CN105274018A - Thermo-sensitive corynebacterium glutamicum, cultivation medium, colloidal particles, preparation method and application - Google Patents
Thermo-sensitive corynebacterium glutamicum, cultivation medium, colloidal particles, preparation method and application Download PDFInfo
- Publication number
- CN105274018A CN105274018A CN201410290140.7A CN201410290140A CN105274018A CN 105274018 A CN105274018 A CN 105274018A CN 201410290140 A CN201410290140 A CN 201410290140A CN 105274018 A CN105274018 A CN 105274018A
- Authority
- CN
- China
- Prior art keywords
- corynebacterium glutamicum
- tct006
- temperature sensitive
- sensitive type
- type corynebacterium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses thermo-sensitive corynebacterium glutamicum, a cultivation medium, colloidal particles, a preparation method and application. The cultivation medium comprises glucose, cane molasses, corn steep liquor, KH<2>PO<4>, MgSO<4> 7H<2>O, FeSO<4>, MnSO<4>, biotin and defoaming agents. A fermentation cultivation method includes inoculating the thermo-sensitive corynebacterium glutamicum TCT006 in the cultivation medium and fermenting and cultivating the thermo-sensitive corynebacterium glutamicum in the cultivation medium. The preparation method for the TCT006 immobilized cell colloidal particles includes (1), sterilizing and melting sodium alginate aqueous solution and then cooling the sodium alginate aqueous solution until the temperature of the sodium alginate aqueous solution reaches the room temperature; (2), mixing fermentation liquor of the thermo-sensitive corynebacterium glutamicum TCT006 and the sodium alginate aqueous solution with each other to obtain mixed liquid; (3), sucking the mixed liquid by the aid of a silicone tube and then squeezing out the mixed liquid; (4), dripping squeezed particles into CaCl<2> aqueous solution and immobilizing the particles. The thermo-sensitive corynebacterium glutamicum, the cultivation medium, the colloidal particles, the preparation method and the application have the advantages that monosodium glutamate can be easily separated and extracted, large-scale continuous production can be facilitated, and the thermo-sensitive corynebacterium glutamicum, the cultivation medium and the colloidal particles are low in production cost.
Description
Technical field
The present invention relates to a kind of temperature sensitive type Corynebacterium glutamicum, substratum, micelle and preparation method and application.Background technology
Glutamate-producing strain membrane structure and specific variations functionally during the key of glutamic acid fermentation is fermentation culture, thus cytolemma is transformed into be conducive to the pattern of L-glutamic acid to film exosmosis.The bacterial strain that current glutamic acid fermentation adopts is generally vitamin H defective type corynebacterium glutamicum strain and responsive to temperature type glutamic acid mutation bacterial strain.Wherein vitamin H defective type corynebacterium glutamicum strain is very high for vitamin H fluctuation susceptibility in fermentation substrate, and the defect that surfactant method fermentation operation is loaded down with trivial details and penicillin method fermentation yield is low.
Relative to the vitamin H defective type corynebacterium glutamicum strain of domestic current use, temperature sensitive mutation corynebacterium glutamicum strain only needs control temperature can realize the production of L-glutamic acid, and the step of its Metabolism control is comparatively easy, has better production application prospect.But research shows: it is comparatively responsive that conventional temperature responsive type corynebacterium glutamicum strain gathers front and back changes in environmental conditions for L-glutamic acid, key point is the nutrient environment influence condition in leavening temperature, fringe time and L-glutamic acid secretion period.In single fermentation system, the operability of the change while of multifactor itself is more difficult, and the uncertain factor simultaneously caused is more
Summary of the invention
Technical problem to be solved by this invention is very high for vitamin H fluctuation susceptibility in fermentation substrate in order to overcome vitamin H defective type corynebacterium glutamicum strain, and surfactant method complex operation and penicillin method yield poorly; And conventional temperature responsive type corynebacterium glutamicum strain to gather front and back changes in environmental conditions for L-glutamic acid comparatively responsive, all be unfavorable for L-glutamic acid continuous prodution, and provide a kind of temperature sensitive type Corynebacterium glutamicum, substratum, fermentation process, micelle and preparation method and application's (preferably consistent with denomination of invention, PLSCONFM) herein.The fermention medium of temperature sensitive type Corynebacterium glutamicum of the present invention and fermentation process thereof can improve the cell density of Corynebacterium glutamicum in the short period of time, thus improve the specific production rate of product, and the preparation method of its micelle adopts immobilization embedded technology, simple to operate, embedding efficiency is higher, in for the continuous prodution of L-glutamic acid, can be good at the fluctuation of elimination growing environment to the impact of L-glutamic acid continuous prodution.
