CN101586135A - High yield gellan gum fermentation preparation - Google Patents
High yield gellan gum fermentation preparation Download PDFInfo
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- CN101586135A CN101586135A CNA2009101439068A CN200910143906A CN101586135A CN 101586135 A CN101586135 A CN 101586135A CN A2009101439068 A CNA2009101439068 A CN A2009101439068A CN 200910143906 A CN200910143906 A CN 200910143906A CN 101586135 A CN101586135 A CN 101586135A
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Abstract
A high yield gellan gum fermentation preparation comprises following steps: 1. culturing the slant strains in scale-up to a seed liquid in a shake flask; 2. inoculating the seed liquid in the shake flask into a sterile culture medium in a first-class seed tank for scale-up culture; 3. inoculating the seed liquid in the first-class seed tank into a sterile culture medium in a second-class seed tank for scale-up culture; 4. inoculating the seed liquid in the second-class seed tank into a sterile culture medium in a third-class seed tank for scale-up culture; 5. collecting the gellan gum fermentation liquid; wherein the strains are the sphingomonaspaucimobilis and the sterile water is added in the culture process in the three-class fermentation tanks. The method can shorten the fermentation period from 50-60 hours to 40-45 hours, improves the coarse polysaccharide content from 1.5-1.6g/ml to 2.0-2.5g/100ml and realizes the low energy and high yield.
Description
Technical field
The present invention relates to the microbial fermentation field, the high yield fermentation preparation of particularly a kind of microbial polysaccharide---gelling gum.
Background technology
Gelling gum be a kind of from the Bulbus Cardiocrini Cathayani separating obtained gram-negative bacteria--sphingomonas paucimobilis (Sphingomonas the paucimobilis)--exocellular polysaccharide that is produced, there is consumption low, the transparency height, the fragrance releasability is strong, acidproof, advantage such as alkaline-resisting.Gelling gum can be extensive use of in varieties of food items, can be described as one of the most excellent thickening material of present performance, stablizer, and has good gel property.It has been widely used in industries such as food, daily use chemicals, medicine.Gelling gum went through in Application in Food in Japan in 1988, its utilization in food of drugs approved by FDA in 1992, the European Community was formally listed it in edible safety code (E-418) table in 1994, China was then listed gelling gum among the GB2760-1996 in 1996, ratify it as food thickener, stablizer, can in varieties of food items, need an amount of the use by ordinary production.
The monose molecular composition of gelling gum is a glucose, rhamnosyl, gluconic acid, acetate fat, L-R-Glyceric acid fat group, main chain are linear tetrose repeating units, by 1, and 3-and 1, the glucose unit that 4-connects, 1, the gluconic acid unit and 1 that 3-connects, the rhamnosyl unit that 4-connects forms.Molecular weight is 0.5 * 10
6Dalton.
At present domestic have many enterprises and scientific research institution at the fermentation research that carries out the gelling gum product, but effect is not very desirable, mainly shows long, the problems such as the product yield is low, production cost height of fermentation time.
Summary of the invention
The present invention selects aerobic gram-negative bacteria for use on existing fermentation manufacturing technique basis--sphingomonas paucimobilis (Sphingomonas paucimobilis), the method for carrying out zymotechnique control.Main process comprises the pH of gelling gum seed liquor, the control of OD value, and benefit is gone into sterilized water during the fermentation, control fermentation culture process mixing speed and advance the temperature of air, reach fermentation period and foreshorten to 40～45h by 50 original～60h, Crude polysaccharides content is increased to 2.0～2.5g/100ml by 1.5～1.6g/100ml, has really realized gelling gum low energy aim of high yield, and has improved the effective rate of utilization of equipment greatly.
The present invention is easy to realize that it is low consume energy, mends method into sterilized water by the fermentation later stage, has solved the low contradiction of later stage dissolved oxygen of fermenting, and has increased and has put tank volume, has improved single jar of output; By the effective control to the whole process inlet air temperature of fermenting, the temperature control of three grade fermemtation jar has been shortened fermentation period 30-34 ℃ of cultivation.Make and put jar Crude polysaccharides concentration and reach 2.0～2.5g/100ml.
