CN101235401A - Fermentation method for preparing L-amino acid - Google Patents

Fermentation method for preparing L-amino acid Download PDF

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CN101235401A
CN101235401A CNA2007100370901A CN200710037090A CN101235401A CN 101235401 A CN101235401 A CN 101235401A CN A2007100370901 A CNA2007100370901 A CN A2007100370901A CN 200710037090 A CN200710037090 A CN 200710037090A CN 101235401 A CN101235401 A CN 101235401A
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brevibacterium flavum
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corynebacterium glutamicum
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CN101235401B (en
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雷耀辉
吴品芳
马兴群
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Shandong Xiang Weiss Medical Technology Co., Ltd.
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SHANGHAI XIANGWEISI CHEMICALS CO Ltd
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Abstract

The invention relates to a method for preparing L-amino acid through fermenting. The method of the invention uses fermentation additive betaine phosphate. With the method of the invention, the acid yield of the L-amino acid can be prominently increased, the cost of manufacture is lowered, and the method is suitable for large-scale industrial production.

Description

The method of fermentation preparation of L-aminoacid
Technical field
The present invention relates to technical field of bioengineering.Be specifically related to a kind of method of fermentation preparation of L-aminoacid.
Background technology
L-amino acid is widely used in fodder industry, protective foods and medicine industry.After Japan consonance fermentation company in 1956 was with fermentative Production L-glutamic acid, amino acid whose fermentative production development was very fast, and up to the present, most amino acid can be used fermentation method and Production by Enzymes.
In recent years, along with expanding economy, the progressively raising of living standards of the people, people award increasing concern to healthy aspect, and the amino acid whose demand of L-is also rising year by year.But the amino acid whose fermentation and acid level of most of L-is lower, and production cost is higher, has suppressed the amino acid whose application of L-.Therefore, improve the amino acid whose acid production rate of L-and have very important realistic meaning.
Summary of the invention
Technical problem to be solved by this invention is to overcome above-mentioned weak point, the method for the fermentation preparation of L-aminoacid that a kind of production cost of research and design is lower.
The invention provides a kind of method of fermentation preparation of L-aminoacid.
To achieve the object of the present invention, the present invention has adopted following technical proposal:
The present invention has used the fermentation assistant betaine phosphate.Betaine phosphate has following array structure and performance:
Figure A20071003709000061
Molecular formula: C 5H 14NO 6P
Molecular weight: 215.14
Physicals: white is little yellow crystalline powder extremely, odorlessness, and sour-puckery flavor, the tool moisture absorption, very easily water-soluble, be dissolved in ethanol.
The use of fermentation assistant betaine phosphate can obviously improve the amino acid whose yield of L-, reduces production costs.
Fermentation preparation of L-aminoacid method provided by the invention specifically comprises the following steps:
(1) seed culture:
Seed culture medium g/dl: glucose 3, corn steep liquor 3, beans are dense 2, K 2HPO 40.15, MgSO 47H 2O 0.04, urea 0.2, pH 7.0-7.2; A certain amount of seed culture medium is put into seeding tank, sterilized 15 minutes down at 115 ℃.Brevibacterium flavum ATCC14067 was inserted in cooling back, in 32 ℃ of following aerated culture 9 hours;
(2) fermentation culture:
Fermention medium g/dl: glucose 14, molasses 0.1, Na 2HPO 40-0.1 KCl 0.12, MgSO 47H 2O0.08, FeSO 44H 2O 0.0002, MnSO 40.0002, vitamins B 120mu g/l;
Add the betaine phosphate of 0.05-0.3g/dl in above-mentioned fermention medium, adjusting pH is 7.0-7.2, and this substratum is put in the fermentor tank, and is standby in 15 minutes sterilization backs of 115 ℃ of heating;
The seed that step (1) is obtained is connected in the fermentor tank, and 34 ℃ of aerobic fermentations are cultivated; In culturing process, adopt ammoniacal liquor to regulate pH and maintain 7.0-7.2, simultaneously, continue to add 70% D/W, make sugared concentration be controlled at 1-2g/dl; Before fermentation ends 2 hours stop to continue to add glucose.
