CN103710398A - Method for reinforcing activity of fermentation strain of 2-keto-L-gulonic acid - Google Patents
Method for reinforcing activity of fermentation strain of 2-keto-L-gulonic acid Download PDFInfo
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- CN103710398A CN103710398A CN201310692592.3A CN201310692592A CN103710398A CN 103710398 A CN103710398 A CN 103710398A CN 201310692592 A CN201310692592 A CN 201310692592A CN 103710398 A CN103710398 A CN 103710398A
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Abstract
The invention discloses a method for reinforcing activity of a fermentation strain of 2-keto-L-gulonic acid, which belongs to the technical field of microbial fermentation. The method comprises the following steps: separately sterilizing a nitrogen source and a carbon source in a 2-keto-L-gulonic acid seeding tank culture medium, and mixing the nitrogen source with the carbon source after cooling to 28-30 DEG C. The separate sterilization method of the nitrogen source and the carbon source provided by the invention can be used for reducing the generation of harmful substances, ensuring little damage of nutritional ingredients in the culture medium, facilitating the production and reproduction of producing strain, reinforcing the activity of the strain and shortening the fermentation period of the 2-keto-L-gulonic acid.
Description
Technical field
The invention belongs to microbial fermentation technology field, more particularly, relate to a kind of method that strengthens KGA fermented bacterium vigor.
Background technology
Vitamins C (VC) is a kind of water-soluble vitamins, has another name called L-xitix, can participate in multiple hydroxylation reaction in body
And redox reaction, be the important cellular metabolism redox compound of a class, in organism, play requisite physiological action.Vitamins C is used for preventing vitamin C deficiency, the assisting therapy of various acute and chronic transmissible diseases and purpura etc., and vitamins C is also widely used in food, feed, makeup.Early 1970s, Yin Guanglin etc. have filtered out mixed strains-bacillus megaterium (Bacillus megaterium) and the ordinary student ketone 2-KLG bacterium (Ketogulonigenium sp.) of the important as precursors KGA (2-KLG) that can directly L-sorbose be converted into synthesise vitamins C, have invented Vitamin C Two-step Fermentation method.The method has simplified by two-step fermentation the synthetic route that Lai Shi method is produced vitamins C greatly, has the outstanding advantage that glucose acid invert ratio is high, and domestic production manufacturer production vitamins C all adopts two-step fermenting at present.The fermentation level of second step mixed fungus fermentation has determined the fermentation level of two-step fermenting.In current ascorbic manufacturing enterprise, the operational path of fermentation and the transformation of reactor research do not have large breakthrough.In the process of fermentative production KGA, improve fermentation manufacturing technique, strengthen spawn activity, shorten fermentation period, it is essential improving fermentation technique.By nitrogen, carbon, divide the enhancing KGA fermented bacterium vigor that disappears also there is no pertinent literature report at present.
Summary of the invention
For the above-mentioned problems in the prior art, the invention provides a kind of method that strengthens KGA fermented bacterium vigor.
In order to address the above problem, the technical solution adopted in the present invention is as follows:
Strengthen a method for KGA fermented bacterium vigor, the nitrogenous source in KGA seed tank culture base and carbon source are separated to sterilization, to be cooledly mix during to 28 ℃-30 ℃.
Further, described nitrogenous source comprises corn steep liquor, yeast extract paste and urea.
Further, described carbon source is L-sorbose.
Seed tank culture based formulas (weight percent): sorbose 1.7%, corn steep liquor 0.5%, yeast extract paste 0.3%, urea 0.2%, magnesium sulfate 0.02%, potassium primary phosphate 0.1%, calcium carbonate 0.5%, PH6.7-7.2.
Seed tank culture condition: culture temperature 28-30 ℃, culturing process is controlled dissolved oxygen 30-50%, inoculum size 10%.
Seed tank culture base disinfection way: the main carbon source L-sorbose of seed tank culture base and other raw material divide and disappear, to be cooled to about 30 ℃ mixing.
Seed tank culture is to logarithmic phase, plants the iodimetry,iodometry that is determined as of KGA in liquid; Plant nectar degree in liquid and be determined as microscopic counting, adopt blood counting chamber to count.
