CN109371072A - A kind of technique for reducing glutamic acid fermentation microbiological contamination and improving fermentation efficiency - Google Patents
A kind of technique for reducing glutamic acid fermentation microbiological contamination and improving fermentation efficiency Download PDFInfo
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- 230000004151 fermentation Effects 0.000 title claims abstract description 80
- 238000000855 fermentation Methods 0.000 title claims abstract description 79
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 title claims abstract description 32
- 235000013922 glutamic acid Nutrition 0.000 title claims abstract description 32
- 239000004220 glutamic acid Substances 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title claims abstract description 23
- 238000011169 microbiological contamination Methods 0.000 title claims abstract description 11
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims abstract description 51
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 claims abstract description 17
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 claims abstract description 17
- 229960000367 inositol Drugs 0.000 claims abstract description 17
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 claims abstract description 17
- 241000186226 Corynebacterium glutamicum Species 0.000 claims description 17
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- 239000000654 additive Substances 0.000 claims description 11
- 230000000996 additive effect Effects 0.000 claims description 11
- 239000002609 medium Substances 0.000 claims description 11
- 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 10
- 239000008103 glucose Substances 0.000 claims description 10
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 8
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 8
- 239000001963 growth medium Substances 0.000 claims description 8
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 7
- 240000008042 Zea mays Species 0.000 claims description 6
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 6
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 6
- 235000005822 corn Nutrition 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 239000000908 ammonium hydroxide Substances 0.000 claims description 4
- 239000004202 carbamide Substances 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 4
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 4
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 4
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 4
- 238000000527 sonication Methods 0.000 claims description 4
- 238000009210 therapy by ultrasound Methods 0.000 claims description 3
- 241000894006 Bacteria Species 0.000 abstract description 19
- 235000001014 amino acid Nutrition 0.000 abstract description 2
- 150000001413 amino acids Chemical class 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 42
- 238000006243 chemical reaction Methods 0.000 description 28
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 16
- 210000004027 cell Anatomy 0.000 description 11
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- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- HHLFWLYXYJOTON-UHFFFAOYSA-N glyoxylic acid Chemical compound OC(=O)C=O HHLFWLYXYJOTON-UHFFFAOYSA-N 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 238000011218 seed culture Methods 0.000 description 4
- 230000003519 ventilatory effect Effects 0.000 description 4
- NAOLWIGVYRIGTP-UHFFFAOYSA-N 1,3,5-trihydroxyanthracene-9,10-dione Chemical compound C1=CC(O)=C2C(=O)C3=CC(O)=CC(O)=C3C(=O)C2=C1 NAOLWIGVYRIGTP-UHFFFAOYSA-N 0.000 description 3
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- 238000002525 ultrasonication Methods 0.000 description 3
- KPGXRSRHYNQIFN-UHFFFAOYSA-N 2-oxoglutaric acid Chemical compound OC(=O)CCC(=O)C(O)=O KPGXRSRHYNQIFN-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- -1 carbon dicarboxylic acids Chemical class 0.000 description 2
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- 238000002474 experimental method Methods 0.000 description 2
- 229930195712 glutamate Natural products 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 description 2
- 230000037353 metabolic pathway Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 229910052700 potassium Inorganic materials 0.000 description 2
- 239000011591 potassium Substances 0.000 description 2
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- 230000002441 reversible effect Effects 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- HWXBTNAVRSUOJR-UHFFFAOYSA-N alpha-hydroxyglutaric acid Natural products OC(=O)C(O)CCC(O)=O HWXBTNAVRSUOJR-UHFFFAOYSA-N 0.000 description 1
- 229940009533 alpha-ketoglutaric acid Drugs 0.000 description 1
- 238000005576 amination reaction Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
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- 230000007613 environmental effect Effects 0.000 description 1
- 230000009123 feedback regulation Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 125000000291 glutamic acid group Chemical group N[C@@H](CCC(O)=O)C(=O)* 0.000 description 1
- 230000003752 improving hair Effects 0.000 description 1
- 238000009655 industrial fermentation Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/14—Glutamic acid; Glutamine
Abstract
The invention belongs to amino acid fermentation technical fields, and disclose a kind of technique for reducing glutamic acid fermentation microbiological contamination and improving fermentation efficiency comprising following steps: fermentation is divided into two stages, wherein it is 3.5-4.0 that the first stage, which controls pH,；It is 5.0 or more that second stage, which controls pH, and inositol and dimethylformamide are added into fermentation medium.Present invention process is able to suppress fermentation miscellaneous bacteria, while improving fermentation efficiency.