In the time of 0,3,8,14 and 18 hours, add the betaine phosphate of 0.15g/dl in fermentation culture respectively, carry out fermentation culture.Cultivate after 1-7 days, stop to add glucose, continue fermentation again after 2 hours, fermentation finishes.
The amino acid whose fermentation condition of L-adopts the conventional production methods of specific substrates among the present invention, and according to the difference of substrate, condition also can change.Leavening temperature is 20-40 ℃ scope, especially 28-40 ℃ scope.The pH value is controlled at neutral 7-7.2.The concentration of fermentation assistant betaine phosphate in fermented liquid is 0.01%-5.0%, and preferable range is 0.05%-1.0%.Fermentation needs to stir or vibration under aerobic conditions.The fermentation time scope was at 1-7 days.For the fermentation amino acid whose separation and purification in back, can adopt conventional ion-exchange and other methods.
The L-amino acid that the inventive method is suitable for comprises: L-L-glutamic acid, L-Methionin, L-arginine, L-Isoleucine, L-Xie Ansuan, the L-Threonine, L-Histidine, L-phenylalanine, L-tryptophane, L-Serine, L-ornithine, L-citrulline, L-tyrosine and L-leucine.
The carbon source that the inventive method is suitable for comprises: carbohydrate such as glucose, sucrose, fructose, maltose, molasses, raw sugar, the starch fluid of saccharification; Fatty acid such as acetate, propionic acid, phenylformic acid; Organic acid such as pyruvic acid, citric acid, succsinic acid, fumaric acid, oxysuccinic acid; Alcohols such as ethanol, butanols.
The nitrogenous source that the inventive method is suitable for comprises: organic nitrogen source such as beans are dense, corn steep liquor, urea, yeast powder, soya-bean cake hydrolyzed solution; Inorganic nitrogen-sourced as ammonium sulfate, ammonium chloride, ammonium nitrate, ammonium acetate, liquefied ammonia, ammoniacal liquor.
The inventive method fermention medium can use following various inorganic salt: phosphoric acid salt, magnesium salts, calcium salt, sylvite, sodium salt, molysite, manganese salt and other trace metal salts.
The inventive method fermention medium can use following various VITAMIN: vitamin H, VitB1, niacinamide etc.
The inventive method is applicable to following various corresponding amino acid whose zymophyte (brevibacterium sp, Corynebacterium, genus bacillus or Colibacter etc.): L-glutamic acid fermentation bacterium such as bifidus bacillus ATCC 14020, brevibacterium flavum ATCC14067, Brevibacterium lactofermentus ATCC 13869, Brevibacterium lactofermentus FERM-P 5012, Corynebacterium acctoacidophlum ATCC13870, Corynebacterium glutamicum ATCC13032 etc.; L-fermenting lysine bacterium such as brevibacterium flavum ATCC14067, brevibacterium flavum FERM-P 1708, Corynebacterium glutamicum FERM-P 1709, Brevibacterium lactofermentus FERM-P 1712, Brevibacterium lactofermentus FERM-P 1711, brevibacterium flavum ATCC 21475, Corynebacterium glutamicum ATCC 12416, Brevibacterium lactofermentus ATCC 1470, vinegar bran acid corynebacteria A TCC 11472 etc.; L-glutaminate zymophyte such as brevibacterium flavum FERM-P 4272, Corynebacterium acctoacidophlum ATCC 13870, brevibacterium flavum FERM-BP 662, vinegar bran acid corynebacteria A TCC 13870, Corynebacterium glutamicum FERM-BP 663 etc.; L-arginine zymophyte such as brevibacterium flavum FERM-P 4948, Corynebacterium glutamicum FERM-P 7274, brevibacterium flavum NRRL 12235, vinegar bran acid coryneform