CN109112175B - Method for constructing microbial co-culture system to produce pleurotus tuber-regium exopolysaccharide at high yield - Google Patents
Method for constructing microbial co-culture system to produce pleurotus tuber-regium exopolysaccharide at high yield Download PDFInfo
- Publication number
- CN109112175B CN109112175B CN201811075059.1A CN201811075059A CN109112175B CN 109112175 B CN109112175 B CN 109112175B CN 201811075059 A CN201811075059 A CN 201811075059A CN 109112175 B CN109112175 B CN 109112175B
- Authority
- CN
- China
- Prior art keywords
- regium
- fermentation
- pleurotus tuber
- culture
- mycelium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a method for constructing a microbial co-culture system to produce pleurotus tuber-regium exopolysaccharide at high yield, and belongs to the technical field of bioengineering. The invention adopts the co-culture of the pleurotus tuber-regium mycelium and the lactic acid bacteria, optimizes the inoculation amount and the co-culture time of the lactic acid bacteria during the co-culture, improves the yield of the polysaccharide and improves the solubility of the polysaccharide. The strains used in the invention are all edible strains, are safe and reliable, and the raw materials of the used culture medium are cheap and easy to obtain, have no pollution and are easy to realize. The method has simple and convenient working procedures and strong operability, and can realize large-scale and industrial production.
Description
Technical Field
The invention relates to a method for constructing a microbial co-culture system to produce pleurotus tuber-regium exopolysaccharide at high yield, and belongs to the technical field of bioengineering.
Background
Pleurotus tuber-regium (Pleurotus tuber-regium), the academic name Pleurotus cornucopiae, also called Pleurotus tuber-regium, Pleurotus cornucopiae, Poria cocos wolf, etc., belongs to the class of Pleurotaceae, Agaricales, and Basidiomycotina, and is a rare edible and medicinal fungus mainly distributed in tropical and subtropical regions. Pleurotus tuber-regium has high nutritive value, is rich in various bioactive components, and has attracted attention in recent years as polysaccharide which is the highest content component. Research shows that the Pleurotus tuber-regium polysaccharide can significantly improve the immune function of the organism and has good application prospect in the fields of medicine, health products, food, drug carriers and the like. The Pleurotus tuber-regium mycelium can produce extracellular polysaccharide through liquid state fermentation, and although the polysaccharide has a basic structure different from sclerotium and fruiting body polysaccharide, the polysaccharide has a highly branched form, so that the polysaccharide has huge potential research and application values. At present, most methods for producing the exopolysaccharide of the pleurotus tuber-regium mycelium only adopt the liquid fermentation of the pleurotus tuber-regium mycelium, but the obtained exopolysaccharide has the defects of low yield, poor solubility and the like. These problems severely limit their further basic and application research.
Lactic acid bacteria are a general term for a group of bacteria capable of producing a large amount of lactic acid using fermentable carbohydrates, widely exist in the intestinal tract of human body, have attracted attention to the regulation effect on the micro-ecology of the intestinal tract of human body and the promotion effect on the health of human body, and have become a research hotspot in recent years. The complex interactions between different microorganisms are often used in food and agricultural production, and corresponding co-culture systems are often constructed for the increase in production of known bioactive substances or for the increase in unknown metabolites. However, no application example of improving the yield of the polysaccharide by using mixed fermentation of lactic acid bacteria and macrofungi is found at present.
Disclosure of Invention
The first purpose of the invention is to provide a method for high yield of Pleurotus tuber-regium extracellular polysaccharide, which is to construct a co-culture system by using two microorganisms, namely lactobacillus and Pleurotus tuber-regium mycelium. The method specifically comprises the following steps:
(1) adding the pleurotus tuber-regium mycelium into a seed culture solution for pre-culture; adding lactobacillus into the liquid culture medium for pre-culture;
(2) inoculating a culture solution obtained after the pre-culture of the pleurotus tuber-regium mycelium into a fermentation culture medium for liquid fermentation for 7 d;
(3) when the fermentation in the step (2) is carried out to the 3 rd to 6 th days, inoculating a culture solution obtained after the lactic acid bacteria are pre-cultured into the Pleurotus tuber-regium mycelium fermentation liquor for co-culture;
(4) and after the fermentation is finished, centrifuging and filtering to obtain filtrate which is fermentation liquor, adding ethanol into the fermentation liquor, stirring, standing, collecting precipitate, dissolving the precipitate, and freeze-drying to obtain the pleurotus tuber-regium extracellular polysaccharide.
