CN109628514A - A kind of method of fermentation method production L-thiamine - Google Patents
A kind of method of fermentation method production L-thiamine Download PDFInfo
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- CN109628514A CN109628514A CN201910023533.4A CN201910023533A CN109628514A CN 109628514 A CN109628514 A CN 109628514A CN 201910023533 A CN201910023533 A CN 201910023533A CN 109628514 A CN109628514 A CN 109628514A
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- 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
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- 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
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
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Abstract
The invention discloses a kind of methods of fermentation method production L-thiamine: brevibacterium flavum access seed culture medium being spread cultivation, seed liquor 1 is obtained；Pseudomonas nitroreducens access seed culture medium is spread cultivation, seed liquor 2 is obtained；Seed liquor 1 and seed liquor 2 are accessed in the fermentation medium containing matcha powder and fermented to obtain fermentation liquid 1；Ethamine is added into fermentation liquid 1, is continued fermented and cultured and is obtained fermentation liquid 2；Ethamine is added into fermentation liquid 2, obtains fermentation liquid 3.The present invention is by combined ferment culture brevibacterium flavum and Pseudomonas nitroreducens, and the glutamine generated to microbial fermentation is directly utilized, and simplifying production technology reduces production cost.The present invention adds ethamine during fermentation in batches, shortens fermentation time, improve the yield of L-thiamine by adding matcha powder in the fermentation medium.
Description
Technical field
The invention belongs to technical field of microbial fermentation, and in particular to a kind of method of fermentation method production L-thiamine.
Background technique
L-thiamine (N- ethyl-gamma-glutamyl amine) is a kind of distinctive amino acid of tealeaves, is not involved in the conjunction of protein
At, existing in a free form, the 40%-70% for accounting for free amino acid total amount in tealeaves is effective taste compound in green tea,
Content directly determines the quality of tealeaves.L-thiamine have blood pressure lowering, it is antitumor, protection nerve, loosen, antidepression, it is antifatigue,
It improves immunity, mitigate the effects of body fat.It may be used as anti-tumor drug, antihypertensive drugs and tranquilizing the mind sedative in field of medicaments
Object can be used as the quality improver of tea beverage in the food industry, improve the additive of flavour of food products and adding for functional food
Add agent.Therefore,
L-thiamine becomes the compound for attracting attention and gazing in tea health-care composition Study.
The manufacturing method of L-thiamine, there are mainly three types of.First, it is the method directly extracted from tealeaves.But due to dry
L-thiamine content in tealeaves only has 1.5% or so, and photosynthesis can make L-thiamine loss a part, cause yield very
It is low.Then, the second method for being suitable for industrialized production has just been invented, i.e., chemical organic synthesis method.But the method may be residual
Toxicant is stayed, and product includes D, L-type theanine isomers, L-type product can just be obtained by being split, and score is made
From purification operations complexity.The third manufacturing method is produced using microbial enzyme method.Majority is generated with microorganism
Enzyme synthesizes theanine using glutamine and ethamine as precursor.The enzyme that L-thiamine synthesis can be catalyzed at present mainly has 3 kinds: tea
Propylhomoserin synzyme, glutaminase, gamma glutamyl transpeptidase, wherein theanine synthetase is the biosynthetic enzyme in tealeaves,
But fail to come out its gene cloning all the time；Other two kinds of enzymes come from microorganism, more convenient to be utilized.
Chinese patent announces CN104561160A and discloses the method that bioanalysis prepares theanine: nitro is restored false unit cell
Bacteria strain is cultivated to obtain bacterium solution, is filtered to obtain thallus to bacterium solution, by obtained thallus be added to L-Glutamine and
Reacted in ethylamine hydrochloride, through decoloration, film filtering and etc. obtain theanine pulvis or theanine crystal.The technique enzyme is anti-
It answers and cultivates ingredient without containing carbon source, nitrogen source etc. in system, reaction solution ingredient is simple for fermentation liquid, can increase substantially
Product purity, while having enzyme source and reaction solution easily separated, post-process that simple, with short production cycle, yield is high, environmental pollution is small and
The advantages that at low cost.But the preparation method needs to add precursor substance L-Glutamine and second simultaneously during reaction
Amine hydrochlorate, this increases production cost to a certain extent.Chinese patent announces CN104830941A and discloses a kind of mixing
The method that microorganism conversion method efficiently synthesizes L-thiamine, this method use brevibacterium flavum and Pseudomonas nitroreducens
Mixed fermentation is carried out, fermentation liquid heats up carry out thallus inactivation treatment after fermentation, ethamine is then added and carries out synthesis conversion.