Fermentor cultivation based formulas: sorbose 8.0%, corn steep liquor 0.5%, urea 0.1%, magnesium sulfate 0.01%, potassium primary phosphate 0.1%, calcium carbonate 0.3%, wherein sorbose adds with liquid form stream in culturing process.Culture temperature 29-31 ℃, culturing process is controlled dissolved oxygen 30-50%, and the sodium carbonate solution with 25% is controlled PH6.7-7.2.
Beneficial effect: adopt technique of the present invention, nitrogen, carbon divide subdues rare harmful substances generation, and medium nutrient content destroys few, is beneficial to and produces bacterium Reproduction, can effectively strengthen spawn activity, shortens KGA fermentation period.
Embodiment
Below in conjunction with specific embodiment, the present invention is further described.
1, seed tank culture based formulas: sorbose 1.7%, corn steep liquor 0.5%, yeast extract paste 0.3%, urea 0.2%, magnesium sulfate 0.02%, potassium primary phosphate 0.1%, calcium carbonate 0.5%, PH6.7-7.2.
2, seed tank culture condition: culture temperature 28-30 ℃, culturing process is controlled dissolved oxygen 30-50%, inoculum size 10%.
3, seed tank culture base disinfection way: the main carbon source L-sorbose of seed tank culture base and other raw material divide and disappear, to be cooled to about 30 ℃ mixing.
4, seed tank culture, to logarithmic phase, is planted the mensuration of KGA in liquid: improved iodimetry,iodometry; Plant bacterium density measurement in liquid: microscopic counting, adopts blood counting chamber to count.
5, fermentor cultivation based formulas: sorbose 8.0%, corn steep liquor 0.5%, urea 0.1%, magnesium sulfate 0.01%, potassium primary phosphate 0.1%, calcium carbonate 0.3%, wherein sorbose adds with liquid form stream in culturing process.Culture temperature 29-31 ℃, culturing process is controlled dissolved oxygen 30-50%, and the sodium carbonate solution with 25% is controlled PH6.7-7.2.
6, implementation example:
Test group is by above-mentioned 1 seed tank culture based formulas configuration 10000L feed liquid, the main carbon source L-sorbose of seed tank culture base and other raw material are divided and disappeared, to be cooled to about 30 ℃ mixing, access bacterium mixed solution 1000L for cultured production, culture temperature 28-30 ℃, culturing process is controlled dissolved oxygen 30-50%, inoculum size 10%.Incubation time 20 hours, the content of measuring KGA is 11.67mg/ml.Control group is by above-mentioned 1 seed tank culture based formulas configuration 10000L feed liquid, the main carbon source L-sorbose of seed tank culture base and other raw material are closed and disappeared, to be cooled to 30 ℃ of bacterium mixed solution 1000L for the cultured production of access, culture temperature 28-30 ℃, culturing process is controlled dissolved oxygen 30-50%, inoculum size 10%.Incubation time 20 hours, the content of measuring KGA is 8.56mg/ml.Test group has improved 3.11mg/ml compared with the content of control group KGA.Measure nectar degree content test group and improve 20% compared with control group.
It is sorbose 8.0% that cultured test group seeding tank and control group seeding tank are accessed respectively to culture medium prescription, corn steep liquor 0.5%, urea 0.1%, magnesium sulfate 0.01%, potassium primary phosphate 0.1%, the large tank of fermentation of calcium carbonate 0.3%, wherein sorbose adds with liquid form stream in culturing process, and the sodium carbonate solution with 25% is controlled PH6.7-7.2, culture temperature 29-31 ℃, culturing process is controlled dissolved oxygen 30-50%, inoculum size 15%.The content of test group fermentation termination fermented liquid KGA is 78.26mg/ml, fermentation period 30 hours, rate of producing acid 2.61mg/mlh.The content of control group fermentation termination fermented liquid KGA is 77.46mg/ml, fermentation period 34 hours, rate of producing acid 2.28mg/mlh.Test group improves 14.47% compared with control group rate of producing acid.
Seed tank culture base is mainly that nitrogenous source corn steep liquor, yeast extract paste, urea separate sterilization in when sterilization so long as by carbon source L-sorbose and other raw material, just belongs among protection domain of the present invention.
Claims (8)
1. a method that strengthens KGA fermented bacterium vigor, is characterized in that: the nitrogenous source in KGA seed tank culture base and carbon source are separated to sterilization, to be cooledly mix during to 28 ℃-30 ℃.
2. a kind of method that strengthens KGA fermented bacterium vigor according to claim 1, is characterized in that: described nitrogenous source comprises corn steep liquor, yeast extract paste and urea.