In one embodiment of the present invention, the pre-culturing of Pleurotus tuber-regium mycelia in step (1) is carried out by adding Pleurotus tuber-regium mycelia into seed culture solution, and pre-culturing at 160-200rpm and 26-32 deg.C for 2-4 days.
In one embodiment of the present invention, the pre-culture of the lactic acid bacteria in step (1) is performed by adding the lactic acid bacteria to MRS medium and pre-culturing at 30-40 ℃ for 18-36h at 100-120 rpm.
In one embodiment of the present invention, the liquid fermentation in step (2) refers to inoculating 5-10% of Pleurotus tuber-regium mycelium culture solution, inoculating into fermentation medium, and fermenting at 25-32 deg.C and 200rpm for 7 d.
In one embodiment of the invention, the seed culture solution of the Pleurotus tuber-regium mycelium in step (1) and the fermentation medium in step (2) are prepared by the following formula: glucose 20-40g/L, yeast extract powder 3-5g/L, and potassium dihydrogen phosphate (KH)2PO4)0.5-1.5g/L, magnesium sulfate heptahydrate (MgSO)4·7H2O)0.4-1.0g/L。
In one embodiment of the present invention, the co-cultivation in step (3) means that the lactobacillus is inoculated into the Pleurotus tuber-regium mycelium fermentation broth at the time of fermentation to 3-6d, the inoculum size is 2.5-10%, and the co-cultivation is carried out at 150-200rpm and 26-35 ℃ for 1-4 d.
In one embodiment of the present invention, the centrifugation in step (4) refers to 3000-5000g centrifugation for 5-10 min.
In one embodiment of the present invention, the method for obtaining exopolysaccharide in step (4) comprises: adding 95% ethanol with volume 3-5 times of the fermentation liquid into the fermentation filtrate after centrifugation and filtration, stirring, standing for 5-8h, dissolving the precipitate with water, and freeze-drying.
Aiming at the problems of low yield, poor solubility and the like of the existing method for producing the exopolysaccharide of the pleurotus tuber-regium, the invention adopts a mode of co-culturing pleurotus tuber-regium mycelium and lactic acid bacteria to improve the yield of the polysaccharide and improve the solubility of the polysaccharide. By adjusting the inoculation amount of lactic acid bacteria during co-culture and the co-culture time, the yield of the pleurotus tuber-regium extracellular polysaccharide can reach 522.60mg/L, and is increased to 3.7 times compared with the yield of a control group 1 (only liquid fermentation of pleurotus tuber-regium mycelium). The co-culture of the two microorganisms can realize mutual beneficial symbiosis, and the yield of the obtained extracellular polysaccharide is obviously improved. The method has simple and convenient working procedures and strong operability, and can realize large-scale and industrial production.
Detailed Description
The method for measuring the polysaccharide content comprises the following steps:
the measurement of polysaccharide content adopts phenol-sulfuric acid method. 5mg of polysaccharide sample was dispersed in 1mL of 12M sulfuric acid solution and reacted in a water bath at 40 ℃ for 10 min. Then 5mL of deionized water is added, the diluted sample solution is placed in a boiling water bath for continuous reaction for 60min, and the temperature is cooled to room temperature after the reaction is finished. Diluting 1mL of reaction solution by 10 times with 2M sulfuric acid, adding 1mL of 5% phenol solution into 1mL of the diluted solution, shaking and mixing uniformly, adding 5mL of concentrated sulfuric acid (18M) solution, reacting for 30min, and measuring the absorbance value at 490 nm. The standard curve was prepared from glucose at linear concentrations of 12.5. mu.g/mL, 25. mu.g/mL, 50. mu.g/mL and 100. mu.g/mL, respectively, dissolved in 2M sulfuric acid and determined in accordance with the samples. After the determination, the absorbance value of the sample is substituted into the standard curve to obtain the polysaccharide concentration, and the total sugar content is calculated.
