CN101942491A - Method for efficiently separating and extracting welan gum from welan gum fermentation liquor - Google Patents
Method for efficiently separating and extracting welan gum from welan gum fermentation liquor Download PDFInfo
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- CN101942491A CN101942491A CN 201010253396 CN201010253396A CN101942491A CN 101942491 A CN101942491 A CN 101942491A CN 201010253396 CN201010253396 CN 201010253396 CN 201010253396 A CN201010253396 A CN 201010253396A CN 101942491 A CN101942491 A CN 101942491A
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- welan gum
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Abstract
The invention relates to a method for efficiently separating and extracting welan gum from welan gum fermentation liquor, belonging to the technical field of bioengineering. The invention discloses a technique for efficiently separating and extracting the welan gum on the premise of maintaining favorable rheologic behaviors of the welan gum. The method comprises the following steps: fermenting with microbes under aerobic conditions to obtain welan gum fermentation liquor, heating to sterilize the fermentation liquor, settling by using alcohol solvent, separating solid from liquid, concentrating by dehydration, and desalting to obtain the welan gum finished product. The techniques of settling and concentrating in the method can be used for removing most pigments, residual sugar and salt ions in the fermentation liquor, thereby improving the color and properties of the welan gum product; and the techniques of salt alcohol settling and acid alcohol settling can be used for obviously reducing the alcohol consumption in the extraction process, thereby greatly saving the energy consumption in the alcohol recycling process. The invention can use basic microbial polysaccharide extraction technique and production equipment, has the advantages of lower energy consumption and alcohol consumption, and less pollution, and can obtain industrial-grade welan gum.
Description
Technical field
It is that fermentation strain obtains the Welan gum fermented liquid that the present invention utilizes Alcaligenes (Alcaligenes sp.) ATCC31555, by changing pH, salt ionic concentration and the pure addition of extracting solution, obtain the Welan gum of technical grade, be a kind of from fermented liquid high efficiency separation extract the method for Welan gum, belong to technical field of bioengineering.
Background technology
From the sixties in 20th century, xanthan gum realizes commercialization for the first time, and microbial polysaccharide has caused the great interest of scientific circles and industry member gradually, has obtained fast development, various new microbial glycocalix development and use.Welan gum is to utilize carbohydrate synthetic mixed polysaccharide by microorganism, is a kind of high molecular polymer, and monomer whose is made up of glucose, glucuronic acid, seminose and rhamnosyl, and side chain is rhamnosyl or seminose.Welan gum has many good and unique physicochemical properties, can be used as thickening material, suspension agent, stablizer etc., is widely used in industries such as building, oil production.The CP.Kelco company of the U.S. has realized the suitability for industrialized production of Welan gum the eighties in 20th century, is mainly used in concrete additive or oil tertiary recovery at present.Japan has also used Welan gum in preparation self-compacting concrete and self-leveling concrete, and is used for foodstuff additive in approval Welan gum in 1994.The domestic main rheological properties of studying Welan gum, strain improvement and fermentation optimization are very few to the Study on extraction in the Welan gum production process.
Except that containing Welan gum, also contain mycelium, residual medium component and other metabolic by-prodss in the Welan gum fermented liquid, these impurity can influence the quality of Welan gum product.Different extraction and separation method has determined performance, quality and the purposes of Welan gum.The separation-extraction technology of Welan gum comprises: the pre-treatment of fermented liquid, separation and purification, dehydrate and process such as pulverizing.In actual production, the differentiation that these processes are not strict, but merge.
CN100529055C discloses a kind of Alcaligenes and the application in the preparation Welan gum thereof, and what wherein extract the Welan gum employing is coarse filtration, ultrafiltration-separating out alcohol method, removes thalline by coarse filtration, and ultrafiltration and concentration adds the alcoholic solvent precipitation and separates the acquisition Welan gum.Do not see other relevant reports in addition about the Welan gum extraction process.
At present, the extraction process of microbial polysaccharide mainly contains the alcohol precipitation method, salt-alcohol precipitation method, and ultrafiltration-alcohol precipitation method, but directly the alcohol consumption of alcohol precipitation method is big, the cost height, and energy consumption is big; Ultrafiltration-separating out alcohol method requires high to production unit, the membrane module cost depletions is big.The purpose of this invention is to provide a kind of simple to operate, efficiently, the Welan gum separating and extracting method of less energy-consumption, conditions such as pH by changing fermented liquid and salt ionic concentration reduce the alcoholic solvent consumption in the leaching process, and the superperformance of maintenance Welan gum, reducing production costs simultaneously, is the good Welan gum separating and extracting method of a kind of economic benefit.
