CN101492705A - Method of preparing pleurotu ostreatus polysaccharide - Google Patents
Method of preparing pleurotu ostreatus polysaccharide Download PDFInfo
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Abstract
The invention relates to a method for preparing pleurotus ostreatus polysaccharide with easy and convenient operation and little equipment investment. The processing steps are: slant strain activation, shake flask seed culture, liquid submerged fermentation, continuous fermentation, separation, concentration and alcohol deposition of ultrafiltration membrane. With the pleurotus ostreatus polysaccharide of the invention adopted, the continuous fermentation can be carried out in a fermentation cylinder, fermentation time can be lengthened, the operation process can be simplified and polysaccharide yield can be improved.
Description
Technical field
The present invention relates to a kind of preparation method of mushroom polysaccharide, especially a kind of method that adopts the fermentative Production mushroom polysaccharide.
Background technology
The general name of flat mushroom pleurotus one class fungi is one of of all times famous edible health bacterium.Mushroom polysaccharide is a kind of polysaccharide of extraction from flat mushroom, separation, purifying, can promote the synthetic of protein, nucleic acid, improves body biological immune power.
At present mushroom polysaccharide mainly extracts from sporophore, and the artificial culture flat mushroom cycle is long, therefrom extracts polysaccharide process and is not easy to control, and the cost height, yield poorly.
Summary of the invention
Technical problem to be solved by this invention provides the preparation method that a kind of easy and simple to handle, equipment drops into little mushroom polysaccharide, when making it produce exocellular polysaccharide, realizes continuously fermenting in fermentor tank, prolongs fermentation time, the flow process that simplifies the operation, raising polysaccharide yield.
For solving the problems of the technologies described above, the present invention adopts following technical proposal:
(1) slant strains activation:
Flat mushroom strain moved be connected on the potato juice synthetic medium inclined-plane, 24 ℃ constant temperature culture 5-10 days;
The potato juice synthetic medium contains the material of following percent weight in volume: glucose 2.0%, potassium primary phosphate 0.1%, sal epsom 0.05%, potato juice 20%, agar 2.0%.
Cultivate fate and be preferably 8 days.
(2) shake-flask seed is cultivated:
Dress seed culture medium in the triangular flask, sterilization connects the female kind in inclined-plane after the cooling, and shaking culture was cultivated 4-6 days altogether;
The prescription of shake-flask seed substratum is: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
The shake-flask seed culture condition is: under 25 ℃, and shaking speed 150r/min, shaking bottled liquid measure is 2/5 of Erlenmeyer flask total volume, the nutrient solution initial pH value is 6.0-7.0.
(3) liquid submerged fermentation:
Shake-flask seed is inserted the ferment tank that substratum is housed, inoculum size 1-10%, leavening temperature is 28-30 ℃, carries out aeration-agitation during fermentation;
The prescription of deep liquid substratum is: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
The condition of liquid submerged fermentation is: the pH variation range is at 5.5-7.0; Cultivate 24h in early stage, air flow is 1: 1, v/v/m, and stirring velocity is 150r/m; 24h is to fermentation ends, and air flow was controlled at 1.2: 1, v/v/m, and stirring velocity is 180r/m.
(4) continuously ferment:
After liquid submerged fermentation 60-75 hour with the fermented liquid pump circulation, millipore filtration by 0.2 μ m, filtyration velocity be controlled at 1 fermentor tank volume/hour, continuous blow-down filtrate under stable state, mother liquor is back to fermentor tank and reuses, simultaneously import the fresh liquid substratum continuously with pump, the constant volume that keeps material in the fermentor tank, keep a jar interior fermentation condition, whole process was kept 30 hours, finished fermentation, collected all filtrates, supernatant liquor and the merging of above-mentioned filtrate are collected in the centrifugal back of fermented liquid remaining in the fermentor tank, promptly get total fermented liquid;
(5) ultra-filtration membrane separates and concentrates:
Is that 4000 ultra-filtration membrane carries out ultrafiltration with above-mentioned total fermented liquid by the molecular weight cutoff value, gets rid of filtrate, and cyclical operation is reduced to original 1/8-1/3 until the volume of concentrated solution;
(6) alcohol precipitation:
95% ethanol that adds 3 times of volumes is to above-mentioned concentrated solution, and 0-4 ℃ left standstill 12-24 hour, collecting precipitation, and washing is dry, promptly.