(II) culture Medium
Pleurotus tuber-regium seed culture solution and liquid fermentation medium: glucose 20-40g/L, yeast extract powder 3-5g/L, and potassium dihydrogen phosphate (KH)2PO4)0.5-1.5g/L, magnesium sulfate heptahydrate (MgSO)4·7H2O)0.4-1.0g/L。
MRS culture medium: peptone 10.0 g/mL; 10.0g/mL of beef extract; 5.0g/mL of yeast extract; diammonium hydrogen citrate [ (NH)4)2HC6H5O7]2.0 g/mL; glucose (C)6H12O6·H2O)20.0 g/mL; tween 801.0 mL/L; sodium acetate (CH)3COONa·3H2O)5.0 g/mL; dipotassium hydrogen phosphate (K)2HPO4·3H2O)2.0 g/mL; magnesium sulfate (MgSO)4·7H2O)0.58 g/mL; manganese sulfate (MnSO)4·H2O)0.25 g/mL; agar 18.0 g/mL; the pH value is 6.2-6.6.
Example 1
Cutting PDA solid culture medium of Pleurotus tuber-regium mycelium into pieces of 1cm2Adding the small blocks into the sterilized seed culture solution, culturing at 30 deg.C and 180rpm for 3d, inoculating into sterilized liquid fermentation medium, and fermenting at 30 deg.C and 180rpm for 7 days with the inoculum size of 5.0%. Wherein the liquid loading of the seed liquid is 100mL/250mL of triangular flask, and the liquid loading of the fermentation liquid is 190mL/500 mL.
The lactobacillus seed liquid is pre-cultured for 18h in MRS culture medium at the speed of 100rpm and the temperature of 37 ℃. Inoculating lactobacillus seed liquid accounting for 5.0 percent of the total volume of the Pleurotus tuber-regium fermentation liquid into the Pleurotus tuber regium fermentation liquid for 6d, wherein the co-culture temperature is 30 ℃, and the co-culture rotation speed is 180 rpm. And finishing the fermentation when the total fermentation time of the Pleurotus tuber-regium is 7d, namely the co-culture time is 1 d.
And after the fermentation is finished, centrifuging at the rotating speed of 3000g for 15min, and filtering by using quantitative filter paper to remove suspended matters to obtain filtrate, namely the pleurotus tuber-regium-lactic acid bacteria co-culture fermentation liquor.
Adding 95% ethanol with volume of 4 times of the obtained fermentation liquor, stirring, standing for 8h, dissolving the precipitate with water with volume of 2 times of the obtained fermentation liquor, and freeze-drying to obtain the extracellular polysaccharide.
Example 2
Cutting PDA solid culture medium of Pleurotus tuber-regium mycelium into pieces of 1cm2Adding the small blocks into the sterilized seed culture solution, culturing at 30 ℃ for 3d at 180rpm, inoculating into sterilized liquid fermentation medium, and fermenting at 30 ℃ for 7 days at 160rpm with the inoculum size of 10%. Wherein the liquid loading of the seed liquid is 90mL/250mL of triangular flask, and the liquid loading of the fermentation liquid is 180mL/500 mL.
The lactobacillus seed liquid is pre-cultured in MRS culture medium for 24h at 37 deg.C and shaking table rotation speed of 100 rpm. Inoculating lactobacillus seed liquid accounting for 2.5 percent of the total volume of the Pleurotus tuber-regium fermentation liquid into the Pleurotus tuber regium fermentation liquid for 5 days, wherein the inoculation time is the Pleurotus tuber regium fermentation time, the co-culture temperature is 30 ℃, and the co-culture rotation speed is 180 rpm. And finishing the fermentation when the total fermentation time of the Pleurotus tuber-regium is 7d, namely, the co-culture time is 2 days.