Description
Technical field
The invention belongs to amino acid fermentation technical fields, and in particular to a kind of reduction glutamic acid fermentation microbiological contamination and improve hair
The technique of ferment efficiency.
Background technique
Currently, preparing the most common method of glutamic acid is microbe fermentation method.Corynebacterium glutamicum is glutamic acid fermentation
Conventional bacterial strain, belongs to facultative aerobe.The trouble that Corynebacterium glutamicum industrial fermentation processes are commonly encountered is pollution microbes, fermentation dye
The yield and quality that bacterium will lead to product greatly declines, and " tank switching " even occurs when serious, cannot get product.For a long time, such as
It is not only complicated but also difficulty a project that, which prevents fermentation microbiological contamination,.For main miscellaneous bacteria type, by adjusting fermentation parameter with
And the methods of addition specified microorganisms inhibitor is the main means for preventing fermentation miscellaneous bacteria.Corynebacterium glutamicum fermentation produces paddy ammonia
In the process of acid, miscellaneous bacteria is mainly lactic acid bacteria, and how lactic acid bacteria inhibiting is the skill that we need to solve to improve fermentation efficiency
Art problem.
Microorganism is different under different environmental conditions, using different substrate utilization approach, purposefully to cell
Metabolic pathway is modified and is transformed, and changes the original metabolic characteristics of cell, the yield and yield of target product can be improved.When
When four carbon dicarboxylic acids are all supplied by the fixed reaction of CO2, highest theory saccharic acid conversion ratio is 81%；And when the fixed reaction of CO2
It does not work completely, four carbon dicarboxylic acids can only be supplied by glyoxalic acid, and highest theoretical yield is 54%.Glutamic acid production technology
Relative maturity is developed, the key technical indexes is aminoglutaric acid concentration 10%-12%, saccharic acid conversion ratio 55%-60%.But with
External advanced zymotechnique compares, and still has biggish room for promotion.Effectively improve conversion ratio, can save raw material at
Originally, the economic benefit of glutamic acid fermentation is promoted.Metabolic by-product when using Corynebacterium glutamicum progress glutamic acid fermentation is not
More, most important by-product is CO2.Therefore, strengthen the fixed reaction of CO2 in Corynebacterium glutamicum metabolic pathway, and allow thus
Glutamic acid synthesizes crucial enzyme system and effectively, mutually acts synergistically, and will be expected to improve the reclamation rate and saccharic acid conversion ratio of CO2, save
Cost of material increases enterprise profit.
Glutamate producing bacterium membrane structure is special with functionally during the key of glutamic acid fermentation is fermented and cultured
Property variation, be transformed into cell membrane and be conducive to glutamic acid to film exosmosis, that is, complete from the non-accumulation type cell of glutamic acid to paddy ammonia
The transformation of acid accumulation type cell.In this way, since final product glutamic acid is constantly discharged extracellularly, prevent intracellular glutamic acid from
The concentration for causing feedback regulation is run up to, glutamic acid will be synthesized preferentially with keeping in the cell, and constantly be penetrated
Cell membrane is secreted into fermentation medium, to largely be accumulated.The substance for adjusting permeability of cell membrane is more, different
Membrane structure differs greatly between bacterial strain, thus it is not regular follow, select suitable regulator to adjust cell membrane
Permeability is also the technical issues that need to address in glutamic acid fermentation technique.
Summary of the invention
The defects of in order to overcome glutamic acid fermentation microbiological contamination in the prior art and low fermentation efficiency, the invention proposes one
The technique that kind reduces glutamic acid fermentation microbiological contamination and improves fermentation efficiency.
The present invention is achieved by the following technical solution:
A kind of technique for reducing glutamic acid fermentation microbiological contamination and improving fermentation efficiency comprising following steps:
Fermentation is divided into two stages, wherein it is 3.5-4.0 that the first stage, which controls pH,；It is 5.0 or more that second stage, which controls pH,
And inositol and dimethylformamide are added into fermentation medium.
Further, the technique includes the following steps:
Corynebacterium glutamicum seed liquor is inoculated into the fermentor containing fermentation medium with 8% inoculum concentration, fermentation time is
48h；Fermentation time is divided into two stages, and first stage 12h, controlling pH by Feeding ammonia water is 3.5-4.0；Second stage is
36h when second stage starts, inositol and dimethylformamide is added into fermentation medium, while being ultrasonically treated；Second
In stage, pH is controlled 5.0 by auto-feeding ammonium hydroxide, and by flowing the glucose solution for adding concentration for 100g/L for residual sugar control
System is being not less than 1.0%；After the completion of second stage, fermentation liquid is collected.
Preferably, the additive amount of the inositol is 0.6-0.8%.
