CN113943761A - Preparation method of micromolecular beta-1, 3-glucan - Google Patents
Preparation method of micromolecular beta-1, 3-glucan Download PDFInfo
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Abstract
The invention relates to the technical field of microbial fermentation, in particular to a preparation method of micromolecule beta-1, 3-glucan. The method takes macromolecular insoluble beta-1, 3-glucan obtained by fermenting rhizobium GL-1803 as a substrate, hydrolyzes through endo-beta-1, 3-glucanase, and obtains micromolecular beta-1, 3-glucan with the purity of more than or equal to 90 percent through filtering and concentration; the preservation number of the rhizobium GL-1803 is as follows: CGMCC No. 23416. The preparation method is simple, low in production cost and environment-friendly, and the obtained micromolecule beta-1, 3-glucan has high purity and provides possibility for large-scale production of the micromolecule beta-1, 3-glucan.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a preparation method of micromolecule beta-1, 3-glucan.
Background
Beta-1, 3-glucan is a polysaccharide whose main chain is connected by beta-1, 3-glycosidic bonds, and usually contains beta-1, 2-glycosidic bonds, beta-1, 4-glycosidic bonds, and beta-1, 6-glycosidic bond-connected branches of different proportions and sizes depending on the source. The beta-1, 3-glucan has the biological activities of enhancing the immune activity, resisting tumors, oxidation, bacteria, viruses, fungi, cholesterol and blood fat, and the like, and is a good biological effect regulator. Meanwhile, the skin care product has the effects of moisturizing, resisting inflammation, resisting aging, removing wrinkles, removing dandruff, removing jaundice, accelerating repair, increasing skin elasticity and the like, and is widely applied to a plurality of fields of medicines, foods, cosmetics, animal feed additives and the like.
At present, the beta-1, 3-glucan is mainly derived from yeast, edible fungi and plants (such as shiitake mushroom, oat and highland barley) and is limited by the complexity of the technology and the high cost, the beta-1, 3-glucan is mainly a crude product with low purity, the production process generates more byproducts, and the yield is limited by the cost of raw materials. The beta-1, 3-glucan is produced by microbial fermentation, and has the advantages of no limitation of seasons, stable and easily obtained raw material sources, no byproduct generation in the process and stable quality among batches.
The linear chain beta-1, 3-glucan produced by microbial fermentation has the characteristics of simple components and single structure, and is suitable for developing beta-1, 3-glucan derivative products. However, due to the special molecular conformation of the high-polymerization-degree linear chain beta-1, 3-glucan, the high-polymerization-degree linear chain beta-1, 3-glucan is insoluble in water and most common organic solvents, and the application range of the beta-1, 3-glucan is limited. The high polymerization degree linear chain beta-1, 3-glucan is mostly used as food additives due to the gel property, and the market scale and the commercial value are limited.
In order to improve the application range of the beta-1, 3-glucan, one method is to prepare a soluble derivative of the beta-1, 3-glucan, and the other method is to degrade the beta-1, 3-glucan to obtain the soluble beta-1, 3-glucan with small molecular weight. The preparation method of the derivative has the problems of complex process, harsh production conditions and the like. According to the existing, patent publications and literature introduction, the degradation preparation method mainly degrades beta-1, 3-glucan with high polymerization degree by physical, chemical and biological methods and the like, and has the problems of complex production process, low degradation efficiency, complex components of degradation products and the like, and the problems of large three-waste output, serious pollution and the like in the physical, chemical and other methods.
