * | 2011-01-20 | 2013-09-25 | 詹尼温生物技术有限责任公司 | Novel fucosyltransferases and their applications |
| CN102898536A (en) * | 2012-10-19 | 2013-01-30 | 中国科学院海洋研究所 | Method for degrading enteromorpha polysaccharide |
| CN103951761A (en) * | 2014-05-13 | 2014-07-30 | 浙江工商大学 | Method for degrading enteromorpha prolifera polysaccharides by enzymic method |
Non-Patent Citations (2)
| Title |
|---|
| 于广利，赵峡: "《糖药物学》", 31 October 2012 * |
| 李玉等: "《功能性低聚糖合成中糖基转移酶研究进展》", 《食品科学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106632712A (en) * | 2016-11-01 | 2017-05-10 | 福建海兴保健食品有限公司 | Process for extracting oligosaccharide by utilizing polysaccharide extracted green algae waste liquor |
| CN107319540A (en) * | 2017-08-18 | 2017-11-07 | 青岛瑞思德生物科技有限公司 | A kind of health food for improving diabetes containing Enteromorpha and preparation method thereof |
| CN111629732A (en) * | 2017-11-20 | 2020-09-04 | 高丽大学校产学协力团 | Process for preparing various fucosyl oligose and its use |
| CN109734761A (en) * | 2019-02-28 | 2019-05-10 | 福建农林大学 | A kind of preparation method and applications of Enteromorpha fucose complex compound |
| CN112715964A (en) * | 2020-12-16 | 2021-04-30 | 中国科学院南海海洋研究所 | Spirulina oligosaccharide and application thereof in preparation of preparation for regulating intestinal health function |
| CN112715964B (en) * | 2020-12-16 | 2022-04-19 | 中国科学院南海海洋研究所 | Spirulina oligosaccharide and application thereof in preparation of preparation for regulating intestinal health function |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105695537B (en) | 2019-11-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105695537A (en) | Preparation method of glycosylation enteromorpha oligosaccharide for reducing glucose level and regulating intestinal flora | |
| CN103284152B (en) | Dietary fiber extracted from Chinese yew, extraction method and use | |
| CN107136506A (en) | A kind of Black Box Tracing piece with blood pressure reduction effect and preparation method thereof | |
| CN102038745B (en) | Hypolipidemic traditional Chinese medicine | |
| CN101664180B (en) | Health-care nutritional complexing agent with health effect and preparation method thereof | |
| CN101856374A (en) | Anti-aging compound | |
| CN107286264A (en) | The deep working method of Chinese date nutrient material separation | |
| CN103768191A (en) | Traditional Chinese medicinal mixture for preventing fatty liver and preparation method and application thereof | |
| CN105031289A (en) | Dendrobium officinale and lucid ganoderma capsules and preparation method thereof | |
| CN108186922A (en) | A kind of preparation method of hypoglycemic composition | |
| CN106581166A (en) | Anti-fatigue food, health product or pharmaceutical composition | |
| KR20130074119A (en) | Ginseng prosapogenin high concentration containing ginseng leaf or stem preparation using sonication and process for thereof | |
| CN102579530A (en) | Preparation method of aralia taibaiensis total saponin having diabetes mellitus resisting effect and medicament | |
| CN112843140B (en) | Medicine for treating thirst and conjunctival congestion caused by diabetes, preparation method and application | |
| CN1565467A (en) | Use of cornel and its extract in preparation alpha-glucosidase inhibitor medicine | |
| CN102309705B (en) | Medicine for reducing serum uric acid, preparation method thereof and purpose thereof | |
| CN107961293A (en) | A kind of sea-buckthorn tomato piece and preparation method thereof | |
| CN104435072B (en) | A kind of extract and preparation method thereof with auxiliary hyperglycemic, reducing blood lipid | |
| CN105125646B (en) | A kind of application of Trichoderma viride in improving maple knotweed prescription medicinal material in Quercetin recovery rate | |
| CN113476561A (en) | Traditional Chinese medicine formula for treating diabetes | |
| CN1332019C (en) | Grifola umbellate zymophyte powder and grifola umbellate polysaccharide producing method and products therefrom | |
| CN102302545A (en) | Dai medicine extract with blood-sugar reducing activity, preparation method, composition and application | |
| CN112754014A (en) | Composition for preventing and treating cardiovascular and cerebrovascular diseases and preparation method thereof | |
| CN101040905B (en) | Medicine made by selfheal for reducing blood sugar | |
| CN102488761A (en) | Traditional Chinese medicine composition for treating diabetes, preparation thereof and its preparation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20210115 Address after: 130000 1st floor, Rongxiang Road Complex Building, medical and Food Industrial Park, Lvyuan Economic Development Zone, Lvyuan District, Changchun City, Jilin Province Patentee after: Qingfeng chain soda beverage (Jilin) Co.,Ltd. Address before: No. 15, Cangshan District, Fujian, Fuzhou, Fujian Patentee before: FUJIAN AGRICULTURE AND FORESTRY University |