CN102174599A - Method for biotransformation of succinic acid by using succinic acid fermentation waste cells - Google Patents
Method for biotransformation of succinic acid by using succinic acid fermentation waste cells Download PDFInfo
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- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 title claims abstract description 123
- 239000001384 succinic acid Substances 0.000 title claims abstract description 61
- 238000000034 method Methods 0.000 title claims abstract description 35
- 239000002921 fermentation waste Substances 0.000 title abstract 2
- 230000036983 biotransformation Effects 0.000 title description 4
- 238000000855 fermentation Methods 0.000 claims abstract description 32
- 230000004151 fermentation Effects 0.000 claims abstract description 32
- 239000012528 membrane Substances 0.000 claims abstract description 22
- 244000005700 microbiome Species 0.000 claims abstract description 21
- 239000002253 acid Substances 0.000 claims abstract description 13
- 239000000843 powder Substances 0.000 claims abstract description 13
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 10
- 239000012510 hollow fiber Substances 0.000 claims abstract description 7
- 239000001888 Peptone Substances 0.000 claims abstract description 6
- 108010080698 Peptones Proteins 0.000 claims abstract description 6
- 235000019319 peptone Nutrition 0.000 claims abstract description 6
- 239000003513 alkali Substances 0.000 claims abstract description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 33
- 239000007788 liquid Substances 0.000 claims description 23
- 239000000835 fiber Substances 0.000 claims description 21
- 241000894006 Bacteria Species 0.000 claims description 19
- 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 claims description 18
- 239000008103 glucose Substances 0.000 claims description 18
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 17
- 235000011121 sodium hydroxide Nutrition 0.000 claims description 11
- 230000008569 process Effects 0.000 claims description 10
- 238000004140 cleaning Methods 0.000 claims description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 8
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 8
- 239000007844 bleaching agent Substances 0.000 claims description 8
- 229910052799 carbon Inorganic materials 0.000 claims description 8
- 239000000460 chlorine Substances 0.000 claims description 8
- 229910052801 chlorine Inorganic materials 0.000 claims description 8
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 6
- 230000002572 peristaltic effect Effects 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 241000606750 Actinobacillus Species 0.000 claims description 5
- 240000008042 Zea mays Species 0.000 claims description 4
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 4
- 230000002000 scavenging effect Effects 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 3
- SRBFZHDQGSBBOR-LECHCGJUSA-N alpha-D-xylose Chemical compound O[C@@H]1CO[C@H](O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-LECHCGJUSA-N 0.000 claims description 3
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 3
- 235000005822 corn Nutrition 0.000 claims description 3
- 230000008878 coupling Effects 0.000 claims description 3
- 238000010168 coupling process Methods 0.000 claims description 3
- 238000005859 coupling reaction Methods 0.000 claims description 3
- 238000012262 fermentative production Methods 0.000 claims description 3
- 235000017550 sodium carbonate Nutrition 0.000 claims description 3
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 3
- 229960003487 xylose Drugs 0.000 claims description 3
- 241000186226 Corynebacterium glutamicum Species 0.000 claims description 2
- 244000068988 Glycine max Species 0.000 claims description 2
- 235000010469 Glycine max Nutrition 0.000 claims description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- SRBFZHDQGSBBOR-QMKXCQHVSA-N alpha-L-arabinopyranose Chemical compound O[C@H]1CO[C@@H](O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-QMKXCQHVSA-N 0.000 claims description 2
- 230000000968 intestinal effect Effects 0.000 claims description 2
- 230000014759 maintenance of location Effects 0.000 claims description 2
- 210000002966 serum Anatomy 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- OGWLTJRQYVEDMR-UHFFFAOYSA-F tetramagnesium;tetracarbonate Chemical compound [Mg+2].[Mg+2].[Mg+2].[Mg+2].[O-]C([O-])=O.[O-]C([O-])=O.[O-]C([O-])=O.[O-]C([O-])=O OGWLTJRQYVEDMR-UHFFFAOYSA-F 0.000 claims description 2
- 241000282414 Homo sapiens Species 0.000 claims 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims 1
- 230000000813 microbial effect Effects 0.000 abstract description 7
- 238000004519 manufacturing process Methods 0.000 abstract description 6
- 239000002699 waste material Substances 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 3
- 238000004134 energy conservation Methods 0.000 abstract description 3
- 230000007613 environmental effect Effects 0.000 abstract description 3
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract 4
- 229910052757 nitrogen Inorganic materials 0.000 abstract 2
- 238000004064 recycling Methods 0.000 abstract 1
- 239000000047 product Substances 0.000 description 8