Disclosure of Invention
In order to solve the problems, the invention provides a preparation method of micromolecular beta-1, 3-glucan. The preparation method of the micromolecule beta-1, 3-glucan takes macromolecule insoluble beta-1, 3-glucan obtained by fermenting rhizobium GL-1803 as a substrate, and hydrolyzes the macromolecule insoluble beta-1, 3-glucan through endo-beta-1, 3-glucanase to obtain micromolecule beta-1, 3-glucan; the preservation number of the rhizobium GL-1803 is as follows: CGMCC No. 23416. The purity of the obtained micromolecule beta-1, 3-glucan is as high as more than 90% (w/w). The preparation method is simple, low in production cost and free of environmental pollution, and provides possibility for large-scale production of the micromolecular beta-1, 3-glucan.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a preparation method of micromolecular beta-1, 3-glucan, which comprises the following steps: carrying out fermentation culture on rhizobium GL-1803; after fermentation, filtering and concentrating the fermentation liquor to obtain concentrated solution containing macromolecular insoluble beta-1, 3-glucan; adding an acid solution into the obtained concentrated solution to adjust the pH value to 4.5-5.5, adding endo-beta-1, 3-glucanase, and reacting at 45-60 ℃ for 1-4 hours; after the reaction is finished, adding an alkali solution to adjust the pH value to 6.5-7.0 to obtain a conversion solution; filtering the conversion solution by a ceramic membrane to obtain a micromolecular beta-1, 3-glucan mixed solution; filtering the mixed solution of the micromolecular beta-1, 3-glucan by using an ultrafiltration membrane to obtain salt-containing micromolecular beta-1, 3-glucan liquid; filtering and concentrating the saliferous micromolecule beta-1, 3-glucan liquid by using a nanofiltration membrane; the preservation number of the rhizobium GL-1803 is as follows: CGMCC No. 23416.
Further, rhizobium GL-1803 is subjected to fermentation culture including seed culture and fermentation culture; seed culture: inoculating rhizobium GL-1803 into a seed culture medium, and carrying out shaking culture at the temperature of 28-32 ℃ and at the speed of 150-240 r/min for 16-24 hours to obtain a seed culture solution; fermentation culture: inoculating the seed culture solution into a fermentation culture medium, performing fermentation culture for 66-72 hours at the initial pH of 7.0 and the stirring speed of 120-200 r/min and the ventilation quantity of 0.35-0.65 vvm at 28-32 ℃.
Still further, the composition of the seed medium is: 1.0-2.0% of cane sugar and (NH)4)2HPO4 0.5%～1.0%，KH2PO4 0.15%～0.2%，MgSO4•7H2O 0.1%，CaCO30.3 percent of corn starch, 0.15 to 0.5 percent of corn starch and pH value adjusted to 7.2; the fermentation medium comprises the following components: sucrose 5.0% (by mass), (NH)4)2HPO40.1%～1.0%，KH2PO4 0.15%～0.25%，MgSO4•7H2O 0.1%，CaCO30.15-0.3 percent of the total weight of the corn steep liquor, 0.1-0.5 percent of the corn steep liquor powder and the pH value is adjusted to 7.0.
Further, after fermentation is finished, carrying out ceramic membrane filtration concentration on the fermentation liquor, controlling the temperature at 30-35 ℃, continuously adding water into the concentrated liquor in the concentration process, stopping adding water when the added water amount reaches 0.5-1.5 times of the initial volume of the fermentation liquor, and continuously concentrating until the content of the solid content of the macromolecular insoluble beta-1, 3-glucan is 40-80g/L, so as to obtain the concentrated liquor containing the macromolecular insoluble beta-1, 3-glucan.
Further, the endo-beta-1, 3-glucanase includes but is not limited to pbBgl64A, and the enzyme adding amount is 0.5-2U/mL.
The pbBgl64A is any one of the proteins a), b) and c) described in the patent PCT/CN 2016/105187.