Compare with from sporophore, extracting polysaccharide process, from submerged fermentation liquid, extract mushroom polysaccharide and have the shortening production cycle, reduce the production floor space, strict control working condition, production process is subjected to advantages such as effect of natural conditions is few.
In the fermentation later stage, there is the feedback inhibition phenomenon in a polysaccharide in jar: when concentration is elevated to a certain degree, the organism polysaccharide synthetic with efflux and will be suppressed; Simultaneously, the polysaccharide that has generated in the fermented liquid also can be consumed by organism.At this moment, fermented liquid is passed through membrane separation unit under the condition of airtight stable state, the solid mycelium is dammed to be continued to utilize in reactor, and polysaccharide is then along with fermented liquid is discharged from, at the filterable fresh medium that replenishes simultaneously in reactor, so just reduce the concentration of polysaccharide in the fermented liquid, broken molecular balance, promoted the growth of organism and the conversion of meta-bolites, realization is continuously fermented, prolong fermentation time, reduce of the consumption of reaction later stage organism simultaneously, improve the productive rate of metabolite polysaccharide.The advantage that the production of continuously fermenting is compared with batch fermentation production embodies by following test:
The comparison with intermittent type fermentation polysaccharide yield of continuously fermenting of test example 1
1, test method
1.1 single-stage zymotechnique
(1) slant strains activation:
Flat mushroom strain moved be connected on the potato juice synthetic medium inclined-plane, 24 ℃ of constant temperature culture 8 days.
Selected potato synthetic medium contains the material of following percent weight in volume: glucose 2.0%, potassium primary phosphate 0.1%, sal epsom 0.05%, potato juice 20%, agar 2.0%.
(2) shake-flask seed is cultivated:
Dress seed culture medium 20ml in the 50ml triangular flask, sterilization connects the female kind in inclined-plane after the cooling, and shaking culture was cultivated 5 days altogether;
The seed culture based formulas: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
The shake-flask seed culture condition is: under 25 ℃, and shaking speed 150r/min, the nutrient solution initial pH value is 6.0-7.0.
(3) single-stage fermentation culture:
The 5L fermentor cultivation:
Shake-flask seed is inserted the ferment tank that the 2L substratum is housed, and inoculum size is 1%, and the condition of fermentation is: leavening temperature is 28-30 ℃; The pH variation range is at 5.5-7.0; Cultivate 24h in early stage, air flow is 1: 1, v/v/m, and stirring velocity is 150r/m; 24h is before put jar, and air flow was controlled at 1.2: 1, v/v/m, and stirring velocity is 180r/m.
The substratum proportioning: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
Put a jar standard: mycelium pellet come-up 1/3, microscopy has clamp connexion, does not have assorted bacterium, and fermentation time is 70 hours altogether.
After the fermentation ends, fermented liquid is centrifugal, collect supernatant liquor.
(4) ultra-filtration membrane separates and concentrates:
Is that 4000 ultra-filtration membrane carries out ultrafiltration with above-mentioned fermented liquid by the molecular weight cutoff value, gets rid of filtrate, and cyclical operation reduces to original 1/5 until the volume of concentrated solution.
(5) alcohol precipitation:
95% ethanol that adds 3 times of volumes is to above-mentioned concentrated solution, and 0-4 ℃ left standstill 18 hours, collecting precipitation, and washing is dry, weighs.
1.2 multistage intermittent type zymotechnique
(1) slant strains activation:
Flat mushroom strain moved be connected on the potato juice synthetic medium inclined-plane, 24 ℃ of constant temperature culture 8 days.
Selected potato synthetic medium contains the material of following percent weight in volume: glucose 2.0%, potassium primary phosphate 0.1%, sal epsom 0.05%, potato juice 20%, agar 2.0%.
(2) shake-flask seed is cultivated:
Dress seed culture medium 20ml in the 50ml triangular flask, sterilization connects the female kind in inclined-plane after the cooling, and shaking culture was cultivated 5 days altogether;
The seed culture based formulas: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
The shake-flask seed culture condition is: under 25 ℃, and shaking speed 150r/min, the nutrient solution initial pH value is 6.0-7.0.
(3) second order fermentation is cultivated:
The 5L fermentor cultivation:
Shake-flask seed is inserted the ferment tank that the 2L substratum is housed, and inoculum size is 1%, and the condition of fermentation is: leavening temperature is 28-30 ℃; The pH variation range is at 5.5-7.0; Cultivate 24h in early stage, air flow is 1: 1, v/v/m, and stirring velocity is 150r/m; 24h is to the commentaries on classics jar, and air flow was controlled at 1.2: 1, v/v/m, and stirring velocity is 180r/m.
