CN104830941A - Microbial conversion method utilizing mixed bacteria to efficiently synthesize L-theanine - Google Patents
Microbial conversion method utilizing mixed bacteria to efficiently synthesize L-theanine Download PDFInfo
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- CN104830941A CN104830941A CN201510211262.7A CN201510211262A CN104830941A CN 104830941 A CN104830941 A CN 104830941A CN 201510211262 A CN201510211262 A CN 201510211262A CN 104830941 A CN104830941 A CN 104830941A
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- theanine
- glutamine
- thiamine
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Abstract
The invention relates to a microbial conversion method utilizing mixed bacteria to efficiently synthesize L-theanine. The method is characterized in that brevibacterium flavum and pseudomonas nitroreducens are adopted together to carry out mixed fermentation, after the fermentation the broth is heated to kill the bacteria, and then ethylamine is added to carry out synthesis and conversion. In the provided method, the glutamine generated during the fermentation process in the broth is directly converted into theanine by glutaminase, the complicated glutamine extraction is saved, thus the yield loss during the glutamine extraction process is also avoided, the production technology is simplified, the production cost is largely reduced, the utilization rate of glutamine is improved, and the production cost is reduced.
Description
Technical field
The invention belongs to technical field of biological fermentation, particularly relate to a kind of method that mixed strains microbe transformation method efficiently synthesizes L-thiamine.
Background technology
Theanine (Theanine) is the characteristic amino acid in tealeaves, is also one of taste compound of tealeaves.It is nineteen fifty by japanese wine family more two youths to reveal tea young sprout from jade and find, and called after theanine (Theanine).In general stem tea, the content of theanine is 1%-2%.From being found to so far, except detecting trace and exist in gill fungus, tea plum, mushroom, oil tea, Camellia etc., not yet find in other plant.The total free aminoacids that in theanine or tea tree, content is the highest, in the young sprout bud-leaf of tea tree, the total free aminoacids of about 70% is theanine.In recent years in the research to tea health-care composition, theanine is the more noticeable compound of one, after its many physiological actions are found, how manufacture, synthesize and extracts theanine and also become the research topic of extensive concern.
Theanine belongs to amides, and its chemistry is called: N-ethyl-γ-L-glutaminate (N-ethyl-γ-L-glutamine), molecular formula is: HOOCCHNH
2cH
2cH
2cONHCH
2cH
3, molecular weight is 174.2.The theanine that nature exists is L-type, and sterling is white, needle-shaped crystals, fusing point 217-218 DEG C (decomposition).Very easily water-soluble, and be insoluble to dehydrated alcohol and ether, and solvability raises with temperature and increases, and has the fragrance of caramel and the fresh refreshing taste of similar monosodium glutamate.Pidolidone and ethamine is generated, ninhydrin reaction purple after 6mol/L hydrochloric acid hydrolysis.
One,
the function of theanine and application
1. the function of theanine
Since theanine self-discovery, widely studied by the main components as tealeaves always.In recent years, some special physiological functions that theanine has constantly are found, thus the another large focus becoming the synthesis of tealeaves functional ingredient and extract, apply, research shows, theanine has many valuable medicinal health effects:
(1) hypotensive activity;
(2) curative effect of antitumor drug is strengthened;
(3) effect such as lax nervous, anxiety;
(4) to cranial nerve cell provide protection;
(5) cerebration is promoted;
(6) premenstrual syndrome effect is improved;
(7) antiobesity action;
(8) antifatigue effect;
(9) immunizing power is improved, defend against computer virus.
2. the application of theanine
The application of 2.1 theanine in field of medicaments
The application of theanine in field of medicaments mainly concentrates on the following aspects:
1) theanine can be used as antitumor drug.Experimentation on animals proves that theanine has retarding action to mouse transferability tumour, can extend its survival time to cases with leukemia mouse.
2) theanine may be used for antihypertensive drugs.Experimentation on animals shows to feed raises high dosage theanine, and systolic pressure, the diastolic pressure of artificial boosting rat, all significantly decrease.
3) theanine can be used in and calm the nerves in downern.In addition theanine is now as the effective constituent in tranquilizer, plays preventive effect to diseases such as Parkinson's disease, senile dementia, afferent nerve dysfunctions.
The application in the food industry of 2.2 theanine
As a kind of emerging foodstuff additive, theanine has excellent processing characteristics and stability, can be widely used in the food such as snack categories, candy and jelly, beverage, chewing gum.Sum up, theanine is mainly used in following field:
1) can be used as the quality improver of tea drink, in production of tea drinks process, add a certain amount of theanine, obviously can improve quality and the local flavor of tea drink.