Further, carrying out ceramic membrane filtration on the conversion solution, continuously adding water into the conversion solution in the filtration process, collecting filtrate, eluting most of micromolecule beta-1, 3-glucan into the filtrate when the conductivity value of the filtrate reaches 500-1500 mu s/cm, stopping adding water, and mixing the filtrates to obtain micromolecule beta-1, 3-glucan mixed solution;
continuously adding water into the micromolecular beta-1, 3-glucan mixed solution in the process of filtering by adopting an ultrafiltration membrane, collecting filtrate, eluting most of micromolecular beta-1, 3-glucan into the filtrate when the conductivity value of the filtrate is 500-1500 mu s/cm, stopping adding water, and mixing the filtrate to obtain the salt-containing micromolecular beta-1, 3-glucan liquid.
Further, continuously adding water into the salt-containing micromolecule beta-1, 3-glucan liquid when the salt-containing micromolecule beta-1, 3-glucan liquid is filtered and concentrated by a nanofiltration membrane, discharging the filtrate outwards, and stopping adding water when the conductivity value of the discharged filtrate is 500-1000 mus/cm; and continuously concentrating until the final mass ratio of the micromolecular beta-1, 3-glucan is more than or equal to 20 percent.
Furthermore, the membrane aperture of the ceramic membrane is 50-200 nm; the cut-off molecular weight of the ultrafiltration membrane is 5000-10000 Da; the cut-off molecular weight of the nanofiltration membrane is 100-200 Da.
Preservation information of the Rhizobium GL-1803:
the rhizobia bacterium (A), (B), (C), (B), (C), (B), (C), (B), (C), (B), (C), (B), (C)Rhizobiumsp.) GL-1803 was deposited at the China general microbiological culture Collection center on the 09.15.2021 year old with the following deposit numbers: CGMCC No.23416, preservation Address: the institute of microbiology, national academy of sciences No. 3, Xilu No. 1, Beijing, Chaoyang, Beijing.
Compared with the prior art, the invention has the following advantages:
the invention adopts rhizobium GL-1803 to carry out fermentation culture to produce macromolecular insoluble beta-1, 3-glucan, and has high yield; concentrating the obtained fermentation liquor, directly using the concentrated fermentation liquor as a substrate, and performing hydrolysis reaction with endo-beta-1, 3-glucanase; after filtration and concentration, the micromolecular beta-1, 3-glucan with the purity of more than or equal to 90 percent can be obtained, and the total extraction yield is more than or equal to 80 percent.
The method has the advantages of simple steps, low production cost and environmental friendliness, and the obtained micromolecule beta-1, 3-glucan has high purity and provides possibility for large-scale production of the micromolecule beta-1, 3-glucan.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular is intended to include the plural unless the context clearly dictates otherwise, and it should be understood that the terms "comprises" and "comprising," when used in this specification, specify the presence of the stated features, steps, operations, and combinations thereof.
In order to make the technical solutions of the present invention more clearly understood by those skilled in the art, the technical solutions of the present invention will be described in detail below with reference to specific embodiments.
Example 1
A preparation method of small molecule beta-1, 3-glucan comprises the following steps:
s1, activating the strain producing beta-1, 3-glucan, inoculating 500mL of seed culture medium for culture, and preparing a seed culture solution;
the strain for producing the beta-1, 3-glucan is rhizobium GL-1803 (CGMCC number 23416).
The composition of the seed culture medium is as follows: sucrose 2.0% (by mass), (NH)4)2HPO4 0.5%，KH2PO40.15%，MgSO4•7H2O 0.1%，CaCO30.3 percent of corn starch, 0.15 percent of corn starch and pH value adjusted to 7.2.
The seed culture temperature is 30 ℃, and the seed culture solution is obtained after shaking culture for 24 hours at 220 r/min.
S2, inoculating the seed culture solution into 6L fermentation medium, and performing fermentation culture.
The fermentation medium comprises the following components: sucrose 5.0% (by mass), (NH)4)2HPO4 0.2%，KH2PO40.2%，MgSO4•7H2O 0.1%，CaCO30.2 percent of corn steep liquor powder and 0.2 percent of corn steep liquor powder, and the pH value is adjusted to 7.0.