This zymotechnique is simple to operate in fulfillment process, and consumed cost is low, and more traditional fermentation process can be saved 20% fermentation and dynamic power, improves single jar of output more than 15%.
High yield gellan gum fermentation preparation of the present invention is divided into three grade fermemtation, and concrete technical process is as follows:
Inclined-plane → shake bottle → first class seed pot → secondary seed jar → three grade fermemtation jar
This preparation method may further comprise the steps:
1) slant strains becomes shake-flask seed liquid through enlarged culturing;
2) shake-flask seed liquid is inserted enlarged culturing in the first class seed pot aseptic culture medium;
3) seed liquor in the first class seed pot is inserted enlarged culturing in the secondary seed jar aseptic culture medium;
4) seed liquor in the secondary seed jar is inserted fermentation culture in the three grade fermemtation jar aseptic culture medium;
5) collect gellan gum fermentation liquid;
Wherein said bacterial classification is a sphingomonas paucimobilis, and adds sterilized water in the culturing process in the three grade fermemtation jar.
The used sphingomonas paucimobilis of the present invention is available from Southern Yangtze University.
More specifically, this method comprises:
1) slant strains becomes shake-flask seed liquid through enlarged culturing, and the shake-flask culture base consists of: peptone: 0.3～0.6, and extractum carnis 0.2～0.4, yeast extract paste 0.05～0.15, all the other are water.
The condition of shaking bottle enlarged culturing is: temperature 28-34 ℃, and cycle 18～30h, shaking speed: 150r/min;
2) cultivate in 0.01～0.03% ratio access first class seed pot of shake-flask seed liquid in the first order seed culture volume with above-mentioned preparation, the substratum of first class seed pot consists of;
Yeast diffusion juice: 0.2～0.5, extractum carnis: 0.3, fish peptone: 0.5, DDFS-303 (food grade defoamer): 0.0003, all the other are water;
The condition of enlarged culturing is in the first class seed pot: culture temperature: 28～34 ℃, and ventilating ratio: 1: 0.5, operating pressure: 0.05～0.08Mpa;
The terminal point of first class seed pot enlarged culturing is: the cycle: 13～16h, pH value be more than 8.2, amino nitrogen more than 350mg/Kg, OD value 0.1 ± 0.02.The OD value detects and is ultraviolet-visible pectrophotometer H0605019, wavelength 520nm.
3) will cultivate in 8～12% ratios access secondary seed jar of the seed liquor in the first class seed pot of above-mentioned preparation in the secondary seed medium volume, the substratum in the secondary seed jar consists of;
White sugar: 2.0～3.0, Yeast diffusion juice: 0.1～0.3, soybean protein powder: 0.2～0.4, KH
2PO
4: 0.05, K
2PHO
4: 0.05, MgSO
4: 0.06, DDFS-303:0.0003, all the other are water;
Condition in the secondary seed jar is: culture temperature: 28～34 ℃, and ventilating ratio: 1: 0.5, operating pressure: 0.05～0.08Mpa;
The terminal point of secondary seed jar enlarged culturing is: cycle: 6～8h, the pH value is 6.0～6.6, amino nitrogen between 40～80mg/Kg, OD value 0.4 ± 0.02.The OD value detects and is ultraviolet-visible pectrophotometer H0605019, wavelength 520nm.