And after the fermentation is finished, centrifuging for 10min at the rotating speed of 4000g, and filtering by adopting quantitative filter paper to remove suspended matters to obtain filtrate, namely the pleurotus tuber-regium-lactic acid bacteria co-culture fermentation liquor.
Adding 95% ethanol with volume of 4 times of the obtained fermentation liquor, stirring, standing for 5h, dissolving the precipitate with water with volume of 2 times of the obtained fermentation liquor, and freeze-drying to obtain the extracellular polysaccharide.
Example 3
The mycelia of Pleurotus tuber-regiumCutting PDA solid culture medium into 1cm2Adding the small blocks into the sterilized seed culture solution, culturing at 30 deg.C and 180rpm for 3d, inoculating into sterilized liquid fermentation medium, and fermenting at 30 deg.C and 180rpm for 7 days with the inoculum size of 5.0%. Wherein the liquid loading of the seed liquid is 100mL/250mL of triangular flask, and the liquid loading of the fermentation liquid is 190mL/500 mL.
The lactobacillus seed liquid is pre-cultured for 18h in MRS culture medium at the speed of 100rpm and the temperature of 37 ℃. Inoculating lactobacillus seed liquid accounting for 5.0 percent of the total volume of the Pleurotus tuber-regium fermentation liquid into the Pleurotus tuber regium fermentation liquid for the 5 th day of the Pleurotus tuber regium liquid fermentation, wherein the co-culture temperature is 30 ℃, and the co-culture rotation speed is 180 rpm. And finishing the fermentation when the total fermentation time of the Pleurotus tuber-regium is 7d, namely the co-culture time is 2 d.
And after the fermentation is finished, centrifuging at the rotating speed of 3000g for 15min, and filtering by using quantitative filter paper to remove suspended matters to obtain filtrate, namely the pleurotus tuber-regium-lactic acid bacteria co-culture fermentation liquor.
Adding 95% ethanol with volume 4 times of the obtained fermentation liquor, stirring, standing for 8h, dissolving the precipitate with water with volume 2 times of the obtained fermentation liquor, and freeze-drying to obtain extracellular polysaccharide.
Example 4
Cutting PDA solid culture medium of Pleurotus tuber-regium mycelium into pieces of 1cm2Adding the small blocks into the sterilized seed culture solution, culturing at 30 ℃ for 3d at 180rpm, inoculating into sterilized liquid fermentation medium, and fermenting at 30 ℃ for 7 days at 160rpm with the inoculum size of 10%. Wherein the liquid loading of the seed liquid is 90mL/250mL of triangular flask, and the liquid loading of the fermentation liquid is 180mL/500 mL.
The lactobacillus seed liquid is pre-cultured in MRS culture medium for 24h at 37 deg.C and shaking table rotation speed of 100 rpm. Inoculating lactobacillus seed liquid accounting for 7.5 percent of the total volume of the Pleurotus tuber-regium fermentation liquid into the Pleurotus tuber regium fermentation liquid for 5 days, wherein the inoculation time is the Pleurotus tuber regium fermentation time, the co-culture temperature is 30 ℃, and the co-culture rotation speed is 180 rpm. And finishing the fermentation when the total fermentation time of the Pleurotus tuber-regium is 7d, namely, the co-culture time is 2 days.
And after the fermentation is finished, centrifuging for 10min at the rotating speed of 4000g, and filtering by adopting quantitative filter paper to remove suspended matters to obtain filtrate, namely the pleurotus tuber-regium-lactic acid bacteria co-culture fermentation liquor.
Adding 95% ethanol with volume of 4 times of the obtained fermentation liquor, stirring, standing for 5h, dissolving the precipitate with water with volume of 2 times of the obtained fermentation liquor, and freeze-drying to obtain the extracellular polysaccharide.
