CN101955549A - Large-headed atractylodes rhizome polysaccharide and preparation method and application thereof - Google Patents
Large-headed atractylodes rhizome polysaccharide and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a method for preparing large-headed atractylodes rhizome polysaccharide and application. The method for preparing the large-headed atractylodes rhizome polysaccharide comprises the following steps of: 1) soaking large-headed atractylodes rhizomes into water, then decocting the rhizomes, collecting filtrate, and concentrating the filtrate to obtain large-headed atractylodes rhizome extract; 2) adding ethanol into the extract, standing the extract at the temperature of 4 DEG C, centrifuging the extract and collecting sediment, dissolving the sediment, removing the ethanol, small molecular sugar, protein and salt by dialysis, and drying the extract to obtain coarse large-headed atractylodes rhizome polysaccharide; and 3) dissolving the large-headed atractylodes rhizome polysaccharide, then adding Saveg liquid in the volume which is 2 times that of the solution, mixing and shaking the solution uniformly, centrifuging the solution and collecting supernate, then removing chloroform and n-butyl alcohol by dialysis from water, and drying the supernate to obtain the large-headed atractylodes rhizome polysaccharide without containing the protein. Proved by experiments, the large-headed atractylodes rhizome polysaccharide can promote proliferation of probiotics of infants, youth, animal bifidobacterium, plant lactobacillus and the like in vitro, can be used as probiotics for production, has the effect of regulating intestinal flora balance, and can be used as novel functional probiotics for development.
Description
Technical field
The present invention relates to a kind of preparation method and its application in regulating the intestinal microflora balance of soluble polysaccharide.
Background technology
Often there are one deck microorganism or microorganism layer to exist in the human body alimentary canal, they are not only harmless to the host, and be useful and essential, this microorganism layer promptly is called normal microflora (normal microbiota) or normal microflora (normal bacteria flora).The growth metabolism activity that these micropopulations (tens billion of, as to account for the 1/50-1/60 of human body weight) enliven in human intestinal has very important physical effect.Closely related with aspects such as human consumption, absorption, immunity, antitumor, anti-ageing, external flora invasion and attack of opposing.Numerous results of study show that age, ethnic group, diet formula, living habit, environment, disease, antibiotic therapy etc. can influence people's intestinal microecology system and structure of community thereof.Bifidus bacillus and Bacterium lacticum are two kinds of most important beneficial bacteria of intestinal tract, and its quantity and composition play keying action to keeping healthy intestinal environment and improving function of immune system.
Bifidus bacillus can be kept the microecological balance of host's enteron aisle, improves the tolerance of human body to lactose; Have the growth of the enteric pathogenic bacteria of inhibition, improve protein metabolism and can synthesize multivitamin, promote human body immune function etc.Bifidus bacillus is in space-time, kind and quantitatively produce corresponding dynamic and change, and reduces with age, thereby directly influences the health of human body.In recent years, the upsurge that has occurred research and development bifidus bacillus product both at home and abroad, at present main product has yoghurt product, capsule etc., but such active bacteria formulation preserves the relatively poor and viable bacteria of validity and take in the condition restriction that is subjected to the hydrochloric acid in gastric juice influence behind the body, and the effect of replenishing bifidus bacillus to body is reduced.Thereby, in body, replenish the bifid somatomedin, adopting the material that promotes bifidus bacillus propagation to come the interior profitable strain of added body will be easier received approach.In the bifid somatomedin, oligosaccharides research all has the proliferation function that promotes bifidus bacillus at most as fructose oligose, soybean oligosaccharide, isomalto Oligosaccharide etc.
Through verification, prebiotics product on the consumption market mainly contains at present: English prebiotics iron zinc calcium glucose, Dumex baby milk powder, Bei Yinmei bovine coloctrum prebiotics milk powder, multidimensional prebiotics bean milk powder, the clear youngster's profit of Heng Shi, the many essences of nest's power or the like, these products invalid diarrhoea of treatment the alteration of intestinal flora diarrhoea, chronic diarrhoea, the antibiotic therapy that cause and constipation etc. the aspect brought into play very big effect, also be the good ancillary drug of some clinical disease simultaneously.They mostly are the product of types of functionality oligose such as adding oligofructose, glucose oligosaccharide, xylo-oligosaccharide, algin oligosaccharide, synanthrin greatly, and are in the majority with milk-product.