Preferably, the additive amount of the dimethylformamide is 0.3-0.4%.
Preferably, the parameter of the ultrasonic treatment are as follows: supersonic frequency is 28 kHz, ultrasonic power density is 100W/L, super
The sonication time is 1h.
Preferably, in the fermentation tank culture medium each component mass percent are as follows: glucose 12%, corn pulp 3%, urea
0.5%, potassium dihydrogen phosphate 0.1%, ferrous sulfate heptahydrate 0.02%, epsom salt 0.02%.
Compared with prior art, the beneficial effect that the present invention obtains mainly includes but is not limited to the following aspects:
The study found that the main miscellaneous bacteria in glutamic acid fermentation process is lactic acid bacteria, the optimal pH of lactic acid bacteria is 5.5-6.0, pH drop
When low 4 or less, the proliferation of lactic acid bacteria obviously slows down, and almost stops growing, but Corynebacterium glutamicum influences and little, the present invention
By at fermentation initial stage by improving acidity, without adding antibiotic, by physical method, the proliferation of lactic acid bacteria inhibiting, thus
So that Corynebacterium glutamicum dominant bacteria increases sharply.
Realize the rational proportion of each nutrient, maximum plays the acid producing ability of thallus, to improve fermentation conversion rate and production
Acid；Glutamate-producing strain proliferation is to maximum value, when glutamic acid generates enzyme system and forms complete, opportune moment be added suitable inositol and
Dimethylformamide can both strengthen the fixed reaction of CO2, and weaken glyoxalic acid circulation, guarantee tricarboxylic acid cycle be not disrupted and
α-ketoglutaric acid is continually supplied, by reduction of amination, glutamic acid is largely accumulated, improves fermentation conversion rate, moreover it is possible to
The enhancing for enough promoting permeability of cell membrane is conducive to substrate molecule and is easier to enter cell contact with biological enzyme, is also beneficial to paddy
Propylhomoserin is secreted into extracellular, not only improves conversion ratio, but also increase glutamic acid yield.
Ultrasonic wave biological mechanism of action is mainly mechanical force caused by cavitation and heat effect, which may cause sky
The breakdown of the cell wall and plasma membrane of cell around steeping or reversible membrane permeability change, and this change is reversible, and cell is certainly
Body can repair wall and cell membrane and cell wall penetrability then can be improved in the breakage of film, and not influence the bioactivity of cell,
Thus ultrasound is temporary to the effect of cell membrane, and after ultrasonication terminates, permeability of cell membranes is returned to originally
State；But excessively high-intensitive ultrasonic wave will cause bacterial strain rupture death, therefore the present invention selects 28 kHz of frequency, power close
It spends 100 W/L and is ultrasonically treated 1 h.
The present invention is pioneeringly by CO2 is fixed and cell-permeant linking is to together, in conjunction with low intensive ultrasonication,
Conversion ratio and glutamic acid yield greatly improved.
Specific embodiment
In order to make those skilled in the art better understand the technical solutions in the application, having below in conjunction with the application
The technical solution of the application is clearly and completely described in body embodiment, it is clear that described embodiment is only this Shen
Please a part of the embodiment, instead of all the embodiments.Based on the embodiment in the application, those of ordinary skill in the art are not having
Every other embodiment obtained under the premise of creative work is made, should fall within the scope of the present invention.
Embodiment 1
A kind of technique for reducing glutamic acid fermentation microbiological contamination and improving fermentation efficiency comprising following steps:
Corynebacterium glutamicum (Corynebacterium glutamicum) ATCC13761 is inoculated into seed culture medium,
32 DEG C, 100rpm shaking table culture 12h, obtained Corynebacterium glutamicum seed liquor；The Corynebacterium glutamicum seed culture medium
Group is divided into (being below mass percent): glucose 6%, corn pulp 3%, potassium dihydrogen phosphate 0.2%, epsom salt
0.02%, manganese sulfate monohydrate 0.01%, 115 DEG C of sterilizing 15min；
Seed liquor is inoculated into the fermentor containing fermentation medium with 8% inoculum concentration, fermentation time 48h；Fermentation time
It is divided into two stages, first stage 12h, fermentation temperature is 32 DEG C, ventilatory capacity 0.3vvm, controls pH by Feeding ammonia water and is
3.5；Second stage is 36h, and fermentation temperature is 32 DEG C, ventilatory capacity 0.4vvm；When second stage starts, into fermentation medium
Adding inositol and dimethylformamide, additive amount is respectively 0.6% and 0.3%, is ultrasonically treated simultaneously, supersonic frequency be 28 kHz,
Ultrasonic power density is 100W/L, sonication treatment time 1h；In second stage, pH is controlled 5.0 by auto-feeding ammonium hydroxide,
And residual sugar is controlled for the glucose solution of 100g/L by stream plus concentration and is being not less than 1.0%；After the completion of second stage, collect
Fermentation liquid；
The group of the fermentation tank culture medium is divided into (mass percent): glucose 12%, corn pulp 3%, urea 0.5%, biphosphate
Potassium 0.1%, ferrous sulfate heptahydrate 0.02%, epsom salt 0.02%.
