CN106868072A - A kind of preparation method of small-molecular-weight Dendrobium officinale polysaccharide - Google Patents
A kind of preparation method of small-molecular-weight Dendrobium officinale polysaccharide Download PDFInfo
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Abstract
The present invention relates to a kind of preparation method of small-molecular-weight Dendrobium officinale polysaccharide, it is characterised in that comprise the following steps：A, bacterial strain are activated and spread cultivation：Under aseptic condition, take appropriate amount of fluid culture medium and prepare fusarium avenaceum bacteria suspension；B, using high pressure Microfluidizer high-pressure homogeneous 5 10 times repeatedly, obtain the homogenate of dendrobium candidum broken wall；C, fermented and cultured：Appropriate amount of fluid culture medium is injected in fermentation tank, is weighed in the homogenate of dendrobium candidum broken wall, fusarium avenaceum seed liquor addition fermentation tank, fermentation completes to obtain zymotic fluid；D, polysaccharide purification：After the completion of fermentation, with decoloration active carbon, standing adsorption 1h is refiltered and removed slag, and obtains filtrate, and vacuum filtration must be precipitated, and ethanol is washed 3 times, is vacuum dried to obtain small-molecular-weight Dendrobium officinale polysaccharide.Small-molecular-weight Dendrobium officinale polysaccharide mean molecule quantity prepared by the present invention is less than 300kDa, can effectively facilitate the propagation of skin probiotics, and can improve skin surface Langerhans cell density, lifts skin immunity and phylactic power defensive power.
Description
Technical field
The present invention relates to active ingredient of natural product extractive technique field, more particularly to a kind of small-molecular-weight dendrobium candidum is more
The preparation method of sugar.
Background technology
Dendrobium candidum（Dendrobium candidum Wall.ex Lindl）It is orchid family（Orchidaceae）Dendrobium
（Dendrobium）The perennial type herbaceous plant that grows nonparasitically upon another plant, rich in dendrobium polysaccharide, dendrobine, amino acid, luxuriant and rich with fragrance class compound and various
Trace element, is the traditional rare traditional Chinese medicine of China.It is used for health products trade in dendrobium candidum, topmost active component is
Dendrobium polysaccharide, it is oral that there is the effects such as strengthening immunity of organisms, antitumor, anti-oxidant and anti-aging.Current cosmetic industry
Also begin to using Dendrobium officinale polysaccharide as cosmetic material, but be mostly to play Dendrobium officinale polysaccharide to possess macromolecule（>
1000kDa）Lasting moisture-keeping efficacy, and for small-molecular-weight dendrobium polysaccharide（Mean molecule quantity is less than 300kDa）In enhancing skin
Efficacy study in terms of immunity then has no document report substantially.
Skin is the ground stayed in that microorganism species are depended on for existence as the ecosystem, and every square centimeter to there are about 1,000,000,000 thin
Bacterium forms the protection shield of Tiny ecosystem in skin surface mutualism.On the surface of human body skin, probiotics, harmful bacteria are there is,
Probiotics mainly has the genus lactubacillus such as Bifidobacterium, lactobacillus acidophilus, and harmful bacteria is then including streptococcus and golden yellow Portugal coccus etc..
Skin health is all right, normal flora balance, probiotics accounts for population advantage, the metabolite on skin（Sebum, sweat）
Utilized by profitable strain mostly, promote pH to decline and formed with skin barrier, harmful bacteria can be suppressed, skin condition is normal；But work as
Skin flora is disorderly, harmful bacteria have the advantage status when, skin pH rises, and skin barrier function disappears, and can cause to be harmful to micro- life
Thing corrodes skin, is inflamed.
Langerhans cell (Langerhans cell, LC) is important antigen presenting cell and monokaryon in immune response
Phagocyte, can capture and process the antigen for invading skin, form Antigenic Peptide-MHC molecule compound, and pass to T cell, can
Make specific T-cell proliferative and activation, trigger immune response.
Therefore, the life of skin surface probiotics can be promoted as long as can prove that the small-molecular-weight dendrobium polysaccharide of present invention preparation
It is long, and the density for improving Langerhans cell in skin, it becomes possible to characterize its effect in terms of cutaneous immunisation power is improved.
The content of the invention
The technical problem to be solved in the present invention, is to provide a kind of small-molecular-weight iron sheet stone of mean molecule quantity less than 300kDa
The preparation method of dry measure used in former times polysaccharide, small-molecular-weight Dendrobium officinale polysaccharide prepared by the method can promote skin proliferation of probiotics, improve
Cutaneous immunisation power.