By domestic and international patent documentation, journal and other document of publishing (as the internet) are retrieved, polysaccharide research mainly concentrates on immunomodulatory, antitumor, antiviral, anti-ageing waiting for a long time, and does not have polysaccharide compound to develop as prebiotics.
The bighead atractylodes rhizome (Rhizoma Atractylodis Macrocephalae) is the dry rhizome of composite family (Compositae) plant bighead atractylodes rhizome Atractylodes macrocephala Koidz.The bighead atractylodes rhizome is a perennial herb, and cultivation history is long, and Dongyang, main product China Zhejiang Province, Pan'an one band are one of famous genuine traditional Chinese medicine materials " Zhejiang eight flavors ", are distributed in Zhejiang, Jiangxi, Hunan, Hubei, Shaanxi, also many cultivations.Soluble polysaccharide, also only limiting to has good action in immunomodulatory and antitumor, reducing blood-fat, not to the relevant report of probiotic bacterium and intestinal microflora effect.
Summary of the invention
The purpose of this invention is to provide a kind of preparation method and its application in regulating the intestinal microflora balance of soluble polysaccharide.
The preparation method of soluble polysaccharide provided by the invention comprises the steps:
1) bighead atractylodes rhizome is soaked in water, decocts then, collect filtrate, concentrate and obtain bighead atractylodes rhizome vat liquor;
2) in the bighead atractylodes rhizome vat liquor that step 1) obtains, add ethanol or ethanolic soln, regulating ethanol final volume percentage concentration is 70%-75% (as 71.25%), place 4 ℃ to leave standstill 24 hours, centrifugal collecting precipitation, to precipitate and use water dissolution, dialysis removes ethanol, small molecular sugar, protein and salt in water then; Concentrate dialysate, vacuum lyophilization obtains bighead atractylodes rhizome Crude polysaccharides;
3) with step 2) bighead atractylodes rhizome Crude polysaccharides water dissolution that obtains, the Saveg liquid of 2 times of volumes of interpolation, mixing shakes up, centrifugal collection supernatant liquor, repeat this step, up to supernatant liquor through ultraviolet determination 260,280nm wavelength place do not have the absorption peak of protein and nucleic acid, this supernatant liquor is the liquid glucose behind the deproteinated; Chloroform and propyl carbinol are removed in dialysis in water then, concentrate dialysate, and vacuum lyophilization obtains nonprotein soluble polysaccharide; Described Saveg liquid is that chloroform and propyl carbinol are 4: 1 mixed mixed solutions by volume;
Also comprise the nonprotein soluble polysaccharide water dissolution that step 3) is obtained in the described method, carry out DEAE-Sepharose Fast Flow anion column chromatography purification, collect the elutriant that contains polysaccharide with 50mM Tris-HCl; Placing the water dialysis to remove the Tris-HCl component elutriant concentrates postlyophilization, obtains soluble polysaccharide.
In order to accelerate dissolved speed, the temperature of described dissolving water is 40-80 ℃, as 60 ℃.
Described step 2) and in the step 3), the molecular weight cut-off of described dialysis is below the 5000Da, to be preferably 3500Da.
Described step 2) and in the step 3), the time of described dialysis is 24-72 hour, is preferably 48 hours.
In the described step 1), the time of described immersion is 12-36 hour, is preferably 24 hours; Described immersion distilled water is the 10ml/g bighead atractylodes rhizome; Described decoction is that the bighead atractylodes rhizome after will soaking directly decocted 30 minutes with its soak solution, filters and collects filtrate, and filter residue was boiled 30 minutes at the decocting that is used for the soak solution equal volume, collects filtrate, merges the filtrate that twice collection obtains.
In the described method, used water is distilled water or deionized water.Such as, above-mentioned steps 1) the immersion bighead atractylodes rhizome water in, step 2) in dissolving bighead atractylodes rhizome Crude polysaccharides water in the dissolution precipitation water, step 3) all can be distilled water, dissolving soluble polysaccharide water can be deionized water before the water for dialysis in the aforesaid method, DEAE-Sepharose Fast Flow anion column chromatography purification.
The soluble polysaccharide of method for preparing is the claimed soluble polysaccharide of the present invention.
The present invention also provides the application of soluble polysaccharide in the balance of propagation that promotes intestinal beneficial bacterium or adjusting intestinal microflora.