3. a kind of method that strengthens KGA fermented bacterium vigor according to claim 1, is characterized in that: described carbon source is L-sorbose.
4. a kind of method that strengthens KGA fermented bacterium vigor according to claim 1, it is characterized in that: seed tank culture based formulas: sorbose 1.7%, corn steep liquor 0.5%, yeast extract paste 0.3%, urea 0.2%, magnesium sulfate 0.02%, potassium primary phosphate 0.1%, calcium carbonate 0.5%, PH6.7-7.2.
5. a kind of method that strengthens KGA fermented bacterium vigor according to claim 1, is characterized in that: seed tank culture condition: culture temperature 28-30 ℃, culturing process is controlled dissolved oxygen 30-50%, inoculum size 10%.
6. a kind of method that strengthens KGA fermented bacterium vigor according to claim 1, it is characterized in that: seed tank culture base disinfection way: the main carbon source L-sorbose of seed tank culture base and other raw material divide and disappear, to be cooled to about 30 ℃ mixing.
7. a kind of method that strengthens KGA fermented bacterium vigor according to claim 1, is characterized in that: with the KGA in iodometric determination kind liquid; With microscopic counting, measure nectar degree in kind of liquid, adopt blood counting chamber to count.
8. a kind of method that strengthens KGA fermented bacterium vigor according to claim 1, it is characterized in that: fermentor cultivation based formulas: sorbose 8.0%, corn steep liquor 0.5%, urea 0.1%, magnesium sulfate 0.01%, potassium primary phosphate 0.1%, calcium carbonate 0.3%, wherein sorbose adds with liquid form stream in culturing process; Culture temperature 29-31 ℃, culturing process is controlled dissolved oxygen 30-50%, and the sodium carbonate solution with 25% is controlled PH6.7-7.2.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510682158.6A CN105274154A (en) | 2013-12-18 | 2013-12-18 | Method for improving activity of 2-keto-L-gulonic acid fermentation strains |
| CN201310692592.3A CN103710398A (en) | 2013-12-18 | 2013-12-18 | Method for reinforcing activity of fermentation strain of 2-keto-L-gulonic acid |
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| CN201310692592.3A CN103710398A (en) | 2013-12-18 | 2013-12-18 | Method for reinforcing activity of fermentation strain of 2-keto-L-gulonic acid |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434830A (en) * | 2016-12-20 | 2017-02-22 | 宁夏启元药业有限公司 | Method for improving 2-keto-L-gluconic acid fermentation efficiency |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111073946A (en) * | 2019-12-23 | 2020-04-28 | 中国科学院沈阳应用生态研究所 | Vc two-step fermentation nutrition optimization method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101113462A (en) * | 2006-07-28 | 2008-01-30 | 中国科学院沈阳应用生态研究所 | Gulonic acid mash quick-hyperfiltration fermentation controlling method |
| CN101736071A (en) * | 2009-12-24 | 2010-06-16 | 安徽丰原发酵技术工程研究有限公司 | Method for producing 2-keto-l-gulonic acid |
| CN102586380A (en) * | 2011-11-01 | 2012-07-18 | 江苏江山制药有限公司 | 2-keto-L-gulonic acid fermentation process |
| CN102586381A (en) * | 2011-11-01 | 2012-07-18 | 江苏江山制药有限公司 | Production process for improving fermentative strength of 2-keto-L-gulonic acid |
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- 2013-12-18 CN CN201310692592.3A patent/CN103710398A/en active Pending
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101113462A (en) * | 2006-07-28 | 2008-01-30 | 中国科学院沈阳应用生态研究所 | Gulonic acid mash quick-hyperfiltration fermentation controlling method |
| CN101736071A (en) * | 2009-12-24 | 2010-06-16 | 安徽丰原发酵技术工程研究有限公司 | Method for producing 2-keto-l-gulonic acid |
| CN102586380A (en) * | 2011-11-01 | 2012-07-18 | 江苏江山制药有限公司 | 2-keto-L-gulonic acid fermentation process |
| CN102586381A (en) * | 2011-11-01 | 2012-07-18 | 江苏江山制药有限公司 | Production process for improving fermentative strength of 2-keto-L-gulonic acid |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434830A (en) * | 2016-12-20 | 2017-02-22 | 宁夏启元药业有限公司 | Method for improving 2-keto-L-gluconic acid fermentation efficiency |
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