In order to solve the above technical problems, the present invention uses following technical scheme：A kind of small-molecular-weight Dendrobium officinale polysaccharide
Preparation method, it is characterised in that comprise the following steps：
A, bacterial strain are activated and spread cultivation：Under aseptic condition, it is 1 × 10 to take appropriate amount of fluid culture medium and prepare strain concentration5-1×106
The bacteria suspension of CFU/mL, the strain is fusarium avenaceum, and then culture presevation CICC 14038 takes 0.2-0.5mL bacterium and hang
Liquid is rule in PDA solid cultures primary surface, and 24-48h is cultivated at a temperature of 20-30 DEG C；The well dispersed single bacterium colony of picking is seeded to
Spread cultivation in fermentation shake flask containing Liquid Culture based component, 20-30 DEG C of cultivation temperature, shaking flask rotating speed 200r/min, oxygen supply
Culture 24-72h, oxygen concentration 0.010-0.030mol/L, obtain fusarium avenaceum fermentation seed liquid, the oat in the fermentation seed liquid
Reaping hook bacteria concentration is up to 1 × 104-1×105CFU/mL；The fluid nutrient medium is sucrose containing 15.0-30.0g/L, 5.0-10.0g/
The MS fluid nutrient mediums of L dusty yeasts；
B, dendrobium candidum broken wall treatment：Under the conditions of 50-52 DEG C, in mass ratio 1:20 mix dendrobium candidum powder and distilled water in advance
Poured into rustless steel container after conjunction, the speed of 15rpm is stirred with scraping, 6-8h is stirred, then using high pressure Microfluidizer
Repeatedly high-pressure homogeneous 5-10 times, homogenization pressure sets 90-130MPa, and 50-52 DEG C of homogenizing temperature obtains the homogenate of dendrobium candidum broken wall；
C, fermented and cultured：Appropriate amount of fluid culture medium is injected in fermentation tank, step B gained dendrobium candidum broken wall homogenate is weighed and is thrown
Enter in fermentation tank, measure dendrobium candidum broken wall and be homogenized in the fusarium avenaceum seed liquor of volume 5.0-10.0% addition fermentation tank, so
After add KH2PO4And Na2HPO4Cushioning liquid regulation fermentation tank in material liquid pH to 5.5-7.0, the oxygen supply at a temperature of 25-35 DEG C
Fermentation 3-5d, oxygen concentration control completes to obtain zymotic fluid in 0.020-0.040mol/L, fermentation；
D, polysaccharide purification：Room temperature is down to after the completion of fermentation, is helped to the diatomite of zymotic fluid quality 0.5-1.0% is added in fermentation tank
The decoloration active carbon of filtering agent and zymotic fluid quality 5.0-8.0%, standing adsorption 1h, then with plate and frame filter press filter and remove residue, must filter
Liquid, then by filtrate and Sevage reagents according to volume ratio 4.0-6.0:1.0 are mixed, and fully vibrate 30min, 3000r/
Min rotating speeds are centrifuged 10min, take supernatant；Again by ethanol in mass ratio 1.0 that supernatant and mass concentration are 95%:1.0-3.0
Mixed, stood 12-36h, vacuum filtration must be precipitated, washed 3 times with the ethanol that mass concentration is 80%, vacuum is done at 60 DEG C
Dry 24h, obtains small-molecular-weight Dendrobium officinale polysaccharide.
The measure of small-molecular-weight polysaccharide content of dendrobium candidum prepared by the present invention：With reference to phend-sulphuric acid；Small-molecular-weight iron
The molecular weight determination of skin dendrobium polysaccharide：Using Sephadex G-200 gel exclusion chromatographies；Small-molecular-weight Dendrobium officinale polysaccharide
Structural characterization need to be measured using high performance liquid chromatograph to polysaccharide after acidolysis.Result shows：Small molecule prepared by the present invention
Amount Dendrobium officinale polysaccharide yield is higher than 32.50%, and mean molecule quantity is less than 300KDa, and its main chain is by D-Glucose, D- galas
Sugar, L- rhamnoses, L-arabinose and D-MANNOSE composition.
Used oat sickle-like bacteria of the present invention（Fusarium avenaceum）It is purchased from the management of Chinese industrial Microbiological Culture Collection
The heart, bacterium numbering：CICC 14038；PDA culture medium used is purchased from Hangzhou Basebio Bio-tech Co., Ltd.；Sucrose used（CAS
57-50-1）It is purchased from Aladdin reagent（Shanghai）Co., Ltd, purity >=99.5%；Used yeast powder（CAS 119-44-8）It is purchased from
Beijing letter profit is precious along Science and Technology Ltd., purity >=99.0%；MS fluid nutrient mediums used are produced for SIGMA companies, the trade mark
M5519；Dendrobium candidum powder used is purchase from 2 years fresh branches of raw dendrobium candidum of Wenzhou District of Zhejiang Province Yueqing City, self-cleaning, is dried in the air
Do, be crushed to the mesh of 50 mesh -100；High pressure Microfluidizer used is purchased from Sea Bream jump precision optical machinery trade Co., Ltd, model
M-110EH；Fermentation tank used is purchased from the moist bioengineering equipment Co., Ltd in Nanjing, model RZY-XGX；Diatomite used is purchased from
Shengzhou Hua Li diatomite products Co., Ltd, the trade mark is CD06, SiO2(%)≥85%；It is gloomy that decoloration active carbon used is purchased from Huzhou
Activated carbon Co., Ltd is contained, the trade mark is ZL-786, methylene blue absorption affinity 12mL/0.1g；Plate and frame filter press used is purchased from Kunshan
Kun Gong environmental protection machinerys Co., Ltd of city, model XAMGZ/30U；Gas bath constant temperature oscillator used is purchased from Jiangsu Jie Ruier electrical equipment to be had
Limit company, model SHZ-82；Centrifuge used is purchased from Yu Jiao Machinery Co., Ltd.s of Zhangjagang City, model PSF-1000；It is used
Vacuum filtration machine is purchased from Shanghai Ke Sheng Instrument Ltd., model ZF-30L；Vacuum drier is purchased from Changzhou day Xiang drying and sets
Standby Co., Ltd, model SZG-350.
