CN101705262A - New process for improving fermentation and acid production rate of glutamic acid temperature sensitive mutant by utilizing glycine betaine - Google Patents
New process for improving fermentation and acid production rate of glutamic acid temperature sensitive mutant by utilizing glycine betaine Download PDFInfo
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- CN101705262A CN101705262A CN200910228945A CN200910228945A CN101705262A CN 101705262 A CN101705262 A CN 101705262A CN 200910228945 A CN200910228945 A CN 200910228945A CN 200910228945 A CN200910228945 A CN 200910228945A CN 101705262 A CN101705262 A CN 101705262A
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Abstract
The invention discloses a new process for improving fermentation and acid production rate of glutamic acid temperature sensitive mutant by utilizing glycine betaine, and relates to an improvement method for producing L-glutamic acid by utilizing temperature sensitive mutant fermentation. According to the principle that in the fermentation process of the glutamic acid, glycine betaine has the effects of methyl provision, regulation of osmotic pressure in thallus cells, thallus fat metabolism promotion and protein synthesis and the like, can improve enzyme activity in thallus, promote thallus to grow, realize greater accumulation of the glutamic in fermentation liquor, thus improving the yield, the invention adopts the method by adding glycine betaine or phosphate glycine betaine in the fermentation medium to effectively improve L-glutamic acid fermentation yield.
Description
[technical field]: the present invention relates to a kind of novel process of utilizing trimethyl-glycine to improve L-glutamic acid responsive to temperature type mutant strain fermentation production rate, belong to technical field of producing amino acid by fermentation.
[background technology]: at present, whole world glutamic acid yield is more than 2,000,000 tons.Chinese glutamic acid yield had reached 1,500,000 tons in 2008, and the output height occupies first place in the world.China has become world's L-glutamic acid and glutamate production center.See that technically China is the production technique of raw material with starch, its main economic and technical indices has entered world's rank of advanced units.Because state of the art improves constantly, production cost descends significantly.Yet, the L-glutamic acid manufacturing enterprise of China but is faced with acid test at present: be that market capacity constantly enlarges on the one hand, L-glutamic acid and monosodium glutamate export volume constantly increase, be on the other hand under the present situation of China's food shortage, rise year by year along with raw and auxiliary material, energy prices, corresponding L-glutamic acid production cost also is greatly improved, and causes the L-glutamic acid productive profit on the low side.
The production level of China fermentative Production L-L-glutamic acid and international counterparts industry still has gap at present.Adopt the low-glucose addition technology at present in the world, produce L-glutamic acid with the cane molasses for the raw material forced fermentation, acid production rate reaches 13%～15%, and glucose acid invert ratio reaches 50%～60%; Produce L-glutamic acid with the amylum hydrolysate of the sugar forced fermentation, acid production rate reaches 16%, and glucose acid invert ratio reaches 65%; And China's monosodium glutamate industry at present many be raw material with amylum hydrolysate of the sugar or beet sirup, adopt vitamin H suboptimal dose method fermentative production L-glutamic acid, average acid production rate 10.3%, average glucose acid invert ratio 58.5%.
Adopting the L-glutamic acid temperature sensitive mutant to carry out forced fermentation, is the present main flow of glutamic acid fermentation in the world.Because the mode of the employing vitamin H suboptimal dose method fermentative production L-glutamic acid that its fermentation control mode and China are existing is different fully, when utilizing its fermentative production L-glutamic acid, do not need to control the vitamin H suboptimal dose, only need just can finish glutamate producing bacterium by of the transformation of growth form cell by physics mode (transformation temperature) to product acid type cell, thereby avoided causing the sour unsettled phenomenon of product because of the influence of raw material, fermentation stability and extensive, fermentation period is short, the plant factor height.In addition, because do not have vitamin H suboptimal dose problem in adopting L-glutamic acid temperature sensitive mutant forced fermentation process, vitamin H can be excessive greatly, thereby can strengthen CO2 fixation reaction in the body, improve the transformation efficiency of L-glutamic acid, reach the purpose of high acid, high conversion sugar.
Trimethyl-glycine is the quaternary ammonium type alkaloid of finding in beet sirup, is the cellular metabolism intermediate product, the methyl of providing is provided, regulates osmotic pressure in the cell, promotes effects such as metabolism of fat and protein synthesis.Microorganism cells all needs stable methyl donor, it is generally acknowledged that cell cultures needs methyl donor.
The molecular weight of trimethyl-glycine is little, and 3 active methyls are arranged, and has also obtained neutralization at the intramolecule positive and negative charge, so it is a methyl donor efficiently.Therefore, trimethyl-glycine can be used as the interior Gelucystine of cell to the methyl donor that methionine(Met) changes, and has the confession methyl usefulness of alternative methionine(Met), thereby has the effect of saving methionine(Met), and gross protein value in the cell, rna content and RNA/DNA ratio are significantly raise.
