JPS5934894A - Preparation of l-proline by fermentation - Google Patents
Preparation of l-proline by fermentationInfo
- Publication number
- JPS5934894A JPS5934894A JP14565482A JP14565482A JPS5934894A JP S5934894 A JPS5934894 A JP S5934894A JP 14565482 A JP14565482 A JP 14565482A JP 14565482 A JP14565482 A JP 14565482A JP S5934894 A JPS5934894 A JP S5934894A
- Authority
- JP
- Japan
- Prior art keywords
- proline
- lactose
- galactose
- medium
- fermentation
- 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
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 title claims abstract description 18
- 238000000855 fermentation Methods 0.000 title claims description 5
- 230000004151 fermentation Effects 0.000 title claims description 5
- 238000002360 preparation method Methods 0.000 title 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims abstract description 13
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims abstract description 11
- 239000008101 lactose Substances 0.000 claims abstract description 11
- 229930182830 galactose Natural products 0.000 claims abstract description 8
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims abstract description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 7
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 claims abstract description 7
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 7
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 6
- 244000005700 microbiome Species 0.000 claims abstract description 5
- 229960002429 proline Drugs 0.000 claims abstract 7
- 229930182821 L-proline Natural products 0.000 claims abstract 4
- 238000000034 method Methods 0.000 claims description 8
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims 1
- 239000005862 Whey Substances 0.000 abstract description 8
- 102000007544 Whey Proteins Human genes 0.000 abstract description 8
- 108010046377 Whey Proteins Proteins 0.000 abstract description 8
- 235000013336 milk Nutrition 0.000 abstract description 6
- 239000008267 milk Substances 0.000 abstract description 6
- 210000004080 milk Anatomy 0.000 abstract description 6
- 244000063299 Bacillus subtilis Species 0.000 abstract description 4
- 235000014469 Bacillus subtilis Nutrition 0.000 abstract description 4
- 244000068988 Glycine max Species 0.000 abstract description 2
- 235000010469 Glycine max Nutrition 0.000 abstract description 2
- 238000012258 culturing Methods 0.000 abstract description 2
- 230000000813 microbial effect Effects 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 6
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- -1 ammonium ions Chemical class 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 2
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 2
- 229910052697 platinum Inorganic materials 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 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 1
- 108010068370 Glutens Proteins 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 235000019687 Lamb Nutrition 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 235000021312 gluten Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 1
- 239000004615 ingredient Substances 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
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 229910017464 nitrogen compound Inorganic materials 0.000 description 1
- 150000002830 nitrogen compounds Chemical class 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 229910001414 potassium ion Inorganic materials 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- WTHDKMILWLGDKL-UHFFFAOYSA-N urea;hydrate Chemical compound O.NC(N)=O WTHDKMILWLGDKL-UHFFFAOYSA-N 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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】 本発明は発酵法によるし一プロリンの製造法に関する。[Detailed description of the invention] The present invention relates to a method for producing monoproline by a fermentation method.
■７−ブロリン（以下、単にプロリンと記す）の発酵法
による製造法に於ては、Ｌ−グルタミン酸生産性細菌か
ら誘導されたアミノ酸要求性のプロリン生産性変異株を
使用する方法や、バチルス属、エンンエリヒア属のし一
ヒスチジン、Ｌ−メチオニン又はＬ−ロイシン要求性変
異株を使用する方法（特公昭４４−’２６９１１号公報
参照）等が知られている。これらプロリン発酵法に於て
はグルコースが炭素源として使用されている。■ In the production method of 7-broline (hereinafter simply referred to as proline) by fermentation, there is a method using a proline-producing mutant strain requiring amino acids derived from L-glutamic acid-producing bacteria, a method using a proline-producing mutant strain of Bacillus spp. , a method using a histidine-, L-methionine-, or L-leucine-requiring mutant strain of the genus Ennerlichia (see Japanese Patent Publication No. 26911/1973), etc. are known. Glucose is used as a carbon source in these proline fermentation methods.
本発明者等は安価で利用価値の少ない乳ホエイを原料と
してプロリンを発酵生産する方法について種々研究を重
ねた結果、自然界から分離したバチルス属に属する細菌
の中に乳ホエイの主成分であるラクトースを資化してプ
ロリンを生産する能力を有する微生物があることを発見
した。The inventors of the present invention have repeatedly conducted various studies on a method of fermenting and producing proline using milk whey, which is inexpensive and has little utility value, as a raw material. As a result, the inventors have found that lactose, the main component of milk whey, is present in bacteria belonging to the genus Bacillus isolated from nature. discovered that there are microorganisms that have the ability to assimilate proline.