bacteria NRRL 12239 etc.; L-phenylalanine zymophyte such as Brevibacterium lactofermentus FERM-P 5417, brevibacterium flavum FERM-P 1916, Corynebacterium glutamicum ATCC 21670, citric acid Arthrobacter ATCC 21422, Brevibacterium lactofermentus ATCC 21420, Corynebacterium acctoacidophlum ATCC21421, brevibacterium flavum NRRL 12269, Corynebacterium glutamicum ATCC 21670; L-Threonine zymophyte such as colon bacillus FERM-P 4900, Corynebacterium glutamicum FERM-P 5835, brevibacterium flavum FERM-P4164, Brevibacterium lactofermentus FERM-P 4180, brevibacterium flavum ATCC 21269, Corynebacterium acctoacidophlum ATCC 21270 etc.; L-isoleucine fermentation bacterium such as brevibacterium flavum FERM-P 3672, Brevibacterium lactofermentus FERM-P 4192, Corynebacterium glutamicum FERM-P 756, Corynebacterium glutamicum FERM-P 757, Corynebacterium glutamicum FERM-P 758, brevibacterium flavum FERM-BP 759, brevibacterium flavum FERM-BP 760 etc.; L-Histidine zymophyte such as brevibacterium flavum FERM-P 2317, Brevibacterium lactofermentus FERM-P 1565, Corynebacterium glutamicum ATCC 14297, Brevibacterium lactofermentus ATCC 21086, Corynebacterium acctoacidophlum ATCC 21407, brevibacterium flavum ATCC 21406, subtilis FERM-BP 217, citric acid Arthrobacter ATCC 21412 etc.; L-Xie Ansuan zymophyte such as Brevibacterium lactofermentus FERM-P 1845, brevibacterium flavum FERM-P 512, Brevibacterium lactofermentus FERM-P 1968, intestinal bacteria NRRL 12285 etc.; L-Serine zymophyte such as Brevibacterium lactofermentus FERM-P 1371; L-ornithine zymophyte such as Brevibacterium lactofermentus FERM-P 5936, Corynebacterium glutamicum FERM-P 5644, citric acid Arthrobacter ATCC 21040, Corynebacterium glutamicum ATCC 13232 etc.; L-citrulline zymophyte such as Corynebacterium glutamicum FERM-P 5643, brevibacterium flavum FERM-P 1645 etc.; L-tyrosine zymophyte such as Corynebacterium glutamicum FERM-P 5836, brevibacterium flavum FERM-P 7914, subtilis FERM-P6758 etc.; L-tryptophane zymophyte such as subtilis FERM-P 5286, Brevibacterium lactofermentus FERM-P7127, brevibacterium flavum FERM-P 2551, Corynebacterium glutamicum FERM-P 7128, brevibacterium flavum ATCC21427, brevibacterium flavum FERM-BP 114, subtilis FERM-BP 208, brevibacterium flavum FERM-BP 475, Corynebacterium glutamicum, subtilis FERM-BP 200 etc.; L-leucine zymophyte such as Brevibacterium lactofermentus FERM-P 1769, brevibacterium flavum FERM-P 420, Corynebacterium glutamicum FERM-P1966, brevibacterium flavum FERM-BP 755, Corynebacterium glutamicum FERM-BP 756, Corynebacterium glutamicum FERM-P 757, brevibacterium flavum FERM-BP 759 etc.
The inventive method can obviously improve the amino acid whose acid production rate of L-, reduces production costs, and helps the suitability for industrialized production of scale.
Embodiment
Embodiment:
Embodiment 1: seed culture medium (g/dl): glucose 3, and corn steep liquor 3, beans are dense 2, K 2HPO 40.15, MgSO 47H 2O 0.04, urea 0.2, and pH transfers to 7.0-7.2, under 115 ℃, heats and sterilizes in 15 minutes.
Brevibacterium flavum ATCC 14067 is inoculated in the 5L seeding tank, under 32 ℃, shaking culture 9h.
In addition, glutamic acid fermentation substratum (g/dl) is: glucose 14, molasses 0.1, Na 2HPO 40～0.1, KCl 0.12, MgSO 47H 2O 0.08, FeSO 44H 2O 0.0002, MnSO 40.0002, vitamins B 120mu g/l.Add the betaine phosphate of 0.05～0.3g/dl in the substratum respectively, adjust pH to 7.0.The 3L substratum is put into 5 liters fermentor tank, under 115 ℃, heat and sterilized in 15 minutes.