Embodiment 2
A kind of technique for reducing glutamic acid fermentation microbiological contamination and improving fermentation efficiency comprising following steps:
Corynebacterium glutamicum (Corynebacterium glutamicum) ATCC13761 is inoculated into seed culture medium,
32 DEG C, 100rpm shaking table culture for 24 hours, obtained Corynebacterium glutamicum seed liquor；The Corynebacterium glutamicum seed culture medium
Group is divided into (being below mass percent): glucose 6%, corn pulp 3%, potassium dihydrogen phosphate 0.2%, epsom salt
0.02%, manganese sulfate monohydrate 0.01%, 115 DEG C of sterilizing 15min；
Seed liquor is inoculated into the fermentor containing fermentation medium with 10% inoculum concentration, fermentation time 48h；Fermentation time
It is divided into two stages, first stage 12h, fermentation temperature is 32 DEG C, ventilatory capacity 0.4vvm, controls pH by Feeding ammonia water and is
3.8；Second stage is 36h, and fermentation temperature is 32 DEG C, ventilatory capacity 0.5vvm, when second stage starts, into fermentation medium
Adding inositol and dimethylformamide, additive amount is respectively 0.8% and 0.4%, is ultrasonically treated simultaneously, supersonic frequency be 28 kHz,
Ultrasonic power density is 100W/L, sonication treatment time 1h；In second stage, pH is controlled 5.0 by auto-feeding ammonium hydroxide,
And residual sugar is controlled for the glucose solution of 100g/L by stream plus concentration and is being not less than 1.0%；After the completion of second stage, collect
Fermentation liquid；
The group of the fermentation tank culture medium is divided into (mass percent): glucose 12%, corn pulp 3%, urea 0.5%, biphosphate
Potassium 0.1%, ferrous sulfate heptahydrate 0.02%, epsom salt 0.02%.
Embodiment 3
Influence of the pH to miscellaneous bacteria content in fermentation process.
Control group: the pH in entire fermentation process is 5.0, remaining is the same as embodiment 1；
Experimental group: embodiment 1.
Other of experimental group and control group fermentation condition are all the same, after detecting the first stage, Lactic Acid from Fermentation Broth bacterium
Quantity.Concrete outcome is shown in Table 1:
Table 1
| Group | Experimental group | Control group |
| Content of lactic acid bacteria (cfu/ml) | It is not detected | 6579 |
Conclusion: the growth of miscellaneous bacteria can obviously be inhibited by adjusting fermentation pH, to promote the proliferation of dominant bacteria.
Embodiment 4
Influence of each factor to conversion ratio and aminoglutaric acid concentration.
Control group is set, in which:
Control group 1: not adding inositol and dimethylformamide, remaining is the same as embodiment 1；
Control group 2: not adding inositol, remaining is the same as embodiment 1；
Control group 3: not adding dimethylformamide, remaining is the same as embodiment 1；
Control group 4: not using ultrasonic treatment, remaining is the same as embodiment 1；
Experimental group is embodiment 1.
Each group conversion ratio and aminoglutaric acid concentration are shown in Table 2.
Table 2
| Group | Conversion ratio % | Aminoglutaric acid concentration % |
| Control group 1 | 48.36 | 10.45 |
| Control group 2 | 54.21 | 12.03 |
| Control group 3 | 55.89 | 11.78 |
| Control group 4 | 57.73 | 12.85 |
| Experimental group | 62.87 | 14.21 |
Conclusion: setting control group, research inositol, dimethylformamide and three kinds of factors of ultrasonic wave are dense to conversion ratio and glutamic acid
The influence of degree, it is found that three kinds of factors have preferable synergy, can significantly improve conversion ratio and aminoglutaric acid concentration.
