CN101712970A - Method for preparing butanedioic acid through fermentation - Google Patents
Method for preparing butanedioic acid through fermentation Download PDFInfo
- Publication number
- CN101712970A CN101712970A CN200910264056A CN200910264056A CN101712970A CN 101712970 A CN101712970 A CN 101712970A CN 200910264056 A CN200910264056 A CN 200910264056A CN 200910264056 A CN200910264056 A CN 200910264056A CN 101712970 A CN101712970 A CN 101712970A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- butanedioic acid
- mixture
- succinic acid
- acid
- 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.)
- Granted
Links
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 title claims abstract description 168
- 238000000855 fermentation Methods 0.000 title claims abstract description 82
- 229960005137 succinic acid Drugs 0.000 title claims abstract description 82
- 230000004151 fermentation Effects 0.000 title claims abstract description 78
- 238000000034 method Methods 0.000 title claims abstract description 61
- 238000004519 manufacturing process Methods 0.000 claims abstract description 18
- 230000000813 microbial effect Effects 0.000 claims abstract description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 60
- 239000001384 succinic acid Substances 0.000 claims description 54
- 239000000203 mixture Substances 0.000 claims description 33
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 13
- 239000002253 acid Substances 0.000 claims description 12
- 238000011218 seed culture Methods 0.000 claims description 6
- 239000003513 alkali Substances 0.000 abstract description 21
- 230000001105 regulatory effect Effects 0.000 abstract description 12
- 238000002360 preparation method Methods 0.000 abstract description 8
- 230000008569 process Effects 0.000 abstract description 8
- 150000001875 compounds Chemical class 0.000 abstract description 6
- ZLNQQNXFFQJAID-UHFFFAOYSA-L magnesium carbonate Chemical compound [Mg+2].[O-]C([O-])=O ZLNQQNXFFQJAID-UHFFFAOYSA-L 0.000 abstract description 5
- 239000001095 magnesium carbonate Substances 0.000 abstract description 5
- 229910000021 magnesium carbonate Inorganic materials 0.000 abstract description 5
- 230000008901 benefit Effects 0.000 abstract description 4
- 238000010170 biological method Methods 0.000 abstract 1
- 239000011734 sodium Substances 0.000 description 27
- 239000003795 chemical substances by application Substances 0.000 description 22
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 20
- 230000003472 neutralizing effect Effects 0.000 description 20
- 239000002585 base Substances 0.000 description 14
- 239000007788 liquid Substances 0.000 description 10
- 238000002156 mixing Methods 0.000 description 10
- 230000000694 effects Effects 0.000 description 9
- 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 8
- 239000008103 glucose Substances 0.000 description 8
- 230000004060 metabolic process Effects 0.000 description 8
- 230000012010 growth Effects 0.000 description 7
- 239000002609 medium Substances 0.000 description 7
- 238000009825 accumulation Methods 0.000 description 6
- 229910019440 Mg(OH) Inorganic materials 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 4
- 235000014380 magnesium carbonate Nutrition 0.000 description 4
- 229960001708 magnesium carbonate Drugs 0.000 description 4
- 150000007524 organic acids Chemical class 0.000 description 4
- 230000035699 permeability Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 241000606750 Actinobacillus Species 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 3
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- 230000002503 metabolic effect Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 241000948980 Actinobacillus succinogenes Species 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 230000031018 biological processes and functions Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- DTBNBXWJWCWCIK-UHFFFAOYSA-N phosphoenolpyruvic acid Chemical compound OC(=O)C(=C)OP(O)(O)=O DTBNBXWJWCWCIK-UHFFFAOYSA-N 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229940040526 anhydrous sodium acetate Drugs 0.000 description 1
- 238000013475 authorization Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000021523 carboxylation Effects 0.000 description 1
- 238000006473 carboxylation reaction Methods 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000012847 fine chemical Substances 0.000 description 1
- 238000005189 flocculation Methods 0.000 description 1
- 230000016615 flocculation Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000005431 greenhouse gas Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000029219 regulation of pH Effects 0.000 description 1
- 230000000452 restraining effect Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Abstract
The invention relates to a method for preparing butanedioic acid through fermentation. A fermentation system is kept to have a proper pH mainly by adding two types of mixed alkaline compound s instead of a single alkaline compound in the process of producing butanedioic acid through microbial fermentation, and the proportion of mixed alkali is regulated so as to ensure that the butanedioic acid produced is at a high level. The method not only can utilize the cheap alkaline compound s, but also can basically reach the level of butanedioic acid production when basic magnesium carbonate is utilized as the only alkaline compound to regulate pH. Therefore, by utilizing a mixed-alkali pH-regulating strategy to optimize a production process, the method has the advantages of reducing production cost of butanedioic acid biological preparation, reducing the difficulty of separating and extracting downstream products, increasing economic benefit and helping to the industrial preparation of butanedioic acid biological method.