Trimethyl-glycine can prevent the violent change of cell intermediate ion concentration as a kind of important osmotic pressure buffer substance in the cell.When in the cell during osmotic pressure violent change, when high, cell begins to produce or absorbs trimethyl-glycine to keep the balance of normal osmotic pressure, prevents the outflow of cell moisture and the invasion of salinity simultaneously as the external penetration voltage rise.
[summary of the invention]: the present invention adopts the method for adding a certain amount of trimethyl-glycine in substratum to improve the productive rate of L-L-glutamic acid temperature sensitive mutant fermentation production process.This method only need be added the significantly raising that an amount of trimethyl-glycine or phosphoric acid salt trimethyl-glycine just can be realized L-L-glutamic acid acid production rate in initial fermention medium, be suitable for suitability for industrialized production.
The objective of the invention is to be achieved through the following technical solutions:
A kind of method that improves the L-glutamic acid fermentation production rate provided by the invention, it is characterized in that: utilize existing L-glutamic acid temperature sensitive mutant, in existing L-glutamic acid fermentation substratum, add trimethyl-glycine or phosphoric acid salt trimethyl-glycine, make its concentration in substratum be 0.01～10g/L, preferred 0.05～7.5g/L, more preferably 0.1～15g/L.
The present invention is according in L-glutamic acid temperature sensitive mutant fermenting process, trimethyl-glycine can be used as the thalline methyl donor, regulates osmotic pressure in the somatic cells, promotes metabolism of fat and protein synthesis etc. in the cell, have effects such as the thalli growth of stimulation, the acid of promotion product, thereby can improve glutamate producing bacterium L-glutamic acid fermentation production rate by the method for in L-glutamic acid fermentation substratum, adding trimethyl-glycine.
[embodiment]:
The present invention is further illustrated below by embodiment, and the cited case does not limit protection scope of the present invention:
Embodiment 1:
The bacterial strain that adopts is L-glutamic acid temperature sensitive mutants such as Corynebacterium glutamicum, brevibacterium flavum; Substratum is fermention medium [glucose 18%, corn steep liquor 2%, the MgSO that generally adopts existing
47H
2O0.15%, K
2HPO
40.45%, MnSO
40.0003%, FeSO
40.0003%, V
B10.00004%] adds the 0.1g/L trimethyl-glycine in; Cultural method: bacterial classification is inserted seed culture medium [glucose 2.5%, corn steep liquor 3%, K
2HPO
43H
2O 0.3%, MgSO
47H
2O 0.1%, 0.0002%, FeSO
40.0002%, V
B10.00002%, urea 0.055%] in, inoculum size is 10%; 30 ℃, pH be 7.0 and dissolved oxygen be to control automatically in 5L under 20% condition to cultivate 12h in the fermentor tank to logarithmic phase, inoculum size by 10% inserts the 30L that contains fermention medium and controls in the fermentor tank automatically, control initial incubation temperature is 30 ℃, as 20 times of concentration OD of fermented liquid dilution
620nmValue is 0.35 o'clock, in 5 minutes, improve temperature to 37 ℃ cultivation, feed suitable air, regulate the agitation as appropriate rotating speed, adopting the control of oxygen supply pattern stage by stage dissolved oxygen: 0～10h is that 20%, 10～32h is 5%, control pH 7.0～7.2 by auto-feeding ammoniacal liquor, add an amount of bubble enemy froth breaking by stream, and to add concentration by stream be that the glucose solution of 800g/L is controlled at 1.5% with residual sugar, fermenting to 32h stops.When putting jar, the output of L-L-glutamic acid is 195.0g/L, and glucose acid invert ratio is 61.5%.Comparison is according to the facts tested (the L-glutamic acid yield is 180.0g/L, and glucose acid invert ratio is 60.0%) and has been improved 8.3% and 2.5% respectively.
Embodiment 2:
The bacterial strain that adopts is L-glutamic acid temperature sensitive mutants such as Corynebacterium glutamicum, brevibacterium flavum; Substratum is to add the 2.0g/L trimethyl-glycine in the existing fermention medium (with embodiment 1) that generally adopts; Cultural method is with embodiment 1.When putting jar, the output of L-L-glutamic acid is 207.0g/L, and glucose acid invert ratio is 63.6%, and comparison is according to the facts tested (the L-glutamic acid yield is 180.0g/L, and glucose acid invert ratio is 60.0%) and improved 15.0% and 6.0% respectively.