In each fermentor tank, insert the seed that 300ml obtains by above-mentioned cultivation, aerated culture, temperature is 34 ℃.In culturing process, adopt ammoniacal liquor to regulate the pH of substratum, make it maintain 7.2, simultaneously, continue to add 70% D/W, make sugared concentration be controlled at 1-2g/dl (pressing glucose calculates).After the fermentation culture 30 hours, stop to add glucose, after 2 hours, fermentation finishes.Measure the L-glutamic acid content that accumulates in the substratum, the results are shown in Table 1.
Table 1.
The addition of betaine phosphate (g/dl) The amount of L-L-glutamic acid (g/dl)
0 12.4
0.05 12.5
0.1 12.3
0.15 13.2
0.2 12.4
0.25 12.6
0.3 11.4
Embodiment 2: seed culture medium (g/dl): glucose 3, and corn steep liquor 3, beans are dense 2, K 2HPO 40.15, MgSO 47H 2O 0.04, urea 0.2, and pH transfers to 7.0-7.2, under 115 ℃, heats and sterilizes in 15 minutes.
Brevibacterium flavum ATCC 14067 is inoculated in the 5L seeding tank, under 32 ℃, shaking culture 9h.
In addition, L-glutamic acid fermentation substratum (g/dl) is: glucose 14, molasses 0.1, Na 2HPO 40～0.1, KCl 0.10, MgSO 47H 2O 0.08, FeSO 44H 2O 0.0002, MnSO 40.0002, vitamins B 120mu g/l adjusts pH to 7.0.The 3L substratum is put into 5 liters fermentor tank, under 115 ℃, heat and sterilized in 15 minutes.
In each fermentor tank, insert the seed that 300ml obtains by above-mentioned cultivation, aerated culture, temperature is 34 ℃.In culturing process, adopt ammoniacal liquor to regulate the pH of substratum, make it maintain 7.2, simultaneously, continue to add 70% D/W, make sugared concentration be controlled at 1-2g/dl (pressing glucose calculates).In fermentation culture 0,3,8,14,18 hours, the betaine phosphate of adding 0.15g/dl was cultivated after 30 hours, stopped to add glucose respectively, and after 2 hours, fermentation finishes.Measure the L-glutamic acid content that accumulates in the substratum, the results are shown in Table 2.
Table 2.
The interpolation time (h) of betaine phosphate The amount of L-L-glutamic acid (g/dl)
0 13.6
3 12.5
8 12.3
14 12.5
18 12.2
Embodiment 3: seed culture medium (g/dl): glucose 2.5, (NH4) 2SO 40.5, KH 2PO 40.1, MgSO 47H 2O 0.05, corn steep liquor 3.5-4.0, CaCO 31.0 pH transfers to 7.0-7.2,0.1Mpa pressure was sterilized 20 minutes down.
Brevibacterium flavum ATCC 14067 is inoculated in the 5L seeding tank, under 31 ℃, shaking culture 12h.
The fermention medium of L-Methionin (g/dl) is: glucose 18, (NH4) 2SO 450, KH 2PO 41, KCl 0.12, MgSO 47H 2O 0.5, corn steep liquor 0.5-1.5, and L-Threonine 0.4, CaCO3 4.0, add the betaine phosphate of 0.05-0.3g/dl in the substratum respectively, adjust pH to 7.0-7.2.The 3.6L substratum is put into 7 liters fermentor tank, and 0.07Mpa pressure was sterilized 8 minutes down.
In each fermentor tank, insert the seed that 360ml obtains by above-mentioned cultivation, aerated culture, temperature is 30 ℃.In culturing process, adopt ammoniacal liquor to regulate the pH of substratum, make it maintain 6.7, simultaneously, continue to add 70% D/W, make sugared concentration be controlled at 3-4g/dl (pressing glucose calculates).Fermentation culture 72 hours, the output of L-Methionin sees Table 3.
Table 3.