The small-molecular-weight Dendrobium officinale polysaccharide prepared in the present invention is to bifidobacterium adolescentis, bifidobacterium infantis, not tally double
The growing multiplication of discrimination bacillus and lactobacillus acidophilus has facilitation, by taking the product prepared in embodiment 1 as an example, tests as follows：
The 4 kinds of bacterial strains that will have been activated are respectively with 1.0% inoculum concentration（Concentration is 106CFU/mL）It is inoculated in and adds small molecule respectively
In the MRS culture mediums of amount Dendrobium officinale polysaccharide mass fraction 0,0.5%, 1.0%, 1.5%, 2.0%, 2.5%, every group of 3 parallel realities
Test, 24h is cultivated under the conditions of 38 DEG C, the absorbance of each nutrient solution at wavelength 600nm is determined using ultraviolet-uisible spectrophotometer
（OD600nm）.Bifidobacterium adolescentis (Bifidobacterium adolesentis) bacterium numbering CICC 6070, baby used
Youngster Bifidobacterium（Bifidobacterium infantis）Bacterium numbering CICC 6069, bifidobacterium bifidum
（Bifidobacterium bifidum）Bacterium numbering CICC 6071, lactobacillus acidophilus（Lactobacillus
acidophilus）Bacterium numbering CICC 6096 is purchased from Chinese industrial Microbiological Culture Collection administrative center；MRS cultures used
Base is purchased from Qingdao Hai Bo Bioisystech Co., Ltd；Ultraviolet-uisible spectrophotometer used is purchased from You Nike（Shanghai）Instrument is limited
Company, model UV-2100PC.
Bifidobacterium and lactobacillus acidophilus OD in various concentrations small-molecular-weight Dendrobium officinale polysaccharide nutrient solution600nmValue is listed in
Table 1, Different adding amount small-molecular-weight Dendrobium officinale polysaccharide is shown in Fig. 1 to the influence that Bifidobacterium and lactobacillus acidophilus grow.
The Bifidobacterium of table 1 and lactobacillus acidophilus OD in various concentrations small-molecular-weight Dendrobium officinale polysaccharide nutrient solution600nmValue
From table 1, Fig. 1, with the increase of small-molecular-weight Dendrobium officinale polysaccharide addition, typical skin probiotics youth bifid
The growth of bacillus, bifidobacterium infantis, bifidobacterium bifidum and lactobacillus acidophilus shows as the trend for increasing, and is in addition
2.0%（Mass percent）When OD600nmValue reaches maximum, when small-molecular-weight Dendrobium officinale polysaccharide addition reaches 2.5%（Quality hundred
Divide ratio）When, growth promoting function has weakened, and this is probably to cause the osmotic pressure of nutrient solution, pH value to be sent out because polysaccharide concentration is higher
Raw change and the accumulation of metabolite are limited caused by the propagation of probiotics.Experimental result is it can be shown that less than 2%（Quality
Percentage）Concentration when, small-molecular-weight Dendrobium officinale polysaccharide plays good facilitation to the growing multiplication of probiotics, is one
Plant effective prebiotics.
The present invention is also by small-molecular-weight Dendrobium officinale polysaccharide to the animal model-spontaneity closest to mankind SLE
The influence of SLE Model Female mouse NZBWF1/J epidermis Langerhans cell density come characterize its improve cutaneous immunisation power in terms of
Effect, by embodiment 2 prepare product as a example by, test it is as follows：
12 2 monthly age NZBWF1/J female mices are divided into dexamethasone（Dexamethasone）Group 6, dexamethasone
The daily intraperitoneal injections of 1.0mg/kg；Small-molecular-weight Dendrobium officinale polysaccharide group 6, small-molecular-weight Dendrobium officinale polysaccharide 50mg/kg
Daily intraperitoneal injection, continuous use 6 months is to mouse August age since the monthly age of mouse 2；SPF grades of C57BL/6 pure lines female is small
Mouse 6, August age.
The same position mouse ear of clip under mouse ether general anesthesia, is separated ear both sides skin using disecting microscope, is scraped
Except two lateral cartilageses, it is put into the biological EDTA separating liquids of pH7.2, is soaked under 38 DEG C of constant temperature and separate true epidermis, table under 2h, microscope
Skin is fixed on 20min in 4 DEG C of acetone, is rinsed 3 times with PBS phosphate buffers, each 3min.It is 1 to be separately added into concentration:50 it is anti-
Mouse Ia antigens OX3,1:In eppendorf miniature tubes, 4 DEG C are overnight for 200 anti-mouse Ia antigens OX4 and mouse skin,
PBS is rinsed, and adds biotinylated sheep anti-mouse igg serum, 38 DEG C of incubation 2h, PBS flushings, plus ABC（Avidin/biotin/
Enzyme）Compound, PBS is rinsed after being incubated 1h.Under disecting microscope, epidermis is laid in slide, plus substrate 3- amino -9- ethyl cards
Bar azoles（AEC）After color development at room temperature is dried, glycerin gelatine mounting.