The fermentation culture temperature is 30 ℃, the initial pH value is 7.0, the stirring speed is 200r/min, the ventilation rate is 0.5vvm, the fermentation culture is carried out for 72 hours, and the fermentation broth is obtained, wherein the content of macromolecular insoluble beta-1, 3-glucan in the fermentation broth is 35 g/L.
And S3, after fermentation is finished, carrying out ceramic membrane filtration and concentration on the fermentation liquor, controlling the operating temperature to be 30 ℃, discharging the filtrate, continuously adding purified water into the concentrated solution in the concentration process, stopping adding the purified water when the adding amount of the purified water reaches 1 time of the initial volume of the fermentation liquor, and continuously concentrating until the solid content of the macromolecular insoluble beta-1, 3-glucan is 80g/L, so as to obtain the concentrated solution containing the macromolecular insoluble beta-1, 3-glucan.
The membrane aperture of the ceramic membrane is 200 nm.
S4, transferring the concentrated solution containing the macromolecular insoluble beta-1, 3-glucan into a conversion tank, adding a hydrochloric acid solution into the concentrated solution, and adjusting the pH value to 5.0;
the concentration of the hydrochloric acid solution is 2 mol/L.
S5, adding an enzyme preparation, heating the conversion solution in the conversion tank to 55 ℃, and stirring for reaction for 2.5 hours.
The enzyme of the enzyme preparation was pbBgl 64A. The pbBgl64A is protein a) described in PCT/CN 2016/105187.
The addition amount of the enzyme preparation is 1U/mL.
And S6, after the conversion is finished, adding a sodium hydroxide solution into the conversion solution, and neutralizing to pH6.8 to obtain the conversion solution.
The concentration of the sodium hydroxide solution is 1 mol/L.
And S7, performing ceramic membrane filtration on the conversion solution, adding purified water into the conversion solution in the filtration process, collecting filtrate, stopping adding the purified water when the conductivity of the filtrate is reduced to 1000 mu S/cm, and mixing the filtrates to obtain the micromolecular beta-1, 3-glucan mixed solution.
The membrane aperture of the ceramic membrane is 50 nm.
S8, filtering the mixed solution of the micromolecule beta-1, 3-glucan by using an ultrafiltration membrane, adding purified water into the mixed solution of the micromolecule beta-1, 3-glucan in the filtering process, collecting filtrate, stopping adding the purified water when the electric conductivity of the filtrate is reduced to 1000 mu S/cm, and mixing the filtrate to obtain the salt-containing micromolecule beta-1, 3-glucan liquid.
The cutoff molecular weight of the ultrafiltration membrane is 10000 Da.
S9, filtering and concentrating the salt-containing micromolecule beta-1, 3-glucan liquid by using a nanofiltration membrane, discharging filtrate, continuously adding purified water into the concentrated solution in the concentration process, stopping adding the purified water when the conductivity of the discharged filtrate is reduced to 600 mu S/cm, and continuously concentrating until the final mass ratio of the micromolecule beta-1, 3-glucan is 20% to obtain the micromolecule beta-1, 3-glucan concentrated solution.
The molecular weight cut-off of the nanofiltration membrane is 200 Da.
S10, drying the micromolecule beta-1, 3-glucan concentrated solution by using a spray drying tower, wherein the air inlet temperature of the spray drying tower is 130 ℃, and the air outlet temperature of the spray drying tower is 80 ℃; obtaining the micromolecular beta-1, 3-glucan powder, and packaging to obtain the micromolecular beta-1, 3-glucan product.
The purity of the obtained micromolecular beta-1, 3-glucan product is 90.45 percent, and the total extraction yield is 81.45 percent.
Example 2
The difference from example 1 is that the fermentation broth after completion of the fermentation in step S3 was concentrated by ceramic membrane filtration until the content of insoluble solids of macromolecular β -1, 3-glucan was 40g/L, and the other steps were the same as in example 1. The purity of the obtained micromolecular beta-1, 3-glucan product is 91.05%, and the total extraction yield is 83.65%.