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 241000948980 Actinobacillus succinogenes Species 0.000 description 4
- 238000013475 authorization Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 229940041514 candida albicans extract Drugs 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000011218 seed culture Methods 0.000 description 3
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- UXFQFBNBSPQBJW-UHFFFAOYSA-N 2-amino-2-methylpropane-1,3-diol Chemical compound OCC(N)(C)CO UXFQFBNBSPQBJW-UHFFFAOYSA-N 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000031018 biological processes and functions Effects 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 125000001477 organic nitrogen group Chemical group 0.000 description 2
- 230000003204 osmotic effect 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
- 229920001817 Agar Polymers 0.000 description 1
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 1
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
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- MSJMDZAOKORVFC-SEPHDYHBSA-L disodium fumarate Chemical compound [Na+].[Na+].[O-]C(=O)\C=C\C([O-])=O MSJMDZAOKORVFC-SEPHDYHBSA-L 0.000 description 1
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- 235000019253 formic acid Nutrition 0.000 description 1
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- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 235000009973 maize Nutrition 0.000 description 1
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- 239000002994 raw material Substances 0.000 description 1
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- 239000011780 sodium chloride Substances 0.000 description 1
- 235000019294 sodium fumarate Nutrition 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a method for biologically converting succinic acid by using succinic acid fermentation waste cells, which comprises the following steps: (1) carrying out normal fermentation for producing succinic acid by the microorganisms; (2) after the fermentation of succinic acid fermentation production by using microorganisms is finished, separating microbial cells in fermentation liquor in a fermentation tank; (3) inoculating the microbial cells separated in the previous step into a fermentation tank for continuous fermentation; (4) adding alkali for many times to neutralize the generated acid; (5) after the fermentation is finished, a succinic acid product is separated. The invention has the beneficial effects that: the fermentation broth after the fermentation is finished is filtered by a hollow fiber membrane or centrifuged to recycle the cells to the fermentation tank for continuous utilization, the method can reduce the use amount of expensive nitrogen sources such as yeast powder, peptone and the like, can realize the recycling of the nitrogen sources in the fermentation process, reduces the production cost of products, reduces the burden of waste treatment, and achieves the social benefits of environmental protection and energy conservation.
Description
Technical field
The present invention relates to the production method of a kind ofization Succinic Acid, be specifically related to a kind of method of utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid.
Background technology
Utilizing microorganism to carry out in the process of Succinic Acid fermentative production, organic nitrogen source as the main component in the raw material to microbial growth metabolism played crucial effects.Most of nitrogenous source participates in the synthetic of microorganism cells, and small portion then remains in the fermented liquid.Usually select the organic nitrogen source that utilized by microorganism easily in the fermentation, for example yeast powder, peptone and corn steep liquor etc., these nitrogenous sources can guarantee the output and the production efficiency of product to the full extent, yet but cost an arm and a leg.
Along with being accompanied by the principal product Succinic Acid in the Succinic Acid fermenting process, the generation of by product formic acid, acetate, lactic acid etc., and need the acid that constantly neutralizes and produce with alkali during the fermentation, therefore the concentration of the metal ion in the fermented liquid also can constantly raise (causing the raising of osmotic pressure), too high concentration of metal ions produces considerable influence to the growth of thalline with product acid, cell reduces significantly at fermentation later stage acid producing ability, and fermentation also stops immediately, and cell just is discharged as waste.Yet still have a complete set of enzyme system that catalytic substrate generates product in the cell, discharging causes these a complete set of enzyme systems with catalytic substrate generation product and the waste of nitrogenous source.
Publication number is the patent application of CN1189854, disclosing a kind of single microorganism that utilizes is 1 with the fermentable carbon source bio-transformation, ammediol, it is 1 with single microorganism with a kind of carbon substrate bio-transformation that this invention provides a kind of, the method of ammediol is not seen relevant disclosing waste cell recycle bio-transformation Succinic Acid and the relevant patent of other organic acids thereof.
