CN103880970A - Method utilizing ion-exchange resin to decolor crude polysaccharide in ganoderma sinensis submerged fermentation mycelium - Google Patents
Method utilizing ion-exchange resin to decolor crude polysaccharide in ganoderma sinensis submerged fermentation mycelium Download PDFInfo
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Abstract
The invention discloses a method utilizing ion-exchange resin to decolor crude polysaccharide in ganoderma sinensis submerged fermentation mycelium. The method comprises the following steps: (a) carrying out a pretreatment on crude polysaccharide in ganoderma sinensis submerged fermentation mycelium so as to obtain refined ganoderma sinensis polysaccharide with a molecular range of 30 KD to 300 KD; (b) transferring the refined ganoderma sinensis polysaccharide obtained in the step (a) to an ion-exchange resin to carrying out a decoloring treatment so as to obtain the discolored ganoderma sinensis polysaccharide, wherein in the decoloring treatment, the conditions are as follows: the decoloring time is 2 to 4 hours, the using amount of resin is 0.25 to 0.75 g/mL, the decoloring temperature is 25 to 35 DEG C, and the pH value is 6 to 7. The actual decoloring rate of the discolored ganoderma sinensis polysaccharide is 85% to 90%, and the recovery rate of polysaccharide is 65% to 70%.
Description
Technical field
The present invention relates to Chemistry for Chinese Traditional Medicine field, be specifically related to a kind of method that adopts ion exchange resin to decolour to purple sesame liquid submerged fermentation mycelium polysaccharides liquid.
Background technology
Purple sesame (Ganoderma sinense) be China rare Chinese medicine glossy ganoderma one (Chinese Pharmacopoeia Commission. Beijing of Pharmacopoeia of People's Republic of China [S] version in 2010: Chinese Medicine science and technology press, 2010:174), be the treasure in motherland's pools of traditional Chinese medicine, have the reputation of " celestial grass ".Pharmacology and clinical study at all times all proves, effect that purple sesame truly has and prevents and cures diseases, promotes longevity.The Compendium of Material Medica of the Shennong's Herbal of the Eastern Han Dynasty, the famous medical scholar's LI Shi-Zhen of the Ming Dynasty, all has detailed very sure record to effect of purple sesame.Modern pharmacology and clinical practice have further confirmed the pharmacological action of purple sesame, large quantity research shows that purple sesame polysaccharide is one of its main active ingredient, there is immunomodulatory, antitumor (Yang Guohong, Yang Yifang, Jin Juandi. the antitumor active site research [J] of purple sesame liquid submerged fermentation liquid. herbal medicine, 2008,39 (6): 877-880), anti-oxidant, antiviral, hypoglycemic and the pharmacological action such as protect the liver, and confirm that purple sesame polysaccharide is that purple sesame is strengthened the body resistance to consolidate the constitution, strengthening by means of tonics, the main component of promoting longevity.
Purple sesame polysaccharide product color is dark, pigment is more, have a strong impact on purity, colourity, the biological activity of polysaccharide, also hindered the further investigation to polysaccharide structures and its Relation between Biological Activity, therefore (Luo Xi should decolour in purge process, Tang Qing nine *, Jinsong ZHANG, etc. ganoderan resin method decoloration process is optimized [J]. Food science, 2011,32 (16): 5-10).Tradition decoloring method generally has charcoal absorption, hydrogen peroxide oxidation method etc., wherein activated carbon method bleaching time is longer, polysaccharide retention rate is low, and hydrogen peroxide is strong oxidizer, easily destroys polysaccharide structures, affect its activity (Yuan Hongbo, Jinsong ZHANG, Jia Wei, etc. utilize the research [J] of macroporous resin to the decolouring of lower molecular weight ganoderan. foodstuffs industry science and technology, 2009,30 (3): 204-206).
