JPS62195291A - Production of fats and oils containing gamma-linolenic ester by tissue culture of evening primrose - Google Patents
Production of fats and oils containing gamma-linolenic ester by tissue culture of evening primroseInfo
- Publication number
- JPS62195291A JPS62195291A JP61036390A JP3639086A JPS62195291A JP S62195291 A JPS62195291 A JP S62195291A JP 61036390 A JP61036390 A JP 61036390A JP 3639086 A JP3639086 A JP 3639086A JP S62195291 A JPS62195291 A JP S62195291A
- Authority
- JP
- Japan
- Prior art keywords
- medium
- acid ester
- evening primrose
- callus
- linolenic acid
- 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.)
- Granted
Links
- 241000219925 Oenothera Species 0.000 title claims abstract description 22
- 235000004496 Oenothera biennis Nutrition 0.000 title claims abstract description 18
- 238000004519 manufacturing process Methods 0.000 title claims description 14
- 239000003921 oil Substances 0.000 title abstract description 40
- 239000003925 fat Substances 0.000 title abstract description 39
- 150000002148 esters Chemical class 0.000 title abstract description 13
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 claims abstract description 13
- UQHKFADEQIVWID-UHFFFAOYSA-N cytokinin Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1CC(O)C(CO)O1 UQHKFADEQIVWID-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000004062 cytokinin Substances 0.000 claims abstract description 10
- 239000003375 plant hormone Substances 0.000 claims abstract description 10
- 229930192334 Auxin Natural products 0.000 claims abstract description 9
- 239000002363 auxin Substances 0.000 claims abstract description 9
- 241000196324 Embryophyta Species 0.000 claims abstract description 8
- FAIXYKHYOGVFKA-UHFFFAOYSA-N Kinetin Natural products N=1C=NC=2N=CNC=2C=1N(C)C1=CC=CO1 FAIXYKHYOGVFKA-UHFFFAOYSA-N 0.000 claims abstract description 6
- QANMHLXAZMSUEX-UHFFFAOYSA-N kinetin Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CO1 QANMHLXAZMSUEX-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229960001669 kinetin Drugs 0.000 claims abstract description 6
- 235000019197 fats Nutrition 0.000 claims description 41
- VZCCETWTMQHEPK-UHFFFAOYSA-N gamma-Linolensaeure Natural products CCCCCC=CCC=CCC=CCCCCC(O)=O VZCCETWTMQHEPK-UHFFFAOYSA-N 0.000 claims description 30
- 235000020664 gamma-linolenic acid Nutrition 0.000 claims description 30
- 229960002733 gamolenic acid Drugs 0.000 claims description 30
- -1 γ-linolenic acid ester Chemical class 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 10
- 235000014113 dietary fatty acids Nutrition 0.000 claims description 8
- 239000000194 fatty acid Substances 0.000 claims description 8
- 229930195729 fatty acid Natural products 0.000 claims description 8
- 150000004665 fatty acids Chemical class 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- VZCCETWTMQHEPK-QNEBEIHSSA-N gamma-linolenic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/CCCCC(O)=O VZCCETWTMQHEPK-QNEBEIHSSA-N 0.000 claims description 5
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 4
- 239000003617 indole-3-acetic acid Substances 0.000 claims description 4
- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 claims description 3
- 210000004027 cell Anatomy 0.000 claims 2
- OVSKIKFHRZPJSS-DOMIDYPGSA-N 2-(2,4-dichlorophenoxy)acetic acid Chemical compound OC(=O)[14CH2]OC1=CC=C(Cl)C=C1Cl OVSKIKFHRZPJSS-DOMIDYPGSA-N 0.000 claims 1
- 210000004748 cultured cell Anatomy 0.000 claims 1
- 206010020649 Hyperkeratosis Diseases 0.000 abstract description 30
- 229920001817 Agar Polymers 0.000 abstract description 6
- 239000008272 agar Substances 0.000 abstract description 6
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 abstract description 5
- 239000007787 solid Substances 0.000 abstract description 5
- 239000000843 powder Substances 0.000 abstract description 3
- 230000035784 germination Effects 0.000 abstract description 2
- YIVXMZJTEQBPQO-UHFFFAOYSA-N 2,4-DB Chemical compound OC(=O)CCCOC1=CC=C(Cl)C=C1Cl YIVXMZJTEQBPQO-UHFFFAOYSA-N 0.000 abstract 1
- 235000019198 oils Nutrition 0.000 description 38
- 239000002609 medium Substances 0.000 description 29