Summary of the invention
The purpose of this invention is to provide a kind of high efficiency separation and extract the method for Welan gum.
Further purpose of the present invention is to provide a kind of maintenance high performance Welan gum extracting method, and this method is simple to operate, and is energy-saving and cost-reducing, can be used for large-scale production process.
The object of the invention can reach by the following technical programs:
The present invention utilizes Alcaligenes (Alcaligenes sp.) ATCC31555, this bacterial strain is at [CompetitiveAdsorption Between an AMPSVR-Based Fluid Loss Polymer and Welan GumBiopolymer in Oil Well Cement, Journal ofAppliedPolymer Science, Vol.116,2913-2919 (2010) VC 2010Wiley Periodicals, Inc.] open, with it is that fermentation strain obtains the Welan gum fermented liquid, by changing the pH of extracting solution, salt ionic concentration and pure addition, the Welan gum of acquisition technical grade.
A kind of from the Welan gum fermented liquid high efficiency separation extract the method for Welan gum, technological process is:
(1) Welan gum fermented liquid preparation:
Alcaligenes (Alcaligenes sp.) ATCC 31555 seed liquor are forwarded to fermention medium with volume ratio 6% inoculum size, obtain yellow thick fermented liquid at 30 ℃, 200r/min bottom fermentation 72h;
Used fermention medium consists of sucrose 40g/L, dregs of beans 5g/L, KH
2PO
40.6g/L, MgSO
47H
2O 0.2g/L, FeCl
20.1g/L, using the tap water constant volume, pH 7.0～7.2;
(2) fermented liquid is heated the microorganism of killing in the fermented liquid, temperature is 80～100 ℃, and the time is 10～45 minutes; And with water quench to room temperature;
(3) with certain density acid solution the pH of Welan gum fermented liquid is transferred to 1.0～5.0, used acid solution is hydrochloric acid, acetate, phosphoric acid or its combination of 1～4mol/L;
And/or add a certain amount of salt ion solution; The salt that adds is selected sodium-chlor, Repone K, ammonia chloride, magnesium chloride, calcium chloride, aluminum chloride or its combination for use, and the amount of adding salt makes that cation concn reaches 0.01～0.5mol/L in the Welan gum fermented liquid;
(4) add a certain amount of alcoholic solvent, make the pure mass concentration of the mixed solution after the interpolation reach 30%～80%, and stir; Used alcoholic solvent is selected from ethanol, Virahol, methyl alcohol or its combination;
(5) solid-liquid separation is removed supernatant liquor, obtains solid substance; Isolating method adopts settlement separate, centrifugation, filtering separation and combination thereof in the solid-liquid separation process;
(6) add the alcoholic solvent of 0.2～1.0 times of the solid substance volume obtain, regulator solution pH is to neutral, press filtration or press dewatering; Perhaps the solid substance that step (5) is obtained directly carries out press filtration or press dewatering;
(7) dry and pulverizing, drying temperature is below 80 ℃, drying means is constant pressure and dry, vacuum-drying or vacuum freezedrying, is dried to product water content＜15%, promptly gets the Welan gum finished product.
The salt ion solution of the described interpolation of step (3), the salt ion of interpolation is CaCl
2, adopting the CaCl2 solution that adds 2.5mol/L to make the final cation concn of fermented liquid is 0.1mol/L.
Beneficial effect of the present invention:
1, the invention provides a kind of easy to operate simply, easily mass-producing enlarges the Welan gum extraction process of producing;
2, this extraction process significantly reduces than the alcohol consumption of direct alcohol precipitation method;
3, the sedimentary water content of wet Welan gum that obtains of this extraction process is starkly lower than the Welan gum precipitation that direct alcohol precipitation method obtains, and follow-up alcohol reclaims and dry required energy consumption obviously reduces;
4 compare with film-alcohol precipitation method, and this extraction process requires lowly to equipment, and equipment maintenance cost is little.
Embodiment
Below in conjunction with embodiment the specific embodiment of the present invention is further described, but to claim of the present invention without limits.
Fermention medium is formed: sucrose 40g/L, dregs of beans 5g/L, KH
2PO
40.6g/L, MgSO
47H
2O0.2g/L, FeCl
20.1g/L pH 7.0～7.2, fermentation condition is: the Alcaligenes seed liquor is forwarded to fermention medium with 6% (v/v) inoculum size, and at 30 ℃, 500r/min bottom fermentation 72h obtains the Welan gum fermented liquid.The Welan gum fermented liquid that obtains is 10～20g/L with Welan gum content in crude product in alcohol precipitation, the dry back mensuration fermented liquid.