Immobilization technology is one of most effectual way improving biological catalyst utilising efficiency and biological reactions rates, contributes to the production process realizing high efficiency serialization and repetition, is more conducive to the separation and Extraction of Sodium Glutamate.Therefore adopt immobilization to complete the growth of the higher glutamate producing bacterium of the microbe density of growth phase, so put into produce L-glutamic acid environment in, the fluctuation of growing environment can be eliminated on the impact of L-glutamic acid continuous prodution.
The present invention finally solves the problems of the technologies described above by the following technical programs.
The invention provides a kind of temperature sensitive type Corynebacterium glutamicum TCT006.
Present invention also offers the substratum of a kind of described temperature sensitive type Corynebacterium glutamicum TCT006, it comprises the component of following masses: the glucose of 8kg, the cane molasses of 2kg, the corn steep liquor of 5kg, the KH of 12kg
2pO
4, the MgSO of 0.24kg
47H
2the FeSO of O, 0.26g
4, the MnSO of 0.26g
4, the vitamin H of 6mg and the defoamer of 30 ~ 50g, to improve quality with the substratum of 12 ~ 14L, the pH value of described fermention medium is 7.0.
Present invention also offers the fermentation culture method of a kind of described temperature sensitive type Corynebacterium glutamicum TCT006, it comprises the following step: described temperature sensitive type Corynebacterium glutamicum TCT006 is inoculated in fermentation culture in described substratum.Wherein, the inoculum size of described temperature sensitive type Corynebacterium glutamicum TCT006 is preferably 15% ~ 18%.During described fermentation culture, the pH value of fermention medium is preferably 7.0 ~ 7.2.The stirring velocity of described fermentation culture is preferably 180 ~ 240r/min; Temperature is preferably 32 ~ 33 DEG C, and the time is preferably 8 ~ 12h.
Present invention also offers a kind of preparation method of temperature sensitive type Corynebacterium glutamicum TCT006 sodium alginate calcium salt immobilized cell micelle, it comprises the following step:
(1) by the sodium alginate aqueous solution (mass percent) of 2% ~ 3%, sterilizing is cooled to room temperature after melting;
(2) bacteria suspension of the temperature sensitive type Corynebacterium glutamicum TCT006 of 20% ~ 40% is mixed with the volume ratio of 1:9 with the sodium alginate aqueous solution in step (1), fully stir and make uniform mixed solution;
(3) squeezed out by another mouth of pipe after mixed solution being sucked by silicone tube with electronic peristaltic pump;
(4) bead squeezed out is instilled the CaCl of 2% ~ 4%
2in the aqueous solution, immobilization 4 ~ 8h.
Present invention also offers the temperature sensitive type Corynebacterium glutamicum TCT006 micelle that a kind of above-mentioned preparation method obtains.
Immobilization temperature sensitive type Corynebacterium glutamicum TCT006 micelle of the present invention, its embedding rate, preferably more than 95%, continuously ferments more than 6 times.
Present invention also offers a kind of described application of temperature sensitive type Corynebacterium glutamicum TCT006 micelle in L-glutamic acid continuous prodution.
The production method of L-glutamic acid of the present invention, it comprises the following step: by above-mentioned immobilization temperature sensitive type Corynebacterium glutamicum TCT006 micelle, adopts pillar fluidized bed aerosol generator to carry out the continuous reaction of L-glutamic acid.Wherein, described fluidized bed aerosol generator conditional controls: the content of glucose is 12 ~ 18g/L, and temperature is preferably 36 ~ 38 DEG C, and pH value is preferably 7.2 ~ 7.4.
