CN108467876B - Fermentation method for increasing yield of curdlan - Google Patents
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Abstract
The invention discloses a novel fermentation method for improving curdlan yield, which comprises the following steps: inoculating the seed culture solution of the strain producing curdlan into a fermentation culture medium for culturing, respectively discharging fermentation liquor and supplementing fresh culture medium in 48 hours and 72 hours of curdlan fermentation, and controlling the dissolved oxygen in the fermentation process to be not less than 30%. Compared with the discharging sugar supplement process, the yield of curdlan is improved by 40 percent and the gel strength of the fermentation liquor is improved by 58 percent by adopting the process. When the dissolved oxygen in the gel production period is controlled to be more than 30 percent, compared with the process in which the dissolved oxygen is less than 10 percent, the yield of curdlan is improved by 86.7 percent, and the gel strength is improved by 91.4 percent. The method of the invention improves the rate of producing curdlan gum in the middle and later stages of fermentation, has simple process control and saves production cost, and provides a new method for producing curdlan gum by microbial fermentation.
Description
Technical Field
The invention belongs to the field of microbial polysaccharide fermentation, and particularly relates to a fermentation method for improving curdlan yield.
Background
Curdlan (Curdlan) is a novel microbial exopolysaccharide, also known as a thermal gel due to its unique property of forming a gel under heat. Chemical structural analysis of Curdlan showed that it is an unbranched, homogeneous polysaccharide polymer of single D-glucose linked by β -1, 3-glycosidic linkages at C1 and C3 positions.
Curdlan fermentation is a typical uncoupled fermentation, and no curdlan is produced during the growth of the strain; after the thalli grow, the thalli begin to secrete curdlan under the condition of nitrogen source limitation. The curdlan is water insoluble and easily soluble in alkali solution and dimethyl sulfoxide. When the temperature is heated to 55 ℃, the low-strength reversible solid curdlan can be obtained; when the temperature is continuously heated to be higher than 80 ℃, irreversible solid curdlan can be formed. Due to the characteristics, the health food is safe and nontoxic, and is widely applied to foods such as noodles, jelly and the like. The FDA in the united states approved curdlan as an edible polysaccharide in 1996, which is the third microbial polysaccharide to be used as a food additive. The curdlan approved by the Chinese food and drug administration in 2006 can be used in the food field.
Curdlan fermentation is a process of synthesizing polysaccharides from small molecular sugars such as glucose and sucrose by Agrobacterium sp or Rhizobium sp. In the fermentation process, a large amount of carbon source is consumed, so that the content of the carbon source is reduced in the later fermentation period, and the requirement of thalli for synthesizing curdlan cannot be met. Liu Hui and the like (influence of different substrate feeding modes on Alcaligenes faecalalis fermentation production thermal gel, biotechnology, 18(4) in 2008: 78-81) optimize curdlan fermentation production by adopting a two-stage method, ammonia water is used for controlling the pH value to be 7.0 in a thallus growth stage, glucose is used for continuous feeding in a curdlan synthesis stage, and the yield of curdlan is improved by 157%. The ATCC31749 strain was used by Korean Lee et al (Production of current use sugar or sugar cane rubbers by two o-step fed-batch culture of Agrobacterium species, Journal of Industrial Microbiology & Biotechnology 1997,18: 255-. Curdlan is exopolysaccharide produced under the condition of nitrogen source limitation, when the nitrogen source is sufficient, the thalli only accumulate biomass, and when the nitrogen source is exhausted, the thalli stop growing and begin to synthesize curdlan (regulating and controlling research on exopolysaccharide synthesis by Agrobacterium sp.atcc 31749 by ntrB gene, industrial microorganism, vol.41, sixth phase, 26-33, 2011).
However, the invention unexpectedly discovers that the culture medium containing the nitrogen source is supplemented in the middle and later stages of fermentation, the yield of the curdlan is not reduced, the yield of the curdlan is improved by about 40%, the quality of the curdlan product is improved, and the gel strength of key performance indexes of the curdlan is increased by 58%.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for producing curdlan by semi-continuous culture and supplement of culture medium components, which can obviously improve the yield of curdlan and is beneficial to reducing the production cost of curdlan. The material feeding time of the invention is divided into two stages in the aspect of material feeding time selection, wherein the material feeding time of the first material feeding is 30-54h, and the material feeding time of the second material feeding is 60-84 h. Discharging and feeding materials in the two time periods can increase the bacterial mass and the curdlan yield. Compared with the discharging sugar supplement process, the yield of curdlan is improved by 40 percent and the gel strength of the fermentation liquor is improved by 58 percent by adopting the process. When the dissolved oxygen in the gel production period is controlled to be more than 30 percent, compared with the process in which the dissolved oxygen is less than 10 percent, the yield of curdlan is improved by 86.7 percent, and the gel strength is improved by 91.4 percent.