- 239000002253 acid Substances 0.000 description 16
- 235000020661 alpha-linolenic acid Nutrition 0.000 description 4
- 238000012136 culture method Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 230000006698 induction Effects 0.000 description 4
- 229960004488 linolenic acid Drugs 0.000 description 4
- KQQKGWQCNNTQJW-UHFFFAOYSA-N linolenic acid Natural products CC=CCCC=CCC=CCCCCCCCC(O)=O KQQKGWQCNNTQJW-UHFFFAOYSA-N 0.000 description 4
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 235000014593 oils and fats Nutrition 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerol Natural products OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 239000010779 crude oil Substances 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 235000013402 health food Nutrition 0.000 description 2
- 230000005802 health problem Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 229940094443 oxytocics prostaglandins Drugs 0.000 description 2
- 238000004161 plant tissue culture Methods 0.000 description 2
- 150000003180 prostaglandins Chemical class 0.000 description 2
- OYHQOLUKZRVURQ-NTGFUMLPSA-N (9Z,12Z)-9,10,12,13-tetratritiooctadeca-9,12-dienoic acid Chemical compound C(CCCCCCC\C(=C(/C\C(=C(/CCCCC)\[3H])\[3H])\[3H])\[3H])(=O)O OYHQOLUKZRVURQ-NTGFUMLPSA-N 0.000 description 1
- HXKWSTRRCHTUEC-UHFFFAOYSA-N 2,4-Dichlorophenoxyaceticacid Chemical compound OC(=O)C(Cl)OC1=CC=C(Cl)C=C1 HXKWSTRRCHTUEC-UHFFFAOYSA-N 0.000 description 1
- 102100034544 Acyl-CoA 6-desaturase Human genes 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010019133 Hangover Diseases 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 108010037138 Linoleoyl-CoA Desaturase Proteins 0.000 description 1
- 206010036618 Premenstrual syndrome Diseases 0.000 description 1
- 241000245063 Primula Species 0.000 description 1
- 235000016311 Primula vulgaris Nutrition 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- DTOSIQBPPRVQHS-PDBXOOCHSA-N alpha-linolenic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(O)=O DTOSIQBPPRVQHS-PDBXOOCHSA-N 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 150000001735 carboxylic acids Chemical class 0.000 description 1
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000004626 essential fatty acids Nutrition 0.000 description 1
- 125000004494 ethyl ester group Chemical group 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 235000020256 human milk Nutrition 0.000 description 1
- 210000004251 human milk Anatomy 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 150000002617 leukotrienes Chemical class 0.000 description 1
- 229940127554 medical product Drugs 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 229940039177 nioxin Drugs 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
3、 の量 な−
(産業上の利用分野)
本発明はγ−リノレン酸エステル含有油脂の製造方法、
特に2組織培養によりγ−リノレン酸エステル含有量の
高い油脂を製造する方法に関する。[Detailed Description of the Invention] 3. Amount of (Industrial Application Field) The present invention provides a method for producing a γ-linolenic acid ester-containing fat or oil,
In particular, the present invention relates to a method for producing fats and oils with a high content of γ-linolenic acid ester by two-tissue culture.
(従来の技術)
γ−リノレン酸(cis、 cis、 cis−6・9
−12−octadecatrienic acid)
はカルボン酸末端から6゜9および12番目の炭素に不
飽和結合を持つ炭素数18のシス型の脂肪酸で、母乳1
月見草種子などにグリセリンエステルとして存在する。(Prior art) γ-linolenic acid (cis, cis, cis-6/9
-12-octadecatrinenic acid)
is an 18-carbon cis-type fatty acid with unsaturated bonds at the 6°9 and 12th carbons from the carboxylic acid terminal, and is found in breast milk.
It exists as a glycerin ester in seeds such as evening primrose.
γ−リルン酸は生体内においては、必須脂肪酸であるリ
ノール酸から不飽和化酵素(Δ−6−desatura
se)により誘導される。γ−リルン酸はビスホモγ−
リノレン酸となり、さらに生体内で多様な生理活性を示
すプロスタグランジンやロイコトリエンへと変換される
。この代謝経路における第1の律速因子は上記不飽和化
酵素である。この酵素の働きは。In the body, γ-lylinic acid is synthesized from linoleic acid, an essential fatty acid, by a desaturase (Δ-6-desaturase).
se). γ-Lilunic acid is bishomoγ-
It becomes linolenic acid, which is further converted in vivo to prostaglandins and leukotrienes that exhibit various physiological activities. The first rate-limiting factor in this metabolic pathway is the desaturase mentioned above. What is the function of this enzyme?