Summary of the invention
Collect thalline after the objective of the invention is to utilize hollow-fibre membrane with filtering fermentation liquor the fermented liquid after the Succinic Acid fermentation, or by centrifugal thalline is turned back in the fermentor tank, only add water and glucose, carbon sources such as stalk hydrolyzed solution transform and generate Succinic Acid.Can reduce the usage quantity of expensive nitrogenous sources such as yeast powder, peptone and the adding of mineral ion, realize the recycle of nitrogenous source in the fermenting process, not only reduce the products production cost but also alleviated the burden of offal treatment, reached green, environmental protection, energy-conservation social benefit.
The scheme of finishing the foregoing invention task is that a kind of method of utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid is characterized in that step is as follows:
(1) microorganism carries out the fermentation of normal succinic acid-producing;
(2) after microorganism to be utilized carries out the fermentation ends of Succinic Acid fermentative production, the microorganism cells in the fermented liquid in the fermentor tank is separated;
(3) microorganism cells that previous step is separated in rapid is transferred into fermentor tank relaying supervention ferment;
(4) repeatedly add in the alkali and the acid that produces;
(5) after the fermentation ends, isolate the Succinic Acid product.
Fermentation condition is as follows:
Actication of culture: bacterial classification carries out plate streaking in slant medium after, activation culture 24h in 37 ℃ of anaerobism incubators; Seed culture: the bacterial classification after the activation culture is transferred in the seed culture medium, cultivates behind 10～12h as seed liquor for 37 ℃;
Fermentor cultivation: in the glucose access fermentor tank that will activate good seed liquor and go out good, inoculum size is 5～10%, and v/v, (volume ratio) mixing speed are 200rpm, 37 ℃ of fermentation culture, C0
2Air flow is 0.25vvm;
Slant medium (g/L): glucose 10 (branch disappears), yeast extract paste 5, NaHCO
310, NaH
2PO
42H
2O 9.6, K
2HPO
43H
2O 15.5, agar 20,7.0,121 ℃ of sterilizations of pH 15min.
Seed culture medium (g/L): glucose 10 (branch disappears), yeast extract paste 5, NaHCO
310, NaH
2PO
42H
2O 9.6, K
2HPO
43H
2O 15.5,7.0,121 ℃ of sterilizations of pH 15min.
Fermention medium (g/L): glucose 60 (branch disappears), yeast extract paste 10, Disodium fumarate 1, KH
2PO 3, MgCl
26H
2O0.3, CaCl
20.3 NaCl 1,7.0,121 ℃ of sterilizations of pH 15min.
Step in the above scheme (1) described " microorganism cells in the fermented liquid in the fermentor tank is separated " mainly comprises 2 kinds of methods:
First method is the recycle that the coupling of hollow-fibre membrane and fermentor tank reaches cell, concrete grammar utilizes peristaltic pump that it is pumped into the fermented liquid of fermentor tank combination (see figure 1) after with fermentation ends to filter in the hollow-fibre membrane for hollow-fibre membrane being carried out aseptic cleaning then, with cell retention in the tubular fibre mould, freshly prepd glucose solution or stalk hydrolyzed solution are being sprung back over from hollow-fibre membrane in the fermentor tank, and cell is got back to fermentor tank relaying supervention ferment again in the recoil process.
Another kind method is after treating fermentation ends, with the fermented liquid in the fermentor tank, transfers to and carries out centrifugal collection bacterium mud in the big centrifuge tube, and these operations are all carried out under sterile state, and fermentor tank relaying supervention ferment is gone in switching then.
The aseptic scavenging solution that carries out aseptic cleaning of hollow-fibre membrane of the present invention comprises: the HCl solution of configuration pH 1.9～2.1, the NaOH solution of configuration pH 9.9～10.1, the chlorine bleach liquor of w/v0.45～0.55%; Each 2.0L, and sterilized water 5.0L.Standby.