Described intestinal beneficial bacterium is bifidobacteria infantis, bifidobacterium adolescentis, animal bifidobacteria or plant lactobacillus.Described intestinal beneficial bacterium is preferably bifidobacteria infantis CICC6069, bifidobacterium adolescentis CICC6070, bifidumbacterium bifidum 1.1852, animal bifidobacteria 1.2268 or plant lactobacillus 1.124.Soluble polysaccharide can be in the propagation of external promotion intestinal beneficial bacterium, and the balance that can regulate intestinal microflora to a certain extent.
The extracting method of soluble polysaccharide of the present invention removes albumen, dialysis, anion column chromatography, the soluble polysaccharide based on very high purity of extraction through alcohol precipitation, Saveg method, purity reaches chromatographically pure, and molecular weight is 5527Da, mainly by pectinose, semi-lactosi, glucose and seminose are formed.
The authentication method and the instrument of the soluble polysaccharide that the method that we adopt is extracted all belong to up-to-date scientific payoffs, detect sensitive and accurate, with a high credibility.High performance liquid chromatography and gel permeation chromatography are mainly adopted in the polysaccharide molecule flow measurement, and present method is utilized laser detector, and be highly sensitive, and need not mark song.It is the pulse Amperometric Detection Coupled that the monose composition mostly adopts liquid chromatography or gas-chromatography, the detector of employing, does not need before the sample column or post-column derivation, the resolving power height, and the result of acquisition is more reliable.
The present invention has verified by experiment that first soluble polysaccharide has promoter action to the propagation of intestinal beneficial bacterium, experiment showed, that this soluble polysaccharide can be in the propagation of probiotic bacteriums such as external promotion baby, youth, animal bifidobacteria and plant lactobacillus.Can be used as a kind of prebiotics production utilization, have the effect of the intestinal microflora of adjusting equilibrated, can be used as the exploitation of new type functional prebiotics.We study soluble polysaccharide to little ecological aspect, remedied this blank on the one hand, and be hopeful to be added in the product the functional prebiotics product of development of new.
Description of drawings
Fig. 1 makes the sample photo of bighead atractylodes rhizome Crude polysaccharides for the inventive method.
Fig. 2 makes the sample photo of the smart polysaccharide of the bighead atractylodes rhizome for the inventive method.
Fig. 3 makes the Sepharose CL-6B gel chromatography elution curve of the smart polysaccharide of the bighead atractylodes rhizome for embodiment 1
Fig. 4 makes the UV scanning figure of the smart polysaccharide of the bighead atractylodes rhizome for embodiment 1
The smart polysaccharide of the different addition bighead atractylodes rhizomes of Fig. 5 is to the influence of bifidobacterium growth
The different addition soluble polysaccharides of Fig. 6 (AMP) are to the influence of growth of lactobacillus
The FT-IR spectrogram of Fig. 7 soluble polysaccharide (AMP)
A is monose mark product (1.0 μ g/mL) HPAEC-PAD color atlass among Fig. 8; B makes the HPAEC-PAD color atlas of the smart polysaccharide of the bighead atractylodes rhizome for embodiment 1
Embodiment
Experimental technique described in the following embodiment if no special instructions, is ordinary method.
The extracting method of embodiment 1, soluble polysaccharide
1, the extracting method of soluble polysaccharide
Get the 20g bighead atractylodes rhizome (available from hundred Wang Shanyaodian of Beijing Tongrentang), add 200mL distilled water immersion 24h, its soak solution directly decocts the 30min after-filtration then, and filter residue adds same amount distilled water (200mL) and decocts 30min again, the merging filtrate evaporation concentration obtains bighead atractylodes rhizome vat liquor to 100mL.
2) the extraction purifying of soluble polysaccharide
Water extraction and alcohol precipitation method prepares bighead atractylodes rhizome Crude polysaccharides: bighead atractylodes rhizome vat liquor is concentrated 3 times of volume ethanol solution of back adding, and (concentration expressed in percentage by volume is 95%, alcoholic acid final volume percentage concentration is 71.25%), place 4 ℃ of refrigerator alcohol precipitation 24h, the centrifugal 10min collecting precipitation of 8000rpm, (40-80 ℃ of hot distilled water of precipitation, common room temperature water gets final product, the water that temperature is high slightly can be accelerated dissolution rate) redissolve after, place dialysis tubing (Oso-T8340, MD34 (3500), U.S.'s associating carbonization) in, dialysis tubing is placed the deionized water 48h that dialyses, remove ethanol, small molecular sugar, protein, salt etc.Concentrate dialysate, vacuum lyophilization gets bighead atractylodes rhizome Crude polysaccharides, and the raw sugar extraction yield is 10% (making raw sugar quality/bighead atractylodes rhizome dry weight), and sample is seen Fig. 1.