糖尿病、過剰のアルコール、老化などが原因となって低
下する。その結果、プロスタグランジンなどの生成が抑
制され、健康障害が起こることが知られている。γ−リ
ノレン酸およびその塩やエステルはこれら健康障害に対
する治療用栄養素として有効である。例えば2月経前症
候群、心臓疾患。Diabetes, excess alcohol, aging, etc. can lead to a decline. As a result, the production of prostaglandins and the like is suppressed, which is known to cause health problems. γ-Linolenic acid and its salts and esters are effective as therapeutic nutrients for these health disorders. For example, premenstrual syndrome, heart disease.
血中コレステロールの低下、高血圧、湿疹、アレルギー
、二日酔などに効果があることが知られている。It is known to be effective in lowering blood cholesterol, high blood pressure, eczema, allergies, and hangovers.
γ−リルン酸は既述のように、自然界においては通常そ
のグリセリンエステルである油脂の形態で存在し、かつ
γ−リルン酸エステル単独では存在しないため、γ−リ
ルン酸エステルをはじめ他の脂肪酸エステルを含有する
油脂が採取・利用されている。このようなγ−リルン酸
エステル含有油脂は1通常、植物種子、特に月見草(マ
ツヨイグサ; 0enothera biennis
)などのマツヨイグサ属植物の種子から抽出される。し
かし。As mentioned above, γ-lylunic acid usually exists in the form of oil and fat, which is its glycerin ester, in the natural world, and γ-lylunic acid ester does not exist alone. Oils and fats containing are collected and used. Such gamma-lyrunic acid ester-containing oils and fats are usually obtained from plant seeds, especially evening primrose (evening primrose).
) is extracted from the seeds of plants of the evening primrose genus. but.
マツヨイグサ属植物の種子中に含有される油脂は微量で
あるうえ、該油脂中のγ−リルン酸エステル含量も低い
。通常、このような油脂には、γ−リノレン酸エステル
を構成するγ−リルン酸゛が総油脂の脂肪酸組成の8〜
13重量%の割合で含まれるにすぎない。そのため、γ
−リノレン酸エステル含有油脂を得るには大量の原料(
種子)と大規模な設備とを必要とする。さらに、マツヨ
イグサの栽培には一年を必要とし9種子の収穫時期も限
られているため安定した生産ができない。種子中の油脂
組成や含有量にバラツキがあるため。The amount of oil and fat contained in the seeds of plants of the genus Evening Primrose is small, and the content of γ-lylunic acid ester in the oil is also low. Normally, in such fats and oils, γ-linolenic acid, which constitutes γ-linolenic acid ester, accounts for 8 to 8% of the total fatty acid composition of fats and oils.
It is contained in a proportion of only 13% by weight. Therefore, γ
-To obtain linolenic acid ester-containing fats and oils, a large amount of raw materials (
seeds) and large-scale equipment. Furthermore, it takes one year to cultivate evening primrose, and the harvest period for the nine seeds is also limited, making stable production impossible. This is because there are variations in the oil and fat composition and content in the seeds.
一定した品質のγ−リノレン酸エステル含有油脂が得ら
れない欠点もある。There is also the drawback that γ-linolenic acid ester-containing fats and oils of constant quality cannot be obtained.
(発明が解決しようとする問題点) 本発明は上記従来の欠点を解決するものであり。(Problem that the invention attempts to solve) The present invention solves the above-mentioned conventional drawbacks.
その目的とするところは、γ−リノレン酸エステル含有
率の高いγ−リノレン酸エステル含有油脂を効果的に製
造する方法を提供することにある。The purpose is to provide a method for effectively producing γ-linolenic acid ester-containing fats and oils with a high γ-linolenic acid ester content.
本発明の他の目的は、マツヨイグサ種子の収穫時期や品
質に関係なく安定した品質のγ−リノレン酸エステル含
有油脂を安価に製造する方法を提供゛することにある0
本発明の他の目的は、上記優れた品質のγ−リノレン酸
エステル含有油脂を組織培養法により製造する方法を提
供することにある。Another object of the present invention is to provide a method for inexpensively producing γ-linolenic acid ester-containing oil and fat of stable quality regardless of the harvest time or quality of evening primrose seeds.
Another object of the present invention is to provide a method for producing the above-mentioned excellent quality γ-linolenic acid ester-containing oil and fat by a tissue culture method.