As follows membrane module is carried out aseptic cleaning: (1) at first utilizes the HCl dilute solution that hollow fiber film assembly is carried out wash cycles 2h, and emptying is washed to neutrality; (2) utilize the NaOH dilute solution that hollow fiber film assembly is carried out wash cycles 2h, emptying is washed to neutrality; (3) utilize chlorine bleach liquor's wash cycles 2h, emptying; (4) utilize sterilized water that membrane module is cleaned (not circulating), can realize the aseptically process of hollow-fibre membrane.
The present invention recommends: described HCl pH value of solution is 2.0, described NaOH pH value of solution is 10.0, described chlorine bleach liquor w/v (mass/volume) is 0.5%.
The concrete operations of collection bacterium mud of the present invention are: at first the tail gas relief outlet of fermentor tank is clamped (see figure 1), fermented liquid is pressed onto in the serum bottle of the bacterium of going out in advance from jar by the pressure in the jar, in super clean bench, the fermented liquid branch is installed in the big centrifuge tube of the bacterium of going out then, carry out centrifugal.Bacterium mud is collected in the centrifugal back that finishes in super clean bench, just can be inoculated in the fermentor tank then.
Beneficial effect of the present invention is:
Utilize method of the present invention, with the fermented liquid after the fermentation ends through the tubular fibre membrane filtration or centrifugal cell is returned in the fermentor tank, continue to utilize, this method not only can reduce the usage quantity of expensive nitrogenous sources such as yeast powder, peptone, and can realize the recycle of nitrogenous source in the fermenting process, not only reduced the products production cost but also alleviated the burden of offal treatment, reached green, environmental protection, energy-conservation social benefit, technical solution of the present invention is effective and feasible.
Description of drawings
Fig. 1 is the equipment structure chart of Succinic Acid fermentation of the present invention with the hollow-fibre membrane coupling.
1,2,3 is peristaltic pump among the figure, 4pH indicating meter 5pH meter 6pH conditioning agent (as yellow soda ash, sodium hydroxide, ammoniacal liquor etc.) 7 glucose solutions, 8 hollow-fibre membranes, 9 fermentor tanks, 10 thief holes, 11 carbonic acid gas.
Embodiment
Following examples describe in detail the present invention, but to application of the present invention and unrestricted.
Embodiment 1: prepare the recycle that realizes nitrogenous source in the process of Succinic Acid at biological process.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.Be that nitrogenous source carries out anaerobically fermenting and prepares Succinic Acid at first with the 10g/L yeast powder, when the glucose in the fermentor tank exhausts by the time, just carrying out fermented liquid separates, concrete device is seen Fig. 1, at first opening peristaltic pump 1 pumps into fermented liquid in the hollow-fibre membrane, hollow-fibre membrane can be retained down thalline, fermented liquid part through hollow-fibre membrane flows out through the below, obtain clarifying fermented liquid, a part in addition will turn back to from above and continue in the fermentor tank to separate, opening peristaltic pump 2 after to be separated the finishing pumps into glucose solution in the hollow-fibre membrane, simultaneously peristaltic pump 1 is broken in the other direction, glucose solution can elute to bring back in the fermentor tank and goes when the hollow-fibre membrane being trapped in thalline in the film like this, at this time had only glucose and cell in the fermentor tank, and do not had the influence of organic acid and osmotic pressure etc., and cell just can transform again efficiently, and conversion results sees Table 1.Transform the Succinic Acid that finally can generate 56.2g/L through 2 times.
Table 1 conversion results
Embodiment 2: present embodiment prepares the recycle that realizes nitrogenous source in the process of Succinic Acid at biological process.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.Be that nitrogenous source carries out anaerobically fermenting and prepares Succinic Acid with the 10g/L yeast powder at first, when the glucose in the fermentor tank exhausted by the time, it was centrifugal just to carry out fermented liquid, collects bacterium mud, transforms succinic acid-producing.Experimental result sees Table two, has become the Succinic Acid of 54.8g/L through 2 centrifugal last symbiosis.
Table 2 conversion results
Embodiment 3: the microbial strains of the succinic acid-producing that 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.Be that nitrogenous source carries out anaerobically fermenting and prepares Succinic Acid at first with the 10g/L yeast powder, when the glucose in the fermentor tank exhausts by the time, just carrying out fermented liquid separates, concrete implementation and operation such as embodiment 1, maize straw hydrolyzed solution (be mainly glucose, wood sugar, mass ratio is approximately 2: 1) is replaced glucose solution to be transformed, the results are shown in Table 3, through the Succinic Acid of 2 living 58.95g/L of centrifugal common property.