The production method of L-glutamic acid of the present invention, glutamic acid yield >=100g/L.
Without prejudice to the field on the basis of common sense, above-mentioned each optimum condition, can arbitrary combination, obtains the preferred embodiments of the invention.
Agents useful for same of the present invention and raw material are all commercially.
Positive progressive effect of the present invention is:
1, fermentation culture method of the present invention, by grasping the growth curve of responsive to temperature type bacterial strain, adopt high-density thalline fermentation technique, the cell density of Corynebacterium glutamicum is improved in short duration, thus improve the specific production rate of product, reduce volume of culture, strengthening downstream separation is extracted, shorten the L-glutamic acid production cycle.
2, utilize Immobilization in Sodium Alginate technology, method is simple, and embedding efficiency is higher, maintains the stability of the multienzyme system of cell, and avoids the feedback inhibition of L-glutamic acid secretion accumulation, is conducive to the separation and Extraction of later stage Sodium Glutamate; The responsive to temperature type Corynebacterium glutamicum micelle prepared has good ventilation property, and by pillar fluidized bed aerosol generator, be conducive to the diffusional interchange of substrate and product molecule and the transmission of oxygen, be very applicable to responsive to temperature type glutamic acid fermentation mutant strain fermentative production L-glutamic acid.
3, fermentation method for producing of the present invention solve vitamin H defective type corynebacterium glutamicum strain in fermentation substrate vitamin H fluctuation susceptibility very high, surfactant method complex operation and penicillin method yield poorly; And conventional temperature responsive type corynebacterium glutamicum strain to gather front and back changes in environmental conditions for L-glutamic acid comparatively responsive, be all unfavorable for a difficult problem for L-glutamic acid continuous prodution.
4, compared with traditional L-glutamic acid temperature sensitive strain fermentation technology, adopt immobilization technology to obtain cell micelle, then carry out glutamic acid fermentation technique, production efficiency improves 20 ~ 30%; Simultaneously because later stage bio-transformation system is comparatively pure, be more conducive to the separation and Extraction of later stage Sodium Glutamate, be conducive to the large-scale continuous production of L-glutamic acid, and production cost be low.
Embodiment
Mode below by embodiment further illustrates the present invention, but does not therefore limit the present invention among described scope of embodiments.The experimental technique of unreceipted actual conditions in the following example, conventionally and condition, or selects according to catalogue.
Embodiment 1
The fermention medium of temperature sensitive type Corynebacterium glutamicum TCT006, it comprises the component of following masses: the glucose of 8kg, the cane molasses of 2kg, the corn steep liquor of 5kg, the KH of 12kg
2pO
4, the MgSO of 0.24kg
47H
2the FeSO of O, 0.26g
4, the MnSO of 0.26g
4, the vitamin H of 6mg and the defoamer of 30 ~ 50g, to improve quality with the fermention medium of 12L, the pH value of fermention medium is 7.0.
The fermentation process of temperature sensitive type Corynebacterium glutamicum TCT006, it comprises the following step: by the temperature sensitive type Corynebacterium glutamicum TCT006 of 15%, under 32 ~ 33 DEG C of conditions with 180r/min, is inoculated in fermentation culture 8h in above-mentioned fermention medium.During fermentation culture, the pH value of fermention medium controls between 7.0 ~ 7.2.
The preparation method of temperature sensitive type Corynebacterium glutamicum TCT006 micelle, it comprises the following step:
(1) take appropriate sodium alginate, add distilled water physiological saline alive, prepare the sodium alginate aqueous solution (mass ratio) into about 2 ~ 3%, sterilizing is cooled to room temperature after melting;
(2) the temperature sensitive type Corynebacterium glutamicum TCT006 bacteria suspension of 20% ~ 40% is mixed with the volume ratio of 1:9 with the sodium alginate aqueous solution in step (1), fully stir and make uniform mixed solution;
(3) extruded by another mouth of pipe after mixed solution being sucked by silicone tube with electronic peristaltic pump, and instill the CaCl of 2%
2in the aqueous solution, immobilization 4h, forms the even immobilization temperature sensitive type Corynebacterium glutamicum TCT006 micelle that diameter is 2mm, with for subsequent use after sterilized water washing.
