CN101250563A - Method for realizing excessive accumulation of alpha-ketoglutarate acid by adding alpha-ketoglutarate acid dehydrogenase inhibitor - Google Patents
Method for realizing excessive accumulation of alpha-ketoglutarate acid by adding alpha-ketoglutarate acid dehydrogenase inhibitor Download PDFInfo
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- CN101250563A CN101250563A CNA2008100200394A CN200810020039A CN101250563A CN 101250563 A CN101250563 A CN 101250563A CN A2008100200394 A CNA2008100200394 A CN A2008100200394A CN 200810020039 A CN200810020039 A CN 200810020039A CN 101250563 A CN101250563 A CN 101250563A
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- alpha
- tong wuersuan
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- fermentation
- ketoglutaric acid
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- 238000009825 accumulation Methods 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 title claims abstract description 15
- 229940124186 Dehydrogenase inhibitor Drugs 0.000 title claims description 6
- 239000002253 acid Substances 0.000 title description 12
- KPGXRSRHYNQIFN-UHFFFAOYSA-L 2-oxoglutarate(2-) Chemical compound [O-]C(=O)CCC(=O)C([O-])=O KPGXRSRHYNQIFN-UHFFFAOYSA-L 0.000 title 2
- 238000000855 fermentation Methods 0.000 claims abstract description 28
- 230000004151 fermentation Effects 0.000 claims abstract description 28
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 26
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 claims abstract description 14
- 241000222126 [Candida] glabrata Species 0.000 claims abstract description 10
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229960000485 methotrexate Drugs 0.000 claims abstract description 7
- 230000000694 effects Effects 0.000 claims abstract description 6
- 235000013343 vitamin Nutrition 0.000 claims abstract description 4
- 239000011782 vitamin Substances 0.000 claims abstract description 4
- 229940088594 vitamin Drugs 0.000 claims abstract description 4
- 229930003231 vitamin Natural products 0.000 claims abstract description 4
- -1 hydroxyl sulphur ammonia Chemical compound 0.000 claims description 8
- 230000001580 bacterial effect Effects 0.000 claims description 4
- 229910052799 carbon Inorganic materials 0.000 claims description 4
- 230000002950 deficient Effects 0.000 claims description 3
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 2
- 101710088194 Dehydrogenase Proteins 0.000 claims description 2
- KPGXRSRHYNQIFN-UHFFFAOYSA-N 2-oxoglutaric acid Chemical compound OC(=O)CCC(=O)C(O)=O KPGXRSRHYNQIFN-UHFFFAOYSA-N 0.000 abstract description 12
- 230000006860 carbon metabolism Effects 0.000 abstract description 7
- 230000033228 biological regulation Effects 0.000 abstract description 3
- 230000002503 metabolic effect Effects 0.000 abstract description 3
- 230000004060 metabolic process Effects 0.000 abstract description 3
- 230000001105 regulatory effect Effects 0.000 abstract description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 abstract description 2
- 239000001963 growth medium Substances 0.000 abstract description 2
- 239000002207 metabolite Substances 0.000 abstract description 2
- 108090000623 proteins and genes Proteins 0.000 abstract description 2
- 102000004169 proteins and genes Human genes 0.000 abstract description 2
- HWXBTNAVRSUOJR-UHFFFAOYSA-N alpha-hydroxyglutaric acid Natural products OC(=O)C(O)CCC(O)=O HWXBTNAVRSUOJR-UHFFFAOYSA-N 0.000 abstract 5
- 229940009533 alpha-ketoglutaric acid Drugs 0.000 abstract 5
- 229940123941 Alpha ketoglutarate dehydrogenase inhibitor Drugs 0.000 abstract 2
- 102000006589 Alpha-ketoglutarate dehydrogenase Human genes 0.000 abstract 2
- 108020004306 Alpha-ketoglutarate dehydrogenase Proteins 0.000 abstract 2
- 239000005708 Sodium hypochlorite Substances 0.000 abstract 1
- 230000015556 catabolic process Effects 0.000 abstract 1
- 238000006731 degradation reaction Methods 0.000 abstract 1
- 125000002349 hydroxyamino group Chemical group [H]ON([H])[*] 0.000 abstract 1
- 230000004102 tricarboxylic acid cycle Effects 0.000 abstract 1
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 description 12
- 239000002609 medium Substances 0.000 description 7
- 229940107700 pyruvic acid Drugs 0.000 description 6
- 239000003112 inhibitor Substances 0.000 description 5