Macroporous resin decolouring technology is often applied to the decolouring of Chinese herbal medicine effective ingredients in recent years.Macroporous resin has that physical and chemical stability is high, specific surface area is large, processing power is large, selectivity is good, rate of adsorption is fast, desorption condition is gentle, manipulation of regeneration is convenient, life cycle is long, the not high plurality of advantages of cost, therefore can utilize the selective adsorption function of resin that the pigment in polysaccharide is removed.Domestic and foreign literature takes orthogonal test model to be optimized ganoderan decoloration process mostly, and based on response surface method (response surface methodology, RSM), the report of purple sesame fermentation polysaccharide decolouring Optimization Technology is not yet occurred.
Summary of the invention
The present invention is on the basis of single factor experiment, utilize response surface analysis method to be optimized purple sesame fermentation polysaccharide decoloration process, and in conjunction with actual production conditions, determine rational decoloration process parameter, the separation and purification that purport is follow-up purple sesame polysaccharide and activity research provide basis, simultaneously for decolouring industrialization provides certain theoretical foundation.
Therefore, the invention provides a kind of method that adopts ion exchange resin to decolour to purple sesame liquid submerged fermentation mycelium Crude polysaccharides, comprise the steps:
A) purple sesame liquid submerged fermentation mycelium Crude polysaccharides is carried out to pre-treatment, obtaining molecular weight ranges is the refining purple sesame polysaccharide of 30KD~300KD;
B) refining purple sesame polysaccharide step a) being obtained is placed in ion exchange resin, at bleaching time 2~4h, resin demand 0.25g~0.75g/10ml, decolours under the decolorization condition of 25~35 DEG C of bleaching temperatures, pH value 6~7, obtains decolouring purple sesame polysaccharide afterwards.
Term used in the present invention " purple sesame liquid submerged fermentation mycelium Crude polysaccharides " refers to the purple sesame Crude polysaccharides that utilizes liquid submerged fermentation technology to extract from mycelium.Concrete preparation method is that the tank liquid of putting that fermentation is completed carries out Plate Filtration, discards extracellular fluid, gets and filters rear mycelium; By mycelium deionized water wash, being washed till sheet frame filtrate is that water white transparency also no longer produces bubble, filtration is weighed, mycelium after washing and filtering is weighed, add appropriate multiple deionized water by mycelium weight, put after steeping tank extracts for several times with certain hour heating and separate in Plate Filtration, through scraper plate, concentrated and single tank is concentrated into 85% ethanol that adds the appropriate multiple quality of this concentrated solution after a certain amount of and places 12h in 4 DEG C extracting solution; Alcohol hypostasis is put to whizzer 1200r/min centrifugation, taking precipitate; To wet after the sabot of alcohol hypostasis and put into box baker, and control vacuum tightness and be heated to 30~40 DEG C and take out alcohol hypostasis after drying 2h, pulverize; Again sabot after pulverizing, then put into baking oven, is slowly warming up to 65 DEG C and dries 2h, finally purple sesame liquid submerged fermentation mycelium Crude polysaccharides.
According to the present invention one preferred embodiment, step a) described pre-treatment comprises the steps:
1) purple sesame liquid submerged fermentation mycelium Crude polysaccharides is dissolved in the water by liquid ratio for 150: 1~250: 1;
2) centrifugal more than twice or twice with the speed of 4000~5000rpm/min, get supernatant liquor, each centrifugal 20~40min;
3) supernatant liquor is crossed 30KD hollow fiber ultrafiltration membrane under 20~40 DEG C of temperature, pressure 0.02~0.06Mpa condition, after trapped fluid dilution, crosses 300KD hollow fiber ultrafiltration membrane, after ultrafiltrated concentrated frozen is dry, must refine purple sesame polysaccharide.
Term used in the present invention " liquid ratio " refers to that the volume of " liquid " is divided by the quality of " material ", and " liquid " refers to water here, the ml of unit, and " material " refers to purple sesame Crude polysaccharides, the g of unit.
According to a particularly preferred embodiment of the present, step 1) described in liquid ratio be 180: 1~220: 1, more preferably 200: 1.
According to a particularly preferred embodiment of the present, step 1) described in water be deionized water.
According to a particularly preferred embodiment of the present, step 2) described in centrifugation rate be 4500~5000rpm/min, more preferably 5000rpm/min; Centrifugation time is 25~35min, more preferably 30min.