4) will cultivate in 5～9% ratios access three grade fermemtation jar of the seed liquor in the secondary seed jar of above-mentioned preparation in the three grade fermemtation culture volume, the substratum of three grade fermemtation jar consists of;
White sugar: 2.5～3.5, soybean protein powder: 0.5～0.8, KH
2PO
4: 0.1, MnSO
4H
2O:0.025, K
2HPO
4: 0.15, MgSO
4: 0.06, DDFS-303:0.0003, H
3BO
3: 0.003, all the other are water;
Culture condition in the three grade fermemtation jar is:
The fermentation culture temperature: 30～34 ℃, operating pressure: 0.05～0.08Mpa;
Fermentation is stirred and is used speed control by frequency variation to stir, and air quantity is controlled according to fermentation time segmentation control air input;
Fermentation is stirred and is used speed control by frequency variation to stir, and the frequency conversion rotating speed of concrete control is as follows:
Air quantity is controlled according to fermentation time segmentation control air input, and concrete controlled variable is as follows;
The fermentation viscosity reaches 4000cp when above, mends sterilized water into 10% in the fermentating liquid volume ratio, divides 3 benefits to go into, and frequency every two hours once.
The terminal point that the three grade fermemtation jar is cultivated is: put jar viscosity at 7000～9000cp, the residual sugar ratio control is 0.2～0.4.
The whole fermentation process from first class seed pot to fermentor tank, intake air temperature is controlled at 25～30 ℃, and the temperature of cultivation and fermentation liquid in the fermentor tank is controlled in the processing requirement scope more accurately, and the growth that is beneficial to microbial polysaccharide is synthetic.
Unless otherwise noted, substratum of the present invention is in weight (g)/volume (L) per-cent.For example, consist of example with the shake-flask culture base: take by weighing peptone 0.3～0.6g, extractum carnis 0.2～0.4g, yeast extract paste 0.05～0.15g, be dissolved in the suitable quantity of water after, water is supplied volume to 1 liter.
Embodiment:
Propose following examples and specify, understanding the present invention better, but this embodiment is nonrestrictive.
Shake bottle with 5L, the 500L first class seed pot, 5T secondary seed jar, 50T three grade fermemtation jar is an example.Practical volume is first class seed pot 300L, secondary seed jar 3T, three grade fermemtation jar 35T, the i.e. culture volume that adds in the jar.
Concrete preparation method may further comprise the steps:
1) inoculating needle is drawn the inclined-plane seed that takes a morsel on clean operator's console, inserts the shake-flask culture base and cultivates.
The shake-flask culture base consists of:
Peptone: 0.5, extractum carnis: 0.3, yeast extract paste: 0.1, all the other are water.
The condition of shake-flask culture is: 30 ℃ of temperature, cycle 24h, shaking speed: 150r/min.
2) cultivate in the 0.017% ratio access first class seed pot of shake-flask seed liquid with above-mentioned preparation in the first order seed culture volume:
The substratum of first class seed pot consists of;
Yeast diffusion juice: 0.4, extractum carnis: 0.3, fish peptone: 0.5, DDFS-303:0.0003, all the other are water;
Culture condition in the first class seed pot is: culture temperature: 30 ℃, and ventilating ratio: 1: 0.5, operating pressure: 0.06Mpa;
The cultivation terminal point of first class seed pot is: cycle: 15h, the pH value is 8.32, amino nitrogen 365mg/Kg, OD value 0.086.The OD value detects and is ultraviolet-visible pectrophotometer H0605019, wavelength 520nm.
3) will cultivate in the 10% ratio access secondary seed jar of the seed liquor in the first class seed pot of above-mentioned preparation in the secondary seed medium volume:
The substratum of secondary seed jar consists of;
White sugar: 2.5, Yeast diffusion juice: 0.2, soybean protein powder: 0.3, KH
2PO
4: 0.05, K
2PHO
4: 0.05, MgSO
4: 0.06, DDFS-303:0.0003, all the other are water;
Culture condition in the secondary seed jar is: culture temperature: 30 ℃, and ventilating ratio: 1: 0.5, operating pressure: 0.06Mpa;
The cultivation terminal point of secondary seed jar is: cycle: 7h, and the pH value is 6.56, and amino nitrogen is at 56mg/Kg, OD value 0.410.The OD value detects and is ultraviolet-visible pectrophotometer H0605019, wavelength 520nm.
