CN101624608A - Method for improving output of pyruvic acid by regulating and controlling oxidation-reduction potential of culturing system - Google Patents
Method for improving output of pyruvic acid by regulating and controlling oxidation-reduction potential of culturing system Download PDFInfo
- Publication number
- CN101624608A CN101624608A CN200910184724A CN200910184724A CN101624608A CN 101624608 A CN101624608 A CN 101624608A CN 200910184724 A CN200910184724 A CN 200910184724A CN 200910184724 A CN200910184724 A CN 200910184724A CN 101624608 A CN101624608 A CN 101624608A
- Authority
- CN
- China
- Prior art keywords
- pyruvic acid
- output
- reduction potential
- cctcc
- regulating
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a method for improving the output of pyruvic acid by regulating and controlling an oxidation-reduction potential of a culturing system, belonging to the technical field of optimizing the fermentation process by a cofactor metabolism control strategy. In a fermentation medium of Torulopsis glabrate (T. glabrate) CCTCC M202019, the oxygen-dissolved concentration of the fermentation medium is controlled to be 20 percent, foreign electronic acceptor potassium ferricyanide is added into the fermentation medium, and the initial oxidation-reduction potential of the medium is improved to 400mV, thereby the intracellular NADH level of the Torulopsis glabrate CCTCC M202019 is reduced, the glycolysis speed is accelerated, and the output of the pyruvic acid is improved. Under the fermentation conditions, the intracellular NADH concentration of CCTCC M202019 is reduced to 45.3 percent, and the output of the pyruvic acid is improved to 61.9g/L from 49.6g/L. The strategy also has instructive meanings to other fermentation products.
Description
Technical field
A kind of redox potential of regulating and control culture system improves the method for torulopsis glabrata (T.glabrata) output of pyruvic acid, relates to cofactor metabolic regulation policy optimization fermenting process technical field.
Background technology
Pyruvic acid claims 2-oxo propionic acid, α-Tong Jibingsuan or ethanoyl formic acid again, is one of most important alpha-oxo-carboxyl acid, in industry such as chemical industry, pharmacy and agrochemicals and scientific research purposes is widely arranged.The T.glabrata of polyauxotroph is at present unique one and is used for the microorganism that the industrial fermentation method is produced pyruvic acid.Pyruvic acid is glucolytic end product, and is closely related with the NADH metabolism.Studies show that, suitably reduce NADH level in the T.glabrata born of the same parents, increase NAD
+Supply with, help quickening glycolysis-, improve output of pyruvic acid.
Utilize regulation and control NADH and NAD that biochemical engineering and metabolic engineering strategy can be easily and effectively
+Form and utilizability.The metabolic engineering strategy comprises: (1) overexpression NADH metabolism involved enzyme class; (2) deletion NADH (NAD
+) competition utilizes approach; (3) introduce NADH (NAD
+) regeneration system rapidly.And the method for biochemical engineering comprises: add (1) external source electron acceptor(EA) in fermented liquid, as acetaldehyde and fumaric acid etc.; (2) (sorbyl alcohol increases the NADH utilizability, and gluconic acid then reduces NADH/NAD to have the carbon source of different oxidation state
+Ratio); (3) NAD
+Precursor is effectively to control NADH (NAD in the born of the same parents
+) mode of concentration.Other biochemistry regulation and control strategy is controlled culture condition in addition in addition, for example increases dissolved oxygen level and regulates born of the same parents' external oxidation reduction potential.
The redox potential of fermention medium is the comprehensive parameters of the various envrionment conditionss of reflection, depends on media components, pH, osmotic pressure, temperature and oxyty etc.Studies show that, can realize the regulation and control of metabolism distributions, stream of electrons NADH level in, plurality of enzymes/proteic biosynthesizing and born of the same parents by the redox potential of regulating fermention medium industrial microorganism.Therefore redox potential often is used as the output that a process parameter improves the target metabolite, citric acid, 1 for example, ammediol, wood sugar and aromatic compounds.But, regulate NADH level in the born of the same parents by redox potential, thereby improve T.glabrata glycolysis-speed and output of pyruvic acid, do not see bibliographical information as yet.