Examine under a microscope, Chinese red astrocyte occur and be the positive.Epidermis Langerhans cell density（LC/
mm2）=per the average LC numbers/epidermis area in the visual field, epidermis area is determined by the eyepiece grid that micrometer is demarcated.Each experimental group is small
Mouse epidermis LC density datas are listed in table 2.Spontaneous SLE Model Females mouse NZBWF1/J used is purchased from Nanjing University-Nanjing life
Thing Medicine Research Academy；SPF grades of C57BL/6 mouse inbred lines used, female, in August age, is purchased from Guangdong Medical Lab Animal Center；
Disecting microscope is purchased from rich the regarding in Shenzhen and reaches optical instrument Co., Ltd, model BD-60T.
Each experimental mice epidermis LC mean density value tables of table 2（LC/mm2）
As shown in Table 2, compared with Dexamethasone group SLE type mouse NZBWF1/J, small-molecular-weight Dendrobium officinale polysaccharide group SLE types are small
Mouse NZBWF1/J epidermis LC density substantially increases, it may be possible to promote the generation of GM-CSF due to small-molecular-weight Dendrobium officinale polysaccharide,
Stimulate immature granulocyte, macrophage differentiation ripe simultaneously, improve the immunologic function of body, dexamethasone is used as immune suppression
Preparation is widely used in the treatment and research of the mankind and animal autoimmune disease；Compared with C57BL/6 mouse inbred lines, small point
Son amount Dendrobium officinale polysaccharide group SLE type mouse NZBWF1/J epidermis LC density also has significantly growth.
It is demonstrated experimentally that small-molecular-weight Dendrobium officinale polysaccharide can promote the raising of skin Langerhans cells density, so that can
Lifting skin immunity, lifts skin constitution.
Check experiment：
Macromolecule Dendrobium officinale polysaccharide is obtained according to following steps：By dendrobium candidum powder and distilled water in mass ratio 1:20 throw
In entering reaction vessel, be heated to 85 DEG C, extract 3h, after the completion of suction filtration, repeat extraction 3 times, collect filtrate, vacuum drying is concentrated into
The 1/5 of original volume, then according to filtrate：Sevage reagent=6.0:1.0（Volume ratio）Mixing, fully vibrates 30min,
3000r/min is centrifuged 10min, supernatant is taken, according still further to supernatant：95% ethanol=1.0:2.0 (mass ratioes) add ethanol, quiet
24h is put, vacuum filtration machine is filtered must be precipitated, washed with 80% ethanol 3 times, 60 DEG C of vacuum drying 24h obtain macromolecule iron sheet stone
Dry measure used in former times polyose.
, between 730-1180kDa, its main chain is by D-Glucose, D- galas for gained macromolecule Dendrobium officinale polysaccharide molecular weight
Sugar, L- rhamnoses, L-arabinose and D-MANNOSE composition.
Research macromolecule Dendrobium officinale polysaccharide is to bifidobacterium adolescentis, bifidobacterium infantis, bifidobacterium bifidum and thermophilic
The growing multiplication effect of Lactobacillus lactis, ibid, experimental result is shown in Table 3 to experimental technique.
The Bifidobacterium of table 3 and lactobacillus acidophilus OD in various concentrations macromolecule Dendrobium officinale polysaccharide nutrient solution600nmValue
From table 3, Fig. 2, with the increase of macromolecule Dendrobium officinale polysaccharide addition, typical skin probiotics youth bifid
The growth of bacillus, bifidobacterium infantis, bifidobacterium bifidum and lactobacillus acidophilus does not have and shows obvious growth trend, real
Test result to show, macromolecule dendrobium polysaccharide (730-1180kDa) does not play the growing multiplication of probiotics obvious promotion
Effect.
It is thin to spontaneous SLE Model Females mouse NZBWF1/J epidermis Langerhans by macromolecule Dendrobium officinale polysaccharide
The influence of born of the same parents' density characterizes its effect in terms of cutaneous immunisation power, and ibid, experimental result is shown in Table 4 to experimental technique.
Each experimental mice epidermis LC mean density value tables of table 4（LC/mm2）
As shown in Table 4, compared with C57BL/6 mouse inbred lines, macromolecule Dendrobium officinale polysaccharide group SLE type mouse NZBWF1/J tables
Skin LC density does not rise appreciably, and experiment shows, it is bright that macromolecule dendrobium polysaccharide (730-1180kDa) can not be obviously promoted skin
The raising of Ge Hansi cell densities, does not almost work to lifting skin immunity.
To sum up, experiment shows by contrast, growing multiplication and skin Langerhans of the macromolecule dendrobium polysaccharide in probiotics
The raising aspect of cell density is without obvious facilitation, it is impossible to enough effectively lifting skin immunities.
The present invention carries out broken wall treatment to dendrobium candidum powder first with high pressure microjet homogeneous method, recycles and is selected from iron
The fusarium avenaceum of skin stem of noble dendrobium endogenetic fungus prepares small-molecular-weight Dendrobium officinale polysaccharide to ferment, and its mean molecule quantity is less than
300KDa.By the growth to small-molecular-weight Dendrobium officinale polysaccharide in probiotics and mouse Langerhans cell（(Langerhans
Cell, LC)）Density in terms of influence studied, as a result show that small-molecular-weight Dendrobium officinale polysaccharide can effectively facilitate skin
The propagation of probiotics, and skin surface Langerhans cell density can be improved, so as to lift skin immunity and phylactic power defensive power, maintain
Skin health state.