4) will cultivate in the 8% ratio access three grade fermemtation jar of the seed liquor in the secondary seed jar of above-mentioned preparation in the fermention medium volume:
The substratum of fermentor tank consists of:
White sugar: 3.0, soybean protein powder: 0.7, KH
2PO
4: 0.1, MnSO
4H
2O:0.025,
K
2HPO
4: 0.15, MgSO
4: 0.06, DDFS-303:0.0003, H
3BO
3: 0.003, all the other are water;
Culture condition in the three grade fermemtation jar is:
The fermentation culture temperature: 32.5 ℃, operating pressure: 0.06Mpa;
Fermentation is stirred and is used speed control by frequency variation to stir, and the frequency conversion rotating speed of concrete control is as follows:
Air quantity is controlled according to fermentation time segmentation control air input, and concrete controlled variable is as follows;
The fermentation viscosity reaches more than the 4000cp, mends into sterilized water in 10% ratio in fermentating liquid volume in the jar, i.e. 3.5T.Divide and mend for 3 times, frequency every two hours once.
Cultivation terminal point in the three grade fermemtation jar is: put jar viscosity at 8500cp, the residual sugar ratio control is 0.29.
In the whole fermentation process from the first class seed pot to the fermentor tank, intake air temperature is controlled at 25～30 ℃, and the temperature of cultivation and fermentation liquid in the fermentor tank is controlled in the processing requirement scope more accurately, and the growth that is beneficial to microbial polysaccharide is synthetic.
After the fermentation ends, collect fermented liquid, get quantitative fermented liquid and shake up with water dissolution, centrifugal again, it is alcohol-pickled to use upper solution after centrifugal to add, and dries behind the suction filtration, must Crude polysaccharides concentration at 2.05g/100ml.The single jar of the highest 1.6g/100ml of Crude polysaccharides concentration than the document record has improved 28%.
Claims (10)
1. the gellan gum fermentation preparation of a high yield, it may further comprise the steps:
1) slant strains becomes shake-flask seed liquid through enlarged culturing;
2) shake-flask seed liquid is inserted enlarged culturing in the first class seed pot aseptic culture medium;
3) seed liquor in the first class seed pot is inserted enlarged culturing in the secondary seed jar aseptic culture medium;
4) seed liquor in the secondary seed jar is inserted fermentation culture in the three grade fermemtation jar aseptic culture medium;
5) collect gellan gum fermentation liquid;
Wherein said bacterial classification is a sphingomonas paucimobilis, and adds sterilized water in the culturing process in the three grade fermemtation jar.
2. the preparation method of claim 1 is characterized in that, the consisting of of shake-flask culture base: peptone: 0.3～0.6, and extractum carnis 0.2～0.4, yeast extract paste 0.05～0.15, all the other are water.
3. the preparation method of claim 1 is characterized in that, the condition of shaking bottle enlarged culturing is: temperature 28-34 ℃, and cycle 18～30h, shaking speed: 150r/min.
4. the preparation method of claim 1 is characterized in that, the substratum in the first class seed pot consists of: Yeast diffusion juice: 0.2～0.5, and extractum carnis: 0.3, fish peptone: 0.5, DDFS-303:0.0003, all the other are water; In weight/volume percent.
5. the preparation method of claim 1 is characterized in that, the condition of enlarged culturing is in the first class seed pot: culture temperature: 28～34 ℃, and ventilating ratio: 1: 0.5, operating pressure: 0.05～0.08Mpa.
6. the preparation method of claim 1 is characterized in that, the terminal point of first class seed pot enlarged culturing is: the cycle: 13～16h, the pH value is more than 8.2, amino nitrogen more than 350mg/Kg, OD520nm value 0.1 ± 0.02.
7. the preparation method of claim 1 is characterized in that, the substratum in the secondary seed jar consists of: white sugar: 2.0～3.0, and Yeast diffusion juice: 0.1～0.3, soybean protein powder: 0.2～0.4, KH
2PO
4: 0.05, K
2PHO
4: 0.05, MgSO
4: 0.06, DDFS-303:0.0003, all the other are water.