Summary of the invention
The purpose of this invention is to provide a kind of NADH level in the redox potential regulation and control born of the same parents of utilizing, improve the method for T.glabrata output of pyruvic acid.
Technical scheme of the present invention: a kind of regulating and controlling oxidation-reduction potential of culturing system improves the method for output of pyruvic acid, in torulopsis glabrata (T.glabrata) CCTCC M202019 fermention medium, the oxyty of control fermention medium is 20%, and in fermention medium, add the external source electron acceptor(EA) Tripotassium iron hexacyanide, the initial oxidation reduction potential that improves substratum is to 400mV, thereby reduce NADH level in the torulopsis glabrata CCTCC M202019 born of the same parents, quicken glycolysis-, improve output of pyruvic acid.
Bacterial strain: torulopsis glabrata (T.glabrata) CCTCC M202019, nicotinic acid (NA), vitamin H (Bio), VitB1 (B
1), pyridoxol four kinds of vitamin defective typies such as (Pdx), and the active composing type of pyruvic carboxylase reduces, and is this research department's seed selection bacterial strain, this bacterial classification has been applied for Chinese patent, patent No. ZL 02113142.2, publication number is CN1392246A.
Beneficial effect of the present invention: utilize the present invention can efficiently improve the output of pyruvic acid.NADH concentration reduces by 45.3% in the T.glabrata born of the same parents, and output of pyruvic acid is brought up to 61.9g/L by 49.6g/L.Redox potential regulation and control strategy of the present invention has certain directive significance to the production of other leavened prod.
Embodiment
Comparative examples
The used bacterial classification of the present invention is torulopsis glabrata (Torulopsis glabrata) CCTCC M202019, changeing the inclined-plane earlier cultivated 24 hours in 30 ℃ of incubators, carried out shake-flask culture 24 hours at 30 ℃ of temperature, rotating speed 200r/min then, in the 7L fermentor tank, ferment at last, fermentation condition is: pH 5.5, mixing speed 400r/min, oxyty 20%.Fermention medium is (g/L): glucose 100, ammonium chloride 7, sodium acetate 6, KH
2PO
45, Mg
2SO
47H
2O 0.8, CaCl
22H
2O 2, FeSO
47H
2O 2, ZnCl
20.5, MnCl
24H
2O 12, CuSO
45H
2O 0.05, nicotinic acid 0.08, and VitB1 0.00015, pyridoxol 0.04, vitamin H 0.004, riboflavin 0.01, pH 5.5.
Nicotinic acid 80mg, VitB1 0.15mg, pyridoxol 40mg, vitamin H 4mg, riboflavin 10mg, tap water is settled to 1L.
Under this fermentation condition, NADH concentration is 7.5mg/g dry cell weight (DCW) in the T.glabrata CCTCC M202019 born of the same parents, and output of pyruvic acid is 49.6g/L.
Embodiment 1
Adding an amount of (100～120 μ mol/L) Tripotassium iron hexacyanide in fermention medium makes the initial oxidation reduction potential of fermention medium reach 400mV, with redox electrode assembly (French Mei Tele company product, model Pt 4085-DXK) redox potential in the mensuration fermention medium, the same comparative examples of all the other conditions.
Under this fermentation condition, NADH concentration is 4.1mg/gDCW in the T.glabrata CCTCC M202019 born of the same parents, and output of pyruvic acid is 61.9g/L.