Example 3
The difference from example 1 is that the fermentation broth after the end of the fermentation in step S3 was concentrated by ceramic membrane filtration until the content of insoluble β -1, 3-glucan solids was 60g/L, and the other steps were the same as in example 1. The purity of the obtained micromolecular beta-1, 3-glucan product is 91.21 percent, and the total extraction yield is 82.34 percent.
Example 4
The difference from example 1 is that the procedure of step S5 was the same as that of example 1 except that the temperature of the conversion solution in the conversion tank was raised to 60 ℃ after the enzyme preparation was added, and the reaction was stirred for 1 hour. The purity of the obtained micromolecular beta-1, 3-glucan product is 90.41 percent, and the total extraction yield is 80.02 percent.
Example 5
The difference from example 1 is that the temperature of the conversion solution in the conversion tank was raised to 45 ℃ after the enzyme preparation was added in step S5, and the reaction was stirred for 4 hours, and the other steps were the same as in example 1. The purity of the obtained micromolecular beta-1, 3-glucan product is 90.37%, and the total extraction yield is 80.72%.
Example 6
The difference from example 1 is that in step S7, when the conductivity of the filtrate decreased to 1400. mu.s/cm, the addition of purified water was stopped, and the other steps were the same as in example 1. The purity of the obtained micromolecule beta-1, 3-glucan product is 90.32 percent, and the total extraction yield is 80.84 percent.
Example 7
The difference from example 1 is that in step S7, when the conductivity of the filtrate decreased to 600. mu.s/cm, the addition of purified water was stopped, and the other steps were the same as in example 1. The purity of the obtained micromolecule beta-1, 3-glucan product is 90.57 percent, and the total extraction yield is 81.82 percent.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (8)
1. A preparation method of small molecular beta-1, 3-glucan is characterized by comprising the following steps:
carrying out fermentation culture on rhizobium GL-1803; after fermentation, filtering and concentrating the fermentation liquor to obtain concentrated solution containing macromolecular insoluble beta-1, 3-glucan; adding an acid solution into the obtained concentrated solution to adjust the pH value to 4.5-5.5, adding endo-beta-1, 3-glucanase, and reacting at 45-60 ℃ for 1-4 hours; after the reaction is finished, adding an alkali solution to adjust the pH value to 6.5-7.0 to obtain a conversion solution; filtering the conversion solution by a ceramic membrane to obtain a micromolecular beta-1, 3-glucan mixed solution; filtering the mixed solution of the micromolecular beta-1, 3-glucan by using an ultrafiltration membrane to obtain salt-containing micromolecular beta-1, 3-glucan liquid; filtering and concentrating the saliferous micromolecule beta-1, 3-glucan liquid by using a nanofiltration membrane;
the preservation number of the rhizobium GL-1803 is as follows: CGMCC No. 23416.
2. The process according to claim 1, wherein the fermentation culture of Rhizobium GL-1803 comprises seed culture and fermentation culture;
seed culture: inoculating rhizobium GL-1803 into a seed culture medium, and carrying out shaking culture at the temperature of 28-32 ℃ and at the speed of 150-240 r/min for 16-24 hours to obtain a seed culture solution;
fermentation culture: inoculating the seed culture solution into a fermentation culture medium, performing fermentation culture for 66-72 hours at the initial pH of 7.0 and the stirring speed of 120-200 r/min and the ventilation quantity of 0.35-0.65 vvm at 28-32 ℃.
3. The method of claim 2, wherein the seed medium has a composition of: 1.0-2.0% of cane sugar and (NH)4)2HPO4 0.5%～1.0%，KH2PO4 0.15%～0.2%，MgSO4•7H2O 0.1%，CaCO30.3 percent of corn starch, 0.15 to 0.5 percent of corn starch and pH value adjusted to 7.2; the fermentation medium comprises the following components: sucrose 5.0% (by mass), (NH)4)2HPO4 0.1%～1.0%，KH2PO4 0.15%～0.25%，MgSO4•7H2O 0.1%，CaCO30.15-0.3 percent of the total weight of the corn steep liquor, 0.1-0.5 percent of the corn steep liquor powder and the pH value is adjusted to 7.0.