Description
Technical field
The invention belongs to technical field of bioengineering, relate to a kind of method of utilizing mixed base to regulate the pH preparing butanedioic acid through fermentation.
Background technology
Succinic Acid has another name called succsinic acid, is a kind of di-carboxylic acid, and the C4 hardware and software platform compound as important is widely used in aspects such as food, medicine, spices, washing composition, fine chemical product, tensio-active agent Biodegradable material.The traditional production method of Succinic Acid is to be that raw material is refined synthetic with chemical method with the oil.And along with petroleum resources are exhausted day by day, the Production by Microorganism Fermentation Succinic Acid with its environment friendly, can utilize depleted biomass resource, fixing greenhouse gases CO
2Etc. advantage, become the heat subject of research, just in depth carrying out in the U.S., Japan and other countries.
CO
2With the pH value be the key factor that influences microbial bacteria bulk-growth and its Succinic Acid output.CO
2In the growth metabolism process of thalline, utilized synthetic purpose product Succinic Acid by thalline as carbon source.
And the change of environment pH value can cause the variation of thalline inside and outside chemical environment and enzyme activity, thereby the pair cell metabolism exerts an influence.The pH value can also influence CO
2Level of dissolution, and HCO
3 -, CO
3 2-Dissociation equilibrium, and then influence the synthetic of Succinic Acid.So in the process of microbial fermentation succinic acid-producing, regulate suitable pH level and select suitable regulative mode that metabolic activity and the acid producing ability that improves thalline had vital role.
In addition, during the fermentation, the organic acid accumulation can cause pH to descend, and also needs to add the neutral level that environment is kept in the pH regulator agent.For producing succsinic acid actinobacillus (Actinobacillus succinogenes), the optimal pH of its fermentation succinic acid-producing is between 6.6～7.0.The US5573931 report, MgCO
3Be neutralizing agent preferably, can present regulating power preferably, such as in fermention medium, adding MgCO
3Solid can maintain suitable level with the pH value of fermentation system, makes thalline show higher metabolic activity during the fermentation; But, MgCO
3Cost is higher, and consumption is bigger, also can further increase the difficulty of derived product separation and Extraction, and in fact it be unfavorable for the industrial production of Succinic Acid biological process.
Summary of the invention
Technical purpose of the present invention is to provide a kind of novel method of utilizing preparing butanedioic acid through fermentation, makes this method to come optimization production technology by selecting suitable pH regulator agent, formulation reasonably to regulate strategy, reduces the biological production cost for preparing Succinic Acid.
Technical purpose of the present invention is to realize by following technical scheme:
A kind of method of preparing butanedioic acid through fermentation comprises actication of culture, seed culture, anaerobic fermentation and acid production three steps, it is characterized in that adding in the step of anaerobic fermentation and acid production the mixture of two kinds of alkaline neutralisers, and the pH value of adjusting fermentation system is 6.6～7.0.
Wherein, the mixture of described two kinds of alkaline neutralisers is ammoniacal liquor, NaOH, Na
2CO
3, NaHCO
3, Ca (OH)
2, CaCO
3, Mg (OH)
2In the arbitrarily mixture of combination in twos;
Further, best of breed is in these combinations:
(1) ammoniacal liquor, Mg (OH)
2
(2)NaOH、Mg(OH)
2；
(3)Na
2CO
3、Mg(OH)
2；
(4)NaHCO
3、Mg(OH)
2；
(5)Ca(OH)
2、Mg(OH)
2；
(6)CaCO
3、Mg(OH)
2。
Wherein, the quality proportioning of two of combination kinds of alkaline neutralisers is 1: 3～3: 1.
Beneficial effect of the present invention is:
1, the present invention utilizes the multiple single alkaline neutraliser of prior art to regulate pH value preparing butanedioic acid through fermentation, thereby filters out the alkaline neutraliser that alkalescence is strong and help fermenting; The alkaline neutraliser that screens has been carried out combination be used to regulate pH value preparing butanedioic acid through fermentation, and determined the optimum proportion of mixed base, made the Succinic Acid of producing a higher level;
2, the present invention's relatively inexpensive alkaline neutraliser that not only utilized some, and can reach substantially and utilize alkali formula MgCO
3Succinic Acid production level when regulating pH as unique alkali formula compound.Therefore, the method for utilizing mixed base to regulate the pH value is come optimization production technology, has reduced the production cost of biological preparation Succinic Acid, has reduced the difficulty of derived product separation and Extraction, has improved economic benefit, helps the industrial preparation of Succinic Acid biological process;
3, because of Mg
2+The key enzyme that produces in the succsinic acid actinobacillus pathways metabolism there is promoter action, though utilize alkali formula MgCO
3The production level of Succinic Acid is the highest when regulating pH as unique alkaline neutraliser, but owing to uses alkali formula MgCO
3Cost higher, and Mg (OH)
2Mg can be provided
2+But its alkalescence is relatively poor, so the present invention determines that one of composition is Mg (OH) in the mixed base
2, select the stronger neutralizing agent of other a kind of alkalescence to mix with it again, realized same outstanding technique effect.