The Saveg method is removed albumen: with above-mentioned dialysis 2) back (40-80 ℃ of hot distilled water of the bighead atractylodes rhizome Crude polysaccharides that obtains, normal temperature gets final product, the water that temperature is high slightly can be accelerated dissolution rate) obtain bighead atractylodes rhizome Crude polysaccharides solution after redissolving, with Saveg liquid (chloroform: propyl carbinol (volume ratio)=4: 1) shake up deproteinated with 2: 1 (volume ratio) mixed with liquid glucose, the centrifugal 10min of 8000rpm collects supernatant liquor, repeat 4 times, to ultraviolet determination 260, the liquid glucose of the 280nm wavelength place absorption peak that do not have protein and nucleic acid after obtaining deproteinated.Liquid glucose behind the deproteinated is placed in the dialysis tubing (Oso-T8340, MD34 (3500), U.S.'s associating carbonization), dialysis tubing is placed the deionized water 48h that dialyses, remove chloroform and propyl carbinol, concentrate, lyophilize obtains nonprotein soluble polysaccharide.
DEAE-Sepharose Fast Flow anion column chromatography purification: will be mixed with 10mg/mL solution with deionized water except that the soluble polysaccharide behind the albumen, 0.45 behind the μ m filtering with microporous membrane, with DEAE-Sepharose Fast Flow anion column (glass ion exchange column: 2.6 * 20em, load DEAE-Sepharose Fast Flow, available from Pharmacia) purifying, behind the last sample of 7mL, with 50mM Tris-HCl eluant solution, flow velocity 4.0mL/min, the 2min/ pipe, collect 40 pipes, Fraction Collector is collected elutriant.With the phenolsulfuric acid colorimetry (Xu Bin, Dong Ying, Lin Lin, etc. improvement phenolsulfuric acid method is measured bitter melon polysaccharide content [J]. Food science, 2005,26 (7): 79-82.) follow the tracks of and detect every pipe polysaccharide content.The result shows that soluble polysaccharide wash-out when effluent volume is 120mL is complete.
Collection contains the effluent liquid at the peak of polysaccharide, place dialysis tubing (Oso-T8340, MD34 (3500), U.S.'s associating carbonization) 24h that in deionized water, dialyses, remove the Tris-HCl component, concentrate postlyophilization, obtain pure product of soluble polysaccharide and called after AMP (Atractylodes macrocephala polysaccharides).The pure product photo of this soluble polysaccharide as shown in Figure 2.
2, purity of polysaccharide is identified
1) normal pressure gel chromatography chromatography (glass gel-filtration column: 1.6 * 80cm, load Sepharose CL-6B, available from Pharmacia): with the AMP freeze-drying sample deionized water dissolving of step 1 acquisition, be mixed with 10mg/mL solution, 0.45 go up sample (10mL), 0.05mol/L NaCl wash-out, flow velocity 30mL/h behind the μ m filtering with microporous membrane, the 12min/ pipe, Fraction Collector is collected elutriant automatically.The phenolsulfuric acid colorimetry is followed the tracks of and is detected every pipe polysaccharide content, curve plotting figure.
The result as shown in Figure 3, the result shows that soluble polysaccharide has reached chromatographically pure.
2) ultraviolet (UV) detects: the pure product AMP that step 1 is obtained is prepared into 0.5mg/mL sample liquid to be measured with deionized water dissolving, with the deionized water is blank, in wavelength 220-580nm scope, utilize ultraviolet spectrophotometer to carry out the ultraviolet full wavelength scanner, to determine whether contain nucleic acid and protein among the AMP.
The result shows that the pure product AMP that step 1 obtains does not have absorption at the 260-280nm place, and no protein and nucleic acid are as Fig. 4.
3, soluble polysaccharide is to the external promotes growth test of intestinal microflora
1) the different concns soluble polysaccharide is to the influence of bifidus bacillus growth in vitro: (bifidobacterium adolescentis CICC6070, bifidobacteria infantis CICC6069, bifidus longum bb CICC6068 purchase in Chinese industrial microbial strains preservation administrative center will to activate 5 good strain bifidus bacilluss; Bifidumbacterium bifidum 1.1852, animal bifidobacteria 1.2268 be available from Chinese common micro-organisms culture presevation administrative center (CGMCC); Respectively with 1% inoculum size (10
6CFU/mL) being inoculated in whole mass percentage concentration that the AMP of step 1 preparation adds is in 0%, 0.1%, 0.5%, 1.0%, 1.5%, 2.0% or 2.5% the MRS substratum, each concentration repeats three groups, behind 37 ℃ of cultivation 24h, measure the OD value of each nutrient solution down in the 600nm wavelength.