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 4
- 150000007513 acids Chemical class 0.000 description 4
- 150000003628 tricarboxylic acids Chemical class 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 229930003756 Vitamin B7 Natural products 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 229960003512 nicotinic acid Drugs 0.000 description 2
- 235000001968 nicotinic acid Nutrition 0.000 description 2
- 239000011664 nicotinic acid Substances 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 description 2
- 229960004172 pyridoxine hydrochloride Drugs 0.000 description 2
- 235000019171 pyridoxine hydrochloride Nutrition 0.000 description 2
- 239000011764 pyridoxine hydrochloride Substances 0.000 description 2
- 230000000452 restraining effect Effects 0.000 description 2
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 2
- 239000011735 vitamin B7 Substances 0.000 description 2
- 235000011912 vitamin B7 Nutrition 0.000 description 2
- LWFUFLREGJMOIZ-UHFFFAOYSA-N 3,5-dinitrosalicylic acid Chemical compound OC(=O)C1=CC([N+]([O-])=O)=CC([N+]([O-])=O)=C1O LWFUFLREGJMOIZ-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- ZJKLSPIEPZUTNW-UHFFFAOYSA-N C(=O)=C(C(=O)O)CCC(=O)O Chemical compound C(=O)=C(C(=O)O)CCC(=O)O ZJKLSPIEPZUTNW-UHFFFAOYSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 108010053763 Pyruvate Carboxylase Proteins 0.000 description 1
- 102100039895 Pyruvate carboxylase, mitochondrial Human genes 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000001773 anti-convulsant effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000012262 fermentative production Methods 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- RNUAEUWXRHCGKX-UHFFFAOYSA-N oxythiamine chloride Chemical compound [Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)NC1=O RNUAEUWXRHCGKX-UHFFFAOYSA-N 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 229940061969 rheumatrex Drugs 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 238000012807 shake-flask culturing Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 235000010288 sodium nitrite Nutrition 0.000 description 1
- PODWXQQNRWNDGD-UHFFFAOYSA-L sodium thiosulfate pentahydrate Chemical compound O.O.O.O.O.[Na+].[Na+].[O-]S([S-])(=O)=O PODWXQQNRWNDGD-UHFFFAOYSA-L 0.000 description 1
- 235000014268 sports nutrition Nutrition 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000001256 tonic effect Effects 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
Abstract
The invention relates to a method for adding alpha-ketoglutarate dehydrogenase inhibitor to realize excessive accumulation of alpha-ketoglutaric acid, which belongs to the technical field of the metabolic regulation optimized fermentation process of the protein level. The method of the invention comprises following steps: utilizing multi - vitamin - auxotrophic yeast of Torulopsis glabrata CCTCC M202019 as producing strains, regulating the activity of alpha-ketoglutarate dehydrogenase through adding the alpha-ketoglutarate dehydrogenase inhibitor: hydrogen peroxide, methotrexate, sodium hypochlorite or hydroxyamino in culture medium, purposively lowing the activity of the alpha-ketoglutarate dehydrogenase, reducing the degradation of the alpha-ketoglutaric acid in the metabolic process, and achieving the aim of the excessive accumulation of the alpha-ketoglutaric acid. The method of the invention cuts off carbon metabolism flow on a node of the alpha-ketoglutaric acid through regulating the stream distribution of carbon metabolism and the carbon metabolism flow, wherein the maximum output of the alpha-ketoglutaric acid reaches 23.2g/L, which is increased by 12.2% compared with the control. The invention provides a new thought for the fermentation research of TCA cycle intermediate metabolite.
Description
Technical field
The present invention relates to a kind of interpolation ketoglurate dehydrogenase (KGDH) inhibitor and realize the method for α-Tong Wuersuan (KG) excess accumulation.By of the influence of research KGDH inhibitor, reach a large amount of accumulation of α-Tong Wuersuan to the metabolism stream flow direction and corresponding metabolic flux.The metabolic regulation that belongs to protein level is optimized the fermenting process technical field.