(問題点を解決するための手段および作用)本発明のγ
−リノレン酸エステル含有油脂の製造方法はマツヨイグ
サ(Oenothera)属植物を組織培養し細胞塊を
得る工程、および該細胞塊を植物ホルモンを含有する有
機培地で培養し、γ−リルン酸エステル含有油脂を生産
させる工程、を包含し、そのことにより上記目的が達成
される。(Means and effects for solving the problems) γ of the present invention
- The method for producing linolenic acid ester-containing fats and oils includes the steps of tissue culturing evening primrose (Oenothera) plants to obtain cell masses, culturing the cell masses in an organic medium containing plant hormones, and producing γ-linolenic acid ester-containing fats and oils. The above-mentioned objective is achieved by the process of producing.
本発明方法によれば、まず9月見草、メマツヨイグサ、
オオマツヨイグサなどのマツヨイグサ(Qenothe
ra)属植物の根、茎9葉などの適当な部分を用いて2
通常の方法により細胞塊(カルス)の誘導を行う。使用
される有機培地は何ら格別である必要はなく、植物組織
培養に通常用いられるムラシゲ−スクーグ(Muras
hige−5koog)の培地、ホワイト(White
)の培地、リンスマイヤー−スクーグ(Linssai
er−Skoog)の培地、ガンスレッド(Ganth
eret)の培地、トレンチ(Tulecke)の培地
、・モーレル(Motel)の培地などが用いられうる
。なかでもMurash ige−Skoogの培地や
Whiteの培地が好適に用いられる。これに、カルス
の誘導をより促進させるために必要に応じて植物ホルモ
ン(オーキシン類、サイトカイニン類など)が添加され
る。オーキシン類には例えば、2・4−ジクロロフェノ
キシ酢酸(2・4−D)、インドール酢酸(IAA)、
ナフタレン酢酸(N、A A )がある。According to the method of the present invention, first, September primrose, evening primrose,
Evening primrose (Qenothe)
2 using appropriate parts such as roots, stems, leaves, etc. of plants of the genus ra).
A cell mass (callus) is induced by a conventional method. The organic medium used does not need to be anything special; Murashige-Skoog, which is commonly used for plant tissue culture,
hige-5koog) medium, White
) culture medium, Linsmayer-Skoog (Linssai)
er-Skoog) medium, Ganth
eret's medium, Tulecke's medium, Motel's medium, etc. may be used. Among them, Murashige-Skoog's medium and White's medium are preferably used. Plant hormones (auxins, cytokinins, etc.) are added to this as necessary to further promote callus induction. Examples of auxins include 2,4-dichlorophenoxyacetic acid (2,4-D), indoleacetic acid (IAA),
There is naphthaleneacetic acid (N, AA).
サイトカイニン類には1例えば、カイネチン、ベンジル
アデニンがある。Examples of cytokinins include kinetin and benzyladenine.
上記誘導用培地に植物ホルモンが含有されない場合には
、得られたカルスに含まれるγ−リノレン酸エズテル含
有油脂は極めて少量である。そこで、このカルスにγ−
リノレン酸エステル含有油脂を生産させるべく9次に、
植物ホルモンを含有する有機培地が用いられる。この培
地はMurashige−Skoogの培地など上記い
ずれの培地も使用可能である。When the above-mentioned induction medium does not contain a plant hormone, the amount of γ-linolenic acid ester-containing oil and fat contained in the obtained callus is extremely small. Therefore, this callus has γ−
In order to produce fats and oils containing linolenic acid ester,
An organic medium containing plant hormones is used. As this medium, any of the above-mentioned media such as Murashige-Skoog's medium can be used.
有機培地へ添加される植物ホルモンの種類は特に限定さ
れるものではなく、1種単独で添加してもあるいは2種
以上を混合して添加してもよい。The types of plant hormones added to the organic medium are not particularly limited, and one type may be added alone or two or more types may be added as a mixture.