Example 5
Cutting PDA solid culture medium of Pleurotus tuber-regium mycelium into pieces of 1cm2Adding the small blocks into the sterilized seed culture solution, culturing at 30 ℃ for 3d at 180rpm, inoculating into sterilized liquid fermentation medium, and fermenting at 30 ℃ for 7 days at 160rpm with the inoculum size of 10%. Wherein the liquid loading of the seed liquid is 90mL/250mL of triangular flask, and the liquid loading of the fermentation liquid is 180mL/500 mL.
The lactobacillus seed liquid is pre-cultured in MRS culture medium for 24h at 37 deg.C and shaking table rotation speed of 100 rpm. Inoculating lactobacillus seed liquid accounting for 10 percent of the total volume of the Pleurotus tuber-regium fermentation liquid into the Pleurotus tuber regium fermentation liquid, wherein the inoculation time is 5 days of the Pleurotus tuber regium fermentation time, the co-culture temperature is 30 ℃, and the co-culture rotation speed is 180 rpm. And finishing the fermentation when the total fermentation time of the Pleurotus tuber-regium is 7d, namely, the co-culture time is 2 days.
And after the fermentation is finished, centrifuging for 10min at the rotating speed of 4000g, and filtering by adopting quantitative filter paper to remove suspended matters to obtain filtrate, namely the pleurotus tuber-regium-lactic acid bacteria co-culture fermentation liquor.
Adding 95% ethanol with volume of 4 times of the obtained fermentation liquor, stirring, standing for 5h, dissolving the precipitate with water with volume of 2 times of the obtained fermentation liquor, and freeze-drying to obtain the extracellular polysaccharide.
Comparative example
To better verify the effect of the co-cultivation system used in the present invention on exopolysaccharide production, we performed the following comparative experiments.
Comparative example 1: the inoculation time of the lactic acid bacteria is the same day of inoculation of the Pleurotus tuber-regium fermentation liquor, namely the co-culture time is 7d, and the rest conditions are the same as those in the embodiment 1 of the invention;
comparative example 2: the inoculation time of the lactic acid bacteria is 1d of the fermentation of the pleurotus tuber-regium, namely the co-culture time is 6d, and the rest conditions are the same as those in the embodiment 1 of the invention;
comparative example 3: the inoculation time of the lactic acid bacteria is 2d of the fermentation of the pleurotus tuber-regium, namely the co-culture time is 5d, and the rest conditions are the same as those in the embodiment 1 of the invention.
Comparative example 4: the inoculation amount of the lactic acid bacteria is 12 percent, and the other conditions are the same as those in the embodiment 2 of the invention;
comparative example 5: the inoculation amount of the lactic acid bacteria is 15 percent, and the other conditions are the same as those in the embodiment 2 of the invention;
comparative example 6: the inoculation amount of lactic acid bacteria was 20%, and the other conditions were the same as in example 2 of the present invention.
TABLE 1
Note: the control group 1 is prepared by liquid fermentation of Pleurotus tuber-regium mycelium only without adding lactobacillus for co-culture; control 2 was a single fermentation using only lactic acid bacteria. The polysaccharide yield is the ratio of the polysaccharide content to the total volume of the fermentation liquor after the content of the polysaccharide in the ethanol precipitate is measured by adopting a phenol-sulfuric acid method.
The method of the comparative example is applied to obtain low yield of exopolysaccharide. As can be seen from Table 1, when the number of days for co-culturing the pleurotus tuber-regium mycelium and the lactic acid bacteria is more than 4 days, the yield of the obtained extracellular polysaccharide is lower than that of the control group 1; when the inoculation amount (volume ratio) of lactobacillus inoculated into the Pleurotus tuber-regium fermentation liquid exceeds 10%, both the extracellular polysaccharide yield and the bacterial amount are seriously reduced.