According to a particularly preferred embodiment of the present, step b) described ion exchange resin is LS-850 anionite-exchange resin.
According to a particularly preferred embodiment of the present, step b) middle bleaching time is 2.5h, and resin demand is 0.75g/10ml, and bleaching temperature is 28 DEG C, and pH value is 6.
Decoloring method of the present invention, for resin and purple sesame polysaccharide soln are inserted in tool plug Erlenmeyer flask, is then placed in Erlenmeyer flask shaking table and carries out.The rotating speed of described shaking table is 100~140r/min, preferably 120r/min.
Obtain decolouring purple sesame polysaccharide afterwards according to the inventive method, its actual percent of decolourization is 85%~90%, and polysaccharide recovery is 65%~70%; Under the best decolorization condition of the present invention, the average percent of decolourization of purple sesame polysaccharide is 87.8%, and polysaccharide recovery is 65%~70%.
Brief description of the drawings
Fig. 1 is different sorts resin described in embodiment 1 decolorizing effect to purple sesame polysaccharide;
Fig. 2 is the impact on purple sesame polysaccharide decolorizing effect of bleaching time described in embodiment 2;
Fig. 3 is the impact on purple sesame polysaccharide decolorizing effect of resin demand described in embodiment 3;
Fig. 4 is the impact on purple sesame polysaccharide decolorizing effect of temperature described in embodiment 4;
Fig. 5 is the impact on purple sesame polysaccharide decolorizing effect of pH value described in embodiment 5.
Embodiment
Starting material and reagent
Be used for the preparation of the purple sesame liquid submerged fermentation mycelium Crude polysaccharides of following examples 1-5: the tank liquid of putting that fermentation is completed carries out Plate Filtration, discards extracellular fluid, get and filter rear mycelium; By mycelium deionized water wash, being washed till sheet frame filtrate is that water white transparency also no longer produces bubble, filtration is weighed, mycelium after washing and filtering is weighed, add appropriate multiple deionized water by mycelium weight, put after steeping tank extracts for several times with certain hour heating and separate in Plate Filtration, through scraper plate, concentrated and single tank is concentrated into 85% ethanol that adds the appropriate multiple quality of this concentrated solution after a certain amount of and places 12h in 4 DEG C extracting solution; Alcohol hypostasis is put to whizzer 1200r/min centrifugation, after taking precipitate sabot, put into box baker, control vacuum tightness and be heated to take out alcohol hypostasis after 30~40 DEG C of oven dry 2h, pulverize; Again sabot after pulverizing, then put into baking oven, is slowly warming up to 65 DEG C and dries 2h, finally purple sesame liquid submerged fermentation mycelium Crude polysaccharides.
Be used for the preparation of the refining purple sesame polysaccharide soln of following examples 1-5: purple sesame liquid submerged fermentation mycelium Crude polysaccharides is dissolved in distilled water by liquid ratio at 200: 1, centrifugal (5000rpm/min) processes twice, each 20 minutes, get supernatant liquor and cross 30KD hollow fiber ultrafiltration membrane, after trapped fluid dilution, cross 300KD hollow fiber ultrafiltration membrane, after ultrafiltrated concentrated frozen is dry, must refine polysaccharide GS-B, measuring relative molecular weight through HPLC is 30KD~300KD; Get above-mentioned purple sesame polysaccharide GS-B 1.0g, be dissolved in 100ml (liquid ratio 100: 1) distilled water, leave standstill 30 minutes, centrifugal (5000rpm/min) processes 30 minutes, gets supernatant liquor and must refine purple sesame polysaccharide soln;
Ion exchange resin D101, AB-8, XAD7-HP, LS-840, LS-850 (Shaanxi Lan Shen Special Resin company limited); Phenol, glucose, sulfuric acid, hydrochloric acid, sodium hydroxide analytical pure (reagent company limited of traditional Chinese medicines group).