Embodiment 1
1, the Welan gum fermented liquid with 500kg is heated to 85 ℃, kept 40 minutes, again with water quench to room temperature, manipulate in order to next step;
2, with the hydrochloric acid soln of 1mol/L the pH of Welan gum fermented liquid is transferred to 3.0;
3, the industrial alcohol solvent of interpolation 95% makes the determining alcohol of mixed solution reach 75% (w/w), and stirs;
4, use link-suspended basket centrifuge centrifugal 10 minutes realization solid-liquid separation under 4500g centrifugal force, collect solid substance, get the Welan gum crude product;
5, with 95% the industrial alcohol dissolving of gained solid substance with 0.5 times of solid substance volume, regulator solution pH is to neutral, filter-press dehydration;
6, under 55 ℃ of conditions constant pressure and dry to the product water ratio less than 13%, pulverize at last and obtain the Welan gum product.
Embodiment 2
1, the Welan gum fermented liquid with 200kg is heated to 100 ℃, kept 15 minutes, again with water quench to room temperature, manipulate in order to next step;
2, the magnesium chloride solution with 2.5mol/L is adjusted to 0.05mol/L with cationic concentration of electric charges in the Welan gum fermented liquid;
3, add Virahol and make the determining alcohol of mixed solution reach 42% (w/w), and stir;
4, use link-suspended basket centrifuge centrifugal 5 minutes realization solid-liquid separation under 4500g centrifugal force, collect solid substance, get the Welan gum crude product;
5, with the Virahol dissolving of gained solid substance with 0.2 times of solid substance volume, regulator solution pH is to neutral, and suction filtration is press dewatering again;
6, under 50 ℃ of conditions constant pressure and dry to the product water ratio less than 14%, pulverize at last and obtain the Welan gum product.
Embodiment 3
1, the Welan gum fermented liquid with 300kg is heated to 90 ℃, kept 25 minutes, again with water quench to room temperature, manipulate in order to next step;
2, the sodium chloride solution with 5mol/L is adjusted to 0.15mol/L with cationic concentration of electric charges in the Welan gum fermented liquid;
3, add methyl alcohol, make the determining alcohol of mixed solution reach 75% (w/w), and stir;
4, use link-suspended basket centrifuge centrifugal 10 minutes realization solid-liquid separation under 8000g centrifugal force, collect solid substance, get the Welan gum crude product;
5, add the methyl alcohol of 1.0 times of solid substance volumes, regulator solution pH is to neutrality, filter-press dehydration;
6, under 70 ℃ of conditions constant pressure and dry to weight loss on drying less than 15%, pulverize at last and obtain the Welan gum product.
Embodiment 4
1, the Welan gum fermented liquid with 250kg is heated to 90 ℃, kept 30 minutes, again with water quench to room temperature, manipulate in order to next step;
2, with the phosphoric acid solution of 3mol/L the pH of Welan gum fermented liquid is transferred to 3.0, the calcium chloride solution with 2.5mol/L is adjusted to 0.05mol/L with cationic concentration of electric charges in the Welan gum fermented liquid again;
3, the industrial alcohol solvent of interpolation 95% makes the determining alcohol of mixed solution reach 65% (w/w), and stirs;
4, standing sedimentation 2h, solid substance is collected in press filtration then, gets the Welan gum crude product;
5, with 95% the industrial alcohol dissolving of gained solid substance with 0.8 times of solid substance volume, regulator solution pH is to neutral, and suction filtration is press dewatering again;
6, under 60 ℃ of conditions constant pressure and dry to the product water ratio less than 13%, pulverize at last and obtain the Welan gum product.
Embodiment 5
1, the Welan gum fermented liquid with 500kg is heated to 90 ℃, kept 40 minutes, again with water quench to room temperature, manipulate in order to next step;
2, with the acetic acid solution of 4mol/L the pH of Welan gum fermented liquid is transferred to 2.0, the magnesium chloride solution with 2.5mol/L is adjusted to 0.2mol/L with cationic concentration of electric charges in the Welan gum fermented liquid again;
3, the industrial alcohol solvent of interpolation 95% makes the determining alcohol of mixed solution reach 58% (w/w), and stirs;
4, vacuumizing filtration is collected solid substance, squeeze the Welan gum crude product;
5, with 95% the industrial alcohol dissolving of gained solid substance with 0.6 times of solid substance volume, regulator solution pH is to neutral, filter-press dehydration;
6, under 60 ℃ of conditions constant pressure and dry to the product water ratio less than 12%, pulverize at last and obtain the Welan gum product.