8. the preparation method of claim 1 is characterized in that, the culture condition of secondary seed jar is: culture temperature: 28～34 ℃, and ventilating ratio: 1: 0.5, operating pressure: 0.05～0.08Mpa.
9. the preparation method of claim 1 is characterized in that, the terminal point of secondary seed jar enlarged culturing is: cycle: 6～8h, the pH value is 6.0～6.6, amino nitrogen between 40～80mg/Kg, OD520nm value 0.4 ± 0.02.
10. the preparation method of claim 1 is characterized in that the substratum in the three grade fermemtation jar consists of: white sugar: 2.5～3.5, and soybean protein powder: 0.5～0.8, KH
2PO
4: 0.1, MnSO
4H
2O:0.025, K
2HPO
4: 0.15, MgSO
4: 0.06, DDFS-303:0.0003, H
3BO
3: 0.003, all the other are water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2009101439068A CN101586135A (en) | 2009-06-03 | 2009-06-03 | High yield gellan gum fermentation preparation |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2009101439068A CN101586135A (en) | 2009-06-03 | 2009-06-03 | High yield gellan gum fermentation preparation |
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| CN101586135A true CN101586135A (en) | 2009-11-25 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421718A (en) * | 2013-08-09 | 2013-12-04 | 浙江大学 | Sphingomonas paucimobilis strain and application thereof |
| CN107674854A (en) * | 2017-11-20 | 2018-02-09 | 河北沣川生物科技有限公司 | A kind of fixed nitrogen Sphingol single-cell and the application in gellan gum is prepared |
| CN107805649A (en) * | 2017-11-10 | 2018-03-16 | 浙江帝斯曼中肯生物科技有限公司 | A kind of novel fermentation technique for producing gellan gum |
| CN108929888A (en) * | 2017-06-29 | 2018-12-04 | 帝斯曼知识产权资产管理有限公司 | A kind of zymotechnique producing gellan gum |
-
2009
- 2009-06-03 CN CNA2009101439068A patent/CN101586135A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 王琴丹等: "结冷胶的生物合成研究进展", 《食品科学》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421718A (en) * | 2013-08-09 | 2013-12-04 | 浙江大学 | Sphingomonas paucimobilis strain and application thereof |
| CN103421718B (en) * | 2013-08-09 | 2015-11-11 | 浙江大学 | A kind of sphingomonas paucimobilis bacterial strain and application thereof |
| CN108929888A (en) * | 2017-06-29 | 2018-12-04 | 帝斯曼知识产权资产管理有限公司 | A kind of zymotechnique producing gellan gum |
| CN107805649A (en) * | 2017-11-10 | 2018-03-16 | 浙江帝斯曼中肯生物科技有限公司 | A kind of novel fermentation technique for producing gellan gum |
| EP3483280A1 (en) * | 2017-11-10 | 2019-05-15 | Zhejiang Dsm Zhongken Biotechnology Co., Ltd | A fermentation method for producing gellan gum |
| US20190144903A1 (en) * | 2017-11-10 | 2019-05-16 | Zhejiang Dsm Zhongken Biotechnology Co. Ltd | Fermentation method for producing gellan gum |
| US11149293B2 (en) | 2017-11-10 | 2021-10-19 | Zhejiang Dsm Zhongken Biotechnology Co. Ltd | Fermentation method for producing gellan gum |
| CN107674854A (en) * | 2017-11-20 | 2018-02-09 | 河北沣川生物科技有限公司 | A kind of fixed nitrogen Sphingol single-cell and the application in gellan gum is prepared |
| CN107674854B (en) * | 2017-11-20 | 2020-08-11 | 河北沣川生物科技有限公司 | Nitrogen-fixing sphingosine monad and application thereof in preparation of gellan gum |
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Application publication date: 20091125 |