This method generates glutamine using brevibacterium flavum fermentation, and Pseudomonas nitroreducens fermentation generates catalyzing enzyme, and not
It need to isolate and purify and add ethamine under conditions of glutamine, enzymatic generates L-thiamine.Before glutamine is omitted in this method
The addition of body, and the glutamine directly generated using microbial fermentation, reduce production cost to a certain extent.But
It is this method production cycle length, L-thiamine low output.
So needing the production method that a kind of production cost is low, with short production cycle and L-thiamine yield is high at present.
Summary of the invention
The present invention provides a kind of method of fermentation method production L-thiamine, this method is short by combined ferment culture yellow
Bacillus and Pseudomonas nitroreducens, the glutamine generated to microbial fermentation are directly utilized, and enzymatic produces L- tea
Propylhomoserin, this method is with short production cycle, L-thiamine yield is high.
Illustrate: 1, total inoculum concentration refers to ratio of the sum of the volume for moving into two kinds of seed liquors with nutrient solution volume after inoculation.
The present invention provides a kind of methods of fermentation method production L-thiamine, the described method comprises the following steps:
(1) brevibacterium flavum access seed culture medium is spread cultivation, obtains seed liquor 1；
(2) Pseudomonas nitroreducens access seed culture medium is spread cultivation, obtains seed liquor 2；
(3) seed liquor 1 and seed liquor 2 are accessed in the fermentation medium containing matcha powder and is fermented to obtain fermentation liquid 1；
(4) content for measuring 1 glutamine of fermentation liquid, according to glutamine: ethamine=0.3M:0.05-0.2M ratio
Example adds ethamine into fermentation liquid 1, continues fermented and cultured and obtains fermentation liquid 2；
(5) content for measuring 2 glutamine of fermentation liquid, according to glutamine: ethamine=0.3M:0.3-1.0M ratio
Example, adds ethamine into fermentation liquid 2, obtains fermentation liquid 3.
Seed culture medium in the step (1) includes: glucose 20-40g/L, yeast powder 2-8g/L, ammonium sulfate 2-8g/
L, Dried Corn Steep Liquor Powder 15-25g/L, potassium dihydrogen phosphate 0.5-3g/L, magnesium sulfate 0.8-1.5g/L；PH is 6.9-7.2.
Inoculum concentration is calculated as 0.1g/L with brevibacterium flavum bacterium powder in the step (1).
Seed culture medium in the step (2) includes: glucose 10-20g/L, yeast powder 0.5-1.5g/L, glutamic acid
Sodium 5-15g/L, magnesium sulfate 0.5-1.5g/L, potassium dihydrogen phosphate 0.5-1.5g/L, iron ethylenediaminetetraacetate receive 0.05-0.2g/L,
Defoaming agent 0.1-0.3g/L；PH is 6.9-7.3.
Inoculum concentration is calculated as 0.1g/L with Pseudomonas nitroreducens bacterium powder in the step (2).
The volume ratio that the seed liquor 1 and seed liquor 2 in fermentation medium are inoculated into the step (3) is 1:0.2-0.5,
Total inoculum concentration is 8-12%.
Fermentation medium contains in the step (3): casein hydrolysate 8-15g/L, yeast extract 3-8g/L, beef extract 3-
8g/L, glucose 18-25g/L, ammonium sulfate 0.5-3g/L, polysorbas20 0.5-1.2mL/L, potassium dihydrogen phosphate 2-10g/L, sulfuric acid
Magnesium 0.3-0.7g/L, manganese sulfate 0.1-0.3g/L, ferric sulfate 0.01-0.05g/L, L-sodium 18-25g/L, matcha powder
0.1-1.2g/L；PH is 6.2-6.8.
The matcha powder is commercial product or conventional method product.
Fermentation time is 5-7 hours in the step (3), and cultivation temperature is 32-35 DEG C, ventilatory capacity 1.2-2.4V/V
Min, speed of agitator 280-450r/min.
Fermentation time is 0.5-2 hours in the step (4), and cultivation temperature is 32-35 DEG C, ventilatory capacity 1.2-2.4V/
Vmin, speed of agitator 280-450r/min.