本発明はこの発見に基づいて完成されたものである。以
下、本発明について説明する。The present invention was completed based on this discovery. The present invention will be explained below.
本発明の方法に於て使用する微生物はガラクトース又は
ラクトースを資化して、プロリンを生産する能力を有す
る微生物であり、代表例としてバチルス・スブチリス
ＡＪ３０１０ＦＥＲＭ−Ｐ ６６７りが挙げられる。The microorganism used in the method of the present invention is a microorganism that has the ability to assimilate galactose or lactose to produce proline, and a representative example is Bacillus subtilis.
AJ3010FERM-P 667 is mentioned.
このようなバチルス属のプロリン生産能を有する［救生
物を培養する際に使用される培地は、ラクトース又はガ
ラクトースを炭素源として含有する以外は、特にかわっ
たものではない。ラクトース及びガラクトース源として
は、これらを含有する乳ホエイ、大豆ホエイ等が使用で
きる。これらの炭素源のほかに、副炭素涼としてグルツ
ース、７ユクロース等が培地に含まれていることもある
。炭素源のほかにはアンモニウムイオン、アンモニアガ
ス、アンモニア水等の通常の窒素源、リン酸イオン、マ
グネ／ラムイオン、カリイオン等の無殿イオン、更に必
要によりビタミン、アミノ酸等の有機微欧栄養呉が培地
中に含まれる。The medium used for culturing such Bacillus species having proline-producing ability is not particularly different except that it contains lactose or galactose as a carbon source. As a source of lactose and galactose, milk whey, soybean whey, etc. containing these can be used. In addition to these carbon sources, the culture medium may also contain supplementary carbon sources such as gluten and 7-ucrose. In addition to carbon sources, ordinary nitrogen sources such as ammonium ions, ammonia gas, and ammonia water, non-deposited ions such as phosphate ions, magne/lamb ions, and potassium ions, and organic micronutrients such as vitamins and amino acids are also used as necessary. Contained in the medium.
培養は好気的条件下で行われる。培養の間、培養温度は
２７ないし３７℃の範囲内のめ当な温度に、培地ｐＨは
、５．５から７．５の範囲の適当なｐ Ｈに、それぞれ
保つのが望ましい。かくして、］ないし４４日も培養を
続ければ培地中にブーリンが生成蓄積される。Cultivation is carried out under aerobic conditions. During cultivation, it is desirable to maintain the culture temperature at an appropriate temperature within the range of 27 to 37°C and the medium pH at an appropriate pH within the range of 5.5 to 7.5. Thus, if the culture is continued for 44 days, bourin will be produced and accumulated in the medium.
培地中に蓄積されたプロリンを採取するには、通常の方
法で行うことができる。Proline accumulated in the medium can be collected by conventional methods.
以下、実施例にて説明する。Examples will be described below.
実施例１
ガラクトース ８ ？／ｄｌ、 Ｎ Ｈ４Ｃ１１，０シ
汝。Example 1 Galactose 8? /dl, N H4C11,0 you.
Ｋｃａ Ｏ，２ｑ７ｔｉ７！、 ＫＨ２ＰＯ４０，Ｉｒ
Ｊ、 ）＃ＳＯ４・７Ｈ２００，０４？／ｄｉ 、
カザミノ酸０ 、４ｆｌ／ｄｌ 、 ＦｅＳＯ４・７［
０１Ｈ９／ ｄ７！、 ＭｎＳＯ４・４Ｈ，ＯＩ ｍｇ
／ｄｌを含み、ｐＨ７，０に調節した培地を５ｏｏ＋Ｍ
！フラスコに２０ｍ１スつ入れ殺菌した。これにバチル
ス・ズブチリスＡＪ ３０１０を１白金耳植えつけ、
３１℃で４８時間培養した。なお、培養途中、４５９／
ｄ７！の尿素水を０．３ｍｌずつ添加し、ｐＨの低下を
防止した。Kca O,2q7ti7! , KH2PO40,Ir
J, )#SO4・7H200,04? /di,
Casamino acids 0, 4 fl/dl, FeSO4・7 [
01H9/d7! , MnSO4・4H, OI mg
/dl and adjusted to pH 7.0 at 5oo+M
! One 20ml flask was placed and sterilized. One platinum loop of Bacillus subtilis AJ 3010 was planted in this,
The cells were cultured at 31°C for 48 hours. In addition, during cultivation, 459/
d7! of urea water was added in 0.3 ml portions to prevent the pH from decreasing.