The substratum proportioning: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
Put a jar standard: mycelium pellet come-up 1/3, microscopy has clamp connexion, does not have assorted bacterium, and fermentation time is 70 hours altogether.
The 50L ferment tank is cultivated:
Seed is inserted the ferment tank that the 20L substratum is housed, and inoculum size is 10%, and the condition of fermentation is: leavening temperature is 28-30 ℃; The pH variation range is at 5.5-7.0; Cultivate 24h in early stage, air flow is 1: 1, v/v/m, and stirring velocity is 150r/m; 24h is before put jar, and air flow was controlled at 1.2: 1, v/v/m, and stirring velocity is 180r/m.
The substratum proportioning: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
Put a jar standard: fermentation time 40 hours, mycelium pellet come-up 1/2, microscopy has clamp connexion, does not have assorted bacterium.
After the fermentation ends, fermented liquid is centrifugal, collect supernatant liquor.
(4) ultra-filtration membrane separates and concentrates:
Is that 4000 ultra-filtration membrane carries out ultrafiltration with above-mentioned fermented liquid by the molecular weight cutoff value, gets rid of filtrate, and cyclical operation reduces to original 1/5 until the volume of concentrated solution.
(5) alcohol precipitation:
95% ethanol that adds 3 times of volumes is to above-mentioned concentrated solution, and 0-4 ℃ left standstill 18 hours, collecting precipitation, and washing is dry, weighs.
2.1.3 continuous fermentation process
(1) slant strains activation:
Flat mushroom strain moved be connected on the potato juice synthetic medium inclined-plane, 24 ℃ of constant temperature culture 8 days.
Selected potato synthetic medium contains the material of following percent weight in volume: glucose 2.0%, potassium primary phosphate 0.1%, sal epsom 0.05%, potato juice 20%, agar 2.0%.
(2) shake-flask seed is cultivated:
Dress seed culture medium 20ml in the 50ml triangular flask, sterilization connects the female kind in inclined-plane after the cooling, and shaking culture was cultivated 5 days altogether;
The seed culture based formulas: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
The shake-flask seed culture condition is: under 25 ℃, and shaking speed 150r/min, the nutrient solution initial pH value is 6.0-7.0.
(3) deep liquid is cultivated:
Shake-flask seed is inserted the ferment tank that the 2L substratum is housed, and inoculum size is 1%, and the condition of fermentation is: leavening temperature is 28-30 ℃; The pH variation range is at 5.5-7.0; Cultivate 24h in early stage, air flow is 1: 1, v/v/m, and stirring velocity is 150r/m; 24h is to the commentaries on classics jar, and air flow was controlled at 1.2: 1, v/v/m, and stirring velocity is 180r/m.
The substratum proportioning: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
(4) continuously ferment:
Liquid submerged fermentation after 70 hours with the fermented liquid pump circulation, millipore filtration by 0.2 μ m, filtyration velocity be controlled at 1 fermentor tank volume/hour, continuous blow-down filtrate under stable state, mother liquor is back to fermentor tank and reuses, simultaneously import the fresh liquid substratum continuously with pump, the constant volume that keeps material in the fermentor tank, keep a jar interior fermentation condition, whole process was kept 30 hours, finished fermentation, collected all filtrates, supernatant liquor and the merging of above-mentioned filtrate are collected in the centrifugal back of fermented liquid remaining in the fermentor tank, promptly get total fermented liquid;
(5) ultra-filtration membrane separates and concentrates:
Is that 4000 ultra-filtration membrane carries out ultrafiltration with above-mentioned total fermented liquid by the molecular weight cutoff value, gets rid of filtrate, and cyclical operation reduces to original 1/5 until the volume of concentrated solution.
(5) alcohol precipitation:
95% ethanol that adds 3 times of volumes is to above-mentioned concentrated solution, and 0-4 ℃ left standstill 18 hours, collecting precipitation, and washing is dry, weighs.
2.2 test-results
Test-results sees Table 2
The comparison of three kinds of fermentation mode fermentation times of table 2 and output
Test-results shows, adopts the production mushroom polysaccharide that continuously ferments, and can prolong the fermentation time in the single-stage fermentor tank, and output is single-stage is fermented under the equal conditions 6 times approximately; Be less than in total fermentation time under the situation of intermittent type fermentation time, output is higher than the latter, can drop into by minimizing equipment, simplifies technical process, reduces production costs.