Description of drawings
Fig. 1 regulates the procedure chart of pH preparing butanedioic acid through fermentation for adopting ammoniacal liquor
Fig. 2 is for adopting (a) Ca (OH) respectively
2(b) CaCO
3Regulate the procedure chart of pH preparing butanedioic acid through fermentation
Fig. 3 is for adopting (a) NaOH (b) Na respectively
2CO
3(c) NaHCO
3Regulate the procedure chart of pH preparing butanedioic acid through fermentation
Fig. 4 is for adopting Mg (OH)
2Regulate the procedure chart of pH preparing butanedioic acid through fermentation
Fig. 5 is the Na that utilizes different ratios
2CO
3And Mg (OH)
2Mix and regulate the influence of pH the preparing butanedioic acid through fermentation result
Fig. 6 is NaOH and the Mg (OH) that utilizes different ratios
2Mix and regulate the influence of pH the preparing butanedioic acid through fermentation result
Fig. 7 is for adopting MgCO
3Regulate the procedure chart of pH preparing butanedioic acid through fermentation
Embodiment
The following examples elaborate to the present invention, but to the present invention without limits.
The microbial strains that the method for preparing Succinic Acid of the present invention can be used any anaerobically fermenting succinic acid-producing in the prior art.The microbial strains of the succinic acid-producing that present embodiment adopted is: produce succsinic acid actinobacillus NJ113 (Actinobacillus succinogenes NJ113), this bacterium patent applied for is also obtained the authorization, and the license notification number is CN100537744C.
Fermention medium of the present invention can use the fermention medium of any anaerobically fermenting succinic acid-producing in the prior art.The substratum that present embodiment adopted is: every liter of volume substratum contains glucose (separately sterilization) 50～100g, yeast powder 15g, corn steep liquor 15mL, anhydrous sodium acetate 1.36g, NaCl 1g, K
2HPO
43g, MgCl
20.2g, CaCl
20.2g.
The method of organism of fermentation strain preparation Succinic Acid of the present invention can be anaerobically fermenting preparation method arbitrarily in the prior art, and these methods generally all comprise actication of culture, seed culture, anaerobic fermentation and acid production three steps.The preparation process of this enforcement is:
(1) actication of culture: the slant medium of actication of culture is the solid slant culture base that carbon source, nitrogenous source and inorganic salt can be provided that contains carbohydrate of pH 6.0～8.0, after during slant culture bacterial strain being carried out plate streaking in slant medium, cultivate in the anaerobism incubator, incubator contains N
2, CO
2And H
2Mixed gas, temperature is 30～40 ℃, activation culture 24～48h is used for the preservation of seed culture medium inoculation and bacterial strain;
(2) seed culture: adding seed culture medium in the 100mL serum bottle is 20～80mL, feeds and contains N
2And CO
2Gas mixture, 115～121 ℃ of sterilization 15～30min, the slant culture bacterial strain is inserted in the cooling back, and culture temperature is 30～40 ℃, shaking speed 100～200r/min, incubation time is 10～16h, is used for the fermention medium inoculation;
(3) the 5L ferment tank is cultivated: 5L fermentation cylinder for fermentation culture volume is 2～4L, and inoculum size is 3～7% (v/v), and temperature is 35～40 ℃, and fermentor tank feeds and contains N
2And CO
2Gas mixture, to keep the anaerobic environment of fermentation system, mixing speed is at 100～300rpm, and fermenting process adopts alkaline neutraliser to comprise ammoniacal liquor, NaOH, Na respectively
2CO
3, NaHCO
3, Ca (OH)
2, CaCO
3, Mg (OH)
2PH is 6.6～7.0 in control, and fermentation time is 30～48h, separating and extracting succinic acid.
Analytical procedure:
HPLC system high efficiency liquid chromatograph: WatersHPLC2010 workstation; Chromatographic column Prevail organic acidcolumn 250mm * 4.6mm; Ultraviolet detection wavelength 215nm; Flow velocity 1mL/min, input 20 μ L, moving phase 25mmol/L KH
2PO
4, pH 2.5; Column temperature: room temperature.Organic acid (Succinic Acid (Fluka), acetate (Sigma), formic acid (Fluka), lactic acid (the Fluka)) standardized solution of redistilled water preparation different concns is according to the concentration relationship production standard curve of peak area and organic acid standardized solution.Fermented sample after centrifugal dilution according to each organic acid content of calculated by peak area.
In the present embodiment, the method for adding alkaline neutraliser in the step of succinic acid-producing separately is:
Add ammoniacal liquor and regulate the method for pH value as alkaline neutraliser.