Table 3 conversion results
Embodiment 4: 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.Be that nitrogenous source carries out anaerobically fermenting and prepares Succinic Acid at first, when the glucose in the fermentor tank exhausts by the time, separate just carry out fermented liquid with the 10g/L yeast powder, concrete implementation and operation such as embodiment 1, the yeast powder add 3g/L in fermentor tank transforms.The results are shown in Table 3, through the Succinic Acid of 3 living 81.75g/L of centrifugal common property.
Table 4 conversion results
Embodiment 5, and is basic identical with the foregoing description, but following change is arranged:
Described microorganism is adopted intestinal bacteria;
In the described aseptic scavenging solution that carries out aseptic cleaning: the pH of HCl solution is 1.9; The pH of NaOH solution is 9.9; Chlorine bleach liquor's w/v is 0.45%;
Described carbon source adopts wood sugar; Described nitrogenous source adopts peptone; Described alkaline neutraliser adopts magnesium basic carbonate.
Embodiment 6, and is basic identical with the foregoing description, but following change is arranged:
Described microorganism is adopted Corynebacterium glutamicum;
In the described aseptic scavenging solution that carries out aseptic cleaning: the pH of HCl solution is 2.1; The pH of NaOH solution is 10.1; Chlorine bleach liquor's w/v is 0.55%;
Described carbon source adopts pectinose sugar; Described nitrogenous source adopts Dried Corn Steep Liquor Powder; Described alkaline neutraliser adopts yellow soda ash.
Embodiment 7, and is basic identical with the foregoing description, but following change is arranged: described microorganism adopts big Man Haimushi to produce the succsinic acid bacterium; Described carbon source adopts sucrose; Described nitrogenous source adopts soybean cake powder; Described alkaline neutraliser adopts sodium bicarbonate.
Embodiment 8, and is basic identical with the foregoing description, but following change is arranged: described carbon source adopts stalk through pretreated mixing sugar; Described alkaline neutraliser adopts sodium hydroxide.
Embodiment 9, and is basic identical with the foregoing description, but described alkaline neutraliser adopts ammoniacal liquor.
Claims (10)
1. method of utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid is characterized in that step is as follows:
(1) microorganism carries out the fermentation of normal succinic acid-producing;
(2) after microorganism to be utilized carries out the fermentation ends of Succinic Acid fermentative production, the microorganism cells in the fermented liquid in the fermentor tank is separated;
(3) microorganism cells that previous step is separated in rapid is transferred into fermentor tank relaying supervention ferment;
(4) repeatedly add in the alkali and the acid that produces;
(5) after the fermentation ends, isolate the Succinic Acid product.
2. the method for utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid according to claim 1 is characterized in that, step (1) described " microorganism cells in the fermented liquid in the fermentor tank is separated " is to adopt one of following 2 kinds of methods:
First method is the recycle that the coupling of hollow-fibre membrane and fermentor tank reaches cell: hollow fiber film assembly is carried out aseptic cleaning utilize peristaltic pump that it is pumped in the hollow-fibre membrane fermented liquid after the fermentation ends with fermentor tank combination then to filter, with cell retention in the tubular fibre mould, freshly prepd glucose solution or stalk hydrolyzed solution are being sprung back over from hollow-fibre membrane in the fermentor tank, and cell is got back to fermentor tank relaying supervention ferment again in the recoil process;
Another kind method is after treating fermentation ends, with the fermented liquid in the fermentor tank, transfers to and carries out centrifugal collection bacterium mud in the big centrifuge tube, and these operations are all carried out under sterile state, and fermentor tank relaying supervention ferment is gone in switching then.
3. the method for utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid according to claim 2, it is characterized in that the described aseptic scavenging solution that carries out aseptic cleaning comprises: the NaOH solution of pH 1.9～2.1HCl solution, pH 9.9～10.1, mass/volume are than 0.45～0.55% chlorine bleach liquor; Each 2.0L, and sterilized water 5.0L.
4. the method for utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid according to claim 2 is characterized in that, described HCl pH value of solution is 2.0, described NaOH pH value of solution is 10.0, described chlorine bleach liquor's mass/volume ratio is 0.5%.