Below in conjunction with embodiment the present invention is further elaborated, but the scope of protection of present invention is not limited to following embodiment.
Embodiment
Embodiment 1
(1) slant strains activation:
Flat mushroom strain moved be connected on the potato juice synthetic medium inclined-plane, 24 ℃ of constant temperature culture 8 days.
Selected potato synthetic medium contains the material of following percent weight in volume: glucose 2.0%, potassium primary phosphate 0.1%, sal epsom 0.05%, potato juice 20%, agar 2.0%.
(2) shake-flask seed is cultivated:
Dress seed culture medium 20ml in the 50ml triangular flask, sterilization connects the female kind in inclined-plane after the cooling, and shaking culture was cultivated 5 days altogether;
The seed culture based formulas: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
The shake-flask seed culture condition is: under 25 ℃, and shaking speed 150r/min, shaking bottled liquid measure is 2/5 of Erlenmeyer flask total volume, the nutrient solution initial pH value is 6.5.
(3) deep liquid is cultivated:
Shake-flask seed is inserted the 5L ferment tank that the 2L substratum is housed, and inoculum size is 1%, and the condition of fermentation is: leavening temperature is 28-30 ℃; The pH variation range is at 5.5-7.0; Cultivate 24h in early stage, air flow is 1: 1, v/v/m, and stirring velocity is 150r/m; 24h is to end, and air flow was controlled at 1.2: 1, v/v/m, and stirring velocity is 180r/m.
The substratum proportioning: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
(4) continuously ferment:
Liquid submerged fermentation after 70 hours with the fermented liquid pump circulation, millipore filtration by 0.2 μ m, filtyration velocity be controlled at 1 fermentor tank volume/hour, continuous blow-down filtrate under stable state, mother liquor is back to fermentor tank and reuses, simultaneously import the fresh liquid substratum continuously with pump, the constant volume that keeps material in the fermentor tank, keep a jar interior fermentation condition, whole process was kept 30 hours, finished fermentation, collected all filtrates, supernatant liquor and the merging of above-mentioned filtrate are collected in the centrifugal back of fermented liquid remaining in the fermentor tank, promptly get total fermented liquid;
(5) ultra-filtration membrane separates and concentrates:
Is that 4000 ultra-filtration membrane carries out ultrafiltration with above-mentioned total fermented liquid by the molecular weight cutoff value, gets rid of filtrate, and cyclical operation reduces to original 1/5 until the volume of concentrated solution.
(6) alcohol precipitation:
95% ethanol that adds 3 times of volumes is to above-mentioned concentrated solution, and 0-4 ℃ left standstill 18 hours, collecting precipitation, and washing is dry, promptly gets the 244.6g mushroom polysaccharide.
Embodiment 2
(1) slant strains activation:
Flat mushroom strain moved be connected on the potato juice synthetic medium inclined-plane, 24 ℃ of constant temperature culture 5 days.
Selected potato synthetic medium contains the material of following percent weight in volume: glucose 2.0%, potassium primary phosphate 0.1%, sal epsom 0.05%, potato juice 20%, agar 2.0%.
(2) shake-flask seed is cultivated:
Dress seed culture medium 20ml in the 50ml triangular flask, sterilization connects the female kind in inclined-plane after the cooling, and shaking culture was cultivated 4 days altogether;
The seed culture based formulas: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
The shake-flask seed culture condition is: under 25 ℃, and shaking speed 150r/min, shaking bottled liquid measure is 2/5 of Erlenmeyer flask total volume, the nutrient solution initial pH value is 6.0.
(3) deep liquid is cultivated:
Shake-flask seed is inserted the 5L ferment tank that the 2L substratum is housed, and inoculum size is 10%, and the condition of fermentation is: leavening temperature is 28-30 ℃; The pH variation range is at 5.5-7.0; Cultivate 24h in early stage, air flow is 1: 1, v/v/m, and stirring velocity is 150r/m; 24h is to end, and air flow was controlled at 1.2: 1, v/v/m, and stirring velocity is 180r/m.