By adding ammoniacal liquor, the pH that regulates fermentation system is between 6.6～7.0 in the ferment tank process, and the fermentation result as shown in Figure 1.NH in the fermented liquid
4 +The restraining effect of cell growth is comparatively strong, and thalline utilizes ability lower to sugar, causes a large amount of glucose residues.To fermentation ends, the Succinic Acid mass concentration is 23.5g/L only.Cytolemma is to NH
4 +Higher permeability is arranged, NH
4 +Infiltration can cause that the pH level changes in the born of the same parents, cell needs more energy with NH
4 +Pump, as seen, higher NH in the environment
4 +Concentration can reduce the utilising efficiency of cell to energy.When the energy undersupply, be subjected to NH in the born of the same parents
4 +Influence, bigger variation takes place in the pH in the born of the same parents, influences cell growth metabolism normally, finally causes death.
Present embodiment adopts and embodiment 1 identical fermentation condition and method, and the method that only changes the independent interpolation alkaline neutraliser in the succinic acid-producing step is:
Add Ca (OH) respectively
2And CaCO
3Regulate the method for pH value as alkaline neutraliser.
In fermentor tank, adopt Ca (OH) respectively
2, CaCO
3Regulate pH respectively and ferment, the results are shown in shown in Figure 2.Adopt Ca (OH)
2Regulate, thalline is grown hardly, and the Succinic Acid mass concentration is less than 5g/L (Fig. 2-a).Adopt and add CaCO in advance
3Solid is regulated pH fermentation, the growth fraction Ca (OH) of thalline
2Good slightly when regulating fermentation, but still be subjected to obvious suppression.CaCO
3Dissolving power relatively poor, pH can not be maintained the level that is fit to thalli growth, pH constantly reduces in the fermenting process, to fermentation ends, pH is that 5.6 (Fig. 2-b), Succinic Acid output only is 17.5g/L.The Ca of higher concentration
2+Can change normal flowability of cytolemma and permeability, cause thalline can not carry out the inside and outside matter energy transmission of normal born of the same parents, so that can't the normal growth metabolism.
Present embodiment adopts and embodiment 1 identical fermentation condition and method, and the method that only changes the independent interpolation alkaline neutraliser in the succinic acid-producing step is:
Add NaOH, Na respectively
2CO
3, NaHCO
3Regulate the method for pH value as alkaline neutraliser.
In fermentor tank, adopt stream to add NaOH, Na respectively
2CO
3, NaHCO
3Mode regulate pH, the fermentation result as shown in Figure 3.Along with Na
+Continuous accumulation in fermented liquid, fermentation middle and later periods thalline flocculation phenomenon is serious, thalline OD
660Be obvious downtrending, sugar consumption rate also slows down gradually.NaOH and Na
2CO
3, NaHCO
3Compare, to fermentation ends, the glucose residual content is more, and the Succinic Acid mass concentration is 43.5g/L (Fig. 3-a).Na
2CO
3, NaHCO
3Extra CO can be provided
3 2-, HCO
3 -, help the thalline synthesizing succinic acid, but solvability and alkalescence being lower, it is bigger to the diluting effect of fermented liquid that stream adds process, with NaHCO
3Particularly serious, though there is glucose residue, the Succinic Acid mass concentration has only 37.8g/L (Fig. 3-b).Adopt Na
2CO
3Regulate, a small amount of residual sugar is arranged, the Succinic Acid mass concentration is 39.9g/L (Fig. 3-c).
In the metabolic process of cell, Na
+Crucial effect is arranged, and it can influence the pH regulation and control level in film pH gradient, Premeabilisation of cells pressure and the born of the same parents of striding.Adopt NaOH, Na
2CO
3Regulate cumulative Na in the fermented liquid
+Concentration reaches 2.1mol/L, 1.9mol/L respectively, adopts NaHCO
3Regulate Na
+Be 1.5mol/L.Na
+Excessively accumulation causes height to ooze environment, and thalline is had bigger negative effect, and acid, acid producing ability rapid drawdown are produced in thalline metabolism normally.The fermentation middle and later periods, Na
+A large amount of accumulation may be to cause thalline OD
660The major cause that sharply descend, acid producing ability reduces.
Present embodiment adopts and embodiment 1 identical fermentation condition and method, and the method that only changes the independent interpolation alkaline neutraliser in the succinic acid-producing step is:
Add Mg (OH) respectively
2, MgCO
3Regulate the method for pH value as alkaline neutraliser.
Mg
2+Be the activator of many enzymes, key enzyme phosphoenolpyruvic acid carboxylation kinases just needs Mg in the Succinic Acid route of synthesis
2+As cofactor.Fig. 4 and Fig. 7 adopt Mg (OH) respectively
2, MgCO
3Regulate the fermentation result of pH.Mg
2+Accumulation in fermented liquid does not have the obvious suppression effect to thalli growth, fermentation middle and later periods, OD
660Descend slowly, thalline still has higher metabolic activity.To fermentation ends, glucose can fully be utilized by thalline, and Succinic Acid concentration is up to 57.4g/L (Fig. 7), and ferment effect is best.Adopt Mg (OH)
2During adjusting, because Mg (OH)
2Obvious to the fermented liquid diluting effect, the Succinic Acid mass concentration only has 42.9g/L (Fig. 4).