The immobilization temperature sensitive type Corynebacterium glutamicum TCT006 micelle that above-mentioned preparation method obtains, its embedding rate, more than 95%, can continuously ferment more than 6 times.
The production method of L-glutamic acid, it comprises the following step: by above-mentioned temperature sensitive type Corynebacterium glutamicum TCT006 micelle, adopts pillar fluidized bed aerosol generator to carry out the continuous reaction of L-glutamic acid.Wherein, in described fluidized bed aerosol generator, the content of glucose is 12 ~ 18g/L, and temperature is 36 DEG C, and pH value is 7.2.The production method of above-mentioned L-glutamic acid, glutamic acid yield >=100g/L.
Claims (7)
1. a temperature sensitive type Corynebacterium glutamicum TCT006.
2. a substratum of temperature sensitive type Corynebacterium glutamicum TCT006 as claimed in claim 1, it is characterized in that, it comprises the component of following masses: the glucose of 8kg, the cane molasses of 2kg, the corn steep liquor of 5kg, the KH of 12kg
2pO
4, the MgSO of 0.24kg
47H
2the FeSO of O, 0.26g
4, the MnSO of 0.26g
4, the vitamin H of 6mg and the defoamer of 30 ~ 50g, to improve quality with the substratum of 12 ~ 14L, the pH value of described fermention medium is 7.0.
3. the method for a temperature sensitive type Corynebacterium glutamicum TCT006 fermentation culture as claimed in claim 1, it is characterized in that comprising the following step: described temperature sensitive type Corynebacterium glutamicum TCT006 is inoculated in fermentation culture in substratum as claimed in claim 2.
4. the method for fermentation culture as claimed in claim 3, it is characterized in that, the inoculum size of described temperature sensitive type Corynebacterium glutamicum TCT006 is 15% ~ 18%; During fermentation culture, the pH value sending out substratum is 7.0 ~ 7.2; The stirring velocity of described fermentation culture is 180 ~ 240r/min; Temperature is 32 ~ 33 DEG C, and the time is 8 ~ 12h.
5. a preparation method for temperature sensitive type Corynebacterium glutamicum TCT006 sodium alginate calcium salt immobilized cell micelle, is characterized in that comprising the following step:
(1) by the sodium alginate aqueous solution of 2% ~ 3%, sterilizing is cooled to room temperature after melting;
(2) bacteria suspension of the temperature sensitive type Corynebacterium glutamicum TCT006 containing 20% ~ 40% is mixed with the volume ratio of 1:9 with the sodium alginate aqueous solution in step (1), fully stir and make uniform mixed solution;
(3) squeezed out by another mouth of pipe after mixed solution being sucked by silicone tube with electronic peristaltic pump;
(4) bead squeezed out is instilled the CaCl of 2% ~ 4%
2in the aqueous solution, immobilization 4 ~ 8h.
6. the temperature sensitive type Corynebacterium glutamicum TCT006 micelle that obtains of a preparation method as claimed in claim 5.