In addition, we measured the mass of the exopolysaccharide obtained by co-cultivation, i.e., the solubility, when it was saturated in 100g of water at 28 ℃. Compared with the exopolysaccharide obtained by the simple fermentation of the mycelia of the Pleurotus tuber-regium (control group), the solubility of the exopolysaccharide obtained by the invention is improved from 0.52g/L to 10.33g/L, and the preliminary analysis may be related to the reduction of the mannose content in the exopolysaccharide.
The monosaccharide composition of the Pleurotus tuber-regium extracellular polysaccharide obtained by the invention is shown in Table 2.
TABLE 2
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (9)
1. A method for producing Pleurotus tuber-regium extracellular polysaccharide is characterized in that a co-culture system is constructed by two microorganisms of lactic acid bacteria and Pleurotus tuber-regium mycelium, and the method comprises the following steps:
(1) adding the pleurotus tuber-regium mycelium into a seed culture solution for pre-culture; adding lactobacillus into the liquid culture medium for pre-culture;
(2) inoculating a culture solution obtained after the pre-culture of the pleurotus tuber-regium mycelium into a fermentation culture medium for liquid fermentation for 7 d;
(3) when the fermentation in the step (2) is carried out to the 3 rd to 6 th days, inoculating a culture solution obtained after the lactic acid bacteria are pre-cultured into the Pleurotus tuber-regium mycelium fermentation liquor for co-culture;
(4) centrifuging and filtering after fermentation is finished, wherein the obtained filtrate is fermentation liquor, adding ethanol into the fermentation liquor, stirring, standing, collecting precipitate, dissolving the precipitate, and freeze-drying to obtain pleurotus tuber-regium extracellular polysaccharide;
the liquid fermentation in the step (2) means that the inoculation amount of the Pleurotus tuber-regium mycelium culture solution is 5-10%, and the Pleurotus tuber-regium mycelium culture solution is inoculated into a fermentation culture medium for fermentation for 7 d;
the co-culture in the step (3) means that when the fermentation is carried out to the 3 rd to 6 th days, the lactobacillus is inoculated into the pleurotus tuber-regium mycelium fermentation liquor, the inoculation amount is 2.5 to 10 percent, and the lactobacillus and the pleurotus tuber-regium are co-cultured.
2. The method as claimed in claim 1, wherein the step (1) of pre-culturing Pleurotus tuber-regium mycelia comprises adding Pleurotus tuber-regium mycelia into seed culture solution, and pre-culturing at 160-200rpm and 26-32 deg.C for 2-4 days.
3. The method according to claim 1, wherein the pre-culturing of the lactic acid bacteria in step (1) is performed by adding the lactic acid bacteria to the MRS medium and pre-culturing at 30-40 ℃ for 18-36h at 100-120 rpm.
4. The method as claimed in claim 1, wherein the liquid fermentation in step (2) is performed by inoculating 5-10% of Pleurotus tuber-regium mycelium culture solution into fermentation medium, and fermenting at 25-32 deg.C and 200rpm for 7 days.
5. The method as claimed in claim 1, wherein the seed culture solution of the Pleurotus tuber-regium mycelium in step (1) and the fermentation medium in step (2) are formulated as follows: glucose 20-40g/L, yeast extract powder 3-5g/L, and potassium dihydrogen phosphate (KH)2PO4)0.5-1.5g/L, magnesium sulfate heptahydrate (MgSO)4•7H2O）0.4-1.0 g/L。
6. The method according to claim 1 or 5, wherein the co-cultivation in step (3) is performed by inoculating 2.5-10% of lactobacillus into the Pleurotus tuber-regium mycelium fermentation broth at the time of fermentation to 3-6d, and co-cultivation is performed at 150-200rpm at 26-35 ℃ for 1-4 d.
7. The method as claimed in claim 1, wherein the centrifugation in step (4) is 3000-5000g centrifugation for 5-10 min.
8. The method according to claim 1 or 7, wherein the method for obtaining exopolysaccharide in step (4) is: adding 95% ethanol with volume 3-5 times of the fermentation liquid into the fermentation filtrate after centrifugation and filtration, stirring, standing for 5-8h, dissolving the precipitate with water, and freeze-drying.