Fermentation time is 13-16 hours in the step (5), and cultivation temperature is 32-35 DEG C, ventilatory capacity 1.2-2.4V/
Vmin, speed of agitator 280-450r/min.
Application of the method in L-thiamine production.
Compared with prior art, the invention has the benefit that
(1) invention improves the yield of L-thiamine by adding matcha powder in the fermentation medium.
(2) present invention shortens fermentation time, improves L- tea ammonia by adding ethamine in batches during fermentation
The yield of acid.
(3) present invention is directly generated using fermentation by the way that brevibacterium flavum and Pseudomonas nitroreducens is used in combination
Glutamine is omitted the purification step of paddy ammonia glutamine, simplifies production technology.
(4) present invention does not carry out sterilization treatment to microorganism, not only simplifies in L-thiamine fermentation production process
Production stage, and the production time of L-thiamine is shortened, improve production efficiency.
Specific embodiment
Unless otherwise defined, all technical and scientific terms used herein have and the technical field of the invention
The normally understood identical meaning of those of ordinary skill.In the examples where no specific technique or condition is specified, according in the art
Conventional technical means carries out.
A kind of method of the fermentation method of embodiment 1 production L-thiamine
(1) brevibacterium flavum access seed culture medium is spread cultivation, obtains seed liquor 1, seed culture medium includes: grape
Sugared 20g/L, yeast powder 8g/L, ammonium sulfate 2g/L, Dried Corn Steep Liquor Powder 25g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 1.5g/L,
PH is 6.9, and inoculum concentration is calculated as 0.1g/L with brevibacterium flavum bacterium powder；
(2) Pseudomonas nitroreducens access seed culture medium is spread cultivation, obtains seed liquor 2, seed culture medium packet
Contain: glucose 20g/L, yeast powder 0.5g/L, sodium glutamate 15g/L, magnesium sulfate 0.5g/L, potassium dihydrogen phosphate 1.5g/L, second two
Amine tetraacethyl iron receives 0.05g/L, defoaming agent 0.3g/L, pH 6.9, and inoculum concentration is calculated as with Pseudomonas nitroreducens bacterium powder
0.1g/L；
(3) seed liquor 1 and seed liquor 2 are accessed in the fermentation medium containing matcha powder and are fermented to obtain fermentation liquid 1,
The volume ratio of the seed liquor 1 and seed liquor 2 that are inoculated into fermentation medium is 1:0.2, and total inoculum concentration is 12%, fermentation medium
Contain: casein hydrolysate 8g/L, yeast extract 8g/L, beef extract 3g/L, glucose 25g/L, ammonium sulfate 0.5g/L, polysorbas20
1.2mL/L, potassium dihydrogen phosphate 2g/L, magnesium sulfate 0.7g/L, manganese sulfate 0.1g/L, ferric sulfate 0.05g/L, L-sodium 18g/
L, matcha powder 0.1g/L, pH 6.2, the matcha powder are the commercially available product of residence good fortune skill brand, and fermentation time is 7 hours, culture temperature
Degree is 32 DEG C, and ventilatory capacity is 2.4 (V/Vmin), speed of agitator 280r/min；
(4) content for measuring 1 glutamine of fermentation liquid, according to glutamine: ethamine=0.3M:0.2M ratio is to hair
Ethamine is added in zymotic fluid 1, is continued fermented and cultured and is obtained fermentation liquid 2, fermentation time is 0.5 hour, and cultivation temperature is 35 DEG C, ventilation
Amount is 1.2 (V/Vmin), speed of agitator 450r/min；
(5) content for measuring 2 glutamine of fermentation liquid, according to glutamine: ethamine=0.3M:0.3M ratio, to
Ethamine is added in fermentation liquid 2, obtains fermentation liquid 3, fermentation time is 16 hours, and cultivation temperature is 32 DEG C, and ventilatory capacity is 2.4 (V/
Vmin), speed of agitator 280r/min.