γ−リノレン酸エステル含有油脂の生産効率を考慮する
とオーキシン類およびサイトカイニン類の両者を添加す
る態様が好適である。この場合にはオーキシン類として
ナフタレン酢酸を、サイトカイニン類としてカイネチン
および/またはベンジルアデニンを用いるのが特に好ま
しい。上記側々の植物ホルモンの添加量は微量でよく9
通常、その合計量が0.01〜1100pp 、好まし
くは0.1〜10ppmの範囲にある。過少にすぎると
カルスが増殖しにり<、γ−リルン酸エステル含有油脂
の生産量が低く、かつ該油脂中のγ−リノレン酸エステ
ル含有率が低い。過剰にすぎてもコスト高となるだけで
γ−リノレン酸エステル含有油脂生産の効果は少ない。Considering the production efficiency of γ-linolenic acid ester-containing fats and oils, it is preferable to add both auxins and cytokinins. In this case, it is particularly preferable to use naphthaleneacetic acid as the auxin and kinetin and/or benzyladenine as the cytokinin. The amounts of the above-mentioned plant hormones added may be small.9
Usually, the total amount is in the range of 0.01 to 1100 ppm, preferably 0.1 to 10 ppm. If the amount is too low, callus will proliferate, the production amount of γ-linolenic acid ester-containing oil and fat will be low, and the γ-linolenic acid ester content in the oil and fat will be low. If it is too much, the cost will only increase and the effect of producing γ-linolenic acid ester-containing fats and oils will be small.
培養法は2例えば、液体培地を用いた振盪培養法であっ
ても固体培地を用いた静置培養法であってもよい。固体
の培地としては1例えば寒天を約0.9%含有するMu
rashige−Skoogの培地などが用いられる。The culture method may be a shaking culture method using a liquid medium or a static culture method using a solid medium. As a solid medium, for example, Mu containing about 0.9% agar
Rashige-Skoog's medium or the like is used.
培養温度は15〜36℃、好ましくは25℃前後である
。このような条件下で培養されたカルスは、γ−リルン
酸エステル含有油脂を高割合で生産するようになる。The culture temperature is 15 to 36°C, preferably around 25°C. Callus cultured under such conditions produces a high proportion of γ-lylunic acid ester-containing fats and oils.
このようにして得られたカルスをすりつぶして有機溶剤
で抽出を行うとγ−リノレン酸エステル含有油脂(粗油
)が得られる。カルスを乾燥し。When the callus thus obtained is ground and extracted with an organic solvent, a γ-linolenic acid ester-containing fat (crude oil) is obtained. Dry the callus.
その含水率を10重量%以下、好ましくは5重量%以下
とし、粉末状とした後に有機溶剤による抽出を行っても
よい。有機溶剤として、は、n−へキサン、アセトン、
エタノール、クロロホルムなどが用いられる。得られた
粗油は常法により精製されうる。このようにして得られ
る油脂中のγ−リルン酸エステル含有率はγ−リルン酸
に換算すると総涜脂の脂肪酸組成の20重量%以上1通
常20〜30重量%である。The water content thereof may be set to 10% by weight or less, preferably 5% by weight or less, and after being made into a powder, extraction with an organic solvent may be performed. As organic solvents, n-hexane, acetone,
Ethanol, chloroform, etc. are used. The obtained crude oil can be purified by conventional methods. The content of γ-lylunic acid ester in the oil thus obtained is usually 20 to 30% by weight of the fatty acid composition of the total fatty acid when converted to γ-lylphinic acid.
(作用)
本発明方法によりマツヨイグサ属植物から誘導したカル
スを植物ホルモンを含有する培地で培養すると、該カル
スによりγ−リノレン酸エステル含量の高い油脂が生産
されるようになる。このカルスは速やかに増殖し、長期
間にわたって継代培養しても、その増殖率や油脂の含有
率は変化しない。油脂の組成も変化しない。このような
方法によれば、マツヨイグサ属植物を自然栽培し、油脂
、を得る方法に比べ、栽培のための広い土地を必要とせ
ず、季節に左右されず、かつ短期間で大量のγ−リノレ
ン酸エステル含有油脂を得ることが可能である。(Function) When callus derived from plants of the genus Evening Primrose by the method of the present invention is cultured in a medium containing plant hormones, the callus produces fats and oils with a high content of γ-linolenic acid ester. This callus proliferates rapidly, and its proliferation rate and fat content do not change even when it is subcultured over a long period of time. The composition of fats and oils also remains unchanged. Compared to the method of naturally cultivating evening primrose plants to obtain oils and fats, this method does not require a large amount of land for cultivation, is not affected by the seasons, and can produce a large amount of γ-linolene in a short period of time. It is possible to obtain acid ester-containing fats and oils.