9. Use of the method according to any one of claims 1 to 8 in the field of bioengineering, food, medicine or health care.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811075059.1A CN109112175B (en) | 2018-09-14 | 2018-09-14 | Method for constructing microbial co-culture system to produce pleurotus tuber-regium exopolysaccharide at high yield |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811075059.1A CN109112175B (en) | 2018-09-14 | 2018-09-14 | Method for constructing microbial co-culture system to produce pleurotus tuber-regium exopolysaccharide at high yield |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109112175A CN109112175A (en) | 2019-01-01 |
| CN109112175B true CN109112175B (en) | 2021-01-29 |
Family
ID=64859500
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811075059.1A Active CN109112175B (en) | 2018-09-14 | 2018-09-14 | Method for constructing microbial co-culture system to produce pleurotus tuber-regium exopolysaccharide at high yield |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109112175B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109762745A (en) * | 2019-03-11 | 2019-05-17 | 河北医科大学第四医院 | A kind of fermentation culture method of Pleurotus tuber-regium and the method that active constituent is extracted from culture |
| CN113185572B (en) * | 2021-05-24 | 2023-03-17 | 江南大学 | Extraction method of Pleurotus tuber-regium cell wall glycoprotein |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102132864A (en) * | 2011-02-22 | 2011-07-27 | 江苏安惠生物科技有限公司 | Edible and medicinal nano fungus fiber for inducing proliferation of probiotics and production method thereof |
| CN104046570A (en) * | 2014-06-13 | 2014-09-17 | 浙江省林业科学研究院 | Method for improving yield and product activity of liquid fermentation product of Paecilomyces cicadae |
| CN104277135A (en) * | 2014-09-23 | 2015-01-14 | 华南农业大学 | Pleurotus tuber-regium polysaccharide and extraction method thereof |
| CN105400725A (en) * | 2015-12-18 | 2016-03-16 | 上海理工大学 | Lactobacillus plantarum strain, and applications thereof |
| CN106520584A (en) * | 2016-12-16 | 2017-03-22 | 广西康佳龙农牧集团有限公司 | Culture medium for saccharomycetes and lactic acid bacteria co-culture and preparation method of culture medium |
| CN108103124A (en) * | 2018-03-01 | 2018-06-01 | 江南大学 | A kind of liquid state fermentation of Antrodia camphorata exocellular polysaccharide and purification process |
| CN108299547A (en) * | 2018-03-01 | 2018-07-20 | 江南大学 | A method of preparing glycoprotein from Sclerotium of Pleurotus tuber regium |
| WO2018154116A1 (en) * | 2017-02-24 | 2018-08-30 | Tvedemose Champignon | A method for the production of mushroom constituents |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6805866B2 (en) * | 2001-09-21 | 2004-10-19 | Gaisford And Miller, Inc. | Oral supplement composition containing a plurality of mushroom strains |
-
2018
- 2018-09-14 CN CN201811075059.1A patent/CN109112175B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102132864A (en) * | 2011-02-22 | 2011-07-27 | 江苏安惠生物科技有限公司 | Edible and medicinal nano fungus fiber for inducing proliferation of probiotics and production method thereof |
| CN104046570A (en) * | 2014-06-13 | 2014-09-17 | 浙江省林业科学研究院 | Method for improving yield and product activity of liquid fermentation product of Paecilomyces cicadae |
| CN104277135A (en) * | 2014-09-23 | 2015-01-14 | 华南农业大学 | Pleurotus tuber-regium polysaccharide and extraction method thereof |
| CN105400725A (en) * | 2015-12-18 | 2016-03-16 | 上海理工大学 | Lactobacillus plantarum strain, and applications thereof |
| CN106520584A (en) * | 2016-12-16 | 2017-03-22 | 广西康佳龙农牧集团有限公司 | Culture medium for saccharomycetes and lactic acid bacteria co-culture and preparation method of culture medium |
| WO2018154116A1 (en) * | 2017-02-24 | 2018-08-30 | Tvedemose Champignon | A method for the production of mushroom constituents |