The substratum proportioning: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
(4) continuously ferment:
Liquid submerged fermentation after 60 hours with the fermented liquid pump circulation, millipore filtration by 0.2 μ m, filtyration velocity be controlled at 1 fermentor tank volume/hour, continuous blow-down filtrate under stable state, mother liquor is back to fermentor tank and reuses, simultaneously import the fresh liquid substratum continuously with pump, the constant volume that keeps material in the fermentor tank, keep a jar interior fermentation condition, whole process was kept 30 hours, finished fermentation, collected all filtrates, supernatant liquor and the merging of above-mentioned filtrate are collected in the centrifugal back of fermented liquid remaining in the fermentor tank, promptly get total fermented liquid;
(5) ultra-filtration membrane separates and concentrates:
Is that 4000 ultra-filtration membrane carries out ultrafiltration with above-mentioned total fermented liquid by the molecular weight cutoff value, gets rid of filtrate, and cyclical operation reduces to original 1/3 until the volume of concentrated solution.
(6) alcohol precipitation:
95% ethanol that adds 3 times of volumes is to above-mentioned concentrated solution, and 0-4 ℃ left standstill 12 hours, collecting precipitation, and washing is dry, promptly gets the 144.4g mushroom polysaccharide.
Embodiment 3
(1) slant strains activation:
Flat mushroom strain moved be connected on the potato juice synthetic medium inclined-plane, 24 ℃ of constant temperature culture 10 days.
Selected potato synthetic medium contains the material of following percent weight in volume: glucose 2.0%, potassium primary phosphate 0.1%, sal epsom 0.05%, potato juice 20%, agar 2.0%.
(2) shake-flask seed is cultivated:
Dress seed culture medium 20ml in the 50ml triangular flask, sterilization connects the female kind in inclined-plane after the cooling, and shaking culture was cultivated 6 days altogether;
The seed culture based formulas: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
The shake-flask seed culture condition is: under 25 ℃, and shaking speed 150r/min, shaking bottled liquid measure is 2/5 of Erlenmeyer flask total volume, the nutrient solution initial pH value is 7.0.
(3) deep liquid is cultivated:
Shake-flask seed is inserted the 5L ferment tank that the 2L substratum is housed, and inoculum size is 5%, and the condition of fermentation is: leavening temperature is 28-30 ℃; The pH variation range is at 5.5-7.0; Cultivate 24h in early stage, air flow is 1: 1, v/v/m, and stirring velocity is 150r/m; 24h is to end, and air flow was controlled at 1.2: 1, v/v/m, and stirring velocity is 180r/m.
The substratum proportioning: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
(4) continuously ferment:
Liquid submerged fermentation after 75 hours with the fermented liquid pump circulation, millipore filtration by 0.2 μ m, filtyration velocity be controlled at 1 fermentor tank volume/hour, continuous blow-down filtrate under stable state, mother liquor is back to fermentor tank and reuses, simultaneously import the fresh liquid substratum continuously with pump, the constant volume that keeps material in the fermentor tank, keep a jar interior fermentation condition, whole process was kept 30 hours, finished fermentation, collected all filtrates, supernatant liquor and the merging of above-mentioned filtrate are collected in the centrifugal back of fermented liquid remaining in the fermentor tank, promptly get total fermented liquid;
(5) ultra-filtration membrane separates and concentrates:
Is that 4000 ultra-filtration membrane carries out ultrafiltration with above-mentioned total fermented liquid by the molecular weight cutoff value, gets rid of filtrate, and cyclical operation reduces to original 1/8 until the volume of concentrated solution.
(6) alcohol precipitation:
95% ethanol that adds 3 times of volumes is to above-mentioned concentrated solution, and 0-4 ℃ left standstill 24 hours, collecting precipitation, and washing is dry, promptly gets the 153.5g mushroom polysaccharide.