Embodiment 5
The present invention also passes through the additive amount of concentration gradient verification experimental verification inositol and dimethylformamide to conversion ratio and glutamic acid
The influence of concentration；It is found by experiment that, with the increase of inositol additive amount, conversion ratio and aminoglutaric acid concentration are stepped up, when reaching
When to 0.6%, aminoglutaric acid concentration reaches highest, continues to improve inositol additive amount, aminoglutaric acid concentration does not obviously increase, conversion ratio
It slightly improves, when additive amount increases to 1.0%, conversion ratio and aminoglutaric acid concentration are declined.As dimethylformamide adds
The increase of dosage, conversion ratio and aminoglutaric acid concentration step up, and when reaching 0.3-0.4%, conversion ratio and aminoglutaric acid concentration are most
Height continues to improve dimethylformamide additive amount, and conversion ratio and aminoglutaric acid concentration are declined.
Analysis of experiments:
The difference of cell physiological metabolic activity and permeability of cell membrane is the major reason for causing transform level different.Wherein,
The present invention is conducive to substrate molecule more so that the permeability of cell membrane of thallus enhances by adding suitable dimethylformamide
It is easily accessible cell to contact with biological enzyme, and then improves conversion ratio.Suitable inositol can strengthen the fixed reaction of CO2, promote
The accumulation of glutamic acid improves fermentation conversion rate.The present invention is fixed by CO2 and cell-permeant linking is arrived together, in conjunction with low intensive
Conversion ratio and glutamic acid yield greatly improved so that the production and transportation of substance combine in ultrasonication.
Listed above is only best specific embodiment of the invention.It is clear that the invention is not restricted to which above embodiments, may be used also
With there are many deformations.All changes that those skilled in the art directly can export or associate from present disclosure
Shape is considered as protection scope of the present invention.
Claims (6)
1. a kind of technique for reducing glutamic acid fermentation microbiological contamination and improving fermentation efficiency comprising following steps:
It is divided into two stages, wherein it is 3.5-4.0 that the first stage, which controls pH,；It is 5.0 or more that second stage, which controls pH, and
Inositol and dimethylformamide are added into fermentation medium.
2. technique according to claim 1, which is characterized in that the technique includes the following steps:
Corynebacterium glutamicum seed liquor is inoculated into the fermentor containing fermentation medium with 8% inoculum concentration, fermentation time is
48h；Fermentation time is divided into two stages, and first stage 12h, controlling pH by Feeding ammonia water is 3.5-4.0；Second stage is
36h when second stage starts, inositol and dimethylformamide is added into fermentation medium, while being ultrasonically treated；Second
In stage, pH is controlled 5.0 by auto-feeding ammonium hydroxide, and is being not less than residual sugar control by stream plus glucose solution
1.0%；After the completion of second stage, fermentation liquid is collected.
3. technique according to claim 2, which is characterized in that the additive amount of the inositol is 0.6-0.8%.
4. technique according to claim 2, which is characterized in that the additive amount of the dimethylformamide is 0.3-0.4%.
5. technique according to claim 2, which is characterized in that the parameter of the ultrasonic treatment are as follows: supersonic frequency is
28kHz, sonication treatment time 1h.
6. technique according to claim 2, which is characterized in that the mass percent of each component in the fermentation tank culture medium
Are as follows: glucose 12%, corn pulp 3%, urea 0.5%, potassium dihydrogen phosphate 0.1%, ferrous sulfate heptahydrate 0.02%, epsom salt
0.02%。
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| CN201811213741.2A CN109371072A (en) | 2018-10-18 | 2018-10-18 | A kind of technique for reducing glutamic acid fermentation microbiological contamination and improving fermentation efficiency |
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Cited By (4)
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| CN109504720A (en) * | 2018-12-23 | 2019-03-22 | 呼伦贝尔东北阜丰生物科技有限公司 | The green production process of glutamic acid |
| CN109943604A (en) * | 2019-04-12 | 2019-06-28 | 卢松 | A method of improving glutamic acid fermentation conversion ratio and recovery rate |
| CN110129385A (en) * | 2019-03-18 | 2019-08-16 | 卢松 | A method of it improving bacterial strain and produces sour efficiency |
| CN112481324A (en) * | 2020-12-30 | 2021-03-12 | 赵兰坤 | Novel amino acid fermentation sterilization process |
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| CN109504720A (en) * | 2018-12-23 | 2019-03-22 | 呼伦贝尔东北阜丰生物科技有限公司 | The green production process of glutamic acid |
| CN110129385A (en) * | 2019-03-18 | 2019-08-16 | 卢松 | A method of it improving bacterial strain and produces sour efficiency |
| CN109943604A (en) * | 2019-04-12 | 2019-06-28 | 卢松 | A method of improving glutamic acid fermentation conversion ratio and recovery rate |
| CN112481324A (en) * | 2020-12-30 | 2021-03-12 | 赵兰坤 | Novel amino acid fermentation sterilization process |
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