Equipment and instrument:
UV-2500PC (Japanese Shimadzu); AB204-S type electronic balance (Switzerland MettlerFoledo); PHS-3C type PH counts (Shanghai Lei Ci Instrument Ltd.); The double-deck high-capacity bottle swingging machine of ZHWY-3122A (Shanghai ZHICHENG Anaiytical Instrument Manufacturing Co., Ltd.); BT01-100 constant flow pump (Baoding LanGe constant flow pump Co., Ltd); BS-100A type automatic collection instrument (Shanghai Hu Xi analytical instrument factory).
The detection of polysaccharide: polysaccharide adopts phenolsulfuric acid method to detect.Accurately take 105 DEG C of dextrose anhydrous 0.2540g that are dried to constant weight, dissolve constant volume and in 1000ml volumetric flask, be made into the glucose standardized solution that concentration is 0.254mg/ml, accurate measuring glucose standardized solution adding distil water to cumulative volume is 300 μ l; Taking the test tube of adding distil water as blank, then respectively add 5% phenol solution 300 μ l, sulfuric acid 4.0ml, shakes up rapidly and at room temperature places 30min, in 488nm place mensuration absorbancy.Taking glucose content (μ is g) as X-coordinate, taking absorbance A as ordinate zou, drawing standard curve, regression equation is A=0.011x+0.081 (R
2=0.9982)
Calculate polysaccharide recovery (R by formula (1)
1).
R
1=M/M
0*100% (1)
In formula:
Polysaccharide content after M-decolouring, g;
M
0polysaccharide content before-decolouring, g.
Percent of decolourization (R
2) mensuration: taking distilled water as blank, polysaccharide soln is carried out to all-wave is long to be detected, selects to absorb maximum (263nm) and locate to measure absorbancy.
R
2=(A
0-A)/A
0*100% (2)
In formula:
A
0absorbancy before-decolouring;
Absorbancy after A-decolouring.
Embodiment 1: Resin sieving selection
Get 5 kinds of each 0.5g of the resin of processing, insert in tool plug Erlenmeyer flask, every bottle adds 10ml to refine purple sesame polysaccharide soln, is placed in shaking table, rotating speed 120r/min, and 30 DEG C of constant temperature, jolting 3h, the results are shown in Figure 1.
Resin mainly carries out in two ways to the absorption of macromolecular substance: the one, and the ion on resin and the ion-exchange of adsorbent are adsorbed; The 2nd, phase patibhaga-nimitta is inhaled, and relies on secondary key (hydrophobic interaction power, Van der Waals force etc.) to adsorb material.Decolorizing effect, except main investigation percent of decolourization, also needs to take into account this index of polysaccharide recovery simultaneously.As shown in Figure 1, LS-850 anionite-exchange resin is better than other type of resin (as shown in Figure 1) greatly to purple sesame polysaccharide decolorizing effect.Pigment anionresin in the form of negatively charged ion by ionic linkage and purple sesame polysaccharide in LS-850, and the group of the nonpolar part of pigment in resinous substrates be combined with hydrophobic force, thus reach decolouring object.Therefore the preferred LS-850 anionite-exchange resin of the present invention decolours to purple sesame polysaccharide.
Following examples 2-5 is taking LS-850 as decolorizing resin research object, and choosing 4 factors is bleaching time, resin demand, bleaching temperature, 4 influence factors of pH value, study its impact on purple sesame polysaccharide decolorizing effect, to determine decolouring factors vary scope and optimal response curved surface investigation scope.
Embodiment 2: the impact of bleaching time
5 parts of the purple sesame polysaccharide soln of accurate measuring 10ml, are under 30 DEG C, the pH value condition that is 6.0 in identical resin demand LS-8500.5g, temperature, investigate different time to the impact of decolouring, and the results are shown in Figure 2.
As shown in Figure 2, along with the prolongation of time, resin decolorization successful strengthens, and LS-850 resin is after 3h, and percent of decolourization starts to increase slowly, and has the trend reducing, and movable the beginning towards running balance future development of sorption and desorption of pigment is described.And polysaccharide yield presents contrary trend with percent of decolourization substantially, after 4h, polysaccharide yield almost no longer declines.For the pigment in polysaccharide is more removed, and reduce as much as possible the loss of polysaccharide, consider two indexs, the bleaching time response surface analysis investigation scope of the inventive method is preferably to 2~4h.