Present embodiment adopts and embodiment 1 identical fermentation condition and method, and the method that only changes the interpolation mixed-alkali neutralizing agent in the succinic acid-producing step is:
Utilize ammoniacal liquor and Mg (OH)
2As the mixed-alkali neutralizing agent, and with different blending ratio adjusting pH values.
Adopt ammoniacal liquor and Mg (OH)
2Regulate pH with different mass mixed (3: 1,2: 1,1: 1,1: 2,1: 3), OH in the control mixed base
-Concentration is that 6mol/L (provides 1mol OH by 1mol ammoniacal liquor
-Meter).The result is as shown in table 1 in fermentation.
The ammoniacal liquor of table 1 different ratios and Mg (OH)
2Mix and regulate pH fermentation result contrast
The ammoniacal liquor ratio is high more in mixed base, because the anti-NH of microorganism
4 +Ability is relatively poor, so ferment effect is also poor more.Along with Mg (OH)
2The continuous increase of addition, the residual sugar amount reduces gradually, and the Succinic Acid mass concentration increases gradually.But because Mg (OH)
2Therefore solubleness is not high, and it is bigger to regulate the volume that pH covers, and makes the concentration of Succinic Acid descend to some extent.
Present embodiment adopts and embodiment 1 identical fermentation condition and method, and the method that only changes the interpolation mixed-alkali neutralizing agent in the succinic acid-producing step is:
Utilize NaHCO
3And Mg (OH)
2As the mixed-alkali neutralizing agent, and with different blending ratio adjusting pH values.
Adopt NaHCO
3And Mg (OH)
2Regulate pH with different mass mixed (3: 1,2: 1,1: 1,1: 2,1: 3), OH in the control mixed base
-Concentration is that 6mol/L (presses 1mol NaHCO
32mol is provided OH
-Meter).The result is as shown in table 2 in fermentation.
The NaHCO of table 2 different ratios
3And Mg (OH)
2Mix and regulate pH fermentation result contrast
Though utilize NaHCO
3Effect was better when the adjusting microbial fermentation prepared Succinic Acid, but because NaHCO
3The not high and easy crystallization of solubleness, the volume that covers when therefore regulating pH is bigger, makes the concentration of Succinic Acid descend to some extent.
Embodiment 7
Present embodiment adopts and embodiment 1 identical fermentation condition and method, and the method that only changes the interpolation mixed-alkali neutralizing agent in the succinic acid-producing step is:
Utilize Na
2CO
3And Mg (OH)
2As the mixed-alkali neutralizing agent, and with different blending ratio adjusting pH values.
Adopt Na
2CO
3And Mg (OH)
2Regulate pH with different mass mixed (3: 1,2: 1,1: 1,1: 2,1: 3), OH in the control mixed base
-Concentration is that 6mol/L (presses 1mol Na
2CO
32mol is provided OH
-Meter).The fermentation result as shown in Figure 5.Along with Mg (OH)
2The continuous increase of addition, the residual sugar amount reduces gradually, and the Succinic Acid mass concentration increases gradually.Work as Na
2CO
3And Mg (OH)
2Mass ratio reaches at 1: 1 o'clock, and glucose does not have residue, and the Succinic Acid mass concentration is the highest, is 55.4g/L, Na in the fermented liquid
+Concentration 0.95mol/L is than adopting NaHCO
3The Na that regulates
+Concentration has reduced by 36.7%.Continue to increase Mg (OH) in the mixed base
2Addition, glucose still can consume fully, but the concentration of Succinic Acid descends to some extent.
Present embodiment adopts and embodiment 1 identical fermentation condition and method, and the method that only changes the interpolation mixed-alkali neutralizing agent in the succinic acid-producing step is:
Utilize NaOH and Mg (OH)
2As the mixed-alkali neutralizing agent, and with different blending ratio adjusting pH values.
Adopt NaOH and Mg (OH)
2Mixing solutions is regulated pH in the fermenting process, NaOH and Mg (OH) as neutralizing agent
2Mass ratio was respectively 3: 1,2: 1,1: 1,1: 2,1: 3, OH in the control mixed base
-Concentration is that 6mol/L (presses 1mol Mg (OH)
22mol is provided OH
-Meter), the result as shown in Figure 6.Along with NaOH adding proportion in the mixed alkali liquor reduces, the residual content of glucose reduces gradually.As NaOH and Mg (OH)
2Mass ratio be 1: 1 o'clock, no glucose residue, the mass concentration of Succinic Acid can reach 69.8g/L, Na in the fermented liquid
+Concentration 0.86mol/L is with employing NaHCO
3The Na that regulates
+Concentration is compared, and has reduced by 42.6%.Continue to reduce the adding proportion of NaOH in the mixed base, thalline still can all consume glucose, but the concentration of Succinic Acid presents downtrending.