The result shows that soluble polysaccharide can have good growth promoting function in the propagation of external promotion bifidobacteria infantis, bifidobacterium adolescentis, animal bifidobacteria, and concrete outcome is seen Fig. 5.
2) the different concns soluble polysaccharide influences the growth in vitro of Bacterium lacticum: will activate good 2 strain Bacterium lacticum (Lactobacterium acidophilum 1.2686, plant lactobacillus 1.124 are available from CGMCC) respectively with 1% inoculum size (about 10
6CFU/mL) being inoculated in AMP, to add whole mass percentage concentration be that each concentration repeats three groups in 0%, 0.1%, 0.5%, 1.0%, 1.5%, 2.0% or 2.5% the MRS substratum, 37 ℃ cultivate 24h after, measure the OD value that the 600nm wavelength is measured each nutrient solution down.
The result shows that soluble polysaccharide can promote the propagation of plant lactobacillus, sees Fig. 6.
3) soluble polysaccharide is to the influence of all the other main intestinal microflora growth in vitro: will activate 5 good big class intestinal microflora representative strain (bifidobacterium adolescentis CICC6070, plant lactobacillus CGMCC1.124, intestinal bacteria ATCC83965, enterococcus faecalis CGMCC1.131, clostridium butylicum CGMCC1.336) respectively with 1% inoculum size (about 10
6CFU/mL) being inoculated in AMP (step 1 preparation soluble polysaccharide), to add whole mass percentage concentration be in 0.1% the appropriate media separately and control group (not adding soluble polysaccharide), each group repeats 3 groups, behind 37 ℃ of cultivation 24h, measure the OD value that the 600nm wavelength is measured each nutrient solution down.
Result's (seeing Table 1) shows, soluble polysaccharide can promote the growth of bifidobacterium adolescentis, plant lactobacillus, facilitation effect to all the other 3 classes (intestinal bacteria, clostridium butylicum, enterococcus faecalis) is not obvious, or restraining effect is arranged, and the effect of the intestinal microflora of adjusting equilibrated is arranged.
Table 1: soluble polysaccharide is to the regulating effect of intestinal microflora
4, the structural analysis of soluble polysaccharide
1) molecular weight determination: multiple angle laser light scattering instrument attached gel permeation chromatography (GPC/MALLS) method detects the AMP weight-average molecular weight.Chromatographic condition: chromatographic column Shodex OHpak SB-806M HR (8.0mm * 300mm); DAWN HELEOS-II type laser light scattering instrument, Optilab rEX type differential refractometer (U.S. Wyatt Technology); The dn/dc value is 0.135; Moving phase: 0.1MNaNO
3Flow velocity: 0.5mL/min; Column temperature: 35 ℃; Sampling volume: 200 μ L; Sample concentration: 0.2mg/mL.The weight-average molecular weight that records soluble polysaccharide is 5527Da, and mass distribution sees Table 2 than homogeneous.
Table 2: the molar mass of soluble polysaccharide AMP
2) Infrared spectroscopy: adopt the KBr pressed disc method, the NEXUS-470FTIR infrared detector is at 400-4000cm
-1Sensing range scanning 32 times.Infrared spectrogram (Fig. 7) proves that this material is polysaccharide compound really.
3) acid-hydrolysis method and high performance anion exchange chromatography-pulse ampere (HPAEC-PAD) detects the monose composition measuring: accurately take by weighing the pure product dry powder of AMP 5mg, add the dense H of 200 μ L
2SO
4Leave standstill, add 800 μ L distilled water, 100 ℃ of constant temperature hydrolysis 2.5h are settled to 2mL after the cooling, and hydrolyzed solution is with solid BaCO
3Neutralization, the centrifugal impurity of removing carries out the monose qualitative detection with distilled water with 20 times of supernatant liquor dilutions and after filtering.Select the mixing monose standard substance of different concns (0.2 μ g/mL, 0.5 μ g/mL, 1.0 μ g/mL, 4.0 μ g/mL and 10.0 μ g/mL), (μ g/mL) is X-coordinate with monosaccharide concentration, and (nC * min) is that the typical curve that ordinate zou is drawn every kind of monose carries out the monose detection by quantitative with peak area.