Claims (6)
1, a kind of preparation method of mushroom polysaccharide is characterized in that described method comprises the following step:
(1) slant strains activation:
Flat mushroom strain moved be connected on the potato juice synthetic medium inclined-plane, 24 ℃ constant temperature culture 5-10 days;
(2) shake-flask seed is cultivated:
Dress seed culture medium in the triangular flask, sterilization connects the female kind in inclined-plane after the cooling, and shaking culture was cultivated 4-6 days altogether;
(3) liquid submerged fermentation:
Shake-flask seed is inserted the ferment tank that substratum is housed, inoculum size 1-10%, leavening temperature is 28-30 ℃, carries out aeration-agitation during fermentation;
(4) continuously ferment:
After liquid submerged fermentation 60-75 hour with the fermented liquid pump circulation, millipore filtration by 0.2 μ m, filtyration velocity be controlled at 1 fermentor tank volume/hour, continuous blow-down filtrate under stable state, mother liquor is back to fermentor tank and reuses, simultaneously import the fresh liquid substratum continuously with pump, the constant volume that keeps material in the fermentor tank, keep a jar interior fermentation condition, whole process was kept 30 hours, finished fermentation, collected all filtrates, supernatant liquor and the merging of above-mentioned filtrate are collected in the centrifugal back of fermented liquid remaining in the fermentor tank, promptly get total fermented liquid;
(5) ultra-filtration membrane separates and concentrates:
Is that 4000 ultra-filtration membrane carries out ultrafiltration with above-mentioned total fermented liquid by the molecular weight cutoff value, gets rid of filtrate, and cyclical operation is reduced to original 1/8-1/3 until the volume of concentrated solution;
(6) alcohol precipitation:
95% ethanol that adds 3 times of volumes is to above-mentioned concentrated solution, and 0-4 ℃ left standstill 12-24 hour, collecting precipitation, and washing is dry, promptly.
2, preparation method according to claim 1, it is characterized in that described potato juice synthetic medium contains the material of following percent weight in volume: glucose 2.0%, potassium primary phosphate 0.1%, sal epsom 0.05%, potato juice 20%, agar 2.0%.
3, preparation method according to claim 1 is characterized in that the time of described slant strains activation constant temperature culture is 8 days.
4, preparation method according to claim 1, it is characterized in that described shake-flask seed substratum and deep liquid substratum contain the material of following percent weight in volume: Semen Maydis powder 3%, starch 1%, wheat bran 1%, yeast 1%, potassium primary phosphate 0.1%, sal epsom 0.05%, VITMAIN B1 and Wei ShengsuB2 each 0.005%.
5, preparation method according to claim 1 is characterized in that described shake-flask seed culture condition is: under 25 ℃, and shaking speed 150r/min, shaking bottled liquid measure is 2/5 of Erlenmeyer flask total volume, the nutrient solution initial pH value is 6.0-7.0.
6, preparation method according to claim 1, it is characterized in that the condition of described liquid submerged fermentation is: the pH variation range is at 5.5-7.0; Cultivate 24h in early stage, air flow is 1: 1, v/v/m, and stirring velocity is 150r/m; 24h is to fermentation ends, and air flow was controlled at 1.2: 1, v/v/m, and stirring velocity is 180r/m.
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| CN105902560A (en) * | 2016-05-06 | 2016-08-31 | 吉林化工学院 | Application of pleutotus ostreatus polysaccharide |
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| CN102396717A (en) * | 2011-11-10 | 2012-04-04 | 保定市春华生物科技有限公司 | Pleurotus stem dietary fiber prepared with actinomucor elegans fermentation method and application thereof |
| CN102396717B (en) * | 2011-11-10 | 2013-01-02 | 保定市春华生物科技有限公司 | Pleurotus stem dietary fiber prepared with actinomucor elegans fermentation method and application thereof |
| CN104988183A (en) * | 2015-08-06 | 2015-10-21 | 河北省微生物研究所 | Preparation method of pleurotus ostreatus fermentation broth and application of fermentation broth in degradation of aflatoxin B1 |
| CN105902560A (en) * | 2016-05-06 | 2016-08-31 | 吉林化工学院 | Application of pleutotus ostreatus polysaccharide |
| CN105902560B (en) * | 2016-05-06 | 2019-04-23 | 吉林化工学院 | The application of mushroom polysaccharide |
| CN112481138A (en) * | 2020-12-07 | 2021-03-12 | 贵州大学 | Preparation method for high-yield polysaccharide based on sweet potato wastewater submerged fermentation |
| CN112481138B (en) * | 2020-12-07 | 2023-06-09 | 贵州大学 | Preparation method of high-yield polysaccharide based on deep fermentation of sweet potato wastewater |
| CN114948987A (en) * | 2022-04-27 | 2022-08-30 | 南京农业大学 | Immunity enhancing effect of edible fungus polysaccharide and application of edible fungus polysaccharide in chicken raising industry |
| CN114948987B (en) * | 2022-04-27 | 2024-03-15 | 南京农业大学 | Immunopotentiation effect of edible fungus polysaccharide and application of edible fungus polysaccharide in chicken raising industry |
| CN116064699A (en) * | 2023-01-19 | 2023-05-05 | 安徽师范大学 | Application of oyster mushroom extracellular polysaccharide in preparation of medicine for removing algae toxin toxicity |
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