JPS58187196A - Preparation of amino acid - Google Patents

Abstract

PURPOSE:To prepare efficiently an amino acid, by cultivating a microorganism, capable of producing the amino acid, and immobilized in a specific carrier. CONSTITUTION:A well-known microorganism capable of producing an amino acid is suspended in an aqueous solution containing a compound having photopolymerizable vinyl groups at both ends, e.g. a resin having no cationic or anionic group in the molecule but photopolymerizable vinyl groups at both ends, and the aqueous solution is then irradiated with ultraviolet light to polymerize the above-mentioned compound and entrap and immobilize the microorganism in the resultant polymer. The resultant immobilized miroorganism is then cultivated by the conventional method to collect the aimed amino acid from the resultant culture.

Description

[Detailed description of the invention]

The present invention relates to a method for producing amino acids. Although various proposals have been made regarding methods for producing amino acids using microorganisms, the present inventors immobilized microorganisms capable of producing amino acids. We conducted repeated research to efficiently produce amino acids using the obtained immobilized material. As a result, they discovered that the objective could be achieved by using a specific photopolymerizable compound as an immobilization carrier, and completed the present invention. The present invention involves suspending microorganisms capable of producing amino acids in an aqueous solution containing a compound having a vinyl group that can be polymerized by irradiating light on both ends, and irradiating this suspension with light to produce the above-mentioned compound. This is a method for producing amino acids that involves polymerizing and enclosing microorganisms capable of producing amino acids in the polymer, adding the resulting polymer to a culture medium, and maintaining the culture medium under aeration and stirring. . Various microorganisms are known that have the ability to produce amino acids, and these microorganisms are exemplified below. (1) Lysine-producing bacterium Micrococcus glutamicus Special Publication No. 36-6499
Same as above 91j 52-
102498 (2) Histidine iisi Brevibacterium flavum Special Publication 11851-93
85 Corynebacterium glutamicum same 5
2-18798 Bacillus subtilis 50-13358 Settsia marcescens
Pl 50-37279 Microbacterium ammoniaphyllum same 4
g-18829 Glutamic acid function ■ Brevibacterium lactofermentum TO01
3869 Previbacterium flavumum TCO140
67 Prepipactericum deivaricatamm TO014
020 Corynebacterium glutamicum TC (31
3032 (4 Tryptophan Aim Brevibacterium flavum Special Publication 1882-1882)
8 Microbacterium ammoniaphyllum 48-18828
Colium 9 Tericum glutamicum same 48-18
8287-Slobacter citreus JP-A-4
9-126893 Enterobacter cloacani
44 51-57888 Bacillus subtilis
Special Publication No. 53-39517 Serratea marsetuscens %1llIfi55-81591 Escherichia coli Same No. 56-92796 Bacillus subtilis Special Publication No. 51-6754 Brevibacterium flavum No. 49-61388 Cretia cateforma Special Publication No. 51-6754
Dragon Lathia marcescens JP-A-52-8729
7 Esierihia Cori Special Publication 1977-381
15 (Phrase Proline Production ■ Brevibacterium 72 Bum Special Publication 1975-1175
1 Corynebacterium glutamicum same 49
-28431 Rosecolobactrum coliforme
44-1193 Bacillus subtilis
44-1198 Escherichia Accori
44-1198 (7) Threonine-producing bacterium Micrococcus glutamicum Special Publication No. 47-3899
6 Esierhia ° Cori same 46-34
193 Aerobacter Aerokenes 48-23
54 Serratia marsetuscens 48-235
4 Corynebacterium glutamicum Special Publication 1977-
34956 Previbacterium flag V
Same 45-26708ta + asparagin Shuno! - Brevibacterium 7 labum I￥f open @5
0-88288η (3) Feralanine production - Corynebacterium glutamicum Special Publication 1982-1
0035 Micrococcus glutamicus 1
wj 37-6345 Bacillus subtilis
i! fJ 37-6345 Previpactericum ammoniagenes 37-6345 Mycobacterium lidoclaus JP-A-49-75785