Contain Na
+Alkaline conditioner with contain Mg
2+Alkaline conditioner is mixed to suitable proportion, can alleviate Na
+Excess accumulation, also can reduce Mg (OH)
2Dilute strength to fermented liquid.With Na
2CO
3And Mg (OH)
2Mix the optimum of regulating the pH fermentation and compare, adopt NaOH and Mg (OH)
2To mix the better effects if of adjusting pH fermentation at 1: 1.
Embodiment 9
Present embodiment adopts and embodiment 1 identical fermentation condition and method, and the method that only changes the interpolation mixed-alkali neutralizing agent in the succinic acid-producing step is:
Utilize Ca (OH)
2And Mg (OH)
2As the mixed-alkali neutralizing agent, and with different blending ratio adjusting pH values.
Adopt Ca (OH)
2And Mg (OH)
2Regulate pH with different mass mixed (3: 1,2: 1,1: 1,1: 2,1: 3), OH in the control mixed base
-Concentration is that 6mol/L (presses 1mol Ca (OH)
22mol is provided OH
-Meter).The result is as shown in table 3 in fermentation.
The Ca of table 3 different ratios (OH)
2And Mg (OH)
2Mix and regulate pH fermentation result contrast
Because Ca (OH)
2Solubleness is very low, and the Ca of higher concentration
2+Can change normal flowability of cytolemma and permeability, cause thalline can not carry out the inside and outside matter energy transmission of normal born of the same parents, so that can't the normal growth metabolism.Therefore utilize Ca (OH)
2And Mg (OH)
2When regulating the pH value as the mixed-alkali neutralizing agent, the concentration of Succinic Acid is very low.
Present embodiment adopts and embodiment 1 identical fermentation condition and method, and the method that only changes the interpolation mixed-alkali neutralizing agent in the succinic acid-producing step is:
Utilize CaCO
3And Mg (OH)
2As the mixed-alkali neutralizing agent, and with different blending ratio adjusting pH values.
Adopt CaCO
3And Mg (OH)
2Regulate pH with different mass mixed (3: 1,2: 1,1: 1,1: 2,1: 3), OH in the control mixed base
-Concentration is that 6mol/L (presses 1mol CaCO
32mol is provided OH
-Meter).The result is as shown in table 4 in fermentation.
The CaCO of table 4 different ratios
3And Mg (OH)
2Mix and regulate pH fermentation result contrast
Because CaCO
3Solubleness is very low, and the Ca of higher concentration
2+Can change normal flowability of cytolemma and permeability, cause thalline can not carry out the inside and outside matter energy transmission of normal born of the same parents, so that can't the normal growth metabolism.Therefore utilize CaCO
3And Mg (OH)
2When regulating the pH value as the mixed-alkali neutralizing agent, the concentration of Succinic Acid is lower.
Embodiment 11
Present embodiment adopts and embodiment 1 identical fermentation condition and method, initial glucose concentration is 100g/L in the fermention medium, regulate the pH value with the alkaline neutraliser optimum mixture ratio example of embodiment 5～10 respectively, and regulate pH fermentation result with independent use magnesiumcarbonate and compare, as shown in table 5.
Table 5 utilizes two kinds of alkaline neutralisers mixing to regulate pH and uses magnesiumcarbonate adjusting pH fermentation result contrast separately
Can obtain by table 5, have only the NaOH of employing and Mg (OH)
2Mixing solutions is 1: 1 o'clock as neutralizing agent and mass ratio, regulate pH in the fermenting process, do not have the glucose residue substantially, the mass concentration of Succinic Acid can reach 69.8g/L, the production level of Succinic Acid is suitable when regulating the pH fermentation with independent use magnesiumcarbonate, therefore utilizes cheap NaOH and Mg (OH)
2Mix neutralizing agent and can substitute expensive magnesiumcarbonate neutralizing agent fully, thereby reduce the production cost of Succinic Acid.
Claims (9)
1. the method for a preparing butanedioic acid through fermentation, comprise actication of culture, seed culture, anaerobic fermentation and acid production three steps, the mixture that it is characterized in that two kinds of alkaline neutralisers of external source interpolation in the step of producing succinic acid by microbial fermentation, and the pH value of adjusting fermentation system is 6.6～7.0.
2. the method for preparing butanedioic acid through fermentation according to claim 1, the mixture that it is characterized in that described two kinds of alkaline neutralisers is ammoniacal liquor, NaOH, Na
2CO
3, NaHCO
3, Ca (OH)
2, CaCO
3, Mg (OH)
2In the arbitrarily mixture of combination in twos.