2) can be used as the additive improving flavour of food products.Research shows, theanine can improve the bitter taste of coffee, cocoa, Radix Ginseng beverage, natsudaidai beverage, beer etc., alleviates astringent taste vinous, therefore, can be used as the modifying agent of these flavour of food products.Further, theanine (as sterilization, pH and heating etc.) under common food-processing condition is more stable, so be of wide application.Current Japan has developed and has added the chocolate of theanine, jelly, pudding, chewing gum, health tea and various refreshment drink.
3) can be used as the additive of functional food.Theanine has α intensity of wave in enhancing brain, makes people produce relaxation and improve the effects such as memory, and passes through human trial.Therefore, can be used as functional ingredient in the future and add in food, develop the functional food alleviating nervous and intelligence development.
Two, the production method of theanine
1. chemical synthesis
Pidolidone dehydrogenation is mainly become pyrrolidone hydroxy acid (PCA) by theanine chemical synthesis at present, then adds pure ethamine (99%, solution-air) and is obtained by reacting theanine.Chemosynthesis means prepare that theanine cost is low, complicated operation, severe reaction conditions.Chemosynthesis manufacture be all D L-type raceme, need to carry out splitting and just can obtain L-type product.
2. tealeaves extraction method
Traditional extracting method is: because the total free aminoacids existed in tealeaves is water-soluble, select hot water extraction.Protein, Polyphenols and the partial pigment in millet paste is removed by plumbic acetate precipitation.Logical H
2s removes superfluous lead, removes trimethyl-xanthine with chloroform extraction.In millet paste, theanine is after ventilation breather salify, crosses and filters other component.Theanine mantoquita dissolves through dilute sulphuric acid, adds H
2s and appropriate Ba (0H)
2suction filtration removes filter residue, concentrated, refining after natural theanine sterling.But this method yield is lower, introduce impurity heavy metal ion Pb
2+, Cu
2+.
3. utilize Callus in Camellia sinensis to cultivate and produce theanine
Callus in Camellia sinensis culture method is using tender tealeaves as explant material, callus is produced with special culture medium induction, optimum culture condition, as illumination, temperature, substratum, adds suitable somatomedin, selects suitable precursor substance can obtain good production effect.It is generally acknowledged that, when the concentration of ethamine in substratum is 25mol/L, the theanine biosynthesizing of tealeaves callus reaches maximum value, in tea callus (dry mass), the content of theanine reaches more than 200mg/g, accounts for about 20% of dry weight.Furuya T etc. are in modified MS medium, add substrate (the elementary amine of 20mM, as 2-cyanogen ethamine, fumaric acid, aniline hydrochloride, aniline hydrochloride, hydrochloric acid phenylethylamine etc.), temperature controls, 25 DEG C, shaking culture 4-5 week in dark, can obtain higher theanine output.The method yields poorly, and separation and purification of products process control difficulty that is complicated and production technique is large etc. limits the industrialization of the method.
4. biological fermentation enzyme process
Along with development and the application of biotechnology, utilize fermentable to produce enzyme, and then utilize the extract of enzyme to be more and more widely used to the production carrying out object.Compared with chemical synthesis, Production by Microorganism Fermentation theanine has diverse characteristic.Due to the high specificity that enzymatic reaction has, so without the need to carrying out ethylamino-to alpha-amino group.But need ATP to be hydrolyzed into ADP to provide energy owing to synthesizing theanine in organism, and the acquisition of ATP to lean on the degraded of glucose to promote with regeneration.Therefore, theanine building-up reactions must with the DeR phase coupling of glucose.The reaction of fermentable synthesis theanine, be actually the L-Glutamine deaminase non-specific responding that (high ethamine concentration and specific pH value) carries out under special condition used in microorganism, and this enzyme is more much bigger than ethamine to the avidity of ammonia under normal conditions.
Have been found that at present can catalysis theanine synthesis enzyme mainly contain 3 kinds: theanine synthetase (Theanine synthetase, EC6.3.1.6), L-Glutamine deaminase (L-Glutaminase, EC 3.5.1.2), gamma glutamyl transpeptidase (γ-Glutamyltranspeptidase, γ-GGT).Wherein, theanine synthetase is the biosynthetic enzyme in tealeaves, but fails by its gene clone out always; Other two kinds of enzymes, from microorganism, more conveniently to utilize.The enzyme being mainly used in fermentative production L-thiamine is at present gamma glutamyl transpeptidase.
Adopt microbe fermentation method directly can obtain L-thiamine, theanine can be produced in a large number, be applicable to suitability for industrialized production.But at present the method for Production by Enzymes theanine mainly with glutamine, ethamine for precursor, produce gamma glutamyl transpeptidase or L-Glutamine deaminase by microorganism culturing, synthesis theanine.