4. The preparation method according to claim 1, wherein after the fermentation is finished, the fermentation liquid is subjected to ceramic membrane filtration concentration, the temperature is controlled to be 30-35 ℃, water is continuously added into the concentrated liquid in the concentration process, when the added water amount reaches 0.5-1.5 times of the initial volume of the fermentation liquid, the water is stopped being added, and the concentration is continued until the content of the macromolecular insoluble beta-1, 3-glucan solid is 40-80g/L, so that the concentrated liquid containing the macromolecular insoluble beta-1, 3-glucan is obtained.
5. The method of claim 1, wherein the endo-beta-1, 3-glucanase is pbBgl64A and the amount of enzyme added is 0.5-2U/mL.
6. The preparation method of claim 1, wherein the conversion solution is subjected to ceramic membrane filtration, water is continuously added into the conversion solution during the filtration process, the filtrate is collected, when the conductivity value of the filtrate reaches 500-1500 μ s/cm, most of the micromolecular beta-1, 3-glucan is eluted into the filtrate, the water addition is stopped, and the filtrates are mixed to obtain micromolecular beta-1, 3-glucan mixed solution;
continuously adding water into the micromolecular beta-1, 3-glucan mixed solution in the process of filtering by adopting an ultrafiltration membrane, collecting filtrate, eluting most of micromolecular beta-1, 3-glucan into the filtrate when the conductivity value of the filtrate is 500-1500 mu s/cm, stopping adding water, and mixing the filtrate to obtain the salt-containing micromolecular beta-1, 3-glucan liquid.
7. The preparation method of claim 1, wherein when the salt-containing small molecular weight beta-1, 3-glucan liquid is subjected to nanofiltration membrane filtration and concentration, water is continuously added into the salt-containing small molecular weight beta-1, 3-glucan liquid, the filtrate is discharged outwards, and when the conductivity value of the discharged filtrate is 500-1000 μ s/cm, the water addition is stopped; and continuously concentrating until the final mass ratio of the micromolecular beta-1, 3-glucan is more than or equal to 20 percent.
8. The preparation method according to claim 1, wherein the ultrafiltration membrane has a molecular weight cut-off of 5000-10000 Da; the cut-off molecular weight of the nanofiltration membrane is 100-200 Da.
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Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1985002626A1 (en) * | 1983-12-14 | 1985-06-20 | Cooperatieve Vereniging Suiker Unie U.A. | Process for the recovery of a gelforming polysaccharide, the recovered polysaccharide as well as the gel formed therewith |
| US4576915A (en) * | 1982-10-14 | 1986-03-18 | Daikin Kogyo Co., Ltd. | Strains of Rhizobium phaseoli and preparation of cyclic (1→2)-β- |
| CN102277319A (en) * | 2011-08-02 | 2011-12-14 | 河北鑫合生物化工有限公司 | Rhizobium radiobacter and method for producing curdian fermentation liquor by using strain |
| CN105420160A (en) * | 2015-12-18 | 2016-03-23 | 东华大学 | Curdlan producing strain and application thereof |
| CN105567779A (en) * | 2016-03-03 | 2016-05-11 | 江南大学 | Fermentation method of high-yield and low-molecular-weight thermal gel |