A kind of method of the fermentation method of embodiment 2 production L-thiamine
(1) brevibacterium flavum access seed culture medium is spread cultivation, obtains seed liquor 1, seed culture medium includes: grape
Sugared 40g/L, yeast powder 2g/L, ammonium sulfate 8g/L, Dried Corn Steep Liquor Powder 15g/L, potassium dihydrogen phosphate 3g/L, magnesium sulfate 0.8g/L, pH
It is 7.2, inoculum concentration is calculated as 0.1g/L with brevibacterium flavum bacterium powder；
(2) Pseudomonas nitroreducens access seed culture medium is spread cultivation, obtains seed liquor 2, seed culture medium packet
Contain: glucose 10g/L, yeast powder 1.5g/L, sodium glutamate 5g/L, magnesium sulfate 1.5g/L, potassium dihydrogen phosphate 0.5g/L, ethylenediamine
Tetraacethyl iron receives 0.2g/L, defoaming agent 0.1g/L, pH 7.3, and inoculum concentration is calculated as 0.1g/L with Pseudomonas nitroreducens bacterium powder；
(3) seed liquor 1 and seed liquor 2 are accessed in the fermentation medium containing matcha powder and are fermented to obtain fermentation liquid 1,
The volume ratio of the seed liquor 1 and seed liquor 2 that are inoculated into fermentation medium is 1:0.5, and total inoculum concentration is 8%, fermentation medium
Contain: casein hydrolysate 15g/L, yeast extract 3g/L, beef extract 8g/L, glucose 18g/L, ammonium sulfate 3g/L, polysorbas20
0.5mL/L, potassium dihydrogen phosphate 10g/L, magnesium sulfate 0.3g/L, manganese sulfate 0.3g/L, ferric sulfate 0.01g/L, L-sodium
25g/L, matcha powder 0.1g/L, pH 6.8, the matcha powder are the commercially available product that Jin Tongyu controls brand, and fermentation time is 5 hours,
Cultivation temperature is 35 DEG C, and ventilatory capacity is 1.2 (V/Vmin), speed of agitator 450r/min；
(4) measure 1 glutamine of fermentation liquid content, according to glutamine: ethamine=0.3M:0.05M ratio to
Ethamine is added in fermentation liquid 1, is continued fermented and cultured and is obtained fermentation liquid 2, fermentation time is 2 hours, and cultivation temperature is 32 DEG C, ventilation
Amount is 2.4 (V/Vmin), speed of agitator 280r/min；
(5) content for measuring 2 glutamine of fermentation liquid, according to glutamine: ethamine=0.3M:1.0M ratio, to
Ethamine is added in fermentation liquid 2, obtains fermentation liquid 3, fermentation time is 13 hours, and cultivation temperature is 35 DEG C, and ventilatory capacity is 1.2 (V/
Vmin), speed of agitator 450r/min.
A kind of method of the fermentation method of embodiment 3 production L-thiamine
(1) brevibacterium flavum access seed culture medium is spread cultivation, obtains seed liquor 1, seed culture medium includes: grape
Sugared 30g/L, yeast powder 6g/L, ammonium sulfate 5g/L, Dried Corn Steep Liquor Powder 22g/L, potassium dihydrogen phosphate 2g/L, magnesium sulfate 1.2g/L, pH
It is 7.1, inoculum concentration is calculated as 0.1g/L with brevibacterium flavum bacterium powder；
(2) Pseudomonas nitroreducens access seed culture medium is spread cultivation, obtains seed liquor 2, seed culture medium packet
Contain: glucose 15g/L, yeast powder 1.1g/L, sodium glutamate 12g/L, magnesium sulfate 0.8g/L, potassium dihydrogen phosphate 1.2g/L, second two
Amine tetraacethyl iron receives 0.1g/L, defoaming agent 0.2g/L, pH 7.1, and inoculum concentration is calculated as 0.1g/ with Pseudomonas nitroreducens bacterium powder
L；
(3) seed liquor 1 and seed liquor 2 are accessed in the fermentation medium containing matcha powder and are fermented to obtain fermentation liquid 1,
The volume ratio of the seed liquor 1 and seed liquor 2 that are inoculated into fermentation medium is 1:0.3, and total inoculum concentration is 10%, fermentation medium
Contain: casein hydrolysate 12g/L, yeast extract 7g/L, beef extract 6g/L, glucose 22g/L, ammonium sulfate 2g/L, polysorbas20
0.8mL/L, potassium dihydrogen phosphate 6g/L, magnesium sulfate 0.6g/L, manganese sulfate 0.2g/L, ferric sulfate 0.03g/L, L-sodium 22g/
L, matcha powder 0.8g/L, pH 6.5, the matcha powder are the commercially available product of residence good fortune skill brand, and fermentation time is 6 hours, culture temperature
Degree is 33 DEG C, and ventilatory capacity is 2.0 (V/Vmin), speed of agitator 300r/min；
(4) measure 1 glutamine of fermentation liquid content, according to glutamine: ethamine=0.3M:0.15M ratio to
Ethamine is added in fermentation liquid 1, continues fermented and cultured and obtains fermentation liquid 2, and fermentation time is 1.1 hours, and cultivation temperature is 33 DEG C, is led to
Tolerance is 1.8 (V/Vmin), speed of agitator 330r/min；
(5) content for measuring 2 glutamine of fermentation liquid, according to glutamine: ethamine=0.3M:0.6M ratio, to
Ethamine is added in fermentation liquid 2, obtains fermentation liquid 3, fermentation time is 15 hours, and cultivation temperature is 34 DEG C, and ventilatory capacity is 2.1 (V/
Vmin), speed of agitator 300r/min.