Claims (2)
- One kind from the Welan gum fermented liquid high efficiency separation extract the method for Welan gum, it is characterized in that technological process is:(1) Welan gum fermented liquid preparation:Alcaligenes (Alcaligenes sp.) ATCC 31555 seed liquor are forwarded to fermention medium with volume ratio 6% inoculum size, obtain yellow thick fermented liquid at 30 ℃, 200r/min bottom fermentation 72h;Used fermention medium consists of sucrose 40g/L, dregs of beans 5g/L, KH 2PO 40.6g/L, MgSO 47H 2O 0.2g/L, FeCl 20.1g/L, using the tap water constant volume, pH 7.0～7.2;(2) fermented liquid is heated the microorganism of killing in the fermented liquid, temperature is 80～100 ℃, and the time is 10～45 minutes; And with water quench to room temperature;(3) with certain density acid solution the pH of Welan gum fermented liquid is transferred to 1.0～5.0, used acid solution is hydrochloric acid, acetate, phosphoric acid or its combination of 1～4mol/L;And/or add a certain amount of salt ion solution; The salt that adds is selected sodium-chlor, Repone K, ammonia chloride, magnesium chloride, calcium chloride, aluminum chloride or its combination for use, and the amount of adding salt makes that cation concn reaches 0.01～0.5mol/L in the Welan gum fermented liquid;(4) add a certain amount of alcoholic solvent, make the pure mass concentration of the mixed solution after the interpolation reach 30%～80%, and stir; Used alcoholic solvent is selected from ethanol, Virahol, methyl alcohol or its combination;(5) solid-liquid separation is removed supernatant liquor, obtains solid substance; Isolating method adopts settlement separate, centrifugation, filtering separation and combination thereof in the solid-liquid separation process;(6) add the alcoholic solvent of 0.2～1.0 times of the solid substance volume obtain, regulator solution pH is to neutral, press filtration or press dewatering; Perhaps the solid substance that step (5) is obtained directly carries out press filtration or press dewatering;(7) dry and pulverizing, drying temperature is below 80 ℃, drying means is constant pressure and dry, vacuum-drying or vacuum freezedrying, is dried to product water content＜15%, promptly gets the Welan gum finished product.
- 2. method according to claim 1 is characterized in that the salt ion solution of the described interpolation of step (3), and the salt ion of interpolation is CaCl 2, adopt the CaCl that adds 2.5mol/L 2It is 0.1mol/L that solution makes the final cation concn of fermented liquid.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102618468A (en) * | 2012-03-28 | 2012-08-01 | 南京工业大学 | Temperature-resistant alcaligenes and application thereof in welan gum production |
| CN103695498A (en) * | 2012-09-27 | 2014-04-02 | 天津科百生物科技研发有限公司 | Method of producing welan gum by tank-dividing method fermentation |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1844155A (en) * | 2006-03-24 | 2006-10-11 | 张禹 | Method for preparing a welan gum |
| CN1888054A (en) * | 2006-07-13 | 2007-01-03 | 南京工业大学 | Alcaligenes and application thereof in preparation of welan gum |
| CN101275154A (en) * | 2008-01-22 | 2008-10-01 | 江南大学 | Production method for microbial polysaccharide welan gum |
-
2010
- 2010-08-10 CN CN 201010253396 patent/CN101942491A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1844155A (en) * | 2006-03-24 | 2006-10-11 | 张禹 | Method for preparing a welan gum |
| CN1888054A (en) * | 2006-07-13 | 2007-01-03 | 南京工业大学 | Alcaligenes and application thereof in preparation of welan gum |
| CN101275154A (en) * | 2008-01-22 | 2008-10-01 | 江南大学 | Production method for microbial polysaccharide welan gum |
Non-Patent Citations (1)
| Title |
|---|
| 《微生物学报》 20090504 黄海东等 合成生物聚合物的重要微生物资源-鞘氨醇单胞菌 561 1-2 第49卷, 第5期 2 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102618468A (en) * | 2012-03-28 | 2012-08-01 | 南京工业大学 | Temperature-resistant alcaligenes and application thereof in welan gum production |
| CN102618468B (en) * | 2012-03-28 | 2013-06-19 | 南京工业大学 | Temperature-resistant alcaligenes and application thereof in welan gum production |
| CN103695498A (en) * | 2012-09-27 | 2014-04-02 | 天津科百生物科技研发有限公司 | Method of producing welan gum by tank-dividing method fermentation |
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Application publication date: 20110112 |