The addition of betaine phosphate (g/dl) The amount of L-Methionin (g/dl)
0 11.3
0.05 11.4
0.1 11.6
0.15 12.1
0.2 12.0
0.25 11.2
0.3 10.9
Embodiment 4: seed culture medium (g/dl): glucose 2.5, (NH 4) 2SO 40.2, urea 0.2, KH 2PO 40.15, MgSO 47H 2O 0.04, FeSO 47H 2O 0.001, MnSO 44H 2O 0.001, VB11 00mu g/l, and vitamin H 300mu g/l adjusts pH to 7.0, sterilization.Brevibacterium flavum FERM-P 4164 is inoculated in the 5L seeding tank cultivates 12h.
The fermention medium of L-Threonine (g/dl) is: glucose 8, KH 2PO 40.25, MgSO 47H 2O 0.04, (NH 4) 2SO 42.0, MnSO 44H 2O 0.001, FeSO 44H 2O 0.001, vitamin H 50mu g/l, vitamins B 15mu g/l.Add the betaine phosphate of 0.05-0.3g/dl in the substratum respectively, adjust pH to 7.0.Under 114 ℃, steam sterilizing 15 minutes.Inoculum size by 1% inserts above-mentioned cultured seed liquid in the 10L fermentor tank, fermentation culture 36h, and the output of L-Threonine sees Table 4.
Table 4
The addition of betaine phosphate (g/dl) The amount of L-Threonine (g/dl)
0 10.9
0.05 11.1
0.1 11.2
0.15 11.8
0.2 11.6
0.25 11.0
0.3 10.5
Embodiment 5: seed culture medium (g/dl): glucose 2.5, (NH 4) 2SO 40.5, corn steep liquor 3, KH 2PO 40.1, MgSO 47H 2O 0.05, CaCO 31, adjust pH to 7.0, sterilization.Corynebacterium glutamicum FERM-P 7274 is inoculated in the 5L seeding tank, cultivates 24h down at 30 ℃.
The arginic fermention medium of L-(g/dl) is: glucose 12.5, (NH 4) 2SO 43.0, KH 2PO 40.15, MgSO 47H 2O 0.05, corn steep liquor 1, CaCO 33.0.Add the betaine phosphate of 0.05-0.3g/dl in the substratum respectively, adjust pH to 7.0.Under 114 ℃, steam sterilizing 15 minutes.Inoculum size by 10% inserts above-mentioned cultured seed liquid in the 10L fermentor tank, fermentation culture 96h, and the arginic output of L-sees Table 5.
Table 5
The addition of betaine phosphate (g/dl) The arginic amount of L-(g/dl)
0 3.81
0.05 3.89
0.1 4.05
0.15 4.20
0.2 4.11
0.25 3.98
0.3 3.78
Embodiment 6: seed culture medium (g/dl): sucrose 3.0, KH 2PO 40.15, MgSO 47H 2O 0.04, FeSO 47H 2O 0.001, MnSO 44H 2O 0.001, ammonium acetate 0.3, urea 0.2, soya-bean cake hydrolyzed solution 0.2, VH 0.2mg, VB 10.3mg, adjust pH to 7.0, the 55ml substratum of in the triangular flask of each 500ml, packing into, 0.075Mpa pressure is sterilization 20min down.Brevibacterium flavum FERM-P 3672 is inoculated in the triangular flask, at 31 ℃ of following shaking culture 45h.
The fermention medium of L-Isoleucine (g/dl) is: glucose 14.5, (NH 4) 2SO 42.5, KH 2PO 40.22, K 2HPO 40.1, MgSO 47H 2O 0.04, FeSO 47H 2O 0.0015, MnSO 44H 2O 0.0015, soya-bean cake hydrolyzed solution 0.2, VH 0.01mg, VB 10.25mg, Met 3mg, the betaine phosphate of adding 0.05-0.3g/dl in the substratum is respectively adjusted pH to 7.1.Under 0.075Mpa pressure, steam sterilizing 20 minutes.Inoculum size by 10% inserts above-mentioned cultured seed liquid in the 5L fermentor tank, fermentation culture 80h, and the output of L-Isoleucine sees Table 6.