| CN106399145A (en) * | 2016-05-17 | 2017-02-15 | 江南大学 | Rhizobium radiobacter and method for producing curdlan gum through fermentation of rhizobium radiobacter |
| CN106434495A (en) * | 2016-12-02 | 2017-02-22 | 张星昊 | Rhizobium pusense and method for preparing beta-1,3 glucan fermenting liquid by rhizobium pusense |
| CN106701885A (en) * | 2017-02-14 | 2017-05-24 | 华东师范大学 | Fermenting production method of curdlan |
| CN108467876A (en) * | 2018-03-16 | 2018-08-31 | 华东师范大学 | A kind of fermentation process improving curdlan yield |
| CN112358985A (en) * | 2020-11-09 | 2021-02-12 | 河北沣川生物科技有限公司 | Pradazobium and application thereof in preparation of water-soluble beta-1, 3 glucan |
| CN112646846A (en) * | 2020-12-23 | 2021-04-13 | 山东国力生物科技有限公司 | Method for producing beta-1, 3-glucan by using non-growth coupling characteristic of bacteria |
-
2021
- 2021-12-22 CN CN202111575703.3A patent/CN113943761B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4576915A (en) * | 1982-10-14 | 1986-03-18 | Daikin Kogyo Co., Ltd. | Strains of Rhizobium phaseoli and preparation of cyclic (1→2)-β- |
| WO1985002626A1 (en) * | 1983-12-14 | 1985-06-20 | Cooperatieve Vereniging Suiker Unie U.A. | Process for the recovery of a gelforming polysaccharide, the recovered polysaccharide as well as the gel formed therewith |
| CN102277319A (en) * | 2011-08-02 | 2011-12-14 | 河北鑫合生物化工有限公司 | Rhizobium radiobacter and method for producing curdian fermentation liquor by using strain |
| CN105420160A (en) * | 2015-12-18 | 2016-03-23 | 东华大学 | Curdlan producing strain and application thereof |
| CN105567779A (en) * | 2016-03-03 | 2016-05-11 | 江南大学 | Fermentation method of high-yield and low-molecular-weight thermal gel |
| CN106399145A (en) * | 2016-05-17 | 2017-02-15 | 江南大学 | Rhizobium radiobacter and method for producing curdlan gum through fermentation of rhizobium radiobacter |
| CN106434495A (en) * | 2016-12-02 | 2017-02-22 | 张星昊 | Rhizobium pusense and method for preparing beta-1,3 glucan fermenting liquid by rhizobium pusense |
| CN106701885A (en) * | 2017-02-14 | 2017-05-24 | 华东师范大学 | Fermenting production method of curdlan |
| CN108467876A (en) * | 2018-03-16 | 2018-08-31 | 华东师范大学 | A kind of fermentation process improving curdlan yield |
| CN112358985A (en) * | 2020-11-09 | 2021-02-12 | 河北沣川生物科技有限公司 | Pradazobium and application thereof in preparation of water-soluble beta-1, 3 glucan |
| CN112646846A (en) * | 2020-12-23 | 2021-04-13 | 山东国力生物科技有限公司 | Method for producing beta-1, 3-glucan by using non-growth coupling characteristic of bacteria |
Non-Patent Citations (5)
| Title |
|---|
| HONGLIANG GAO ET AL.: "Characterization and rheological properties analysis of the succinoglycan produced by a high-yield mutant of Rhizobium radiobacter ATCC 19358", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
| RIADH BEN SALAH ET AL.: "Fermentation of date palm juice by curdlan gum production from Rhizobium radiobacter ATCC 6466 Purification, rheological and physico-chemical characterization", 《LWT - FOOD SCIENCE AND TECHNOLOGY》 * |
| VINAY CHAUDHARI ET AL.: "Therapeutic and Industrial Applications of Curdlan With Overview on Its Recent Patents", 《FRONTIERS IN NUTRITION》 * |
| XIAO-YU-ZHU WANG ET AL.: "Enhanced curdlan production with nitrogen feeding during polysaccharide synthesis by Rhizobium radiobacter", 《CARBOHYDRATE POLYMERS》 * |
| 王萧玉竹等: "可得然胶生产菌种的筛选及发酵条件优化", 《食品与生物技术学报》 * |
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