5. the method for utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid according to claim 2, it is characterized in that, it is described that to carry out aseptic cleaning be as follows hollow fiber film assembly to be carried out aseptic cleaning: a. at first to utilize the HCl dilute solution that hollow fiber film assembly is carried out wash cycles 2h, emptying is washed to neutrality; B. utilize the NaOH dilute solution that hollow fiber film assembly is carried out wash cycles 2h, emptying is washed to neutrality; C. utilize chlorine bleach liquor's wash cycles 2h, emptying; D. utilize sterilized water that membrane module is cleaned, can realize the aseptically process of hollow-fibre membrane.
6. the method for utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid according to claim 2, it is characterized in that, the concrete operations of described collection bacterium mud are: at first the tail gas relief outlet of fermentor tank is clamped, fermented liquid is pressed onto in the serum bottle of the bacterium of going out in advance from jar by the pressure in the jar, in super clean bench, the fermented liquid branch is installed in the big centrifuge tube of the bacterium of going out then, carry out centrifugal; Bacterium mud is collected in the centrifugal back that finishes in super clean bench, just can be inoculated in the fermentor tank then.
7. according to the described method of utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid of one of claim 3～6, it is characterized in that the condition of described configuration sterilized water is: 121 ℃, sterilization 15min.
8. according to the described method of utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid of one of claim 3～6, it is characterized in that described microorganism is selected from: produce the succsinic acid actinobacillus, intestinal bacteria, Corynebacterium glutamicum, or Man Haimushi produces succsinic acid bacterium etc.
9. the method for utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid according to claim 7 is characterized in that,
Described carbon source is selected from: glucose, wood sugar, pectinose, sucrose, or stalk etc. is through pretreated mixing sugar;
Described nitrogenous source is selected from: yeast powder, peptone, corn steep liquor or soybean cake powder.
10. the method for utilizing the fermented abandoned cell biological of Succinic Acid to transform Succinic Acid according to claim 7 is characterized in that,
Described alkaline neutraliser is selected from: magnesium basic carbonate, yellow soda ash, sodium bicarbonate, sodium hydroxide or ammoniacal liquor.
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| CN102352383A (en) * | 2011-11-09 | 2012-02-15 | 南京工业大学 | Method for preparing succinic acid by fermenting bagasse |
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| CN102352384A (en) * | 2011-11-11 | 2012-02-15 | 南京工业大学 | Chemical synthesis culture medium for producing succinic acid by fermentation and application thereof |
| CN102718579A (en) * | 2012-07-18 | 2012-10-10 | 寇建国 | Compound microbial liquid drip irrigation fertilizer and preparation method thereof |
| WO2014127636A1 (en) * | 2013-02-21 | 2014-08-28 | 中国科学院过程工程研究所 | Method and device for two-stage dry digestion of wastes |
| CN103205468A (en) * | 2013-04-24 | 2013-07-17 | 南京工业大学 | Method for preparing succinic acid by utilizing sucrose fermentation |
| CN103497890A (en) * | 2013-09-26 | 2014-01-08 | 北京华都诗华生物制品有限公司 | Perfusion culture system and method for bacteria fermentation tank |
| CN105861288A (en) * | 2015-01-23 | 2016-08-17 | 中国科学院微生物研究所 | Method and apparatus for preparing N-acetylneuraminic acid |
| CN105948834A (en) * | 2016-06-22 | 2016-09-21 | 广东惜福环保科技有限公司 | Kitchen waste composting primary fermentation equipment filtrate autocycle reinjection system |
| CN107236769A (en) * | 2017-06-26 | 2017-10-10 | 南京工业大学 | Method for preparing L-ornithine and succinic acid by stages by utilizing membrane circulation bioreactor |
| CN107236769B (en) * | 2017-06-26 | 2021-01-26 | 南京工业大学 | Method for preparing L-ornithine and succinic acid by stages by utilizing membrane circulation bioreactor |
| CN107988137A (en) * | 2017-12-15 | 2018-05-04 | 武汉新华扬生物股份有限公司 | A kind of highly concentrated clostridium butyricum gemma production method and highly concentrated clostridium butyricum gemma product |
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