3. the method for preparing butanedioic acid through fermentation according to claim 2, the mixture that it is characterized in that described two kinds of alkaline neutralisers is ammoniacal liquor and Mg (OH)
2Mixture.
4. the method for preparing butanedioic acid through fermentation according to claim 2, the mixture that it is characterized in that described two kinds of alkaline neutralisers is NaOH and Mg (OH)
2Mixture.
5. the method for preparing butanedioic acid through fermentation according to claim 2, the mixture that it is characterized in that described two kinds of alkaline neutralisers is Na
2CO
3And Mg (OH)
2Mixture.
6. the method for preparing butanedioic acid through fermentation according to claim 2, the mixture that it is characterized in that described two kinds of alkaline neutralisers is NaHCO
3And Mg (OH)
2Mixture.
7. the method for preparing butanedioic acid through fermentation according to claim 2, the mixture that it is characterized in that described two kinds of alkaline neutralisers is Ca (OH)
2And Mg (OH)
2Mixture.
8. the method for preparing butanedioic acid through fermentation according to claim 2, the mixture that it is characterized in that described two kinds of alkaline neutralisers is CaCO
3And Mg (OH)
2Mixture.
9. according to the method for the described preparing butanedioic acid through fermentation of one of claim 1 to 8, the quality proportioning that it is characterized in that described two kinds of alkaline neutralisers is 1: 3～3: 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910264056A CN101712970B (en) | 2009-12-29 | 2009-12-29 | Method for preparing butanedioic acid through fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910264056A CN101712970B (en) | 2009-12-29 | 2009-12-29 | Method for preparing butanedioic acid through fermentation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101712970A true CN101712970A (en) | 2010-05-26 |
| CN101712970B CN101712970B (en) | 2012-09-12 |
Family
ID=42416968
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910264056A Expired - Fee Related CN101712970B (en) | 2009-12-29 | 2009-12-29 | Method for preparing butanedioic acid through fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101712970B (en) |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101955978A (en) * | 2010-08-31 | 2011-01-26 | 南京工业大学 | Method for enhancing concentration and production intensity of succinic acid by adding osmoprotectant |
| CN102352384A (en) * | 2011-11-11 | 2012-02-15 | 南京工业大学 | Chemical synthesis culture medium for producing succinic acid through fermentation and applications thereof |
| CN103361383A (en) * | 2012-04-10 | 2013-10-23 | 中国石油化工股份有限公司 | Fermentation method for preparing succinic acid by adding divalent metal compound |
| CN104131040A (en) * | 2014-07-28 | 2014-11-05 | 南京工业大学 | Method for production of disodium succinate through microbial fermentation |
| EP2778227A4 (en) * | 2011-11-11 | 2014-12-17 | Mitsubishi Chem Corp | Method for producing succinic acid |
| CN104372037A (en) * | 2014-09-11 | 2015-02-25 | 南京工业大学 | Method for producing succinic acid by multistage continuous fermentation |
| CN105567748A (en) * | 2015-12-02 | 2016-05-11 | 南京工业大学 | Method for producing succinic acid by means of fermentation |
| CN107354180A (en) * | 2016-05-09 | 2017-11-17 | 中国石化扬子石油化工有限公司 | A kind of regulation and control method of Actinobacillus succinogenes production butanedioic acid |
| CN110241147A (en) * | 2019-07-05 | 2019-09-17 | 安徽丰原发酵技术工程研究有限公司 | A kind of method of fermenting and producing L MALIC ACID coproduction succinic acid |
| CN110804632A (en) * | 2019-11-18 | 2020-02-18 | 保龄宝生物股份有限公司 | Method for improving erythritol conversion rate by adding mixed magnesium salt |
| CN110964754A (en) * | 2019-12-31 | 2020-04-07 | 广西科学院 | Method for reducing proportion of succinic acid fermentation by-products of actinobacillus succinogenes |
| CN117385106A (en) * | 2023-12-12 | 2024-01-12 | 山东飞扬化工有限公司 | Rapid calculation method of feeding amount, feeding method and feeding system |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100537744C (en) * | 2006-06-14 | 2009-09-09 | 南京工业大学 | A kind of bacterial strain of succinic acid-producing and screening method thereof and application |
| CN100445257C (en) * | 2006-07-18 | 2008-12-24 | 南京工业大学 | Process of separating and extracting succinic acid from anaerobic fermented liquid |
-
2009
- 2009-12-29 CN CN200910264056A patent/CN101712970B/en not_active Expired - Fee Related
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101955978A (en) * | 2010-08-31 | 2011-01-26 | 南京工业大学 | Method for enhancing concentration and production intensity of succinic acid by adding osmoprotectant |