The HPAEC-PAD chromatographic condition: the glycan analysis post (CarboPacTM PA10,4mm * 250mm); The sugar guard column (CarboPacTM PA10,4mm * 50mm); Moving phase: H
2O and 200mmol/L NaOH, linear gradient elution; The ED50A electrochemical detector; Sampling volume: 25 μ L; Flow velocity: 1.0mL/min; Column temperature: 30 ℃.
The result shows that soluble polysaccharide is made up of pectinose, semi-lactosi, glucose, seminose, and the content mol ratio is 13.8: 11.8: 70.2: 4.1; The results are shown in Figure 8.
Claims (10)
1. the preparation method of a soluble polysaccharide comprises the steps:
1) bighead atractylodes rhizome is soaked in water, decocts then, collect filtrate, concentrate and obtain bighead atractylodes rhizome vat liquor;
2) in the bighead atractylodes rhizome vat liquor that step 1) obtains, add ethanol or ethanolic soln, to ethanol final volume percentage concentration be 70%-75%, place 4 ℃ to leave standstill centrifugal collecting precipitation 24 hours, to precipitate and use water dissolution, ethanol, small molecular sugar, protein and salt are removed in dialysis in water then; Concentrate dialysate, vacuum lyophilization obtains bighead atractylodes rhizome Crude polysaccharides;
3) with step 2) bighead atractylodes rhizome Crude polysaccharides water dissolution that obtains, add the Saveg liquid of 2 times of volumes then, mixing shakes up, centrifugal collection supernatant liquor, repeat this step, up to supernatant liquor through ultraviolet determination 260 and 280nm wavelength place do not have the absorption peak of protein and nucleic acid, this supernatant liquor is the liquid glucose behind the deproteinated; Chloroform and propyl carbinol are removed in dialysis in water then, concentrate dialysate, and vacuum lyophilization obtains nonprotein soluble polysaccharide; Described Saveg liquid is that chloroform and propyl carbinol are 4: 1 mixed mixed solutions by volume.
2. method according to claim 1, it is characterized in that: also comprise the nonprotein soluble polysaccharide water dissolution that step 3) is obtained in the described method, carry out DEAE-Sepharose Fast Flow anion column chromatography purification, with 50mM Tris-HCl wash-out, collect the elutriant that contains polysaccharide; The elutriant that will contain polysaccharide places the deionized water dialysis to remove the Tris-HCl component and concentrates postlyophilization, obtain soluble polysaccharide.
3. method according to claim 1 and 2 is characterized in that: described step 2) and in the step 3), the molecular weight cut-off of described dialysis is below the 5000Da, to be preferably 3500Da.
4. according to any described method among the claim 1-3, it is characterized in that: described step 2) and in the step 3), the time of described dialysis is 24-72 hour, is preferably 48 hours.
5. according to any described method among the claim 1-4, it is characterized in that: in the described step 1), the time of described immersion is 12-36 hour, is preferably 24 hours; The consumption of described immersion water is the 10ml/g bighead atractylodes rhizome; Described decoction is that the bighead atractylodes rhizome after will soaking directly decocted 30 minutes with its soak solution, filters and collects filtrate, and filter residue was boiled 30 minutes at the decocting that is used for the soak solution equal volume, collects filtrate, merges the filtrate that twice collection obtains.
6. any described method among the claim 1-5, it is characterized in that: in the described method, used water is distilled water or deionized water.
7. the soluble polysaccharide of any described method preparation among the claim 1-6.
8. the application of the described soluble polysaccharide of claim 7 in propagation that promotes intestinal beneficial bacterium and/or adjusting intestinal microflora balance.
9. application according to claim 8 is characterized in that: described intestinal beneficial bacterium is bifidobacteria infantis, bifidobacterium adolescentis, animal bifidobacteria or plant lactobacillus; Described intestinal beneficial bacterium is preferably bifidobacteria infantis CICC6069, bifidobacterium adolescentis CICC6070, bifidumbacterium bifidum CGMCC1.1852 or plant lactobacillus CGMCC1.124.
10. the described soluble polysaccharide of claim 7 promotes the propagation of intestinal beneficial bacterium and/or the application in the adjusting intestinal microflora equilibrium product in preparation.
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| CN107814852A (en) * | 2017-11-08 | 2018-03-20 | 仲恺农业工程学院 | A kind of extracting method of soluble polysaccharide |
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