As Chinese patent " coli strain of efficiently converting glutamine to synthesize L-theanine and application thereof " CN200910035096.4 [P] .2010-02-10, this invents the intestinal bacteria of said Efficient Conversion L-glutaminate and ethamine, is intestinal bacteria (Escherichia coli) Nan Shi CCTCC No.M 209166.This bacterial strain high yield theanine synthetase, efficiently converting glutamine and ethamine are L-thiamine and succeed in commercial process; To the conversion of glutamine more than 80%.(Jiangsu, Nanjing Normal University, Changzhou A Ge los biosynthesis Science and Technology Ltd. .).
As Chinese patent " method of biosynthesizing theanine by immobilized gene-engineering strain ", CN200810162598.9 [P] .2009-06-03, provide a kind of immobilized gene-engineering strain, called after: colon bacillus Escherichia coli, at the preserving number at Chinese microorganism strain preservation center be: CGMCC No.2714, gene constructed by gamma glutamyl transpeptidase (γ-GGT) in clone E.coli DH5 α, containing recombinant plasmid pET32a-GGT, plasmid size 7563bp, GGT clip size 1743bp, there is amicillin resistance, isopropylthio-β-D-galactoside abduction delivering gamma glutamyl transpeptidase, can be L-thiamine (Tea Inst., Chinese Academy of Agricultural Sciences .) by glutamine Synthesis.
Above-mentioned this be that the conversion method cost of substrate is higher with glutamine, complex process, step is more, and yield is lower, causes cost higher.
Summary of the invention
The object of the invention is to the defect overcoming above-mentioned prior art, provide one to meet industrial production, technique is simple, and production cost is low, and yield is high, and the method that product purity is high, the measured mixed strains microbe transformation method of matter efficiently synthesizes L-thiamine.
Technical scheme taked for achieving the above object is:
A kind of mixed strains microbe transformation method efficiently synthesizes the method for L-thiamine, it is characterized in that adopting brevibacterium flavum and Pseudomonas nitroreducens to carry out mixed fermentation, after fermentation ends, fermented liquid is heated up and carry out thalline inactivation treatment, then add ethamine and carry out synthesis conversion.
Described mixed fermentation take glucose as substrate, be aided with that substratum based on nitrogenous source and trace element carries out, first by basic medium 115 DEG C ~ 121 DEG C moist heat sterilizations 15 minutes, after basic medium is down to 32 DEG C ~ 34 DEG C, access the strain inclined plane lawn of cultured brevibacterium flavum and Pseudomonas nitroreducens.
Described mixed fermentation control condition is: mixing speed 200rpm ~ 400rpm, and aeration volume is than 1:1 ~ 1:5, and constant temperature culture under 32 DEG C ~ 34 DEG C conditions, by regulating mixing speed and ventilation in fermenting process, makes dissolved oxygen maintain 10%-20%.
Starting sampling after 8 hours in fermentation culture and detect pH, OD, glutamine content and content of glutamic acid in fermented liquid, when detecting that glutamine content no longer increases, and when microscopy thalline is aging, stopping cultivating.
Described thalline inactivation treatment refers to and fermented liquid is warming up to 60 DEG C ~ 80 DEG C, maintains 5 minutes ~ 10 minutes.
In described synthesis conversion process, by glutamine: ethamine=0.3M:1.5M ~ 0.3M:2.0M adds ethamine, then at 30 DEG C ~ 34 DEG C, under mixing speed 200rpm ~ 400rpm condition, maintain 16 ~ 24 hours.
Mixed strains microbe transformation method of the present invention efficiently synthesizes the method for L-thiamine, be by the glutamine that produces in fermenting process in fermented liquid through the direct Synthesis theanine of L-Glutamine deaminase effect, eliminate the extraction step of glutamine complexity, avoid the yield losses caused in leaching process, thus simplify production technique, greatly reduce production cost, improve the utilization ratio of glutamine, reduce production cost.
Embodiment
Embodiment 1
For tank volume 10L fermentor tank, take the present invention by two kinds of microorganism mixed fermentations, conversion of glucose is synthesized the technique of L-thiamine, step is as follows:
1. the preparation of fermention medium
Fermention medium is composed of the following components:
Casein hydrolyzate 10 g/L, yeast extract paste 5 g/L, extractum carnis 5 g/L, glucose 20 g/L, dibasic ammonium citrate 2 g/L, tween 80 1.0 mL/L, KH
2pO
46 g/L, MgSO
47 H
2o 0.58 g/L, MnSO44H
2o 0.15 g/L, FeSO
47H
2o 0.03 g/L, L-sodium 25g/L.