A kind of method of the fermentation method of comparative example 1 production L-thiamine
Difference with embodiment 3 is only that the addition that matcha powder is omitted in the fermentation medium of step (3).
A kind of method of the fermentation method of comparative example 2 production L-thiamine
(1) brevibacterium flavum access seed culture medium is spread cultivation, obtains seed liquor 1, seed culture medium includes: grape
Sugared 30g/L, yeast powder 6g/L, ammonium sulfate 5g/L, Dried Corn Steep Liquor Powder 22g/L, potassium dihydrogen phosphate 2g/L, magnesium sulfate 1.2g/L, pH
It is 7.1, inoculum concentration is calculated as 0.1g/L with brevibacterium flavum bacterium powder；
(2) Pseudomonas nitroreducens access seed culture medium is spread cultivation, obtains seed liquor 2, seed culture medium packet
Contain: glucose 15g/L, yeast powder 1.1g/L, sodium glutamate 12g/L, magnesium sulfate 0.8g/L, potassium dihydrogen phosphate 1.2g/L, second two
Amine tetraacethyl iron receives 0.1g/L, defoaming agent 0.2g/L, pH 7.1, and inoculum concentration is calculated as 0.1g/ with Pseudomonas nitroreducens bacterium powder
L；
(3) seed liquor 1 and seed liquor 2 are accessed in the fermentation medium containing matcha powder and are fermented to obtain fermentation liquid 1,
The volume ratio of the seed liquor 1 and seed liquor 2 that are inoculated into fermentation medium is 1:0.3, and total inoculum concentration is 10%, fermentation medium
Contain: casein hydrolysate 12g/L, yeast extract 7g/L, beef extract 6g/L, glucose 22g/L, ammonium sulfate 2g/L, polysorbas20
0.8mL/L, potassium dihydrogen phosphate 6g/L, magnesium sulfate 0.6g/L, manganese sulfate 0.2g/L, ferric sulfate 0.03g/L, L-sodium 22g/
L, matcha powder 0.8g/L, pH 6.5, the matcha powder are the commercially available product of residence good fortune skill brand, and fermentation time is 7.1 hours, culture
Temperature is 33 DEG C, and ventilatory capacity is 2.0 (V/Vmin), speed of agitator 300r/min；
(4) content for measuring 1 glutamine of fermentation liquid, according to glutamine: ethamine=0.3M:0.7M ratio is to hair
Ethamine is added in zymotic fluid 1, fermentation time is 15 hours, and cultivation temperature is 34 DEG C, and ventilatory capacity is 2.1 (V/Vmin), and stirring turns
Speed is 300r/min.
A kind of method of the fermentation method of comparative example 4 production L-thiamine
Chinese patent announces experimental method shown in embodiment 1 in CN104830941 A.
Mass percentage concentration of the high-efficient liquid phase analysis detection L-thiamine in final fermentation liquid is carried out using C18 column.
Table 1
| Experiment numbers | L-thiamine mass percentage concentration/% |
| Embodiment 1 | 13 |
| Embodiment 2 | 14 |
| Embodiment 3 | 16 |
| Comparative example 1 | 9 |
| Comparative example 2 | 10 |
| Comparative example 3 | 7.1 |
Experimental data in table 1 is shown, is produced L-thiamine using method of the invention, can be obtained much higher than existing skill
The yield of art.The experimental data of embodiment 3 and comparative example 1 in further comparison sheet 1, it can be found that matcha powder adds in the present invention
Add, the yield of L-thiamine can be obviously improved.The experimental data of embodiment 3 and comparative example 2 then shows, in the present invention by
Substep addition ethamine, is able to ascend the yield of L-thiamine during fermented and cultured.