Esierihia Sri 55-165
797-valine production ■ Micrococcus glutamicus Special Publication 1977-1
692 Bacillus subtilis same 37-
12448 Essielichia coli IIQ
37-12448 Brevibacterium Ketoglutamicum 51-46836 Corynepactericum
Glutamicum 52-116 Serratia marsetuscens Special Publication 1986-24274 (b) Isoleucine producing bacterium Micrococcus glutamicus Special Publication 1977-3
8995 Brevibacterium flava i 51-6237 Corynebacterium glutamicum
JP-A-51-73188 Serratia marcescens
Special Publication Showa 51-7755 Bacillus subtilis
52-30593 (b) Cystine, cystine production ■ Brevibacterium flavum Special Publication 1977-1
4637 Esierihia coli JP-A-1973
-80490115 Psiella pneumoniae
rlill 54-80490 Bacillus subtilis 54-80490 (b) Citrulline producing bacterium Bacillus subtilis 811840-16
756 ditto l1i11
111150-18@88 Bacillus megaterium
111850-100288) Homoserine production ■ Corynebacterium glutamicum Special Publication 51-6238) Previbacterium flavum
36356) Glutamine production ■ Brevibacterium flavum time af45
3-17675 Same as above JP 54-62388 Corynebacterium glutamicum Same 50-89590 Escherichia coli
56-23893 (b) Tyrosine production ■ Brevibacterium flavum Special Publication 1977-3
8837 Corynenozatherium glutamicum 55-46717 Bacillus subtilis
49-38837-Serine production ■ Brevibacterium lactofermentum JP-A-1987-8
7083 Corynepacterium glutamicum (Special Publication No. 51-33188 Corynepactericum glucinopherum ATOO21341-Leucine Production- Brevibacterium flavum Special Publication No. 1977-2)
4275 Corynebacterium glutamicum % opening 50-123877 Serratia marsetuscens
% extra H47-26313 Micrococcus glutamicum 18347 Escherichia coli 43-8711 Previbacterium lactofermentum 43-
8712 Bacillus subtilis ljl
43-10996 Corynebacterium glutamicum Special-Sho 53-24096φ Heart Medeonine Raw Baking Soda Protaminobacter Candidus JP-A-Sho 50-3
1092 Corynebacterium glutamicum Special Publication 1984-40240 Escherichia coli M
Introduction 56-35992 Next, the photopolymerizable compound used in the present invention refers to a compound having a vinyl group that can be polymerized by irradiating both ends with light. No., pp. 620-628 discloses a tree k (A
There is. This resin is expressed by the following general formula, and specific examples include:

[Hydrophilic 1) IT-1000, INT-2000 with polyethylene glycol as the main chain
,INT-4000゜IN'! '-6000'? Hydrophobic INTP-1 with a propylene glycol main chain
000,11NTP-2000゜1N? ? -3000 etc. Note that these numbers indicate the approximate molecular weight of the main chain. / 8 / / Furthermore, in the present invention, the photopolymerizable resin has a cationic/or anionic group in the molecule represented by the following general formula and a photopolymerizable vinyl group at both ends (6). It can be used in combination with resin (4). (1) Carboxylic acid type anion resin (]INT-207)
(Here, n is a positive integer from 1 to 20.) (Here, n is a positive integer from 1 to 20.)
It is a positive integer of 9. ) (3) Quaternary ammonium salt type cation #! IkCEkJT-425) (where n)i1
It is a positive integer of ~9. ) The microorganism having the ability to produce a 7-mino acid is suspended in the aqueous solution of the photopolymerizable compound. Here, as the microorganism, a culture chamber containing microbial cells obtained by culturing in a usual manner may be used as it is, or microbial cells separated by an operation such as centrifugation, or a washed version thereof, etc. may be used. . A suspension containing a photopolymerizable compound and a microorganism is irradiated with light to polymerize the compound. As a method for enclosing microbial cells in a photopolymerizable compound, the microorganisms may be suspended in an aqueous solvent, thoroughly mixed with the photopolymerizable compound, and then irradiated with light to polymerize. Rays are 250-600
Most preferred are wavelengths in the nm range. Due to the polymerization of the photopolymerizable compound, microbial cells are incorporated into the polymer.
A nationally defined microbial cell is obtained. In order to produce the desired amino acid using the immobilized polymer of the microorganism with amino acid-producing ability obtained in this way, it is necessary to increase the production amount of the components and amino acids necessary for the growth of the microorganism. The polymer may be added to a conventional medium containing ingredients such as those mentioned above, and the medium may be maintained under conventional aeration and agitation. Furthermore, the pIK of the medium to which the polymer has been added may be adjusted to an appropriate value taking into account the type of microorganism, the type of amino acid produced, etc., but it is usually about 678. This medium is heated at 20-40'C, preferably 25-35'C.
A sufficient amount of amino acids is produced and accumulated by holding for 0 to 100 hours, preferably 48 to 72 hours. According to the present invention, microorganisms capable of producing amino acids are tightly encapsulated and immobilized in the polymer, so that the amount of microorganisms that leak out from the polymer is extremely small, stable activity continues, and amino acid production is efficiently carried out. can be manufactured. In the present invention, by selecting a specific photopolymerizable compound as the carrier, the amount of amino acids accumulated is higher than when known immobilized carriers are used, and the method is highly practical. Next, the present invention will be explained in detail with reference to examples. Example 1 Citabrevibacterium lactofermentum (FgRM P-4) was cultured on bouillon agar medium at 30°C for 24 hours.
One platinum loop of No. 547) was seeded with the following composition (lj) A medium Km, and cultured at 30°C for 24 hours. Glucose 4g/dJ Soybean protein hydrolyzate 150~/dZ (as total nitrogen) (NHa) 80. 4.5 9
/dlKH,Po, 0.1
#Mg80.・7)1. OO,041 F0++'2ppm Mn++ 2Biotin
500 r/e vitamin B,
2007r near
40 aid/di nicotinamide
5 Adenine lO l Sodium glutamate 10 1CaC0,
4% (separate sterilization) (pki8.0.115℃, 10 minutes sterilization) After completion of culture, culture 12dtJ! :1a The bacterial cells obtained by separating the hearts were used in the following experiments. Photopolymerizable compound 19. Benzoinoethyl ether 10■
was added to 1.5 ILt of 0.85'A saline solution, and the above microorganism cells (mixed 1 body) suspended in 1 d of the same saline solution were added.
Mix and spread this mixture on a sheet, 300 to 40
0r] IIl was irradiated with near-ultraviolet light for 3 minutes. The obtained polymer was cut into a size Vζ of 5×5u, and
．． Washed with 85% saline. ) Immobilized microbial cells were thus obtained. Next, the immobilized microbial cells were transferred to B medium 20 with the following composition.
The mixture was added to a 500 d shoulder flask containing au and kept at 30° C. for 72 hours with aeration and stirring. B medium glucose 8 9/dt (N
H4) m80i 4.5 g
Phantom 1 @PO4Q, l M Biotin 500 17g Vitamin B r 200 Alania
40 ~7'tLl Nicotinamide
5 Adenine 10 l Sodium glutamate 10 After completion of the culture, the amount of lysine produced and the turbidity of the culture solution (absorbance at 56'l nm was measured for a 26-fold dilution of the culture solution) were determined. The results are shown in Table 1. Comparative Example 1 A mixture of 4% alginic acid 11dK and wet bacterial cells suspended in 085% saline IILtK, which is the same as in Example 1, was mixed with 15% Ca
Cl] to obtain immobilized microbial cells (approximately 5 cods in diameter). After washing this with 0.85% saline, it was added to medium B in the same manner as in Example 1, and the same procedure was carried out thereafter. The results are shown in Table 1. Comparative Example 2 The wet bacterial cells of Example 1 were suspended in 1 ml of 0.85% saline solution and kept at 7.40°C, and this was mixed with 1 m of 4% Nippon Guinan kept at 50°C. Solidified at ℃. The obtained solid was cut into a size of 5); 5' x 5 U, and this was poured into a 15% KCl solution to solidify the gel. The obtained immobilized microbial cells were reduced to 0.85% KO.
After washing with solution J, add to medium B in the same manner as in Example 1, and proceed in the same manner. The results are shown in Table 1. Comparative Example 3 The wet product of Example 1 was suspended in 2 xxl of 0.85% saline, and 0.759 of acrylamide and 30~ of methylenebisacrylamide were added thereto. This mixture was mixed with 0.5d of 5% β-dimethylamine propionitrile and 2.5% Ks.
The mixture was mixed with a solution containing 0.5 N of SsOs and allowed to gel at room temperature. The obtained gel was cut into a size of 5×5×5M and washed with 0.85% saline to obtain immobilized microbial cells. This was added to medium B in the same manner as in Example 1, and the same procedure was carried out thereafter. The results are shown in Table 1. Comparative Example 4 In this example, the wet bacterial cells of Example 1 were added to medium B without being immobilized, and the same procedure as in Example 1 was carried out. Results first
Shown in the table. 48 hour culture 68 hour culture kNT-2000 1.05 0.05 1
．． 70 U, 05 real j! NT-40000,770
,031,50U 03SHENT-60001280,
042030,07 Alkynoic acid 0.6 0
．． 418 1.13 0.20 ratio carrageen dog 0.58 0.08 0.8
6 0.70 comparison acrylamide 0.38 0.07
0.52 0.15 Example - 1,590,792820,88 As is clear from the table 1, according to the present invention, the leakage of microbial cells from the carrier is extremely small, and the amount of lysine produced is lower than that of the known carrier. Example 2 Brevibacterium lactofermentum FBI'tM cultured on bouillon agar medium for 30"C-C for 24 hours
P-1565) 1 platinum loop is 3 g/'
dj. (NH4)*SOa 1.09. 'cu, KH1I
Inoculated into a medium containing PO41,59/di, m@so4・7H20 and yeast extract 1.097di,
The cells were cultured at 0°C for 24 hours. After the culture was completed, the culture solution 2d was centrifuged and the resulting product was used in the following experiment. 1 g of photopolymerizable compound, lO of benzoin ethyl ether
1.5 m of 085% saline solution [dissolved, add 1.5 m of the same saline solution]
The above-mentioned microbial cells (wet cells) suspended in d were mixed, this mixture was spread on a sheet, and near-ultraviolet light having a wavelength of 300 to 400 nm was irradiated for 3 minutes on each side of the sheet. The obtained polymer was cut into a size of 5 x 5 u and washed with 0.85% saline. In this way, immobilized microbial cells were obtained. Next, this immobilized microbial cell was added to 20 m of C medium having the following composition.
The mixture was added to a 500 au shouldered flask containing 300 au and kept at 30°C for 72 hours with aeration and stirring. C medium glucose 10 Q/dl (N
lI4) 5045 KIJ *P04 0.1
g□ Mg804-711.0 0.
04 FFe+'+2ppm Mn++2 #Biotene 500 μVl Vitamin B 15 Q z Arginine 50mf// (11 After completion of culture, measure the generated charge of histidine and the turbidity of the culture solution (absorbance at 562 t]m for a 26-fold dilution of the culture solution) The results are shown in Table 2. Comparative Example 5 4% alginic acid 11LtK A mixture of wet bacterial cells dissolved in the same 0.85% saline IILt as in Example 2 was added to 15%
The solution was dropped into Ca1l to obtain immobilized microbial cells (about 5 mm in diameter). After washing this with 0.85% saline, Example 2
I did the same thing. The results are shown in Table 2. Comparative Example 6 One wet body of Example 2 was mixed with one part of 0.85% saline solution! After mixing with 4% carrageenan IJI7 kept at 50°C and solidifying at 10"C, the obtained solid was suspended in 5), 5x-50 This was cut into pieces of 15% KCj liquid-resistant solution to solidify the gel.
After washing with KOj solution, the same procedure as in Example 2 was carried out. -The results are shown in Table 2. Comparative Example 7 The wet bacterial cells of Example 2 were added to 0.85% saline solution 2aJ14j%!
To turbidity 1, 0.759% of acrylamide and 30~ of methylenebisacrylamide were added. Add this mixture to 5% β
-Dimethylamine propionitrile 0°51 and 2.5%
It was mixed with a solution containing K15ses O,5id and allowed to gel at room temperature. Obtained gel Y, <5>, 5
After cutting into a size of 5 m x 5 m, the pieces were washed with 0.85% saline to obtain immobilized microbial cells. This was carried out in the same manner as in Example 2 at 10°C. The results are shown in Table 2. Comparative Example 8 The wet bacterial cells of Example 2 were added to C medium without immobilization,
The following steps were carried out in the same manner as in Example 2. The results are shown in Table 2. Table 2 ENT-20000290,06 ENT-40000,41 (J, 12 Real ENT-6000 (1580,11 Alginic acid 0.43 0.4
9 ratio carrageenan 11.32 Q
, 32 comparison acrylamide 0.11 (
1,H) Example-0920,90 Example 3 Brevibacterium lactofermentum AJ11445 cultured on bouillon agar medium at 30"C for 24 hours
One platinum loopful of (FERMP-5162) was inoculated into D medium having the following composition, and cultured at 30°C for 24 hours. D medium glucose 1-su 5 9/di 肚, i'04
0.1 /' Mn++2y Vitamin =, 200 r/e Soybean protein hydrolyzate 36~/a (as total nitrogen) Urea
0.4g/J (separately killed l1ii+) (pH7,
Sterilize at 9,115"C for 10 minutes) After culturing, add 20
Bacterial cells obtained by centrifuging aJ were used for experiments. Dissolve 1 g of the photopolymerized compound and 10 da of benzoin ethyl ether in 1.5111 AK of 0.85% saline, mix with this the microbial acid body (fi bacterial cell) of 1F suspended in 1 d of the same saline, and add this. Spread the mixture into ten sheets. It was then irradiated with near ultraviolet light with a wavelength of 300 to 400 nm for 5 minutes. The obtained 1-go mixture was cut into 5×5 brocade pieces, and
．． Washed with 85% saline. Immobilized microbial cells were thus obtained. Next, the immobilized microbial cells were transferred to medium 2 with the same composition except that the amount of glucose added was changed to 3.6 g/j and the amount of urea added was changed by 0.297 dlK.
500 Iu of waste containing 0al17 was added to Fufusco, and a shaking test was conducted at 30°C. Add 05% S of urea to the wave at 8 o'clock! The cells were added in reverse order (total 0.7%) and the culture was continued for an additional 16 hours. After the completion of the culture (24 hours), the amount of glutamic acid produced and the turbidity of the culture solution (552 n for a 26-fold dilution of medium II)
The absorbance of m) was determined. The results are shown in Table 3. In addition, at the end of the culture, the temporarily immobilized bacterial cells were recovered and used in a new medium.The results were also shown in Table 3. Comparative Example 9 Immobilized microbial cells to be used as an alginic acid carrier were obtained in the same manner as in Comparative Example 1 except that the same wet cells as in Example 3 were used. The results are shown in Table 3. Comparative Example 10 Immobilized microorganisms using carakeenan as a carrier were prepared in the same manner as in Comparative Example 2, except that the same wet cells as in Example 3 were used. This immobilized bacterial cell was used in the same manner as in Example 3. The results are shown in Table 3. Comparative Example 11 Comparative Example 3 except that the same wet bacterial cells as in Example 3 were used. Immobilized microbial cells using acrylamide as a carrier were obtained in the same manner as in Example 3. Using these immobilized cells, the same procedure as in Example 3 was carried out. The results are shown in Table 3. Comparative Example 12 Example 3 In addition to the culture medium without immobilizing wet bacterial cells,
The following steps were carried out in the same manner as in Example 3. The results are shown in Table 3. 1st 2nd Memo INT-20000, 240, 060, 300, 03
SINT-40000,090,060,150,03
Example I -6000 0.24 0.06 0.23
0.03 Alginic acid 0.21 0.0
7 0.40 0.20 ratio carrageenan 0.03 0.10
0.03 0.11 Acrylamide 0.03 0.10
0.03 0.10 cases - 1,200,490,720,46 Example 4 Broth agar medium [Haf cultured at 30°C for 24 hours
)k Eacllus 5ubti
One loopful of P-5289 was inoculated into E medium having the following composition and cultured at 30°C for 24 hours. E medium glucose 3 9,1dlNli40
7 2 LCI 0.2 #肚、po
, 0.1 #M Shizu SO4・7H
,00,04# Fe3O4・7ki@0 0.01 lMn
3O4・4H,00,011 Casamino acid 1.0I Oa00. 4 # (Separate assassination-
) After the completion of the culture, the culture solution 2d was centrifuged and the resulting product was used in the following experiment. Photopolymerizable compound 19. Benzoin ethyl ether lO~
was dissolved in 1.5 at of 0.85% saline, mixed with the above-mentioned microorganisms (y1 bacterial cells) suspended in the same saline, spread this mixture on a sheet, and
The front and back surfaces were irradiated with near ultraviolet light having a wavelength of 0.00 nm for 3 minutes each. The obtained polymer was cut into a size of 575 cm, and the size of 0.8
Washed with 5% saline. In this way, immobilized microbial cells were obtained. Next, the immobilized microbial cells were transferred to 500 volumes of medium 20a7 with the same composition except that the amount of glucose added was changed to 1097 di and the amount of casamino acid added was changed to 0.497'dl K in the Σ medium. It was added to a flask with a shoulder and cultured with shaking at 30°C for 90 hours. After the cultivation was completed, the tryptophan production lid and the turbidity of the culture solution (absorbance at 562 nm was measured using a 26-fold dilution of the culture solution) were determined. The results are shown in Table 4. Comparative Example 13 Immobilized microbial cells to be used as an alginic acid carrier were obtained in the same manner as in Comparative Example 1 except that the same wet cells as in Example 4 were used. The results are shown in Table 4. Comparative Example 14 Using the same wet microbial cells as in Example 4, immobilized microbial cells using carrageenan as a carrier were prepared in the same manner as in Comparative Example 2 except that The same procedure as in Example 4 was carried out using the immobilized cells. The results are shown in Table 4. Comparative Example 15 Same as Comparative Example 3 except that the same wet cells as in Example 4 were used. A solidified microbial cell using acrylamide as a carrier was obtained. Using this solidified body, the same procedure as in Example 4 was carried out. The results are shown in Table 4. Comparative Example 16 Wet bacteria of Example 4 The cells were added to the culture medium without immobilization, and the following procedure was carried out in the same manner as in Example 4.The results are shown in Table 4. 12 real ΣNT-60
000,440,13 ENT-4000: し'ENT207 (8・2) 0°28 0.05 example, BNT-40002IN'i'22o (8:2) 0°39
°°09, EN'l'-4000" BNT425 (8:2) "19 007
Alginic acid 0.37 0
．． 42 ratio carrageenan 0.26
0.34 Comparative example Acrylamide 0.15
0.29-0.80
Escherichia coli (Cedar 9) cultured on 0.75 bouillon agar medium at 30°C for 24 hours.
(0ha) AJ11335 (FIRM P-490
1) Inoculate No. 1 platinum loop onto y#I ground with the following composition, and
The cells were cultured at ℃ for 24 hours. ! Medium glucose 3% (NH4) 1
804 1 'Kk1. P0.
0.1 #Mg80.・7H,00
,1y Fe804' 7 to 1@O0,00! 1jin 80
4 ・4H* OO,001ttRNmu
02 L-aspartic acid 100
MVd/L-Nororin 45 〃L
-Inleucine 10 N L-Methionine 10
B, 100 tn (i/dl yeast extract 05%0aCO14% (
Separately provided (m) (pH 7, 0, 1O fractionated at 115° C.) After completion of the culture, the culture solution 1d was centrifuged and the cells obtained were used in the following experiment. 1 g of photopolymerizable compound, lO of benzoin ethyl ether
1, dissolved in 5 ml of 085 saline solution, mixed with 1 of the above microorganisms (wet bacterial cells) suspended in 11 snack water, and spread this mixture on a sheet. Near ultraviolet rays having a wavelength of 300 to 400 nm were irradiated for 3 minutes each on the front and back sides. The obtained polymer was cut into 5×5 III pieces and washed with 0.85% saline. In this way, immobilized microbial cells were obtained. Next, let's look at this immobilized microorganism as described above! 500 containing 2olLt of medium with the composition excluding 05% yeast extract from the medium
The cells were placed in a d capacity flask with a shoulder and cultured at 30'C for 48 hours. After the culture was completed, the amount of threonine produced and the turbidity of the culture solution (absorbance at 562 nm was measured for a 26-fold dilution of the culture solution) were determined. The results are shown in Table 5. Comparative Example 17 Immobilized microbial cells to be used as an alginic acid carrier were obtained in the same manner as in Comparative Example 1 except that the same wet cells as in Example 5 were used. The same procedure as in Example 5 was carried out using this immobilized bacterial cell. The results are shown in Table 5. Comparative Example 18 Immobilized microbial cells using carrageenan as a carrier were obtained in the same manner as in Comparative Example 2, except that the same wet cells as in Example 5 were used. The same procedure as in Example 5 was carried out using this immobilized bacterial cell. The results are shown in Table 5.1 Comparative Example 19 Immobilized microbial cells using acrylamide as a carrier were obtained in the same manner as in Comparative Example 3 except that the same wet cells as in Example 5 were used. Using this immobilized microbial cell (this was carried out in the same manner as in Example 5. The results are shown in Table 5. Comparative Example 20 The wet cells of Example 5 were added to the medium without immobilization,
The following steps were carried out in the same manner as in Example 5. The results are shown in Table 5. Table 5 Utsugi NT-20000,620,06 Shima NT-40000,350,06 夾hNT-jooo O,40o,os Alginic acid 0.32 0.18
Specific carrageenan 0.43 0.
Comparison 19-1,480,30 Example 6 Brevibacterium lactofermentum FIRMP-52 cultured on bouillon agar medium at 30°C for 24 hours
48-1 platinum loop with 3g of glucose, (NH,), 8
Q41, 09/di, KH*PO41, 59/di
, Mg80n 7H@O and seeded at 30°C for 2
It was cultured for 4 hours. After the culture was completed, the culture solution 2d was centrifuged and the obtained bacterial cells were used in the following experiment. 1 g of photopolymerizable compound, 10 ~ benzoin ethyl ether
Dissolved in 0.85% physiological saline (1.5 - liters), mixed with the above microbial image (wet one) suspended in 1 d of the same saline solution, and spread/spread this mixture on a sheet. 300-400
Near ultraviolet light having a wavelength of nm was irradiated for 5 minutes. The resulting polymer was cut into a size of 5 x 5, and the immobilized microbial cells were placed in a 500 ILt shoulder flask containing 20-G medium with the following composition, and cultured at 30°C for 72 hours. . Gjl ground glucose 3 9yal(NH4)1
1804 1,0 //String 1PO41,5#
'Mg8Q, 7H*0 0.04 'Biotin
5.0 μg 7cu Thiamine Hydrochloride 20.0 μg/a1P 80.・711m
0 1 ~/dtMn80. -4H601
M L-Tyrosine 40 1DL-Methionine 40 l Soybean protein hydrolyzate
3.0114/llu fumaric acid - 1.2 g/d/ Vinegar # 0.3''/'' After culturing, measure the amount of phenylalanine produced and the turbidity of the culture solution (absorbance at 562 nm for a 26-fold dilution of the culture solution) ) was sought. The results are shown in Table 6. Comparative Example 21 Immobilized microbial cells to be used as an alginic acid carrier were obtained in the same manner as in Comparative Example 1, except that the same wet cells as in Example 6 were used. The same procedure as in Example 6 was carried out using this immobilized body. The results are shown in Table 6. Comparative Example 22 Immobilized microbial cells using carrageenan as a carrier were obtained in the same manner as in Comparative Example 2, except that the same wet cells as in Example 6 were used. The same procedure as in Example 6 was carried out using this immobilized body. The results are shown in Table 6. Comparative Example 23 Immobilized microbial cells using acrylamide as a carrier were obtained in the same manner as in Comparative Example 3 except for using the same wet cells as in Example 6. The procedure was carried out in the same manner as in Example 6. The results are shown in Table 6. Comparative Example 24 The wet bacterial cells of Example 6 were added to the culture medium without immobilization, and the procedure was carried out in the same manner as in Example 6. The results are shown in Table 6. It is shown in the table. Table 6 gnT-20000,820,08 EN'l'-40001,000,18 Actual ENT-60
001,020,16 Alginic acid 0.93 0
．． 48 ratio carrageenan 0.78 0
．． 42 comparison acrylamide 0.51 0
．． Example 31 - 2, 201, 10 Example 7 Corynebacterium glycinoelum (Coryae-Sigy) cultured on bouillon agar medium at 30°C for 24 hours.
terium glyc4nophelum )mu'f
One platinum loop of '0021341 was inoculated into M medium with the following composition and incubated at 30'C in 24Ii medium Glucose 0.5% Yeast extract 1, 0〃 Peptone 1-θ' Na0j Q, 5 tt (pH 7, 0, 115") After incubation #1), the culture solution was centrifuged for 10 minutes and the resulting bacterial cells were subjected to the following experiment. Photopolymerizable compound 19. Benzoin ethyl ether 10η
Dissolve in 085% food fortune water 15-, then add 7X salt.
Mix the above microorganism cells (1 wet cell) suspended in IILt, spread this mixture on a sheet,
Near ultraviolet rays with a wavelength of 100 nm were irradiated for 3 minutes each on the front and back sides. The obtained polymer was cut into a size of 5×5fi and 0.85
% saline solution. After obtaining the identified microorganisms in this manner, the immobilized microorganisms were transferred to I medium 201 with the following composition.
17 into a 500 d shouldered flask and heated to 30°C.
The cells were incubated for 48 hours and cultured. ■Medium glucose 5% glycine 21 CNHa) *80a 0.3 #Kti,
PO40,1# Mg804・7H,00,051 Biotin 0.2 μ07d Thiamine 11 Folic acid 11 Nicotinic acid amide 2.51 Yeast extract 0.5% (3a00 Hatake 2.5% (separate i[1) (pH7,
Killed in 10 minutes at 0,115℃ 11! After the completion of the culture, the amount of serine produced and the turbidity of the culture solution (absorbance at 562 nm was measured for a 26-fold dilution of the culture solution) were determined. The results are shown in Table 7. In addition, the immobilized bacterial cells after completion of the culture were collected aseptically and subjected to the production of serine again in the same manner as above using a new medium. The results at this time are also shown in Table 7. Comparative Example 25 Immobilized microbial cells to be used as an alginic acid carrier were obtained in the same manner as in Comparative Example 1, except that the same wet cells as in Example 7 were used. Examples 7 and 11 were carried out using this immobilized bacterial cell. The results are shown in Table 7. Comparative Example 26 Immobilized microbial cells using carrageenan as a carrier were obtained in the same manner as in Comparative Example 2, except that the same wet cells as in Example 7 were used. The same procedure as in Example 7 was carried out using this immobilized bacterial cell. The results are shown in Table 7. Comparative Example 27 Immobilized microbial cells using acrylamide as a carrier were obtained in the same manner as Comparative N3 except that the same wet body as in Example 7 was used. The same procedure as in Example 7 was carried out using this immobilized bacterial cell. The results are shown in Table 7. Comparative Example 28 The same procedure as in Example 7 was carried out except that the wet material of Example 7 was added to the medium without immobilization. The results are shown in Table 7. Table 7 nNT-20000, 200, 060, 300, 211
NT-40000, 160, 060, 220, 22 Real E
NT-60000, 260, 150, 270, 19 (
INT-4000゜ENT207 (8:2) 0.14 0.10
0.43 0.10 cases, FiN'l"-4000: B, T220 (8,, 0,160,060,510,0
3, ]iiN'r-4000: , , , 425 (8,, 0,150,040,200,
03 Alginic acid 0.20 0.36 0
,38 0.47 ratio carrageenan 0.20 0.35 0.4
0 0.37 Acrylamide 0.08 0°10 0,
12 0.35 cases - 0,50,680,30,70 Patent applicant Ajinomoto Co., Inc.

Claims (1)

[Claims] A microorganism capable of producing amino acids is suspended in an aqueous S* containing a compound having a vinyl group that can be polymerized by irradiating light at both ends, and this suspension is irradiated with 11EK light to polymerize the above compound. 1. A method for producing amino acids, which comprises: incorporating the microorganism capable of producing amino acids in a polymer; adding the obtained polymer to a culture medium; and maintaining the culture medium under aeration and stirring.