Sterilising conditions is 115 DEG C of moist heat sterilizations 15 minutes.
2. brevibacterium flavum and Pseudomonas nitroreducens mixed fermentation
After prepared by fermention medium, from strain inclined plane in picking one ring lawn access 5L fermention medium, pH value is 6.0, and culture temperature is 32 DEG C, and air flow is 1.0(v/vmin), mixing speed is 220r/min.
3. the end of fermenting and thalline deactivation
Start sampling after 8 hours in fermentation culture and detect pH, OD, glutamine content and content of glutamic acid in fermented liquid, after 38 hours, detect that glutamine content no longer increases, stop fermentation.Glutamine content is 5%.
Fermented liquid is warming up to 60 DEG C, maintains 5 minutes.
4. add ethamine Synthesis L-thiamine
By glutamine: ethamine=0.3M:1.5M adds ethamine, then temperature 30 DEG C, mixing speed 200rpm
Condition under maintain 20 hours.
5. the termination of biotransformation
6. bio-transformation is after 20 hours, and L-thiamine content no longer increases, and biotransformation stops.L-thiamine content is 7%.
Embodiment 2
For tank volume 10L fermentor tank, take the present invention by two kinds of microorganism mixed fermentations, conversion of glucose is synthesized the technique of L-thiamine, step is as follows:
1. the preparation of fermention medium
Fermention medium is composed of the following components:
Casein hydrolyzate 9 g/L, yeast extract paste 5 g/L, extractum carnis 5 g/L, glucose 30 g/L, dibasic ammonium citrate 2 g/L, tween 80 1.0 mL/L, KH
2pO
46 g/L, MgSO
47 H
2o 0.58 g/L, MnSO44H
2o 0.15 g/L, FeSO
47H
2o 0.03 g/L, L-sodium 20g/L.
Sterilising conditions is 120 DEG C of moist heat sterilizations 20 minutes.
2. brevibacterium flavum and Pseudomonas nitroreducens mixed fermentation
After prepared by fermention medium, from strain inclined plane in picking one ring lawn access 5L fermention medium, pH value is 6.2, and culture temperature is 33 DEG C, and air flow is 1.2(v/vmin), mixing speed is 300r/min.
3. the end of fermenting and thalline deactivation
Start sampling after 8 hours in fermentation culture and detect pH, OD, glutamine content and content of glutamic acid in fermented liquid, after 40 hours, detect that glutamine content no longer increases, stop fermentation.Glutamine content is 5.5%.
Fermented liquid is warming up to 65 DEG C, maintains 8 minutes.
4. add ethamine Synthesis L-thiamine
By glutamine: ethamine=0.3M:1.8M adds ethamine, then temperature 32 DEG C, mixing speed 250rpm
Condition under maintain 22 hours.
5. the termination of biotransformation
6. bio-transformation is after 22 hours, and L-thiamine content no longer increases, and biotransformation stops.L-thiamine content is 6.5%.
Embodiment 3
For tank volume 10L fermentor tank, take the present invention by two kinds of microorganism mixed fermentations, conversion of glucose is synthesized the technique of L-thiamine, step is as follows:
1. the preparation of fermention medium
Fermention medium is composed of the following components:
Casein hydrolyzate 12 g/L, yeast extract paste 5 g/L, extractum carnis 5 g/L, glucose 22 g/L, dibasic ammonium citrate 2 g/L, tween 80 1.0 mL/L, KH
2pO
46 g/L, MgSO
47 H
2o 0.58 g/L, MnSO44H
2o 0.15 g/L, FeSO
47H
2o 0.03 g/L, L-sodium 21g/L.
Sterilising conditions is 119 DEG C of moist heat sterilizations 13 minutes
2. brevibacterium flavum and Pseudomonas nitroreducens mixed fermentation
After prepared by fermention medium, from strain inclined plane in picking one ring lawn access 5L fermention medium, pH value is 6.5, and culture temperature is 34 DEG C, and air flow is 2.0(v/vmin), mixing speed is 400r/min.
3. the end of fermenting and thalline deactivation
Start sampling after 8 hours in fermentation culture and detect pH, OD, glutamine content and content of glutamic acid in fermented liquid, after 42 hours, detect that glutamine content no longer increases, stop fermentation.Glutamine content is 6%.
Fermented liquid is warming up to 75 DEG C, maintains 8 minutes.
4. add ethamine Synthesis L-thiamine
By glutamine: ethamine=0.3M:1.9M adds ethamine, then temperature 33 DEG C, mixing speed 300rpm
Condition under maintain 24 hours.