Embodiment 3: the impact of resin demand
5 parts of the purple sesame polysaccharide soln of accurate measuring 10ml, are that 3h, temperature are under 30 DEG C, the pH value condition that is 6.0 at identical bleaching time, investigate different resins consumption to the impact of decolouring, and the results are shown in Figure 3.
As shown in Figure 3, along with the increase of resin demand, percent of decolourization presents ascendant trend, but after resin demand reaches 0.5g, the trend that percent of decolourization increases obviously slows down, and now polysaccharide recovery still continues to drop.For follow-up scientific research or industrial further deep processing processing, when increasing decolorizing effect as far as possible, take into account the more much higher sugared rate of recovery, reduce polysaccharide wastage rate, therefore the resin demand response surface analysis investigation scope of the inventive method is preferably 0.25g~0.75g.
Embodiment 4: the impact of bleaching temperature
5 parts of the purple sesame polysaccharide soln of accurate measuring 10ml, are that 3h, resin demand are under 0.5g, the pH value condition that is 6.0 at identical bleaching time, investigate the impact of different bleaching temperatures on decolouring, the results are shown in Figure 4.
As shown in Figure 4, temperature is higher, and the velocity of diffusion of pigment molecular is faster in theory, and polysaccharide soln viscosity also declines, and this is conducive to the absorption of pigment.As shown in Figure 4, temperature percent of decolourization in the time of 35 DEG C is the highest, and temperature polysaccharide yield in the time of 25 DEG C is the highest, considers two indexs, and the bleaching temperature response surface analysis investigation scope of the inventive method is preferably to 25~35 DEG C.
The impact of embodiment 5:pH value
7 parts of the purple sesame polysaccharide soln of accurate measuring 10ml, are that 3h, resin demand are that 0.5g, temperature are under the condition of 30 DEG C at identical bleaching time, investigate different pH values to the impact of decolouring, and the results are shown in Figure 5.
PH value can affect the charged situation of molecules in solution, thereby affects the size of they and resin reactive force, and suitable pH value can be impelled the loss of the more pigment of resin absorption and minimizing polysaccharide.Purple sesame polysaccharide is acid polysaccharide, and pH value is approximately 6 after testing, is slightly acidic, as shown in Figure 5, in the time that pH value is 6-7, can take into account preferably two indexs of percent of decolourization and polysaccharide recovery.Consider as reducing experiment number to subsequent response surface analysis, reduce by an investigation factor, preferably the pH value of solution is defined as to 6, do not change the pH value of purple sesame polysaccharide soln.
Embodiment 6:
The tank liquid of putting that fermentation is completed carries out Plate Filtration, discards extracellular fluid, gets and filters rear mycelium; By mycelium deionized water wash, being washed till sheet frame filtrate is that water white transparency also no longer produces bubble, filtration is weighed, mycelium after washing and filtering is weighed, add appropriate multiple deionized water by mycelium weight, put after steeping tank extracts for several times with certain hour heating and separate in Plate Filtration, through scraper plate, concentrated and single tank is concentrated into 85% ethanol that adds the appropriate multiple quality of this concentrated solution after a certain amount of and places 12h in 4 DEG C extracting solution; Alcohol hypostasis is put to whizzer 1200r/min centrifugation, after taking precipitate sabot, put into box baker, control vacuum tightness and be heated to take out alcohol hypostasis after 30~40 DEG C of oven dry 2h, pulverize; Again sabot after pulverizing, then put into baking oven, is slowly warming up to 65 DEG C and dries 2h, finally purple sesame liquid submerged fermentation mycelium Crude polysaccharides; Get Crude polysaccharides is dissolved in distilled water by liquid ratio at 200: 1, centrifugal (5000rpm/min) processes twice, each 20 minutes, get supernatant liquor and cross 30KD hollow fiber ultrafiltration membrane, after trapped fluid dilution, cross 300KD hollow fiber ultrafiltration membrane, after ultrafiltrated concentrated frozen is dry, must refine polysaccharide GS-B, measuring relative molecular weight through HPLC is 30KD~300KD; Get above-mentioned purple sesame polysaccharide GS-B 1.0g, be dissolved in 100ml (liquid ratio 100: 1) distilled water, leave standstill 30 minutes, centrifugal (5000rpm/min) processes 30 minutes, gets supernatant liquor and must refine purple sesame polysaccharide soln; 1 part of the purple sesame polysaccharide soln of accurate measuring 10ml, resin LS-8500.75g and purple sesame polysaccharide soln are inserted in tool plug Erlenmeyer flask, then control condition parameter is the rotating speed 120r/min of 30 DEG C of temperature, pH value 6.0, time 2.5h, shaking table, Erlenmeyer flask is placed in to shaking table and starts jolting decolouring, after finishing, adopt polysaccharide recovery and percent of decolourization measuring method, recording polysaccharide recovery is 67.3%, and percent of decolourization is 87.1%.