The small-molecular-weight Dendrobium officinale polysaccharide preparation method that the present invention is provided, it is main using being fermented using fusarium avenaceum, make
Standby small molecule Dendrobium officinale polysaccharide, average yield is higher than 32.50%.
In sum, the small-molecular-weight Dendrobium officinale polysaccharide mean molecule quantity that prepared by the present invention is less than 300kDa, Ke Yiyou
Effect promotes the propagation of skin probiotics, and can improve skin surface Langerhans cell density, thus lifted skin immunity and
Phylactic power defensive power, maintains skin health state.
Brief description of the drawings
The influence that Fig. 1 Different adding amount small-molecular-weight Dendrobium officinale polysaccharides grow to Bifidobacterium and lactobacillus acidophilus；
The influence that Fig. 2 Different adding amount macromolecule Dendrobium officinale polysaccharides grow to Bifidobacterium and lactobacillus acidophilus.
Specific embodiment
The present invention is further described with reference to specific embodiment, but the present invention is not limited in these embodiments.
Embodiment 1：
A, bacterial strain are activated and spread cultivation：Under aseptic condition, draw appropriate amount of fluid culture medium with aseptic dropper and instill equipped with dendrobium candidum
It is 1 × 10 that strain concentration is configured in the strain tube of endogenetic fungus fusarium avenaceum freeze-dried powder6The bacteria suspension of CFU/mL, then
Take 0.2mL bacteria suspensions to be rule in PDA solid cultures primary surface, 48h is cultivated under the conditions of 25 DEG C, the well dispersed single bacterium colony of picking connects
Plant into the fermentation shake flask containing Liquid Culture based component and spread cultivation, 25 DEG C, oxygen supply culture 48h, oxygen under the conditions of 200r/min
Concentration 0.030mol/L, is obtained fermentation seed liquid, and wherein fusarium avenaceum concentration is 1 × 105CFU/mL, Liquid Culture used
Base is 30.0g/L containing sucrose, the MS fluid nutrient mediums of dusty yeast 8.0g/L；
B, dendrobium candidum broken wall treatment：Under the conditions of 50 DEG C, poured into after 3kg dendrobium candidums powder is pre-mixed with 60L distilled water
In rustless steel container, the speed of 15rpm is stirred with scraping, stir 6h, then using high pressure Microfluidizer, pressure is set to
90MPa, temperature is 50 DEG C, high-pressure homogeneous 8 times repeatedly, obtains the homogenate of dendrobium candidum broken wall；
C, fermented and cultured：Gained dendrobium candidum broken wall in step B is homogenized in input fermentation tank, by connecing for homogenate volume 7.0%
The amount of kind is inoculated with into fusarium avenaceum seed liquor, the synchronous rapid A of Liquid Culture based component in fermentation tank, and adds KH2PO4And Na2HPO4
Cushioning liquid adjusts material liquid pH=7.0, the oxygen supply fermentation 3d, oxygen concentration 0.040mol/L at 35 DEG C of temperature；
D, polysaccharide purification：Room temperature is down to after the completion of fermentation, to the super-cell that zymotic fluid quality 0.5% is added in fermentation tank
And the decoloration active carbon of zymotic fluid quality 7.0%, standing adsorption 1h, then with plate and frame filter press filter and remove residue, obtain filtrate；Then press
According to filtrate：Sevage reagent=4.0:1.0（Volume ratio）Mixing, fully vibrates 30min, 3000r/min centrifugation 10min, takes supernatant
Liquid, according still further to supernatant：95% ethanol=1.0:2.0 (mass ratioes) add ethanol, stand 24h, and vacuum filtration machine is filtered must sink
Form sediment, washed with 80% ethanol 3 times, 60 DEG C of vacuum drying 24h obtain small-molecular-weight Dendrobium officinale polysaccharide product 1005.6g.