Table 6
The addition of betaine phosphate (g/dl) The amount of L-Isoleucine (g/dl)
0 2.21
0.05 2.25
0.1 2.33
0.15 2.51
0.2 2.39
0.25 2.30
0.3 2.23
Embodiment 7: seed culture medium (g/dl): glucose 3.0, corn steep liquor 3, yeast powder 0.5, KH 2PO 40.1, MgSO 47H 2O 0.04, MnSO 44H 2O 0.001, VH 0.01mg, VB 10.02mg, adjust pH to 7.0, the 30ml substratum of in the triangular flask of each 500ml, packing into, 0.1Mpa pressure is sterilization 15min down.Brevibacterium flavum FERM-P 512 is inoculated in the triangular flask, at 32 ℃ of following shaking culture 16h.
The fermention medium of L-Xie Ansuan (g/dl) is: glucose 8, (NH 4) 2SO 40.6, KH 2PO 40.12, MgSO 47H 2O 0.04, MnSO 44H 2O 0.001, methionine(Met) 0.05, soya-bean cake hydrolyzed solution 2, corn steep liquor 2.5, VH0.01mg, VB 10.02mg the betaine phosphate of adding 0.05-0.3g/dl in the substratum is respectively adjusted pH to 7.0.Under 0.075Mpa pressure, steam sterilizing 15 minutes.Inoculum size by 10% inserts above-mentioned cultured seed liquid in the 10L fermentor tank, fermentation culture 72h, and the output of L-Xie Ansuan sees Table 7.
Table 7
The addition of betaine phosphate (g/dl) The amount of L-Xie Ansuan (g/dl)
0 5.02
0.05 5.09
0.1 5.16
0.15 5.35
0.2 5.30
0.25 5.21
0.3 5.04
Embodiment 8: seed culture medium (g/dl): glucose 3.0, (NH 4) 2SO 40.5, corn steep liquor 4.0, soya-bean cake hydrolyzed solution 2.0, KH 2PO 40.1, MgSO 47H 2O 0.04, FeSO 47H 2O 0.001, MnSO 44H 2O 0.001, VH 0.02mg, VB 10.03mg, adjust pH to 7.0,0.075Mpa pressure is sterilization 15min down.Corynebacterium glutamicum FERM-P 7128 is inoculated in the 500ml triangular flask, at 32 ℃ of following shaking culture 16h.
The fermention medium of L-tryptophane (g/dl) is: glucose 13, (NH 4) 2SO 44, KH 2PO 41.0, MgSO 47H 2O 0.04, MnSO 44H 2O 0.001, FeSO 47H 2O 0.001, soya-bean cake hydrolyzed solution 2.5, corn steep liquor 2.2, VH 0.005mg, VB 10.01mg the betaine phosphate of adding 0.05-0.3g/dl in the substratum is respectively adjusted pH to 7.0.Under 0.075Mpa pressure, steam sterilizing 15 minutes.Inoculum size by 10% inserts above-mentioned cultured seed liquid in the 5L fermentor tank, fermentation culture, and the output of L-tryptophane sees Table 8.
Table 8
The addition of betaine phosphate (g/dl) The amount of L-tryptophane (g/dl)
0 1.01
0.05 1.12
0.1 1.18
0.15 1.29
0.2 1.20
0.25 1.08
0.3 0.98

Claims (10)

1. the method for a fermentation preparation of L-aminoacid is characterized in that this method comprises the following steps:
(1) seed culture:
Seed culture medium g/dl:
Glucose 3, corn steep liquor 3, beans are dense 2, K 2HPO 40.15, MgSO 47H 2O 0.04, urea 0.2, pH7.0-7.2;
Seed culture medium is put into seeding tank, and 115 ℃ of down sterilizations 15 minutes, brevibacterium flavum ATCC14067 was inserted in the cooling back, in 32 ℃ of following aerated culture 9 hours;
(2) fermentation culture:
Fermention medium g/dl:
Glucose 14, molasses 0.1, Na 2HPO 40-0.1 KCl 0.12, MgSO 47H 2O 0.08, FeSO 44H 2O0.0002, MnSO 40.0002, vitamins B 120mu g/l;
Add the betaine phosphate of 0.05-0.3g/dl in above-mentioned fermention medium, adjusting pH is 7.0-7.2, and this substratum is put in the fermentor tank, and is standby in 15 minutes sterilization backs of 115 ℃ of heating;
The seed that step (1) is obtained is connected in the fermentor tank, and 34 ℃ of aerobic fermentations are cultivated; In culturing process, adopt ammoniacal liquor to regulate pH and maintain 7.0-7.2, simultaneously, continue to add 70% D/W, make sugared concentration be controlled at 1-2g/dl; Before fermentation ends 2 hours stop to continue to add glucose, and in the time of 0,3,8,14 and 18 hours, add the betaine phosphate of 0.15g/dl in fermentation culture respectively, carry out fermentation culture.