Embodiment 7:
The tank liquid of putting that fermentation is completed carries out Plate Filtration, discards extracellular fluid, gets and filters rear mycelium; By mycelium deionized water wash, being washed till sheet frame filtrate is that water white transparency also no longer produces bubble, filtration is weighed, mycelium after washing and filtering is weighed, add appropriate multiple deionized water by mycelium weight, put after steeping tank extracts for several times with certain hour heating and separate in Plate Filtration, through scraper plate, concentrated and single tank is concentrated into 85% ethanol that adds the appropriate multiple quality of this concentrated solution after a certain amount of and places 12h in 4 DEG C extracting solution; Alcohol hypostasis is put to whizzer 1200r/min centrifugation, after taking precipitate sabot, put into box baker, control vacuum tightness and be heated to take out alcohol hypostasis after 30~40 DEG C of oven dry 2h, pulverize; Again sabot after pulverizing, then put into baking oven, is slowly warming up to 65 DEG C and dries 2h, finally purple sesame liquid submerged fermentation mycelium Crude polysaccharides; Get Crude polysaccharides is dissolved in distilled water by liquid ratio at 200: 1, centrifugal (5000rpm/min) processes twice, each 20 minutes, get supernatant liquor and cross 30KD hollow fiber ultrafiltration membrane, after trapped fluid dilution, cross 300KD hollow fiber ultrafiltration membrane, after ultrafiltrated concentrated frozen is dry, must refine polysaccharide GS-B, measuring relative molecular weight through HPLC is 30KD~300KD; Get above-mentioned purple sesame polysaccharide GS-B 1.0g, be dissolved in 100ml (liquid ratio 100: 1) distilled water, leave standstill 30 minutes, centrifugal (5000rpm/min) processes 30 minutes, gets supernatant liquor and must refine purple sesame polysaccharide soln; The purple sesame polysaccharide soln of accurate measuring 10ml1 part, resin LS-8500.75g and purple sesame polysaccharide soln are inserted in tool plug Erlenmeyer flask, then control condition parameter is the rotating speed 120r/min of 28 DEG C of temperature, pH value 6.0, time 2.5h, shaking table, Erlenmeyer flask is placed in to shaking table and starts jolting decolouring, after finishing, adopt polysaccharide recovery and percent of decolourization measuring method, recording polysaccharide recovery is 66.1%, and percent of decolourization is 89.2%.