Claims (10)
1. a kind of method of fermentation method production L-thiamine, which comprises the following steps:
(1) brevibacterium flavum access seed culture medium is spread cultivation, obtains seed liquor 1；
(2) Pseudomonas nitroreducens access seed culture medium is spread cultivation, obtains seed liquor 2；
(3) seed liquor 1 and seed liquor 2 are accessed in the fermentation medium containing matcha powder and is fermented to obtain fermentation liquid 1；
(4) measure 1 glutamine of fermentation liquid content, according to glutamine: ethamine=0.3M:0.05-0.2M ratio to
Ethamine is added in fermentation liquid 1, is continued fermented and cultured and is obtained fermentation liquid 2；
(5) content for measuring 2 glutamine of fermentation liquid, according to glutamine: ethamine=0.3M:0.3-1.0M ratio, to
Ethamine is added in fermentation liquid 2, obtains fermentation liquid 3.
2. according to the method described in claim 1, it is characterized by: the seed culture medium in the step (1) includes: glucose
20-40g/L, yeast powder 2-8g/L, ammonium sulfate 2-8g/L, Dried Corn Steep Liquor Powder 15-25g/L, potassium dihydrogen phosphate 0.5-3g/L, sulfuric acid
Magnesium 0.8-1.5g/L；PH is 6.9-7.2.
3. according to the method described in claim 1, it is characterized by: inoculum concentration is in the step (1) with brevibacterium flavum bacterium powder
It is calculated as 0.1g/L.
4. according to the method described in claim 1, it is characterized by: the seed culture medium in the step (2) includes: glucose
10-20g/L, yeast powder 0.5-1.5g/L, sodium glutamate 5-15g/L, magnesium sulfate 0.5-1.5g/L, potassium dihydrogen phosphate 0.5-
1.5g/L, iron ethylenediaminetetraacetate receive 0.05-0.2g/L, defoaming agent 0.1-0.3g/L；PH is 6.9-7.3.
5. according to the method described in claim 1, it is characterized by: being inoculated into the kind in fermentation medium in the step (3)
The volume ratio of sub- liquid 1 and seed liquor 2 is 1:0.2-0.5, and total inoculum concentration is 8-12%.
6. according to the method described in claim 1, it is characterized by: fermentation medium contains in the step (3): casein hydrolysis
Object 8-15g/L, yeast extract 3-8g/L, beef extract 3-8g/L, glucose 18-25g/L, ammonium sulfate 0.5-3g/L, polysorbas20 0.5-
1.2mL/L, potassium dihydrogen phosphate 2-10g/L, magnesium sulfate 0.3-0.7g/L, manganese sulfate 0.1-0.3g/L, ferric sulfate 0.01-0.05g/
L, L-sodium 18-25g/L, matcha powder 0.1-1.2g/L；PH is 6.2-6.8.
7. according to the method described in claim 1, it is characterized by: in the step (3) fermentation time be 5-7 hour, cultivate
Temperature is 32-35 DEG C, ventilatory capacity 1.2-2.4V/Vmin, speed of agitator 280-450r/min.
8. according to the method described in claim 1, it is characterized by: in the step (4) fermentation time be 0.5-2 hour, train
Supporting temperature is 32-35 DEG C, ventilatory capacity 1.2-2.4V/Vmin, speed of agitator 280-450r/min.
9. according to the method described in claim 1, it is characterized by: in the step (5) fermentation time be 13-16 hour, train
Supporting temperature is 32-35 DEG C, ventilatory capacity 1.2-2.4V/Vmin, speed of agitator 280-450r/min.
10. application of the method according to claim 11 in L-thiamine production.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104830941A (en) * | 2015-04-29 | 2015-08-12 | 宁夏诚志万胜生物工程有限公司 | Microbial conversion method utilizing mixed bacteria to efficiently synthesize L-theanine |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104830941A (en) * | 2015-04-29 | 2015-08-12 | 宁夏诚志万胜生物工程有限公司 | Microbial conversion method utilizing mixed bacteria to efficiently synthesize L-theanine |
Non-Patent Citations (1)
| Title |
|---|
| 华萍 等: ""不同培养条件对悬浮培养茶叶细胞生长及茶氨酸合成的影响"", 《云南植物研究》 * |
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Application publication date: 20190416 |