2. the method for a kind of fermentation preparation of L-aminoacid according to claim 1 is characterized in that this method used the fermentation assistant betaine phosphate of time array structure and performance,
Figure A2007100370900002C1
Molecular formula: C 5H 14NO 6P
Molecular weight: 215.14
Physicals: white is little yellow crystalline powder extremely, odorlessness, and sour-puckery flavor, the tool moisture absorption, very easily water-soluble, be dissolved in ethanol.
3. the method for a kind of fermentation preparation of L-aminoacid according to claim 1 is characterized in that the concentration of wherein said fermentation assistant betaine phosphate in fermented liquid is 0.01%-5%, and preferable range is 0.05%-1.0%.
4. the method for an a kind of fermentation preparation of L-aminoacid as claimed in claim 1 is characterized in that fermentation under aerobic conditions, stirs or vibrates, and the fermentation time scope was at 1-7 days.
5. the method for a kind of fermentation preparation of L-aminoacid according to claim 1 is characterized in that described L-amino acid is L-L-glutamic acid, L-Methionin, the L-arginine, L-Isoleucine, L-Xie Ansuan, the L-Threonine, the L-Histidine, L-phenylalanine, L-tryptophane, the L-Serine, L-ornithine, L-citrulline, L-tyrosine or L-leucine.
6. the method for a kind of fermentation preparation of L-aminoacid according to claim 1 is characterized in that the carbon source carbohydrate that uses in this method starch fluid as glucose, sucrose, fructose, maltose, molasses, raw sugar or saccharification; Lipid acid is acetate, propionic acid, phenylformic acid; Organic acid, pyruvic acid, citric acid, succsinic acid, fumaric acid or oxysuccinic acid; Alcohol is ethanol or butanols.
7. the method for a kind of fermentation preparation of L-aminoacid according to claim 1, the nitrogenous source that it is characterized in that substratum in this method are that the organic nitrogen source beans are dense, corn steep liquor, urea, yeast powder or soya-bean cake hydrolyzed solution or inorganic nitrogen-sourcedly be ammonium sulfate, ammonium chloride, ammonium nitrate, ammonium acetate, liquefied ammonia or ammoniacal liquor.
8. the method for a kind of fermentation preparation of L-aminoacid according to claim 1 is characterized in that the inorganic salt that the fermention medium in this method uses are phosphoric acid salt, magnesium salts, calcium salt, sylvite, sodium salt, molysite, manganese salt or trace metal salts.
9. the method for a kind of fermentation preparation of L-aminoacid according to claim 1 is characterized in that the VITAMIN that the fermention medium in this method uses is vitamin H, VitB1 or niacinamide.