Claims (1)
1, a kind of regulating and controlling oxidation-reduction potential of culturing system improves the method for output of pyruvic acid, it is characterized in that in torulopsis glabrata (T.glabrata) CCTCC M202019 fermention medium, the oxyty of control fermention medium is 20%, and in fermention medium, add the external source electron acceptor(EA) Tripotassium iron hexacyanide, the initial oxidation reduction potential that improves substratum is to 400mV, thereby reduce NADH level in the torulopsis glabrata CCTCC M202019 born of the same parents, quicken glycolysis-, improve output of pyruvic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910184724A CN101624608A (en) | 2009-08-20 | 2009-08-20 | Method for improving output of pyruvic acid by regulating and controlling oxidation-reduction potential of culturing system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910184724A CN101624608A (en) | 2009-08-20 | 2009-08-20 | Method for improving output of pyruvic acid by regulating and controlling oxidation-reduction potential of culturing system |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101624608A true CN101624608A (en) | 2010-01-13 |
Family
ID=41520600
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910184724A Pending CN101624608A (en) | 2009-08-20 | 2009-08-20 | Method for improving output of pyruvic acid by regulating and controlling oxidation-reduction potential of culturing system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101624608A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102864179A (en) * | 2012-08-29 | 2013-01-09 | 太仓市茂通化建有限公司 | Method for preparing pyruvic acid by utilizing torulopsis glabrata efficiently |
| CN102864178A (en) * | 2012-08-29 | 2013-01-09 | 太仓市茂通化建有限公司 | Method for producing pyruvic acid by utilizing torulopsis glabrata |
-
2009
- 2009-08-20 CN CN200910184724A patent/CN101624608A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102864179A (en) * | 2012-08-29 | 2013-01-09 | 太仓市茂通化建有限公司 | Method for preparing pyruvic acid by utilizing torulopsis glabrata efficiently |
| CN102864178A (en) * | 2012-08-29 | 2013-01-09 | 太仓市茂通化建有限公司 | Method for producing pyruvic acid by utilizing torulopsis glabrata |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bafana et al. | New approaches for itaconic acid production: bottlenecks and possible remedies | |
| San et al. | Metabolic engineering through cofactor manipulation and its effects on metabolic flux redistribution in Escherichia coli | |
| Du et al. | Use of oxidoreduction potential as an indicator to regulate 1, 3-propanediol fermentation by Klebsiella pneumoniae | |
| Hong et al. | Importance of redox balance on the production of succinic acid by metabolically engineered Escherichia coli | |
| Jin et al. | Ultrahigh‐cell‐density heterotrophic cultivation of the unicellular green alga Chlorella sorokiniana for biomass production | |
| Hu et al. | Current strategies and future prospects for enhancing microbial production of citric acid | |
| Olajuyin et al. | Efficient production of succinic acid from Palmaria palmata hydrolysate by metabolically engineered Escherichia coli | |
| San-Valero et al. | Influence of electron acceptors on hexanoic acid production by Clostridium kluyveri | |
| US8691552B2 (en) | Microaerobic cultures for converting glycerol to chemicals | |
| Liu et al. | Fermentation and redox potential | |
| US20100297715A1 (en) | Method for producing succinic acid | |
| Qi et al. | A protocol for optimization vinegar fermentation according to the ratio of oxygen consumption versus acid yield | |
| Wang et al. | R-acetoin accumulation and dissimilation in Klebsiella pneumoniae | |
| da Silva et al. | The industrial versatility of Gluconobacter oxydans: current applications and future perspectives | |
| Gallardo et al. | Influence of nutritional and operational parameters on the production of butanol or 1, 3-propanediol from glycerol by a mutant Clostridium pasteurianum | |
| Han et al. | Production of 2, 3-butanediol by a low-acid producing Klebsiella oxytoca NBRF4 | |
| Guo et al. | Efficient production of 2, 3-butanediol from cheese whey powder (CWP) solution by Klebsiella pneumoniae through integrating pulsed fed-batch fermentation with a two-stage pH control strategy | |
| CN102154339A (en) | Construction method of gene engineering strain for producing succinic acid escherichia coli | |
| Goel et al. | Suppressed acid formation by cofeeding of glucose and citrate in Bacillus cultures: emergence of pyruvate kinase as a potential metabolic engineering site | |
| Shanmugam et al. | Engineering biocatalysts for production of commodity chemicals | |
| Basso et al. | Insufficient uracil supply in fully aerobic chemostat cultures of Saccharomyces cerevisiae leads to respiro-fermentative metabolism and double nutrient-limitation | |
| Martin et al. | Performance and stability of ethanologenic Escherichia coli strain FBR5 during continuous culture on xylose and glucose | |
| CN103184243A (en) | Fermentation production method for xylitol | |
| Sridhar et al. | Influence of redox potential on product distribution in Clostridium thermosuccinogenes | |
| Crognale et al. | Fed-batch gluconic acid production from Penicillium variabile P16 under different feeding strategies |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20100113 |