Embodiment 8:
The tank liquid of putting that fermentation is completed carries out Plate Filtration, discards extracellular fluid, gets and filters rear mycelium; By mycelium deionized water wash, being washed till sheet frame filtrate is that water white transparency also no longer produces bubble, filtration is weighed, mycelium after washing and filtering is weighed, add appropriate multiple deionized water by mycelium weight, put after steeping tank extracts for several times with certain hour heating and separate in Plate Filtration, through scraper plate, concentrated and single tank is concentrated into 85% ethanol that adds the appropriate multiple quality of this concentrated solution after a certain amount of and places 12h in 4 DEG C extracting solution; Alcohol hypostasis is put to whizzer 1200r/min centrifugation, after taking precipitate sabot, put into box baker, control vacuum tightness and be heated to take out alcohol hypostasis after 30~40 DEG C of oven dry 2h, pulverize; Again sabot after pulverizing, then put into baking oven, is slowly warming up to 65 DEG C and dries 2h, finally purple sesame liquid submerged fermentation mycelium Crude polysaccharides; Get Crude polysaccharides is dissolved in distilled water by liquid ratio at 200: 1, centrifugal (5000rpm/min) processes twice, each 20 minutes, get supernatant liquor and cross 30KD hollow fiber ultrafiltration membrane, after trapped fluid dilution, cross 300KD hollow fiber ultrafiltration membrane, after ultrafiltrated concentrated frozen is dry, must refine polysaccharide GS-B, measuring relative molecular weight through HPLC is 30KD~300KD; Get above-mentioned purple sesame polysaccharide GS-B 1.0g, be dissolved in 100ml (liquid ratio 100: 1) distilled water, leave standstill 30 minutes, centrifugal (5000rpm/min) processes 30 minutes, gets supernatant liquor and must refine purple sesame polysaccharide soln; The purple sesame polysaccharide soln of accurate measuring 10ml1 part, resin LS-8500.75g and purple sesame polysaccharide soln are inserted in tool plug Erlenmeyer flask, then control condition parameter is the rotating speed 120r/min of 35 DEG C of temperature, pH value 6.0, time 3h, shaking table, Erlenmeyer flask is placed in to shaking table and starts jolting decolouring, after finishing, adopt polysaccharide recovery and percent of decolourization measuring method, recording polysaccharide recovery is 68.7%, and percent of decolourization is 85.3%.
Claims (10)
1. a method that adopts ion exchange resin to decolour to purple sesame liquid submerged fermentation mycelium Crude polysaccharides, comprises the steps:
A) purple sesame liquid submerged fermentation mycelium Crude polysaccharides is carried out to pre-treatment, obtaining molecular weight ranges is the refining purple sesame polysaccharide of 30KD~300KD;
B) refining purple sesame polysaccharide step a) being obtained is placed in ion exchange resin, be 2~4h at bleaching time, resin demand is 0.25g~0.75g/10ml, and bleaching temperature is to decolour under 25~35 DEG C, the pH value decolorization condition that is 6~7, obtains the purple sesame polysaccharide after decolouring.
2. method according to claim 1, is characterized in that the pre-treatment described in step a) comprises the steps:
1) purple sesame liquid submerged fermentation mycelium Crude polysaccharides is dissolved in the water by liquid ratio for 150: 1~250: 1;
2) centrifugal more than twice or twice with the speed of 4000~5000rpm/min, get supernatant liquor, each centrifugal 20~40min;
3) supernatant liquor is crossed 30KD hollow fiber ultrafiltration membrane under 20~40 DEG C of temperature, pressure 0.02~0.06Mpa condition, after trapped fluid dilution, crosses 300KD hollow fiber ultrafiltration membrane, after ultrafiltrated concentrated frozen is dry, must refine purple sesame polysaccharide.
3. method according to claim 2, is characterized in that step 1) described in liquid ratio be 180: 1~220: 1, preferably 200: 1.
4. method according to claim 2, is characterized in that step 1) described in water be deionized water.
5. method according to claim 2, is characterized in that step 2) described in centrifugation rate be 4500~5000rpm/min, preferably 5000rpm/min; Centrifugation time is 25~35min, preferably 30min.
6. method according to claim 1, is characterized in that the ion exchange resin described in step b) is LS-850 anionite-exchange resin.
7. method according to claim 1, is characterized in that described in step b) that bleaching time is 2.5h, and described resin demand is 0.75g/10ml, and described bleaching temperature is 28 DEG C, and described pH value is 6.
8. method according to claim 1, it is characterized in that step b) in decolorization in shaking table, carry out.
9. method according to claim 8, the rotating speed that it is characterized in that described shaking table is 100~140r/min.
10. method according to claim 9, the rotating speed that it is characterized in that described shaking table is 120r/min.
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