Background technology
α-Tong Wuersuan, claim α-glue ketone acid again, 2-oxopentanedioic acid or alpha-carbonyl pentanedioic acid are one of important intermediate product in tricarboxylic acid (TCA) circulation, playing an important role in the metabolism of microorganism cells, also is synthetic multiple amino acids, proteinic important precursor.Its structural formula is:
The α-Tong Wuersuan structural formula
α-Tong Wuersuan has important application prospects in fields such as medicine, organic synthesis, nutrition-fortifying agents, and main application fields is at present: as the composition of sports nutrition beverage; Organic intermediate; The matched reagent of biochemical reagents and survey liver function; Physique strengthens tonic; Reduce postoperative patient and patients on long-term's body loss; At the precursor of brain as tyrosine and L-glutamic acid; Research simultaneously shows that also α-Tong Wuersuan has anti-cyanogen effect, is used with Sodium Nitrite, Sulfothiorine and can improves anti-cyanogen ability, and anticonvulsant action is arranged.α-Tong Wuersuan is mainly consumed at present in medical institutions, is used for diagnosing and sacred disease being treated.
Hydrogen peroxide, methotrexate, clorox and hydroxyl sulphur ammonia are extensive to the research of restraining effect in human nerve and heart disease of KGDH, and the rarely seen report of the research in microorganism especially yeast.Originally determined the restraining effect of these four kinds of materials to KGDH, simultaneously clear and definite inhibition concentration and interpolation time are for the fermentation research of TCA intercycle meta-bolites provides a new thinking.
Summary of the invention
The purpose of this invention is to provide a kind of method that the ketoglurate dehydrogenase inhibitor is realized the α-Tong Wuersuan excess accumulation of adding.Utilize torulopsis glabrata (T.glabrata) the CCTCCM202019 fermentative production KG of polyauxotroph, add an amount of KGDH inhibitor during the fermentation, autotelic inhibition KGDH activity reaches the purpose of a large amount of accumulation KG.
Technical scheme of the present invention: a kind of method of adding ketoglurate dehydrogenase inhibitor realization α-Tong Wuersuan excess accumulation, be that torulopsis glabrata (T.glabrata) the CCTCC M202019 that utilizes multiple vitamin defective type is the production bacterial strain, by add the KGDH inhibitor in substratum: hydrogen peroxide, methotrexate, clorox or hydroxyl sulphur ammonia are regulated the KGDH activity, the degraded of autotelic minimizing KG, carbon stream is terminated on this node of KG, realize the α-Tong Wuersuan excess accumulation; Regulate and control method is:
Fermentation 40h, when the interpolation hydrogen peroxide reached 5mM in fermention medium, the α-Tong Wuersuan accumulation volume reached 23.2g/L, and comparison is according to having improved 12.2%;
Or fermentation initial stage 0h, when the interpolation methotrexate reached 0.08 μ M in fermention medium, the α-Tong Wuersuan accumulation volume reached 22.7g/L, and comparison is according to having improved 10.1%;
Or fermentation initial stage 0h, when the interpolation clorox reached 0.4mM in fermention medium, the α-Tong Wuersuan accumulation volume reached 22.2g/L, and comparison is according to having improved 7.5%;
Or fermentation initial stage 0h, when interpolation hydroxyl sulphur ammonia reached 0.48 μ M in fermention medium, the α-Tong Wuersuan accumulation volume reached 22.0g/L, and comparison is according to having improved 6.1%.
1, bacterial classification
Torulopsis glabrata (T.glabrata) CCTCC NO.M 202019, be nicotinic acid, vitamin H, VitB1,4 kinds of vitamin deficient strain of pyridoxine hydrochloride, and the active composing type of pyruvic carboxylase reduces, and is this research department's seed selection bacterial strain, the patent No.: ZL 02113142.2.
2, substratum and cultural method
Seed culture medium: seed and slant medium (g/L): glucose 30, peptone 10, KH
2PO
41, MgSO
47H
2O 0.5; Agar 20 (slant medium), pH 5.5.
Fermention medium: glucose 100g/L; NH
4Cl 7g/L; KH
2PO
45g/L; MgSO
47H
2O 0.8g/L; Nicotinic acid 8mg/L; Pyridoxine hydrochloride 0.4mg/L; VitB1 0.02mg/L; Vitamin H 0.04mg/L; CaCO
360g/L; Add the KGDH inhibitor of different concns according to requirement of experiment.