Small-molecular-weight Dendrobium officinale polysaccharide yield manufactured in the present embodiment be 33.52%, mean molecule quantity 221.60kDa, its
Main chain is made up of D-Glucose, D- galactolipins, L- rhamnoses, L-arabinose and D-MANNOSE, and mol ratio is 3.112:
2.013 : 1.561: 1.034 : 2.487。
Embodiment 2：
A, bacterial strain are activated and spread cultivation：Under aseptic condition, draw appropriate amount of fluid culture medium with aseptic dropper and instill equipped with dendrobium candidum
It is 8.5 × 10 that strain concentration is configured in the strain tube of endogenetic fungus fusarium avenaceum freeze-dried powder5The bacteria suspension of CFU/mL, then
Take 0.4mL bacteria suspensions to be rule in PDA solid cultures primary surface, 36h is cultivated under the conditions of 20 DEG C, the well dispersed single bacterium colony of picking connects
Plant into the fermentation shake flask containing Liquid Culture based component and spread cultivation, 20 DEG C, oxygen supply culture 72h, oxygen under the conditions of 200r/min
Concentration 0.010mol/L, is obtained fermentation seed liquid, and wherein fusarium avenaceum concentration is 6.5 × 104CFU/mL, Liquid Culture used
Base is 18.0g/L containing sucrose, the MS fluid nutrient mediums of dusty yeast 10.0g/L；
B, dendrobium candidum broken wall treatment：Under the conditions of 52 DEG C, poured into after 3kg dendrobium candidums powder is pre-mixed with 60L distilled water
In rustless steel container, the speed of 15rpm is stirred with scraping, stir 8h, then using high pressure Microfluidizer, pressure is set to
130MPa, temperature is 52 DEG C, high-pressure homogeneous 10 times repeatedly, obtains the homogenate of dendrobium candidum broken wall；
C, fermented and cultured：Gained dendrobium candidum broken wall in step B is homogenized in input fermentation tank, by connecing for homogenate volume 10.0%
The amount of kind is inoculated with into fusarium avenaceum seed liquor, the synchronous rapid A of Liquid Culture based component in fermentation tank, and adds KH2PO4And Na2HPO4
Cushioning liquid adjusts material liquid pH=5.5, the oxygen supply fermentation 5d, oxygen concentration 0.030mol/L at 25 DEG C of temperature；
D, polysaccharide purification：Room temperature is down to after the completion of fermentation, to the super-cell that zymotic fluid quality 0.8% is added in fermentation tank
And the decoloration active carbon of zymotic fluid quality 8.0%, standing adsorption 1h, then with plate and frame filter press filter and remove residue, obtain filtrate；Then according to
Filtrate：Sevage reagent=5.0:1.0（Volume ratio）Mixing, fully vibrates 30min, 3000r/min centrifugation 10min, takes supernatant
Liquid, according still further to supernatant：95% ethanol=1.0:3.0 (mass ratioes) add ethanol, stand 36h, and vacuum filtration machine is filtered must sink
Form sediment, washed with 80% ethanol 3 times, 60 DEG C of vacuum drying 24h obtain small-molecular-weight Dendrobium officinale polysaccharide product 1035.6g.
Small-molecular-weight Dendrobium officinale polysaccharide yield manufactured in the present embodiment is 34.52%, mean molecule quantity 1.5kDa, its master
Chain is made up of D-Glucose, D- galactolipins, L- rhamnoses, L-arabinose and D-MANNOSE, and mol ratio is 2.452: 1.025
: 2.231: 3.154 : 1.562。
Embodiment 3：
A, bacterial strain are activated and spread cultivation：Under aseptic condition, draw appropriate amount of fluid culture medium with aseptic dropper and instill equipped with dendrobium candidum
It is 1 × 10 that strain concentration is configured in the strain tube of endogenetic fungus fusarium avenaceum freeze-dried powder5The bacteria suspension of CFU/mL, Ran Houqu
0.5mL bacteria suspensions are rule in PDA solid cultures primary surface, and 24h, the well dispersed single bacterium colony inoculation of picking are cultivated under the conditions of 30 DEG C
Spread cultivation into the fermentation shake flask containing Liquid Culture based component, 30 DEG C, oxygen supply culture 24h under the conditions of 200r/min, oxygen is dense
Degree 0.020mol/L, is obtained fermentation seed liquid, and wherein fusarium avenaceum concentration is 1 × 104CFU/mL, fluid nutrient medium used is
The MS fluid nutrient mediums of 15.0g/L containing sucrose, dusty yeast 10.0g/L；
B, dendrobium candidum broken wall treatment：Under the conditions of 51 DEG C, poured into after 3kg dendrobium candidums powder is pre-mixed with 60L distilled water
In rustless steel container, the speed of 15rpm is stirred with scraping, stir 6.5h, then using high pressure Microfluidizer, pressure is set to
100MPa, temperature is 51 DEG C, high-pressure homogeneous 5 times repeatedly, obtains the homogenate of dendrobium candidum broken wall；
C, fermented and cultured：Gained dendrobium candidum broken wall in step B is homogenized in input fermentation tank, by connecing for homogenate volume 5.0%
The amount of kind is inoculated with into fusarium avenaceum seed liquor, the synchronous rapid A of Liquid Culture based component in fermentation tank, and adds KH2PO4And Na2HPO4
Cushioning liquid adjusts material liquid pH=6.0, the oxygen supply fermentation 4d, oxygen concentration 0.020mol/L at 30 DEG C of temperature；
D, polysaccharide purification：Room temperature is down to after the completion of fermentation, to the super-cell that zymotic fluid quality 0.7% is added in fermentation tank
And the decoloration active carbon of zymotic fluid quality 5.0%, standing adsorption 1h, then with plate and frame filter press filter and remove residue, obtain filtrate；Then according to
Filtrate：Sevage reagent=6.0:1.0（Volume ratio）Mixing, fully vibrates 30min, 3000r/min centrifugation 10min, takes supernatant
Liquid, according still further to supernatant：95% ethanol=1.0:1.0 (mass ratioes) add ethanol, stand 24h, and vacuum filtration machine is filtered must sink
Form sediment, washed with 80% ethanol 3 times, 60 DEG C of vacuum drying 24h obtain small-molecular-weight Dendrobium officinale polysaccharide product 975.0g.