培養終了時におけるし一プロリンの蓄積量は１２０ ｍ
ｇ／ｄｅであった。The amount of Shiichiproline accumulated at the end of the culture was 120 m
g/de.
実施例２
下記の組成の酸沈・乳ホエイを乳糖として４ ？ ／
ｄｌ、 （ＮＩ−１４）２ Ｓｏイ ２．５？／
ｄ＆ ＫＨ，ＰＯイ ０．２？／ｄｌ。Example 2 Acid-precipitated milk whey with the following composition was used as lactose.4? /
dl, (NI-14)2 Soi 2.5? /
d & KH, PO i 0.2? /dl.
ＭｇＳＯ４・７Ｈ２００、１？／ｄｉ １ 酵母エキ
ス０−０５ ｔｙ’ｄｌ。MgSO4・7H200,1? /di 1 yeast extract 0-05 ty'dl.
ＦｅＳＯ４１Ｈ，，０１ｍ９／ｄｅ、 ＭｎＳＯ４・
４Ｈ２０１Ｍ９／ｄｉ及び炭酸カル／ラム２．５？／ｄ
／、（別殺添加）を含み、Ｋ ＯＨでｐ Ｈ７，２に調
節した培地を調製し５００ｍ１フラスコンこ２０ｍ１ず
つ入れ、殺菌した。これにバチルス・ズブチリスＡＪ３
０１０１白金耳植えつけ、３１℃で７２時間培養した。FeSO41H,,01m9/de, MnSO4・
4H201M9/di and Cal/Rum carbonate 2.5? /d
/, (separately added), a medium adjusted to pH 7.2 with KOH was prepared, placed in 20ml each of 500ml flasks, and sterilized. In this, Bacillus subtilis AJ3
0101 platinum loops were planted and cultured at 31°C for 72 hours.
培養終了時におけるし一プロリンの蓄積量は、１１０１
１９／ｄ７！であった。The amount of proline accumulated at the end of the culture was 1101
19/d7! Met.
尚、酸沈・乳ホエイの成分は、水分９３．９％、乳糖４
．５％、全固形分６．１％、窒素化合物０．８０％、灰
分０．７０９６、脂肪０．１％であった。In addition, the ingredients of acid precipitated milk whey are 93.9% water and 4% lactose.
．． 5%, total solids 6.1%, nitrogen compounds 0.80%, ash 0.7096, and fat 0.1%.
特許出願人 味の素株式会社Patent applicant: Ajinomoto Co., Inc.
Claims (1)
りＬ−プロリンを生産する能力を有する微生物を、ラク
トース又はガラクトースを炭素源として含有する液体培
地中で培養し、培養液中にＬ−プロリンを生成せしめ、
該プロリンを採取することを特徴とする発酵法によるし
一プロリンの製造法。A microorganism belonging to the genus Bacillus that has the ability to produce L-proline from lactose or lacto-X is cultured in a liquid medium containing lactose or galactose as a carbon source, and L-proline is produced in the culture solution. Seshime,
A method for producing proline by a fermentation method, which comprises collecting the proline.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14565482A JPS5934894A (en) | 1982-08-23 | 1982-08-23 | Preparation of l-proline by fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14565482A JPS5934894A (en) | 1982-08-23 | 1982-08-23 | Preparation of l-proline by fermentation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS5934894A true JPS5934894A (en) | 1984-02-25 |
Family
ID=15390007
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP14565482A Pending JPS5934894A (en) | 1982-08-23 | 1982-08-23 | Preparation of l-proline by fermentation |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5934894A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6252344U (en) * | 1985-09-20 | 1987-04-01 |
-
1982
- 1982-08-23 JP JP14565482A patent/JPS5934894A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6252344U (en) * | 1985-09-20 | 1987-04-01 | ||
| JPH0545874Y2 (en) * | 1985-09-20 | 1993-11-29 |
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