10. the method for a kind of fermentation preparation of L-aminoacid according to claim 1 is characterized in that this method is used corresponding to amino acid whose zymophyte and is bifidus bacillus ATCC 14020, brevibacterium flavum ATCC 14067, Brevibacterium lactofermentus ATCC 13869, Brevibacterium lactofermentus FERM-P 5012, Corynebacterium acctoacidophlum ATCC13870 or Corynebacterium glutamicum ATCC13032 as L-glutamic acid fermentation bacterium; L-fermenting lysine bacterium brevibacterium flavum ATCC 14067, brevibacterium flavum FERM-P 1708, Corynebacterium glutamicum FERM-P 1709, Brevibacterium lactofermentus FERM-P 1712, Brevibacterium lactofermentus FERM-P 1711, brevibacterium flavum ATCC21475, Corynebacterium glutamicum ATCC 12416, Brevibacterium lactofermentus ATCC 1470 or vinegar bran acid corynebacteria A TCC 11472; L-glutaminate zymophyte is brevibacterium flavum FERM-P 4272, Corynebacterium acctoacidophlum ATCC 13870, brevibacterium flavum FERM-BP 662, vinegar bran acid corynebacteria A TCC 13870 or Corynebacterium glutamicum FERM-BP 663; L-arginine zymophyte is brevibacterium flavum FERM-P 4948, Corynebacterium glutamicum FERM-P 7274, brevibacterium flavum NRRL 12235 or vinegar bran acid coryneform bacteria NRRL 12239; L-phenylalanine zymophyte is Brevibacterium lactofermentus FERM-P 5417, brevibacterium flavum FERM-P 1916, Corynebacterium glutamicum ATCC 21670, citric acid Arthrobacter ATCC 21422, Brevibacterium lactofermentus ATCC21420, Corynebacterium acctoacidophlum ATCC21421, brevibacterium flavum NRRL 12269 or Corynebacterium glutamicum ATCC 21670; L-Threonine zymophyte is colon bacillus FERM-P 4900, Corynebacterium glutamicum FERM-P 5835, brevibacterium flavum FERM-P 4164, Brevibacterium lactofermentus FERM-P 4180, brevibacterium flavum ATCC 21269 or Corynebacterium acctoacidophlum ATCC 21270; L-isoleucine fermentation bacterium is brevibacterium flavum FERM-P 3672, Brevibacterium lactofermentus FERM-P 4192, Corynebacterium glutamicum FERM-P 756, Corynebacterium glutamicum FERM-P 757, Corynebacterium glutamicum FERM-P 758, brevibacterium flavum FERM-BP 759 or brevibacterium flavum FERM-BP 760; L-Histidine zymophyte is brevibacterium flavum FERM-P 2317, Brevibacterium lactofermentus FERM-P 1565, Corynebacterium glutamicum ATCC 14297, Brevibacterium lactofermentus ATCC21086, Corynebacterium acctoacidophlum ATCC 21407, brevibacterium flavum ATCC 21406, subtilis FERM-BP 217 or citric acid Arthrobacter ATCC 21412; L-Xie Ansuan zymophyte is Brevibacterium lactofermentus FERM-P 1845, brevibacterium flavum FERM-P 512, Brevibacterium lactofermentus FERM-P 1968 or intestinal bacteria NRRL 12285; L-Serine zymophyte is Brevibacterium lactofermentus FERM-P 1371; L-ornithine zymophyte is Brevibacterium lactofermentus FERM-P 5936, Corynebacterium glutamicum FERM-P 5644, citric acid Arthrobacter ATCC 21040 or Corynebacterium glutamicum ATCC 13232; L-citrulline zymophyte is Corynebacterium glutamicum FERM-P 5643 or brevibacterium flavum FERM-P 1645; L-tyrosine zymophyte is Corynebacterium glutamicum FERM-P 5836, brevibacterium flavum FERM-P 7914 or subtilis FERM-P 6758; L-tryptophane zymophyte subtilis FERM-P 5286, Brevibacterium lactofermentus FERM-P 7127, brevibacterium flavum FERM-P 2551, Corynebacterium glutamicum FERM-P 7128, brevibacterium flavum ATCC 21427, brevibacterium flavum FERM-BP 114, subtilis FERM-BP208, brevibacterium flavum FERM-BP475, Corynebacterium glutamicum or subtilis FERM-BP 200; L-leucine zymophyte is Brevibacterium lactofermentus FERM-P 1769, brevibacterium flavum FERM-P 420, Corynebacterium glutamicum FERM-P 1966, brevibacterium flavum FERM-BP 755, Corynebacterium glutamicum FERM-BP 756, Corynebacterium glutamicum FERM-P 757 or brevibacterium flavum FERM-BP 759.
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