| CN108103124A (en) * | 2018-03-01 | 2018-06-01 | 江南大学 | A kind of liquid state fermentation of Antrodia camphorata exocellular polysaccharide and purification process |
| CN108299547A (en) * | 2018-03-01 | 2018-07-20 | 江南大学 | A method of preparing glycoprotein from Sclerotium of Pleurotus tuber regium |
Non-Patent Citations (2)
| Title |
|---|
| "Proteomic insights into the stimulatory effect of Tween 80 on mycelial growth and exopolysaccharide production of an edible mushroom Pleurotus tuber-regium";Bo-Bo Zhang et al.;《Biotechnology Letters 》;20120620;第34卷;第1863-1867页 * |
| 虎奶菇菌丝体胞外多糖深层发酵培养基的优化;孟凡杰;《浙江化工》;20101231;第41卷(第2期);第14-17页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109112175A (en) | 2019-01-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2023097375A (en) | Method for preparation of schizophyllan and ergothioneine by co-fermentation of two strains | |
| CN101880700B (en) | Liquid fermentation method capable of improving yield of ganoderma iucidum polysaccharide | |
| CN106978465B (en) | Fermentation method for improving fermentation yield of total triterpenoids in inonotus obliquus | |
| CN109504725B (en) | Method for preparing high-purity hericium erinaceus polysaccharide by fermenting hericium erinaceus and fermentation culture medium | |
| CN112760271B (en) | Process for producing clostridium butyricum through high-density fermentation under negative pressure condition and application | |
| CN109112175B (en) | Method for constructing microbial co-culture system to produce pleurotus tuber-regium exopolysaccharide at high yield | |
| CN102517354A (en) | Preparation method of water-soluble active polysaccharide in boletus edulis, and application thereof | |
| CN103750448A (en) | Method for fermenting mulberries | |
| CN105361161A (en) | Making method of cordyceps militaris kiwi enzyme capable of enhancing immunity | |
| CN110904156B (en) | Method for increasing yield of triterpenoids in phellinus igniarius liquid state fermentation | |
| CN113215206B (en) | Preparation and purification method of grifola frondosa polysaccharide with high antioxidant activity | |
| CN111437224B (en) | Method for extracting antioxidant component from cannabis sativa leaves by using microorganisms and application of antioxidant component | |
| CN111635924B (en) | Liquid fermentation method for increasing yield of ganoderma lucidum extracellular polysaccharide | |
| CN106755182B (en) | Method for promoting ganoderma lucidum liquid fermentation to produce extracellular polysaccharide | |
| CN102703542B (en) | Method for synthesizing lactobionic acid by ultrasound field assisted microbial transformation | |
| CN1141392C (en) | Process for preparing ganoderic polyose and ganoderic acid by fermentation during which raw materials are supplemented | |
| CN102071165A (en) | Method for improving biomass of lactic acid bacteria at low pH by adding glutamic acid | |
| CN110964761A (en) | Tremella and application thereof | |
| CN109673393A (en) | A kind of production method of peacilomyce hepiahi bacterium filament | |
| CN106755164A (en) | A kind of method for improving aspergillus oryzae kojic acid yield | |
| CN105087402B (en) | A kind of method of liquid submerged fermentation production ganoderma lucidum mycelium | |
| CN114790438A (en) | Method for improving yield and oxidation resistance of antrodia camphorata exopolysaccharide | |
| CN102041287B (en) | Method for producing polysaccharide by crossly culturing tremella fuciformis basidiospores by stages | |
| CN104705764A (en) | Solid-liquid separation method for fermentation product | |
| CN113046407B (en) | Method for producing ganoderma triterpene in large scale through liquid state fermentation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |