JPH01144987A - Production of l-phenylalanine by fermentation - Google Patents
Production of l-phenylalanine by fermentationInfo
- Publication number
- JPH01144987A JPH01144987A JP30519287A JP30519287A JPH01144987A JP H01144987 A JPH01144987 A JP H01144987A JP 30519287 A JP30519287 A JP 30519287A JP 30519287 A JP30519287 A JP 30519287A JP H01144987 A JPH01144987 A JP H01144987A
- Authority
- JP
- Japan
- Prior art keywords
- phenylalanine
- tyrosine
- citrobacter
- growth
- medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 title claims abstract description 40
- 229960005190 phenylalanine Drugs 0.000 title claims abstract description 22
- 238000000855 fermentation Methods 0.000 title claims description 8
- 230000004151 fermentation Effects 0.000 title claims description 8
- 238000004519 manufacturing process Methods 0.000 title claims description 5
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims abstract description 24
- 229960004441 tyrosine Drugs 0.000 claims abstract description 13
- 241000588923 Citrobacter Species 0.000 claims abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 9
- 238000000034 method Methods 0.000 claims description 9
- 244000005700 microbiome Species 0.000 claims description 6
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims description 3
- 239000002243 precursor Substances 0.000 abstract description 3
- 241000588919 Citrobacter freundii Species 0.000 abstract description 2
- 238000012258 culturing Methods 0.000 abstract description 2
- 230000000813 microbial effect Effects 0.000 abstract 2
- 150000001875 compounds Chemical class 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 11
- 229920001817 Agar Polymers 0.000 description 6
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 6
- 239000008272 agar Substances 0.000 description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- -1 succinic acid, alcohols Chemical class 0.000 description 3
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 description 2
- 150000003863 ammonium salts Chemical class 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- QDGAVODICPCDMU-UHFFFAOYSA-N 2-amino-3-[3-[bis(2-chloroethyl)amino]phenyl]propanoic acid Chemical compound OC(=O)C(N)CC1=CC=CC(N(CCCl)CCCl)=C1 QDGAVODICPCDMU-UHFFFAOYSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- 239000004254 Ammonium phosphate Substances 0.000 description 1
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 0.000 description 1
- 101000614400 Homo sapiens Serine/threonine-protein phosphatase 2A regulatory subunit B'' subunit alpha Proteins 0.000 description 1
- 241000192130 Leuconostoc mesenteroides Species 0.000 description 1
- VZUNGTLZRAYYDE-UHFFFAOYSA-N N-methyl-N'-nitro-N-nitrosoguanidine Chemical compound O=NN(C)C(=N)N[N+]([O-])=O VZUNGTLZRAYYDE-UHFFFAOYSA-N 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 101710148027 Ribulose bisphosphate carboxylase/oxygenase activase 1, chloroplastic Proteins 0.000 description 1
- 102100040446 Serine/threonine-protein phosphatase 2A regulatory subunit B'' subunit alpha Human genes 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 229910000148 ammonium phosphate Inorganic materials 0.000 description 1
- 235000019289 ammonium phosphates Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000003729 cation exchange resin Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 235000013985 cinnamic acid Nutrition 0.000 description 1
- 229930016911 cinnamic acid Natural products 0.000 description 1
- 229960004106 citric acid Drugs 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 235000012907 honey Nutrition 0.000 description 1
- 229910001410 inorganic ion Inorganic materials 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- CNFDGXZLMLFIJV-UHFFFAOYSA-L manganese(II) chloride tetrahydrate Chemical compound O.O.O.O.[Cl-].[Cl-].[Mn+2] CNFDGXZLMLFIJV-UHFFFAOYSA-L 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 1
- 238000002816 microbial assay Methods 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 229960000999 sodium citrate dihydrate Drugs 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
〈産業上の利用分野〉
本発明は発酵法によるＬ−フェニルアラニンの製造法に
関する。DETAILED DESCRIPTION OF THE INVENTION <Industrial Application Field> The present invention relates to a method for producing L-phenylalanine by a fermentation method.
〈従来の技術〉
従来、シトロバクタ−属の微生物を用いるＬ−フェニル
アラニンの製造法としては、桂皮酸とアンモニアを原料
とする酵素法（特開昭６１−２７１９９８号公報）など
が知られている。<Prior Art> Conventionally, as a method for producing L-phenylalanine using microorganisms of the genus Citrobacter, an enzymatic method using cinnamic acid and ammonia as raw materials (Japanese Patent Application Laid-open No. 271998/1983) is known.
〈発明が解決しようとする問題点〉
しかしながら、従来の方法は、高価な前駆体を使用する
ものであり、経済的に極めて不利な方法であった。<Problems to be Solved by the Invention> However, the conventional method uses an expensive precursor and is economically extremely disadvantageous.
く問題点を解決するための手段および作用〉そこで本発
明者らは、前記問題点を解決すべく鋭意研究の結果、発
酵法によってＬ−フェニルアラニンが著量蓄積すること
を見い出し、本発明に到達した。Means and Effects for Solving the Problems In order to solve the problems, the present inventors conducted intensive research and found that L-phenylalanine accumulates in significant amounts by fermentation, and thus arrived at the present invention. did.
すなわち、本発明はシトロバクタ−属に属し、生育にＬ
−チロシンを要求し、かつＬ−フェニルアラニン生産能
を有する微生物を、液体培地中で好気的に培養してＬ−
フェニルアラニンを培養液中に生成・蓄積せしめ、該培
養液よりＬ−フェニルアラニンを採取することを！徴と
する発酵法によるＬ−フェニルアラニンの製造法である
。That is, the present invention belongs to the genus Citrobacter, and the growth
- A microorganism that requires tyrosine and has the ability to produce L-phenylalanine is cultured aerobically in a liquid medium to produce L-phenylalanine.
Phenylalanine is produced and accumulated in the culture solution, and L-phenylalanine is collected from the culture solution! This is a method for producing L-phenylalanine using a fermentation method.
次に本発明の詳細な説明する。Next, the present invention will be explained in detail.
本発明で用いられる微生物はシトロバクタ−属に属し、
生育にＬ−チロシンを要求し、かつＬ−フェニルアラニ
ン生産能を有する微生物であれば特に限定されない。The microorganism used in the present invention belongs to the genus Citrobacter,
The microorganism is not particularly limited as long as it requires L-tyrosine for growth and has the ability to produce L-phenylalanine.
ここでチロシンの要求性は１ｅａｋｙ型要求性であって
も本発明の範囲に含まれる。Here, even if the requirement for tyrosine is 1eaky type requirement, it is included in the scope of the present invention.
本発明で用いられる微生物の代表的なものとしてはたと
えば以下のものがある。シトロバクタ−・７ｏインデ（
−（ｃｌｔｒｏｂａｃｔｅｒ ｆｒｅｕｎｄｉｉ）ＴＹ
ＲＡ−１（微工研菌寄第９６１８号）。Typical microorganisms used in the present invention include, for example, the following. Citrobacter 7oinde (
-(cltrobacter freundii)TY
RA-1 (Feikoken Bibori No. 9618).
この菌株はシトロバクタ−・フロインディー０ＦＰＲ７
２（微工研菌寄第９５３７号）より誘導された変異株で
生育にＬ−チロシンを要求する変異株である。変異株の
誘導は通常の変異株処理法によって比較的容易に行うこ
とができる。This strain is Citrobacter freundii 0FPR7
This is a mutant strain derived from No. 2 (Keikoken Bibori No. 9537) that requires L-tyrosine for growth. Mutant strains can be induced relatively easily by conventional mutant strain treatment methods.
本発明におけるＬ−フェニルアラニン生産用の液体培地
は炭素源、窒素源、無機イオン、生育に必要なＬ−チロ
シンおよび必要に応じてその他の有機微量成分を含有す
る通常の液体培地である。炭素源としては、グルコース
、フラグドースやでん粉およびセルロースの加水分解物
、ＷＪ蜜などの糖類、フマール酸、クエン酸、コハク酸
などのごとき有機酸、グリセロールのごときアルコール
類などを２〜１５％、窒素源として、酢酸アンモニウム
のごとき有機アンモニウム塩、ａｈアンモニウム、塩化
アンモニウム、リン酸アンモニウム、硝酸アンモニウム
のごとき無機アンモニウム塩、アンモニアガス、アンモ
ニア水、尿素などを０．５〜４．０％、これらの他にリ
ン酸カリウム、硫酸マグネシウム、硫酸第１鉄７水和物
、塩化マンガン４水和物、Ｌ−チロシンなどが少量添加
される。The liquid medium for producing L-phenylalanine in the present invention is a conventional liquid medium containing a carbon source, a nitrogen source, inorganic ions, L-tyrosine necessary for growth, and other organic trace components as necessary. Carbon sources include glucose, hydrolysates of flagose, starch, and cellulose, sugars such as WJ honey, organic acids such as fumaric acid, citric acid, and succinic acid, alcohols such as glycerol, and 2 to 15% nitrogen. Sources include organic ammonium salts such as ammonium acetate, inorganic ammonium salts such as ah ammonium, ammonium chloride, ammonium phosphate, ammonium nitrate, ammonia gas, aqueous ammonia, urea, etc., in an amount of 0.5 to 4.0%, in addition to these. Potassium phosphate, magnesium sulfate, ferrous sulfate heptahydrate, manganese chloride tetrahydrate, L-tyrosine, etc. are added in small amounts.
培養は好気的条件で行う。培養の間、液体培地のｐＨは
５から９に、温度は２４〜３７°Ｃに調節し、４８〜１
２０時間振盪または通気培養すれば好ましい結果が得ら
れる。Cultivation is performed under aerobic conditions. During the cultivation, the pH of the liquid medium was adjusted to 5 to 9, the temperature was adjusted to 24 to 37 °C, and the temperature was adjusted to 48 to 1.
Favorable results can be obtained by incubating with shaking or aeration for 20 hours.
培養液からＬ−フェニルアラニンは、イオン交換樹脂法
、活性炭吸着法など公知方法によって採取することがで
きる。L-phenylalanine can be collected from the culture solution by a known method such as an ion exchange resin method or an activated carbon adsorption method.
〈実施例〉 以下実施例により、本発明を具体的に説明する。<Example> The present invention will be specifically described below with reference to Examples.
実験例１
（Ｌ−チロシン要求変異株の取得）
５ ｍｌのし培地（トリプトン１％、酵母エキス０．５
％、塩化ナトリウム０．５％、ｐＨ７，５＞中で、３時
間培養した対数期のシトロバクタ−・フロインデイ０Ｆ
ＰＲ７２に通常の方法でＮ −メチル−Ｎ−−ニトロ−
Ｎ−ニトロソグアニジン処理＜ １５０μｒ／ｍｌ、３
７℃、１５分）し、Ｌ−チロシン要求変異株ＴＹＲＡ−
１（ｆｌ工研菌寄第９６１８号）を取得した。Experimental Example 1 (Obtaining an L-tyrosine-requiring mutant strain) 5 ml rice culture medium (tryptone 1%, yeast extract 0.5
%, sodium chloride 0.5%, pH 7.5> for 3 hours in log phase Citrobacter freundei 0F
PR72 was added with N-methyl-N-nitro-
N-nitrosoguanidine treatment < 150 μr/ml, 3
7°C, 15 minutes) and L-tyrosine-requiring mutant strain TYRA-
1 (fl Koken Bibori No. 9618).
実験例２
（Ｌ−チロシン要求変異株の検定）
下記第１表に示す各菌株を、Ｌ寒天培地で２４時間培養
し、その菌体なごく微量かきとり、Ｌ−チロシン無添加
およびＬ−チロシン２０μｇ ／ ｍｌを添加した下記
組成の合成寒天平板培地に薄く塗布し、３７℃で４日間
培養し、その生育の有無を観察した。Ｌ−チロシン無添
加寒天平板培地で生育できず、Ｌ−チロシン添加寒天平
板培地で生育するしのをＬ−チロシン要求変異株とした
。Experimental Example 2 (Analysis of L-Tyrosine-requiring Mutant Strains) Each strain shown in Table 1 below was cultured on L-agar medium for 24 hours, and a very small amount of the cells were scraped off. /ml was applied onto a synthetic agar plate medium having the following composition, cultured at 37°C for 4 days, and the presence or absence of growth was observed. A strain that could not grow on an agar plate medium without L-tyrosine and grew on an agar plate medium supplemented with L-tyrosine was designated as an L-tyrosine auxotrophic mutant.
合成寒天培地
グルコース ０．２％クエン酸
ナトリウム２水和物 ０．０５％ＫＨ２ＰＯ４０，２
％
に２ＨＰＯ４０，７％
（ＮＨ４）２３０４ ０．１％Ｍ ｇＳ
Ｏ４・７Ｈ２００，０１％
寒 天 １．
５％結果は第１表に示すとおりであり、本発明方法で使
用するＬ−チロシン要求性シトロバクター・フロインデ
イＴＹＲＡ−１は親株のシトロバクタ−・フロインデイ
０ＦＰＲ７２との比做により明らかにＬ−チロシン要求
性を獲得している。Synthetic agar medium Glucose 0.2% Sodium citrate dihydrate 0.05% KH2PO40,2
%2HPO40.7% (NH4)2304 0.1%M gS
O4.7H200.01% Agar 1.
The 5% results are shown in Table 1, and the L-tyrosine-requiring Citrobacter freundei TYRA-1 used in the method of the present invention is clearly L-tyrosine-requiring by comparison with the parent strain Citrobacter freundei 0FPR72. has acquired sexuality.
第 １ 表
実験例３
下記の組成の発酵用培地４Ｑｍｌを１１容三角フラスコ
に入れ、１１５°Ｃで１５分蒸気滅菌しな。これに第１
表に示す各菌株−白金耳植菌し、３０°Ｃで９６時間回
転振盪培培養た。Table 1 Experimental Example 3 Put 4Qml of fermentation medium with the following composition into an 11 volume Erlenmeyer flask and steam sterilize at 115°C for 15 minutes. This is the first
Each bacterial strain shown in the table was inoculated with a platinum loop and cultured in a rotary shaking medium at 30°C for 96 hours.
発酵用培地組成
グルコース １０％（ＮＨ４）２
ＳＯ４２，５％
ＫＨ２ＰＯ４０，０４％
ＭｇＳＯ４・７Ｈ２００，０４％
ＦｅＳＯ４・７Ｈ２０１Ｑｐｐｎ
ＭｎＣＪ ４ ・４Ｈ２０７ｐｐｎ
Ｌ−チロシン ０．１％ＣａＣＯ３４％
ｐＨ７，０（ＫＯＨで中和）
培養終了後、菌体、炭酸カルシウムを除去した液中のＬ
−フェニルアラニン濃度をロイコノストック・メセンテ
ロイデスＡＴＣＣ８０４２を用いる微生物定量法で定量
したところ第２表に示すような結果を得た。Fermentation medium composition Glucose 10% (NH4)2
SO42,5% KH2PO40,04% MgSO4・7H200,04% FeSO4・7H201Qppn MnCJ 4・4H207ppn L-tyrosine 0.1%CaCO34% pH 7,0 (neutralized with KOH) After culturing, remove bacterial cells and calcium carbonate L in the liquid
- Phenylalanine concentration was determined by a microbial assay using Leuconostoc mesenteroides ATCC 8042, and the results shown in Table 2 were obtained.
第 ２ 表
また、発酵終了後の液を強酸性陽イオン交換樹脂゛′ダ
イヤイオン”５Ｋ−１０４（Ｈ＋型）（三菱化成社製）
に通し、Ｌ−フェニルアラニンを吸着させ、該樹脂を水
洗後、０．５Ｎアンモニア水で溶出した溶出液を濃縮し
、活性炭処理して脱色し冷却後培養液１１あたり１．２
ｇのＬ−フェニルアラニンの結晶を得た。Table 2 Also, after the completion of fermentation, the liquid was treated with strongly acidic cation exchange resin "Diaion" 5K-104 (H+ type) (manufactured by Mitsubishi Kasei Corporation).
After passing the resin through water to adsorb L-phenylalanine, the resin was washed with water, the eluate was concentrated with 0.5N ammonia water, and treated with activated carbon to decolorize it.
g of L-phenylalanine crystals were obtained.
〈発明の効果〉
本発明の方法によれば、高価な前駆体を使用することな
く、かつＬ−フェニルアラニンを著量蓄積せしめること
ができるため、工業的に極めて有利である。<Effects of the Invention> According to the method of the present invention, it is possible to accumulate a significant amount of L-phenylalanine without using an expensive precursor, and therefore it is extremely advantageous industrially.
特許出顎人 東し株式会社Patent Jaw Man Toshi Co., Ltd.
Claims (1)
生育にＬ−チロシンを要求し、かつＬ−フェニルアラニ
ン生産能を有する微生物を、液体培地中で好気的に培養
してフェニルアラニンを培養液中に生成・蓄積せしめ、
該培養液よりＬ−フェニルアラニンを採取することを特
徴とする発酵法によるＬ−フェニルアラニンの製造法。Belongs to the genus Citrobacter,
A microorganism that requires L-tyrosine for growth and has the ability to produce L-phenylalanine is cultivated aerobically in a liquid medium to produce and accumulate phenylalanine in the culture solution,
A method for producing L-phenylalanine by a fermentation method, which comprises collecting L-phenylalanine from the culture solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP30519287A JPH01144987A (en) | 1987-12-01 | 1987-12-01 | Production of l-phenylalanine by fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP30519287A JPH01144987A (en) | 1987-12-01 | 1987-12-01 | Production of l-phenylalanine by fermentation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH01144987A true JPH01144987A (en) | 1989-06-07 |
Family
ID=17942157
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP30519287A Pending JPH01144987A (en) | 1987-12-01 | 1987-12-01 | Production of l-phenylalanine by fermentation |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01144987A (en) |
-
1987
- 1987-12-01 JP JP30519287A patent/JPH01144987A/en active Pending
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