| CN102352384A (en) * | 2011-11-11 | 2012-02-15 | 南京工业大学 | Chemical synthesis culture medium for producing succinic acid through fermentation and applications thereof |
| EP2778227A4 (en) * | 2011-11-11 | 2014-12-17 | Mitsubishi Chem Corp | Method for producing succinic acid |
| CN103361383A (en) * | 2012-04-10 | 2013-10-23 | 中国石油化工股份有限公司 | Fermentation method for preparing succinic acid by adding divalent metal compound |
| CN104131040A (en) * | 2014-07-28 | 2014-11-05 | 南京工业大学 | Method for production of disodium succinate through microbial fermentation |
| CN104372037A (en) * | 2014-09-11 | 2015-02-25 | 南京工业大学 | Method for producing succinic acid by multistage continuous fermentation |
| CN105567748A (en) * | 2015-12-02 | 2016-05-11 | 南京工业大学 | Method for producing succinic acid by means of fermentation |
| CN105567748B (en) * | 2015-12-02 | 2019-07-26 | 南京工业大学 | A kind of method of fermentation production of succinic acid |
| CN107354180A (en) * | 2016-05-09 | 2017-11-17 | 中国石化扬子石油化工有限公司 | A kind of regulation and control method of Actinobacillus succinogenes production butanedioic acid |
| CN110241147A (en) * | 2019-07-05 | 2019-09-17 | 安徽丰原发酵技术工程研究有限公司 | A kind of method of fermenting and producing L MALIC ACID coproduction succinic acid |
| CN110804632A (en) * | 2019-11-18 | 2020-02-18 | 保龄宝生物股份有限公司 | Method for improving erythritol conversion rate by adding mixed magnesium salt |
| CN110964754A (en) * | 2019-12-31 | 2020-04-07 | 广西科学院 | Method for reducing proportion of succinic acid fermentation by-products of actinobacillus succinogenes |
| CN110964754B (en) * | 2019-12-31 | 2021-09-07 | 广西科学院 | Method for reducing proportion of succinic acid fermentation by-products of actinobacillus succinogenes |
| CN117385106A (en) * | 2023-12-12 | 2024-01-12 | 山东飞扬化工有限公司 | Rapid calculation method of feeding amount, feeding method and feeding system |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101712970B (en) | 2012-09-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101712970B (en) | Method for preparing butanedioic acid through fermentation | |
| CN1884484A (en) | Succinic acid-producing strain and its screening method and uses | |
| CN101709263B (en) | Chemostat continuous cultivation device | |
| CN104487582A (en) | Method for preparing organic acid by batch-feeding carbon source substrate and base | |
| CN103602714A (en) | Method for producing tetracycline by fermentation of streptomyces aureus | |
| CN100467607C (en) | Method for producing 1,3-propylene of using glycerol anaerobic fermentation | |
| CN101705261A (en) | Preparation method of Gamma-aminobutyric acid | |
| CN101153294B (en) | Immobilized cell single-tank high-strength continuous fermentation process for succinic acid | |
| CN103952447B (en) | Method for producing succinic acid by virtue of fermentation under anaerobic conditions | |
| CN101182555B (en) | Method for producing amber acid through regulating and controlling anaerobic fermentation by oxidation reduction potential | |
| CN101955978A (en) | Method for enhancing concentration and production intensity of succinic acid by adding osmoprotectant | |
| RU2447143C2 (en) | METHOD FOR SUBMERGED CULTIVATION OF Bacillus brevis FOR PRODUCING GRAMICIDIN S | |
| CN102352382A (en) | Method producing malic acid through double-phase fermentation | |
| CN103773691B (en) | A kind of method of closed fast culture microalgae | |
| CN116426577A (en) | Calcium hydroxide as neutralizer combined with CO 2 Method for producing succinic acid by pulse feedback feed supplement fermentation | |
| CN106399387A (en) | Nitrogen defect control method for production of ethanol through fermentation of synthesis gas with mixed bacteria | |
| CN103088081B (en) | Biotin supplemented fermentation method for producing glutamic acid | |
| CN101497871B (en) | Alcohol fermentation anaerobic high temperature bacterium culture medium, preparation and use thereof | |
| CN102747113A (en) | Process for producing 2-keto-D-gluconic acid by high-concentration fermentation | |
| CN103773819A (en) | Method for producing lactic acid and simultaneously realizing coupling preparation of GABA (gamma-aminobutyric acid) | |
| CN102352384B (en) | Chemical synthesis culture medium for producing succinic acid through fermentation and applications thereof | |
| CN110004202B (en) | Method for synthesizing hexanoic acid by catalyzing carbohydrate through microbial co-culture | |
| CN101555498A (en) | Technique for producing 2-keto-D-gluconic acid by fermentation of gluconate | |
| CN101698836B (en) | Method for supplementing material in the fermentation process of transglutaminase | |
| CN108060192B (en) | Fermentation medium for improving fermentation level of meleumycin and feeding method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120912 Termination date: 20171229 |