7. one kind the application of temperature sensitive type Corynebacterium glutamicum TCT006 micelle in L-glutamic acid continuous prodution as claimed in claim 6.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410290140.7A CN105274018A (en) | 2014-06-24 | 2014-06-24 | Thermo-sensitive corynebacterium glutamicum, cultivation medium, colloidal particles, preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410290140.7A CN105274018A (en) | 2014-06-24 | 2014-06-24 | Thermo-sensitive corynebacterium glutamicum, cultivation medium, colloidal particles, preparation method and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105274018A true CN105274018A (en) | 2016-01-27 |
Family
ID=55143858
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410290140.7A Pending CN105274018A (en) | 2014-06-24 | 2014-06-24 | Thermo-sensitive corynebacterium glutamicum, cultivation medium, colloidal particles, preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105274018A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108715823A (en) * | 2018-06-15 | 2018-10-30 | 南京中医药大学 | A kind of bacterial strain being used to prepare mangiferin glucosides and its application |
| CN113444655A (en) * | 2020-03-26 | 2021-09-28 | 吉林中粮生化有限公司 | Corynebacterium glutamicum, temperature-sensitive strain with high glutamic acid yield, acquisition method and application of temperature-sensitive strain and glutamic acid fermentation method |
-
2014
- 2014-06-24 CN CN201410290140.7A patent/CN105274018A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108715823A (en) * | 2018-06-15 | 2018-10-30 | 南京中医药大学 | A kind of bacterial strain being used to prepare mangiferin glucosides and its application |
| CN108715823B (en) * | 2018-06-15 | 2021-08-06 | 南京中医药大学 | Strain for preparing mangiferin glycoside and application thereof |
| CN113444655A (en) * | 2020-03-26 | 2021-09-28 | 吉林中粮生化有限公司 | Corynebacterium glutamicum, temperature-sensitive strain with high glutamic acid yield, acquisition method and application of temperature-sensitive strain and glutamic acid fermentation method |
| CN113444655B (en) * | 2020-03-26 | 2023-05-16 | 吉林中粮生化有限公司 | Corynebacterium glutamicum, temperature-sensitive strain with high glutamic acid yield, obtaining method and application thereof, and glutamic acid fermentation method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Panesar et al. | Biotechnological approaches for the value addition of whey | |
| CN101838672B (en) | Method for producing gamma-amino butyric acid by using immobilized lactobacillus plantarum | |
| Häggström et al. | Calcium alginate immobilized cells of Clostridium acetobutylicum for solvent production | |
| CN103224965B (en) | Method for producing pyrroloquinoline quinine through microbial fermentation and fermentation medium used in same | |
| CN103409476B (en) | A kind of fermentation of L isoleucines and isolation and purification method | |
| CN109486876A (en) | A method of threonine is extracted and is purified in fermentation | |
| CN104694612A (en) | Method for highly producing L-tryptophan employing industrial fermentation | |
| CN102643770A (en) | Colibacillus capable of generating succinic acid by utilizing synthetic medium pure anaerobic growth and application thereof | |
| CN103614428B (en) | A kind of method of fermenting and producing L tryptophans | |
| CN104372037A (en) | Method for producing succinic acid by multistage continuous fermentation | |
| CN103627739A (en) | Method for producing L-lactic acid by fermenting bagasse cellulose | |
| CN105219661B (en) | The special strain therefore of synthesis of oligonucleotides galactolipin and method with its synthesis of oligonucleotides galactolipin | |
| CN104830918A (en) | Novel production method of sodium gluconate | |
| CN102146415A (en) | Gene knockout bacterium of gluconobacter oxydans and preparation method thereof | |
| CN105274018A (en) | Thermo-sensitive corynebacterium glutamicum, cultivation medium, colloidal particles, preparation method and application | |
| CN101153294B (en) | Immobilized cell single-tank high-strength continuous fermentation process for succinic acid | |
| CN104212851B (en) | A kind of multistage continuously ferment production L-phenylalanine method | |
| CN103642854A (en) | Method for producing succinic acid from corynebacterium glutamicum through immobilized repeated batch fermentation | |
| CN103243128B (en) | High-yield production method of GABA (gamma amino butyric acid) through mixed fermentation of brevibacterium tianjinese and lactobacillus plantarum | |
| CN104593306B (en) | A kind of E bacterial strain HY 05C high-density cultivation methods | |
| CN103773819B (en) | A kind of lactic acid of producing is coupled simultaneously and prepares the method for GABA | |
| CN101153297A (en) | Novel single-tank hemicontinuous high-strength ferment high optical purity L- lactic acid technique for rhizopus oryzae bacterium ball | |
| CN103205368A (en) | Domestication method for high temperature-resistant ethanol-tolerant aroma-producing yeast | |
| CN105255954A (en) | Fermentation method for producing erythritol | |
| CN104673767A (en) | Method for producing feruloyl esterase |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160127 |