Embodiment 3:
The bacterial strain that adopts is L-glutamic acid temperature sensitive mutants such as Corynebacterium glutamicum, brevibacterium flavum; Substratum is to add the 7.5g/L trimethyl-glycine in the existing fermention medium (with embodiment 1) that generally adopts; Cultural method is with embodiment 1.When putting jar, the output of L-L-glutamic acid is 214.3g/L, and glucose acid invert ratio is 63.9%, and comparison is according to the facts tested (the L-glutamic acid yield is 180.0g/L, and glucose acid invert ratio is 60.0%) and improved 19.1% and 6.5% respectively.
Embodiment 4:
The bacterial strain that adopts is L-glutamic acid temperature sensitive mutants such as Corynebacterium glutamicum, brevibacterium flavum; Substratum is to add the 15g/L trimethyl-glycine in the existing fermention medium (with embodiment 1) that generally adopts; Cultural method is with embodiment 1.When putting jar, the output of L-L-glutamic acid is 205.0g/L, and glucose acid invert ratio is 61.7%, and comparison is according to the facts tested (the L-glutamic acid yield is 180.0g/L, and glucose acid invert ratio is 60.0%) and improved 13.9% and 2.8% respectively.
Claims (3)
1. novel process that improves the L-glutamic acid fermentation production rate, its key step is: adopt the L-glutamic acid temperature sensitive mutant to obtain L-L-glutamic acid through fermentation, it is characterized in that, in existing fermention medium, add trimethyl-glycine or phosphoric acid salt trimethyl-glycine, make its concentration in substratum be 0.01～15g/L.
2. method according to claim 1 is characterized in that: wherein the concentration of said trimethyl-glycine is 0.05～7.5g/L.
3. method according to claim 1 is characterized in that: wherein the concentration of said trimethyl-glycine is 0.1～15g/L.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102660520A (en) * | 2012-05-02 | 2012-09-12 | 潍坊祥维斯化学品有限公司 | Fermentative nutrition aid, application of nutrition aid to preparation of xylanase and application method for nutrition aid |
| CN103243131A (en) * | 2013-05-28 | 2013-08-14 | 山东祥维斯生物科技有限公司 | Method for preparing L-glutamic acid by fermentation |
| CN104357586A (en) * | 2014-11-11 | 2015-02-18 | 河北欣港药业有限公司 | Fermentation production method of rifamycin SV based on phosphate glycine betaine concentration as control parameter |
| CN106701855A (en) * | 2017-03-01 | 2017-05-24 | 中粮生化能源（龙江）有限公司 | Method for fermenting temperature-sensitive strains by phosphoric acid to produce glutamic acid |
| CN112094874A (en) * | 2020-09-16 | 2020-12-18 | 内蒙古格林特制药有限责任公司 | Culture medium for producing griseofulvin through fermentation |
| CN112695061A (en) * | 2020-11-04 | 2021-04-23 | 呼伦贝尔东北阜丰生物科技有限公司 | L-glutamic acid total nutrient fed-batch high-density fermentation method |
| 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 |
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2009
- 2009-12-03 CN CN200910228945A patent/CN101705262A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102660520A (en) * | 2012-05-02 | 2012-09-12 | 潍坊祥维斯化学品有限公司 | Fermentative nutrition aid, application of nutrition aid to preparation of xylanase and application method for nutrition aid |
| CN102660520B (en) * | 2012-05-02 | 2013-05-08 | 潍坊祥维斯化学品有限公司 | Fermentative nutrition aid, application of nutrition aid to preparation of xylanase and application method for nutrition aid |
| CN103243131A (en) * | 2013-05-28 | 2013-08-14 | 山东祥维斯生物科技有限公司 | Method for preparing L-glutamic acid by fermentation |
| CN103243131B (en) * | 2013-05-28 | 2015-06-10 | 山东祥维斯生物科技有限公司 | Method for preparing L-glutamic acid by fermentation |
| CN104357586A (en) * | 2014-11-11 | 2015-02-18 | 河北欣港药业有限公司 | Fermentation production method of rifamycin SV based on phosphate glycine betaine concentration as control parameter |
| CN104357586B (en) * | 2014-11-11 | 2015-11-04 | 河北欣港药业有限公司 | It is the fermentation method for producing of the Rifamycin Sodium of controling parameters based on phosphoric acid betaine concentration |
| CN106701855A (en) * | 2017-03-01 | 2017-05-24 | 中粮生化能源（龙江）有限公司 | Method for fermenting temperature-sensitive strains by phosphoric acid to produce glutamic acid |
| 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 |
| CN112094874A (en) * | 2020-09-16 | 2020-12-18 | 内蒙古格林特制药有限责任公司 | Culture medium for producing griseofulvin through fermentation |
| CN112695061A (en) * | 2020-11-04 | 2021-04-23 | 呼伦贝尔东北阜丰生物科技有限公司 | L-glutamic acid total nutrient fed-batch high-density fermentation method |
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Open date: 20100512 |