Shake-flask culture: the seed of 30 ℃, 200rpm being cultivated 24h down changes fermentation culture over to 10% inoculum size and cultivates 72h based under 30 ℃, 200rpm condition.
3, analytical procedure
Determination of glucose: 3,5-dinitrosalicylic acid method.
The mensuration of pyruvic acid (PYR) and α-Tong Wuersuan (KG): adopt high performance liquid chromatography (HPLC) to measure.With reference to Agillent organic acid measuring method.Chromatographic condition is: ZORBAX SB-Aq reversed-phase column, and column temperature: 35 ℃, detector: UV 210nm, moving phase: 19.7g/L Na
2HPO
4+ 1.2%H
3PO
4+ 1% acetonitrile, pH2.0, flow velocity: 1.0mL/min, sampling volume: 5 μ L.
Dry cell weight (DCW) is measured: according to dry cell weight and corresponding OD value drawing standard curve,
1OD＝0.29g/L?DCW。
Beneficial effect of the present invention: the present invention utilizes carbon metabolism and the distribution of the carbon metabolism flow flow direction in the regulation and control industrial microorganism, makes carbon metabolism flow terminate in that wherein the KG maximum production reaches 23.2g/L on this node of KG, and comparison is according to having improved 12.2%.
Embodiment
Embodiment 1 adds hydrogen peroxide to the influence to T.glabrata CCTCC NO.M 202019 fermentation and acids
The fermentation initial stage is added hydrogen peroxide, and along with the increase of hydrogen peroxide addition, output of pyruvic acid presents ascendant trend, and when the hydrogen peroxide addition was 5mM, KG output reached and is 21.8g/L to the maximum, and comparison is shone has increased by 5.6%, at this moment C
KG/ C
PYRValue is 0.73.Investigate the fermentation different steps and add the 5mM hydrogen peroxide to producing the influence of acid, KG produces acid initial stage (40h) and adds hydrogen peroxide, and the accumulation volume of KG increases maximum, is 23.2g/L, and comparison is according to having improved 12.2%.
The optimum addition that draws hydrogen peroxide thus is 5mM, and the interpolation time is that KG produces the acid initial stage (40h).
Embodiment 2 adds the influence of methotrexate to CCTCC NO.M 202019 fermentation and acids
The fermentation initial stage is added different concns Rheumatrex (MTX), increase along with the MTX addition, PYR and KG output all present the trend of falling after rising, when MTX concentration in the nutrient solution reaches 0.08 μ M, PYR and KG output all reach maximum value, be respectively 30.4g/L and 22.7g/L, comparison is shone has increased by 8.9% and 10.1% respectively, at this moment C
KG/ C
PYRValue is 0.75.Investigate the fermentation different steps and add MTX to producing the influence discovery of acid, along with the carrying out of fermentation, the influence that MTX produces KG to fermentation diminishes.
The optimum addition that draws MTX thus is 0.08 μ M, and the interpolation time is fermentation initial stage (0h).
Embodiment 3 adds the influence of clorox to CCTCC NO.M 202019 fermentation and acids
Add clorox and find that when interpolation concentration reached 0.4mM, KG output was 22.2g/L in substratum, comparison is according to increasing by 7.5%, C
Pyr/ C
KgValue has descended 6%.Investigate the fermentation different times and add 0.4mMNaClO
3The influence of producing acid is found fermentation initial (0h) is the best interpolation time.
The optimum addition that draws clorox thus is 0.4mM, and the interpolation time is fermentation initial stage (0h).
Embodiment 4 adds the influence of hydroxyl sulphur ammonia to CCTCC NO.M 202019 fermentation and acids
Add hydroxyl sulphur ammonia and find that when interpolation concentration reached 0.48 μ M, KG output was 22.0g/L in substratum, comparison is shone increases by 6.1%, and PYR output has descended 13.7%, C
Pyr/ C
KgValue is tending towards definite value, shows the increase along with oxythiamine concentration, and the KG fractional yield increases, and entering TCA round-robin carbon metabolism flow flow increases.
The optimum addition that draws hydroxyl sulphur ammonia thus is 0.48 μ M, and the interpolation time is fermentation initial stage (0h).