The following is a detailed description of the technical solution of the present invention.
The invention provides a method for producing curdlan by fermentation, which comprises the steps of inoculating a seed culture solution of a strain producing curdlan into a fermentation culture medium for culture, feeding materials during fermentation, controlling pH, and controlling dissolved oxygen concentration DO to be not less than 30% until the end of fermentation culture.
The method comprises the following specific steps:
(1) scraping a ring of bacteria from the inclined plane of the strain producing curdlan, inoculating the bacteria into a seed culture medium, and performing shaking culture to obtain activated seed liquid of the strain producing curdlan;
(2) inoculating the seed liquid obtained in the step (1) into a fermentation culture medium;
(3) discharging fermentation liquor when the fermentation is carried out for 48 hours, then feeding the culture medium with the same volume in the step (2) for one time, continuing the fermentation, and controlling the dissolved oxygen concentration DO;
(4) when the fermentation time is up to 72 hours, discharging fermentation liquor, then feeding the culture medium in the step (2) with the same volume once, continuing fermentation, and controlling the dissolved oxygen concentration DO;
(5) and (3) adding alkali into the fermentation liquor after the fermentation is finished to dissolve, centrifuging, regulating the pH value of the supernatant by using hydrochloric acid, centrifuging to obtain a precipitate, washing the precipitate by using distilled water for three times, and drying to obtain the curdlan.
In the step (1), the curdlan producing strain comprises one or more of agrobacterium ATCC31749, rhizobium ATCC 31750 or ATCC21680, or other strains which can produce curdlan and are publicly reported; preferably, it is Agrobacterium ATCC31749, Rhizobium ATCC 31750 or ATCC 21680.
In the step (1), the seed culture medium comprises: contains (g/L) in each liter of culture medium: 10-30 of cane sugar, (NH)4)2HPO43-8，KH2PO41-3 parts of corn steep liquor, 2-4 parts of MgSO4.7H2O 0.5-2，CaCO31-4; preferably, the culture medium contains (g/L): sucrose 20, (NH)4)2HPO4 5，KH2PO41.5, corn steep liquor 3, MgSO4.7H2O 1，CaCO3 3。
In the step (1), the temperature of the shaking culture is 28-32 ℃; preferably, it is 30 ℃.
In the step (1), the rotation speed of the shaking culture is 150-; preferably, it is 250 r/min.
In the step (1), the shaking culture time is 16-20 h; preferably 18 h.
In the step (2), the fermentation medium comprises the following components: contains (g/L) in each liter of culture medium: glucose 40-100, (NH)4)2HPO41 to 3 portions of yeast extract, 0.5 to 1 portion of KH2PO4 0.5-2、MgSO4·7H2O 0.1-1、FeSO4·7H2O 0.01-1、MnSO4·H2O 0.01-0.05、CoCl2·6H2O 0.01、ZnCl20.01、CaCO31-5; preferably, the culture medium contains (g/L): glucose 50, (NH)4)2HPO41.5, yeast extract 1, KH2PO41、MgSO4·7H2O 0.5、FeSO4·7H2O 0.05、MnSO4·H2O 0.02、CoCl2·6H2O 0.01、ZnCl20.01 and CaCO33。
In the step (2), the fermentation conditions are preferably: the rotating speed is 500r/min, the ventilation volume is 1vvm, and the fermentation is 96 h.
In the step (2), the inoculation volume is 5-10% (v/v); preferably, it is 10% (v/v).
In the step (3), the volume of the discharged materials accounts for 10% -30% of the total volume of the culture medium; preferably, it is 25%.
In the step (3), the volume of the feed supplement accounts for 10-30% of the total volume of the culture medium; preferably, it is 25%;
wherein the emptying volume is the same as the feeding volume.
Wherein the components of the culture medium are as follows: contains (g/L) in each liter of culture medium: glucose 40-100, (NH)4)2HPO41 to 3 portions of yeast extract, 0.5 to 1 portion of KH2PO40.5-2、MgSO4·7H2O 0.1-1、FeSO4·7H2O 0.01-1、MnSO4·H2O 0.01-0.05、CoCl2·6H2O 0.01、ZnCl20.01、CaCO31-5; preferably, the culture medium contains (g/L): glucose 50, (NH)4)2HPO41.5, 1g of yeast extract and KH2PO41、MgSO4·7H2O 0.5、FeSO4·7H2O 0.05、MnSO4·H2O 0.02、CoCl2·6H2O 0.01、ZnCl20.01 and CaCO33。
In the step (3), the dissolved oxygen concentration DO in the fermentation liquor is not lower than 30%; preferably, it is 35%.
In the step (4), the volume of the discharged materials accounts for 10-30% of the total volume of the culture medium; preferably, it is 12.5%.
In the step (4), the primary flow volume accounts for 10-30% of the total volume of the culture medium; preferably, it is 12.5%.
Wherein the emptying volume is the same as the primary flow volume.
Wherein the components of the culture medium are as follows: contains (g/L) in each liter of culture medium: glucose 40-100, (NH)4)2HPO41 to 3 portions of yeast extract, 0.5 to 1 portion of KH2PO40.5-2、MgSO4·7H2O 0.1-1、FeSO4·7H2O 0.01-1、MnSO4·H2O 0.01-0.05、CoCl2·6H2O 0.01、ZnCl20.01、CaCO31-5; preferably, the culture medium contains (g/L): glucose 50, (NH)4)2HPO41.5, yeast extract 1, KH2PO41、MgSO4·7H2O 0.5、FeSO4·7H2O 0.05、MnSO4·H2O 0.02、CoCl2·6H2O 0.01、ZnCl20.01 and CaCO3 3。
In the step (4), the dissolved oxygen concentration DO in the fermentation liquor is not lower than 30%; preferably, it is 35%.
In the step (5), the alkali is NaOH, KOH, Ca (OH)2One or more of the following; preferably, it is NaOH.
In the step (5), the concentration of the alkali liquor is 0.5-3 mol/L; preferably, it is 1 to 3 mol/L; further preferably, it is 1 mol/L.
In the step (5), the concentration of the hydrochloric acid is 1-3 mol/L; preferably, it is 3 mol/L.
In the step (5), the volume of distilled water added for each time of water washing is 50-150 ml; preferably 150 ml.
In the step (5), the drying temperature is 50-100 ℃; preferably, it is 60 ℃.
In the step (5), the pH value of the supernatant is adjusted to 5.0-8.0 by hydrochloric acid; preferably, it is 7.0.
The invention has the beneficial effects that: according to the fermentation characteristics of curdlan, the yield of curdlan is improved from 40g/L to 56g/L by a semi-continuous fermentation method, and is improved by 40%. The obtained curdlan meets the requirements of national standards on curdlan and can meet the industrial production requirements of curdlan. The method of the invention improves the rate of producing curdlan gum in the middle and later stages of fermentation, has simple process control and saves production cost, and provides a new method for producing curdlan gum by microbial fermentation.
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FIG. 1 shows the effect of different feeding methods on curdlan fermentation bacteria amount
Detailed Description
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.
Example 1 (first feeding carbon source)
(1) Scraping one ring from the slant of ATCC31749 strain preserved in laboratory into 500ml triangular flask containing 100ml seed culture medium, and shake culturing at 30 deg.C and 250r/min for 18h to obtain seed solution; wherein the seed culture medium comprises the following components in g/L: sucrose (20 g), (NH)4)2HPO4 5g，KH2PO41.5g, corn steep liquor 3g, MgSO4.7H2O 1g，CaCO3 3g。
(2) Inoculating the seed solution of the ATCC31749 strain of the step (1) into 8L of fermentation medium, wherein the inoculation volume is 10% (v/v). After inoculation, the ventilation capacity is 1vvm at 30 ℃ and 500r/min, and the culture is carried out for 96 h; wherein the fermentation medium comprises the following components in g/L: glucose 50g, (NH)4)2HPO41.5g, yeast extract 1g, KH2PO4 1g、MgSO4·7H2O 0.5g、FeSO4·7H2O 0.05g、MnSO4·H2O 0.02g、CoCl2·6H2O 0.01g、ZnCl20.01g of calcium carbonate and CaCO3 3g。
(3) Fermenting for 48h, wherein the volume of discharged materials is 25% of the total volume, and then adding glucose solution with the same volume and the concentration of 50g/L for one time. And (5) continuing fermenting for 96h, and finishing the fermentation.
(4) Dissolving 10g of the fermentation liquor in 150ml of alkali, centrifuging, adjusting the pH of supernatant to 7.0 by hydrochloric acid, centrifuging to obtain precipitate, washing the precipitate with distilled water for three times, drying at 60 ℃ to obtain a curdlan sample, and weighing to calculate the yield of curdlan.
(5) And (4) measuring the gel strength of the sample obtained in the step (4). The method for measuring the gel strength comprises the following steps: according to the national standard method, 0.3g of sample is taken, 15ml of water is added, then high-speed homogenization is carried out for 10min at 13000r/min, the homogenized suspension is transferred into a test tube with the diameter of 18mm multiplied by 180mm, water bath heating at 95 ℃ is carried out for 10min, tap water is cooled to room temperature, the part of the test tube, which is 10mm-15mm away from the bottom, is taken for flat cutting, a stainless steel piston type cylindrical probe with the diameter of 0.5cm is selected, and the probe moves at the speed of 250 mm/min.
Example 2 (continuous flow Medium)
(1) The fermentation process was the same as in example 1 for the 0-48h stage.
(2) Fermenting for 48h, wherein the volume of discharged materials is 25% of the total volume, slowly adding culture medium with the same volume, and continuously adding culture medium for 24 h. And when the fermentation is carried out for 72 hours, continuously discharging the materials, wherein the discharge volume accounts for 12.5 percent of the total volume, slowly supplementing the culture medium with the same volume, and continuously supplementing for 12 hours. Fermenting for 96 h. The dissolved oxygen concentration is controlled to be more than 30 percent by adjusting the stirring speed and the ventilation quantity in the fermentation process. The fed-batch culture medium comprises the following components: contains (g/L) in each liter of culture medium: (NH)4)2HPO41.5g, yeast extract 1g, KH2PO41g、MgSO4·7H2O 0.5g、FeSO4·7H2O 0.05g、MnSO4·H2O 0.02g、CoCl2·6H2O 0.01g、ZnCl20.01g。
(3) Curdlan yield was calculated and gel strength was measured according to the method of example 1.
Example 3 (one-time feeding medium)
(1) The process was the same as in example 1 during the fermentation period 0-48 h.
(2) Fermenting for 48h, wherein the volume of discharged materials is 25% of the total volume, and then supplementing the culture medium with the same volume once. When the fermentation time is 72 hours, discharging is continued, the discharged material accounts for 12.5 percent of the total volume, and then the culture medium with the same volume is replenished once. The fermentation was continued for 96 h. The dissolved oxygen concentration is controlled to be more than 30 percent by adjusting the stirring speed and the ventilation quantity in the fermentation process. The feed medium composition was the same as in example 2.
(3) Curdlan yield was calculated and gel strength was measured according to the method of example 1.
Example 4 (controlling dissolved oxygen at the gel production stage by 20% -30%)
(1) The fermentation process was the same as in example 2 for the 0-48h stage.
(2) Fermenting for 48h, wherein the volume of the discharged materials is 25% of the total volume, slowly adding culture medium with the same volume, and continuously adding culture medium for 24 h. When the fermentation time is up to 72h, the fermentation is continued, the volume of the discharged materials accounts for 12.5 percent of the total volume, and then the culture medium with the same volume is slowly supplemented for 12 h. The fermentation was continued for 96 h. The dissolved oxygen concentration is controlled to be 20-30% by adjusting the stirring speed and the ventilation quantity in the fermentation process. The feed medium composition was the same as in example 2.
(3) Curdlan yield was calculated and gel strength was measured according to the method of example 1.
Example 5 (controlling dissolved oxygen 10% -20% in the gel production period)
(1) The fermentation process was the same as in example 2 for the 0-48h stage.
(2) Fermenting for 48h, wherein the volume of the discharged materials is 25% of the total volume, slowly adding culture medium with the same volume, and continuously adding culture medium for 24 h. And (3) continuously discharging when fermenting for 72h, wherein the discharge volume accounts for 12.5% of the total volume, and then slowly supplementing the culture medium with the same volume for 12 h. The fermentation was continued for 96 h. The concentration of dissolved oxygen is controlled to be 10-20% by adjusting the stirring speed and the ventilation rate in the fermentation process. The feed medium composition was the same as in example 2.
(3) Curdlan yield was calculated and gel strength was measured according to the method of example 1.
Example 6 (controlling dissolved oxygen at gel production time to 10% or less)
(1) The fermentation process was the same as in example 2 for the 0-48h stage.
(2) Fermenting for 48h, wherein the volume of the discharged materials is 25% of the total volume, slowly adding culture medium with the same volume, and continuously adding culture medium for 24 h. And (3) continuously discharging when fermenting for 72h, wherein the discharge volume accounts for 12.5% of the total volume, and then slowly supplementing the culture medium with the same volume for 12 h. The fermentation was continued for 96 h. The concentration of dissolved oxygen is controlled to be below 10 percent by adjusting the stirring speed and the ventilation rate in the fermentation process. The feed medium composition was the same as in example 2.
(3) Curdlan yield was calculated and gel strength was measured according to the method of example 1.
TABLE 1 Effect of different feeding regimes on fermentation
TABLE 2 Cola fermentation 96 yields and gel strengths tabulations
As can be seen from tables 1 and 2 and FIG. 1, after discharging, the culture medium with the same volume is supplemented once, and compared with the simple supplement of the carbon source after discharging, the yield of curdlan can be improved, the yield of curdlan is respectively improved by 40% and 38% in examples 2 and 3, and the gel strength is respectively improved by 54.5% and 58.2%. The production rate of curdlan is improved by 30% compared with the process of simply supplementing the carbon source after supplementing the culture medium.
In the middle and later stages of fermentation of the curdlan, the viscosity is increased along with the increase of the yield, the thalli are gradually cracked and die, the carbon source cannot meet the synthesis requirement of the curdlan gradually, and the carbon source is supplemented generally. In curdlan fermentation, supplementation with supplemental carbon sources (sucrose, glucose) has been reported [ Zhan North-of-the Alcaligenes faecalis, continuous culture under different substrate-limiting conditions, 2008 ]. The invention adopts a semi-continuous culture method, namely discharging materials in the middle and later stages of fermentation, and then supplementing a carbon source, but the problems of reduction of the bacterial mass, dilution of nutritional factors and the like are caused. During fermentation of other products, a certain amount of nitrogen source is supplemented, but curdlan is synthesized under the condition of nitrogen source limitation, so that no report of nitrogen source supplementation in the middle and later stages of fermentation is seen at present.
However, the invention unexpectedly discovers that a certain culture medium is added in the middle and later stages of fermentation of the curdlan, so that a carbon source required by fermentation can be provided, the viscosity of fermentation liquor is reduced, nutritional factors in a fermentation system can be increased, the bacterial mass can be increased, the yield of the curdlan in the later stage is increased, and the gel strength of a curdlan product is improved.
TABLE 3 Effect of different dissolved oxygen control strategies on curdlan fermentation
As can be seen from Table 3, the yield of curdlan produced by the process with dissolved oxygen controlled by more than 30% in the gel production period is higher than that produced by the process with dissolved oxygen controlled by 10% -20% and that produced by the process with dissolved oxygen controlled by less than 10%, the yield is improved by 86.7%, and the gel strength of example 4 is improved by 91.4% compared with that of example 6.
The protection of the present invention is not limited to the above embodiments. Variations and advantages that may occur to those skilled in the art may be incorporated into the invention without departing from the spirit and scope of the inventive concept, and the scope of the appended claims is intended to be protected.
Claims (5)
1. A method for producing curdlan by fermentation is characterized in that strain seed culture solution for producing curdlan is inoculated into a fermentation culture medium for culture, feeding and supplementing materials are carried out during the fermentation process, and the pH and the dissolved oxygen concentration DO are controlled to be not less than 30% until the fermentation culture is finished; the DO is not less than 30 percent, namely the DO is 30 to 35 percent; the fermentation medium comprises the following components: comprises per liter of fermentation medium (g/L): 40-50 parts of glucose, (NH)4)2HPO4 1 to 3 portions of yeast extract, 0.5 to 1 portion of KH2PO40.5-2、MgSO4·7H2O 0.1-1、FeSO4·7H2O 0.01-1、MnSO4·H2O 0.01-0.05、CoCl2·6H2O 0.01、ZnCl2 0.01、CaCO3 1-5；
The steps are specifically as follows:
(1) scraping a ring of bacteria from the inclined plane of the strain producing curdlan, inoculating the bacteria into a seed culture medium, and performing shaking culture to obtain activated seed liquid of the strain producing curdlan;
(2) inoculating the seed liquid obtained in the step (1) into a fermentation culture medium; the inoculation volume is 5-10% (v/v);
(3) discharging fermentation liquor when the fermentation is carried out for 48 hours, then feeding the fermentation culture medium with the same volume in the step (2) for one time, continuing the fermentation, and controlling the dissolved oxygen concentration DO; the volume of the discharged materials accounts for 10% -30% of the total volume of the culture medium; the volume of the supplementary material accounts for 10-30% of the total volume of the culture medium; wherein the emptying volume is the same as the feeding volume;
(4) when the fermentation time is up to 72 hours, discharging fermentation liquor, then feeding the fermentation medium in the step (2) with the same volume once, continuing the fermentation, and controlling the dissolved oxygen concentration DO; the volume of the discharged materials accounts for 10% -30% of the total volume of the culture medium; the volume of the supplementary material accounts for 10-30% of the total volume of the culture medium; wherein the emptying volume is the same as the feeding volume;
(5) adding alkali into the fermentation liquor after the fermentation is finished to dissolve, centrifuging, regulating the pH value of the supernatant by hydrochloric acid, centrifuging to obtain a precipitate, washing the precipitate by distilled water for three times, and drying to obtain the curdlan;
the curdlan producing strain is selected from agrobacterium ATCC 31749.
2. The method of claim 1, wherein in step (1), the culture medium of the strain seed consists of: contains (g/L) in each liter of culture medium: 10-30 of cane sugar, (NH)4)2HPO43-8，KH2PO41-3 parts of corn steep liquor, 2-4 parts of MgSO4.7H2O 0.5-2，CaCO31-4。
3. The method according to claim 1, wherein in the step (1), the temperature of the shaking culture is 28-32 ℃; and/or the rotation speed of the shaking culture is 150-250 r/min; and/or the shaking culture time is 16-20 h.
4. The method of claim 1, wherein in step (2), the fermentation conditions are: the rotating speed is 500r/min, the ventilation volume is 1vvm, and the fermentation is 96 h.
5. The method of claim 1, wherein in step (5), the base is NaOH, KOH, Ca (OH)2One or more of the above; and/or, the concentration of the alkali liquor is 0.5-3 mol/L; and/or the concentration of the hydrochloric acid is 1-3 mol/L; and/or the volume of distilled water added for each time of water washing is 50-150 ml; and/or the drying temperature is 50-100 ℃; and/or, the pH value of the supernatant is adjusted to 5.0-8.0 by hydrochloric acid.
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| CN109837231B (en) * | 2019-04-18 | 2021-11-02 | 山东省农业科学院农产品研究所 | Alcaligenes faecalis and method for preparing curdlan with different molecular weights by using same |
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| CN113957024B (en) * | 2021-12-22 | 2022-04-19 | 山东国力生物科技有限公司 | Rhizobium GL-1803 and application thereof in preparation of insoluble beta-glucan |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1057862A3 (en) * | 1996-12-17 | 2001-02-07 | Warner-Lambert Company | Polymer film compositions for capsules |
| 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 |
| CN104745656A (en) * | 2015-03-20 | 2015-07-01 | 江南大学 | Method for directly producing beta-1,3-glucooligosaccharides by virtue of thermal gel fermentation liquor |
| CN105420159A (en) * | 2015-12-18 | 2016-03-23 | 东华大学 | Curdlan strain and application thereof |
| 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 |
| CN106701885A (en) * | 2017-02-14 | 2017-05-24 | 华东师范大学 | Fermenting production method of curdlan |
-
2018
- 2018-03-16 CN CN201810219817.6A patent/CN108467876B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1057862A3 (en) * | 1996-12-17 | 2001-02-07 | Warner-Lambert Company | Polymer film compositions for capsules |
| 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 |
| CN104745656A (en) * | 2015-03-20 | 2015-07-01 | 江南大学 | Method for directly producing beta-1,3-glucooligosaccharides by virtue of thermal gel fermentation liquor |
| CN105420159A (en) * | 2015-12-18 | 2016-03-23 | 东华大学 | Curdlan strain and application thereof |
| 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 |
| CN106701885A (en) * | 2017-02-14 | 2017-05-24 | 华东师范大学 | Fermenting production method of curdlan |
Non-Patent Citations (2)
| Title |
|---|
| Hong-Tao Zhang 等.Improved curdlan fermentation process based on optimization of dissolved oxygen combined with pH control and metabolic characterization of Agrobacterium sp. ATCC 31749.《Appl Microbiol Biotechnol》.2011,第93卷(第1期), * |
| Improved curdlan fermentation process based on optimization of dissolved oxygen combined with pH control and metabolic characterization of Agrobacterium sp. ATCC 31749;Hong-Tao Zhang 等;《Appl Microbiol Biotechnol》;20110708;第93卷(第1期);第368页右段Materials and methods部分-第369页左栏第1-3段、图1 * |
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