Small-molecular-weight Dendrobium officinale polysaccharide yield manufactured in the present embodiment be 32.50%, mean molecule quantity 110.50kDa, its
Main chain is made up of D-Glucose, D- galactolipins, L- rhamnoses, L-arabinose and D-MANNOSE, and mol ratio is 1.547:
1.751: 2.323: 2.872: 1.310。
Embodiment 4：
A, bacterial strain are activated and spread cultivation：Under aseptic condition, draw appropriate amount of fluid culture medium with aseptic dropper and instill equipped with dendrobium candidum
It is 4.2 × 10 that strain concentration is configured in the strain tube of endogenetic fungus fusarium avenaceum freeze-dried powder5The bacteria suspension of CFU/mL, then
Take 0.3mL bacteria suspensions to be rule in PDA solid cultures primary surface, 24h is cultivated under the conditions of 28 DEG C, the well dispersed single bacterium colony of picking connects
Plant into the fermentation shake flask containing Liquid Culture based component and spread cultivation, 28 DEG C, oxygen supply culture 30h, oxygen under the conditions of 200r/min
Concentration 0.020mol/L, is obtained fermentation seed liquid, and wherein fusarium avenaceum concentration is 3 × 104CFU/mL, fluid nutrient medium used
It is 18.0g/L containing sucrose, the MS fluid nutrient mediums of dusty yeast 5.0g/L；
B, dendrobium candidum broken wall treatment：Under the conditions of 50 DEG C, poured into after 3kg dendrobium candidums powder is pre-mixed with 60L distilled water
In rustless steel container, the speed of 15rpm is stirred with scraping, stir 6.5h, then using high pressure Microfluidizer, pressure is set to
110MPa, temperature is 50 DEG C, high-pressure homogeneous 7 times repeatedly, obtains the homogenate of dendrobium candidum broken wall；
C, fermented and cultured：Gained dendrobium candidum broken wall in step B is homogenized in input fermentation tank, by connecing for the homogenate % of volume 8.5
The amount of kind is inoculated with into fusarium avenaceum seed liquor, the synchronous rapid A of Liquid Culture based component in fermentation tank, and adds KH2PO4And Na2HPO4
Cushioning liquid adjusts material liquid pH=6.8, the oxygen supply fermentation 4d, oxygen concentration 0.030mol/L at 28 DEG C of temperature；
D, polysaccharide purification：Room temperature is down to after the completion of fermentation, to the super-cell that zymotic fluid quality 0.6% is added in fermentation tank
And the decoloration active carbon of zymotic fluid quality 6.5%, standing adsorption 1h, then with plate and frame filter press filter and remove residue, obtain filtrate；Then according to
Filtrate：Sevage reagent=4.5:1.0（Volume ratio）Mixing, fully vibrates 30min, 3000r/min centrifugation 10min, takes supernatant
Liquid, according still further to supernatant：95% ethanol=1.0:2.0 (mass ratioes) add ethanol, stand 12h, and vacuum filtration machine is filtered must sink
Form sediment, washed with 80% ethanol 3 times, 60 DEG C of vacuum drying 24h obtain small-molecular-weight Dendrobium officinale polysaccharide product 988.5g.
Small-molecular-weight Dendrobium officinale polysaccharide yield manufactured in the present embodiment is 32.95%, mean molecule quantity 300kDa, its master
Chain is made up of D-Glucose, D- galactolipins, L- rhamnoses, L-arabinose and D-MANNOSE, and mol ratio is 3.005: 2.645
: 1.006: 2.336 : 1.496。
Embodiment 5：
A, bacterial strain are activated and spread cultivation：Under aseptic condition, draw appropriate amount of fluid culture medium with aseptic dropper and instill equipped with dendrobium candidum
It is 7.8 × 10 that strain concentration is configured in the strain tube of endogenetic fungus fusarium avenaceum freeze-dried powder5The bacteria suspension of CFU/mL, then
Take 0.25mL bacteria suspensions to be rule in PDA solid cultures primary surface, 30h, the well dispersed single bacterium colony of picking are cultivated under the conditions of 23 DEG C
It is seeded in the fermentation shake flask containing Liquid Culture based component and is spread cultivation, 23 DEG C, oxygen supply culture 72h under the conditions of 200r/min,
Oxygen concentration 0.030mol/L, is obtained fermentation seed liquid, and wherein fusarium avenaceum concentration is 5.3 × 104CFU/mL, liquid training used
Foster base is 28.0g/L containing sucrose, the MS fluid nutrient mediums of dusty yeast 6.0g/L；
B, dendrobium candidum broken wall treatment：Under the conditions of 52 DEG C, poured into after 3kg dendrobium candidums powder is pre-mixed with 60L distilled water
In rustless steel container, the speed of 15rpm is stirred with scraping, stir 6-8h, then using high pressure Microfluidizer, pressure is set to
120MPa, temperature is 52 DEG C, high-pressure homogeneous 8 times repeatedly, obtains the homogenate of dendrobium candidum broken wall；
C, fermented and cultured：Gained dendrobium candidum broken wall in step B is homogenized in input fermentation tank, by connecing for homogenate volume 9.0%
The amount of kind is inoculated with into fusarium avenaceum seed liquor, the synchronous rapid A of Liquid Culture based component in fermentation tank, and adds KH2PO4And Na2HPO4
Cushioning liquid adjusts material liquid pH=6.0, the oxygen supply fermentation 4d, oxygen concentration 0.035mol/L at 28 DEG C of temperature；
D, polysaccharide purification：Room temperature is down to after the completion of fermentation, to the super-cell that zymotic fluid quality 1.0% is added in fermentation tank
And the decoloration active carbon of zymotic fluid quality 7.5%, standing adsorption 1h, then with plate and frame filter press filter and remove residue, obtain filtrate；Then according to
Filtrate：Sevage reagent=5.5:1.0（Volume ratio）Mixing, fully vibrates 30min, 3000r/min centrifugation 10min, takes supernatant
Liquid, according still further to supernatant：95% ethanol=1.0:2.5 (mass ratioes) add ethanol, stand 30h, and vacuum filtration machine is filtered must sink
Form sediment, washed with 80% ethanol 3 times, 60 DEG C of vacuum drying 24h obtain small-molecular-weight Dendrobium officinale polysaccharide product 1026.0g.