Claims (1)
1, a kind of method of adding ketoglurate dehydrogenase inhibitor realization α-Tong Wuersuan excess accumulation, torulopsis glabrata (T.glabrata) CCTCCM202019 that it is characterized in that utilizing multiple vitamin defective type is for producing bacterial strain, by in substratum, adding the ketoglurate dehydrogenase inhibitor: hydrogen peroxide, methotrexate, clorox or hydroxyl sulphur ammonia, adjusting ketoglurate dehydrogenase activity, the degraded of autotelic minimizing α-Tong Wuersuan, carbon stream is terminated on this node of α-Tong Wuersuan, realize the α-Tong Wuersuan excess accumulation; Regulate and control method is:
Fermentation 40h, when the interpolation hydrogen peroxide reached 5mM in fermention medium, the α-Tong Wuersuan accumulation volume reached 23.2g/L;
Or fermentation initial stage 0h, when the interpolation methotrexate reached 0.08 μ M in fermention medium, the α-Tong Wuersuan accumulation volume reached 22.7g/L;
Or fermentation initial stage 0h, when the interpolation clorox reached 0.4mM in fermention medium, the α-Tong Wuersuan accumulation volume reached 22.2g/L;
Or fermentation initial stage 0h, when interpolation hydroxyl sulphur ammonia reached 0.48 μ M in fermention medium, the α-Tong Wuersuan accumulation volume reached 22.0g/L.
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|---|---|---|---|
| CN2008100200394A CN101250563B (en) | 2008-03-20 | 2008-03-20 | Method for realizing excessive accumulation of alpha-ketoglutarate acid by adding alpha-ketoglutarate acid dehydrogenase inhibitor |
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| CN101250563B CN101250563B (en) | 2010-11-03 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102269737A (en) * | 2010-06-07 | 2011-12-07 | 北京嘉事联博医药科技有限公司 | HPLC (high performance liquid chromatography) detection method of arginine ketoglutarate |
| CN103911400A (en) * | 2014-04-02 | 2014-07-09 | 江南大学 | Method for efficiently producing alpha-oxoglutarate by adopting whole-cell transformation |
| CN105177065A (en) * | 2015-09-11 | 2015-12-23 | 浙江树人大学 | Method for synthesizing alpha-ketoglutaric acid by biological conversion method |
| CN110404074A (en) * | 2019-08-20 | 2019-11-05 | 中国医学科学院基础医学研究所 | Application of the OGDH mortifier in treatment disease of viral infection |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1259425C (en) * | 2003-11-13 | 2006-06-14 | 江南大学 | Method for microbial fermentation synthesis of alpha- ketoglutaric acid |
-
2008
- 2008-03-20 CN CN2008100200394A patent/CN101250563B/en not_active Expired - Fee Related
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102269737A (en) * | 2010-06-07 | 2011-12-07 | 北京嘉事联博医药科技有限公司 | HPLC (high performance liquid chromatography) detection method of arginine ketoglutarate |
| WO2011153884A1 (en) * | 2010-06-07 | 2011-12-15 | 北京嘉事联博医药科技有限公司 | Method for detecting l-arginine-alpha-ketoglutarate using high performance liquid chromatography |
| CN102269737B (en) * | 2010-06-07 | 2013-01-30 | 北京嘉事联博医药科技有限公司 | HPLC (high performance liquid chromatography) detection method of arginine ketoglutarate |
| CN103911400A (en) * | 2014-04-02 | 2014-07-09 | 江南大学 | Method for efficiently producing alpha-oxoglutarate by adopting whole-cell transformation |
| CN103911400B (en) * | 2014-04-02 | 2016-04-27 | 江南大学 | A kind of method adopting resting cell to produce α-ketoglutaric acid |
| CN105177065A (en) * | 2015-09-11 | 2015-12-23 | 浙江树人大学 | Method for synthesizing alpha-ketoglutaric acid by biological conversion method |
| CN105177065B (en) * | 2015-09-11 | 2019-07-23 | 浙江树人大学 | A kind of method of biotransformation method synthesis α-ketoglutaric acid |
| CN110404074A (en) * | 2019-08-20 | 2019-11-05 | 中国医学科学院基础医学研究所 | Application of the OGDH mortifier in treatment disease of viral infection |
| CN110404074B (en) * | 2019-08-20 | 2021-07-13 | 中国医学科学院基础医学研究所 | Use of OGDH inhibitors in the treatment of viral infectious diseases |
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| Publication number | Publication date |
|---|---|
| CN101250563B (en) | 2010-11-03 |
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