5. the termination of biotransformation
6. bio-transformation is after 24 hours, and L-thiamine content no longer increases, and biotransformation stops.L-thiamine content is 8.5%.
The invention is not restricted to the scheme that above-mentioned embodiment provides.
Claims (6)
1. a mixed strains microbe transformation method efficiently synthesizes the method for L-thiamine, it is characterized in that adopting brevibacterium flavum and Pseudomonas nitroreducens to carry out mixed fermentation, after fermentation ends, fermented liquid is heated up and carry out thalline inactivation treatment, then add ethamine and carry out synthesis conversion.
2. efficiently synthesize the method for L-thiamine according to mixed strains microbe transformation method according to claim 1, it is characterized in that described mixed fermentation is is substrate with glucose, is aided with that substratum based on nitrogenous source and trace element carries out.
3. the method for L-thiamine is efficiently synthesized according to the mixed strains microbe transformation method described in claim 1 or 2, it is characterized in that described mixed fermentation control condition is: mixing speed 200 ~ 400rpm, aeration volume is than 1:1 ~ 1:5, constant temperature culture under 30 DEG C ~ 34 DEG C conditions, in fermenting process, control dissolved oxygen at 10%-20%.
4. the method for L-thiamine is efficiently synthesized according to mixed strains microbe transformation method according to claim 1, it is characterized in that: start sampling after 8 hours in fermentation culture and detect pH, OD, glutamine content and content of glutamic acid in fermented liquid, when detecting that glutamine content no longer increases, and microscopy thalline aging time, stop cultivate.
5. efficiently synthesize the method for L-thiamine according to mixed strains microbe transformation method according to claim 1, it is characterized in that described thalline inactivation treatment refers to that just fermented liquid is warming up to 60 DEG C ~ 80 DEG C, maintain 5 minutes ~ 10 minutes.
6. the method for L-thiamine is efficiently synthesized according to mixed strains microbe transformation method according to claim 1, it is characterized in that in described synthesis conversion process, by glutamine: ethamine=0.3M:1.5M ~ 0.3M:2.0M adds ethamine, then at 30 DEG C ~ 34 DEG C, under mixing speed 200rpm ~ 400rpm condition, maintain 16 ~ 24 hours.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108517292A (en) * | 2018-04-27 | 2018-09-11 | 宜昌市汇宜兴新能源科技有限公司 | A method of theanine is extracted from tealeaves automatically |
| CN109628514A (en) * | 2019-01-10 | 2019-04-16 | 内蒙古拜克生物有限公司 | A kind of method of fermentation method production L-thiamine |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1225946A (en) * | 1998-12-28 | 1999-08-18 | 山东大学 | Glutamine fermentation production process |
| CN102051391A (en) * | 2009-10-27 | 2011-05-11 | 浙江博信药业有限公司 | Fermentation method for producing glutamine |
| CN103667382A (en) * | 2013-12-24 | 2014-03-26 | 山东民强生物科技股份有限公司 | Method for producing L-glutamine by fermentation of microorganisms |
| CN104087535A (en) * | 2014-07-10 | 2014-10-08 | 南通励成生物工程有限公司 | Method for preparing theanine by biological process |
-
2015
- 2015-04-29 CN CN201510211262.7A patent/CN104830941A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1225946A (en) * | 1998-12-28 | 1999-08-18 | 山东大学 | Glutamine fermentation production process |
| CN102051391A (en) * | 2009-10-27 | 2011-05-11 | 浙江博信药业有限公司 | Fermentation method for producing glutamine |
| CN103667382A (en) * | 2013-12-24 | 2014-03-26 | 山东民强生物科技股份有限公司 | Method for producing L-glutamine by fermentation of microorganisms |
| CN104087535A (en) * | 2014-07-10 | 2014-10-08 | 南通励成生物工程有限公司 | Method for preparing theanine by biological process |
Non-Patent Citations (2)
| Title |
|---|
| 王春晖: "茶氨酸生物合成研究", 《中国优秀博硕士学位论文全文数据库（硕士） 工程科技I辑》 * |
| 赵伟睿等: "微生物细胞通透性改善方法与策略", 《中国生物工程杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108517292A (en) * | 2018-04-27 | 2018-09-11 | 宜昌市汇宜兴新能源科技有限公司 | A method of theanine is extracted from tealeaves automatically |
| CN109628514A (en) * | 2019-01-10 | 2019-04-16 | 内蒙古拜克生物有限公司 | A kind of method of fermentation method production L-thiamine |
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Application publication date: 20150812 |