Small-molecular-weight Dendrobium officinale polysaccharide yield manufactured in the present embodiment be 34.20%, weight average molecular weight 98.74kDa, its
Main chain is made up of D-Glucose, D- galactolipins, L- rhamnoses, L-arabinose and D-MANNOSE, and mol ratio is 1.894:
2.620 : 3.128: 1.772: 2.315。
Claims (1)
1. a kind of preparation method of small-molecular-weight Dendrobium officinale polysaccharide, it is characterised in that comprise the following steps：
A, bacterial strain are activated and spread cultivation：Under aseptic condition, it is 1 × 10 to take appropriate amount of fluid culture medium and prepare strain concentration5-1×106
The bacteria suspension of CFU/mL, the strain is fusarium avenaceum, and then culture presevation CICC 14038 takes 0.2-0.5mL bacterium and hang
Liquid is rule in PDA solid cultures primary surface, and 24-48h is cultivated at a temperature of 20-30 DEG C；The well dispersed single bacterium colony of picking is seeded to
Spread cultivation in fermentation shake flask containing Liquid Culture based component, 20-30 DEG C of cultivation temperature, shaking flask rotating speed 200r/min, oxygen supply
Culture 24-72h, oxygen concentration 0.010-0.030mol/L, obtain fusarium avenaceum fermentation seed liquid, the oat in the fermentation seed liquid
Reaping hook bacteria concentration is up to 1 × 104-1×105CFU/mL；The fluid nutrient medium is sucrose containing 15.0-30.0g/L, 5.0-10.0g/
The MS fluid nutrient mediums of L dusty yeasts；
B, dendrobium candidum broken wall treatment：Under the conditions of 50-52 DEG C, in mass ratio 1:20 mix dendrobium candidum powder and distilled water in advance
Poured into rustless steel container after conjunction, the speed of 15rpm is stirred with scraping, 6-8h is stirred, then using high pressure Microfluidizer
Repeatedly high-pressure homogeneous 5-10 times, homogenization pressure sets 90-130MPa, and 50-52 DEG C of homogenizing temperature obtains the homogenate of dendrobium candidum broken wall；
C, fermented and cultured：Appropriate amount of fluid culture medium is injected in fermentation tank, step B gained dendrobium candidum broken wall homogenate is weighed and is thrown
Enter in fermentation tank, measure dendrobium candidum broken wall and be homogenized in the fusarium avenaceum seed liquor of volume 5.0-10.0% addition fermentation tank, so
After add KH2PO4And Na2HPO4Cushioning liquid regulation fermentation tank in material liquid pH to 5.5-7.0, the oxygen supply at a temperature of 25-35 DEG C
Fermentation 3-5d, oxygen concentration control completes to obtain zymotic fluid in 0.020-0.040mol/L, fermentation；
D, polysaccharide purification：Room temperature is down to after the completion of fermentation, is helped to the diatomite of zymotic fluid quality 0.5-1.0% is added in fermentation tank
The decoloration active carbon of filtering agent and zymotic fluid quality 5.0-8.0%, standing adsorption 1h, then with plate and frame filter press filter and remove residue, must filter
Liquid, then by filtrate and Sevage reagents according to volume ratio 4.0-6.0:1.0 are mixed, and fully vibrate 30min, 3000r/
Min rotating speeds are centrifuged 10min, take supernatant；Again by ethanol in mass ratio 1.0 that supernatant and mass concentration are 95%:1.0-3.0
Mixed, stood 12-36h, vacuum filtration must be precipitated, washed 3 times with the ethanol that mass concentration is 80%, vacuum is done at 60 DEG C
Dry 24h, obtains small-molecular-weight Dendrobium officinale polysaccharide.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104844723A (en) * | 2015-06-05 | 2015-08-19 | 云南金九地生物科技有限公司 | Preparation method and application of dendrobium officinale extract |
-
2017
- 2017-03-13 CN CN201710145503.1A patent/CN106868072B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104844723A (en) * | 2015-06-05 | 2015-08-19 | 云南金九地生物科技有限公司 | Preparation method and application of dendrobium officinale extract |
Non-Patent Citations (4)
| Title |
|---|
| TAO-BIN HE等: "Structural characterization and immunomodulating activity of polysaccharide from Dendrobium officinale", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
| 付伟丽等: "铁皮石斛多糖研究进展", 《内江师范学院学报》 * |
| 张秒高等: "铁皮石斛多糖原料特性和加工方面的研究进展", 《安徽农业科学》 * |
| 罗秋莲等: "铁皮石斛多糖的分离纯化及其结构研究", 《广西大学学报（自然科学版）》 * |
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