カルスから抽出して得たγ−リノレン酸エステル含有油
脂は1例えば、不飽和化酵素の不活性化による健康障害
の解消などを目的とした健康食品として利用される。γ
−リルン酸エステル含有油脂をケン化してγ−リルン酸
とした後、メチルエステルやエチルエステルとしビタミ
ンFの製造原料に用いるなど、医療品原料としても有用
である。カルスからγ−リルン酸エステル含有油脂を抽
出する以外にも乾燥カルスを必要に応じて粉砕した後そ
のまま、もしくは他の食品に混合して利用することもで
きる。γ-Linolenic acid ester-containing fats and oils extracted from callus are used as health foods, for example, to eliminate health problems caused by inactivation of desaturases. γ
- It is also useful as a raw material for medical products, such as by saponifying lylunic acid ester-containing fats and oils to produce γ-lylunic acid, and then converting it to methyl ester or ethyl ester, which is used as a raw material for producing vitamin F. In addition to extracting γ-lyrunic acid ester-containing fats and oils from callus, dried callus can also be used as it is or mixed with other foods after pulverizing if necessary.
(実施例) 本発明を実施例について説明する。(Example) The present invention will be described with reference to examples.
!施■土
(A)カルスの誘導および培養: Murashige
−Skoogの培地にオーキシンとして2・4−ジクロ
ロフェノキシ酢酸(2・4−D)を1 ppm+サイト
カイニンとしてカイネチンをo、tppmとなるように
添加した。これに寒天粉末を0.9wt%となるように
加えて寒天溶液とし、オートクレーブで125℃にて1
5分間殺菌した。この寒天溶液で固体培地を調製した。! Soil application (A) Callus induction and culture: Murashige
-2,4-dichlorophenoxyacetic acid (2,4-D) as auxin was added to Skoog's medium at 1 ppm + kinetin as cytokinin at o, tppm. Add agar powder to 0.9wt% to make an agar solution, and autoclave at 125℃ for 1 hour.
Sterilized for 5 minutes. A solid medium was prepared with this agar solution.
別に、無菌的に発芽させたマツヨイグサ(Oenoth
era biennis)を発芽後3日目に採取した
。Separately, aseptically germinated evening primrose (Oenoth)
era biennis) were collected on the third day after germination.
マツヨイグサの茎の径1〜51−の部分を長さ約51−
に切断し、上記の固体培地に置床した。これを暗所にて
25℃に保ち2〜3週間培養すると、茎の切断面から白
色カルスが発生した。カルスの発生したマツヨイグサの
茎を上記と同様の新たな培地に移植し、同条件で培養を
続けてさらにカルスを増殖させた。次に、増殖したカル
スのみを新たな培地に移植し、12代0まで継代培養を
行った。Measure the part of the evening primrose stem with a diameter of 1 to 51 mm to a length of approximately 51 mm.
It was cut into pieces and placed on the solid medium described above. When this was cultured in the dark at 25° C. for 2 to 3 weeks, white callus was generated from the cut surface of the stem. The evening primrose stems in which callus had developed were transplanted to a new medium similar to the above, and culture was continued under the same conditions to further propagate callus. Next, only the proliferated calli were transplanted to a new medium and subcultured until generation 12.
(B) γ−リノレン酸エステル含有油脂を含むカル
スの培養および油脂の抽出: Murashige−
Skoogの培地にオーキシンとしてNAAを2ppm
+ サイトカイニンとしてカイネチンを0.lppm+
の割合で添加した。これを(A)項と同様の方法で殺菌
した。(B) Culture of callus containing γ-linolenic acid ester-containing oil and fat extraction: Murashige-
2 ppm NAA as auxin in Skoog's medium
+ Kinetin as cytokinin 0. lppm+
It was added at a ratio of This was sterilized in the same manner as in section (A).
この培地300++1を11の三角フラスコに入れ。Pour 300++1 of this medium into 11 Erlenmeyer flasks.
これに(A)項で継代培養した12代0まカルスをこの
培地に移植し、暗所にて30℃で往復式振盪機(90ス
トローク/sin、)により振盪培養を4週間行った。The 12th generation callus subcultured in section (A) was then transplanted into this medium, and cultured with shaking in the dark at 30°C using a reciprocating shaker (90 strokes/sin) for 4 weeks.
培養液を濾過してカルスを得、これを60℃で24時間
乾燥したところ・0.6gの乾燥カルスが得られた。こ
れを乳鉢で粉砕しn−へキサンを加えて抽出を行ったと
ころ、 0.02gの油脂が得られた。The culture solution was filtered to obtain callus, which was dried at 60° C. for 24 hours. 0.6 g of dry callus was obtained. When this was crushed in a mortar and extracted by adding n-hexane, 0.02 g of oil and fat was obtained.
これを常法によりメチルエステル化し、油脂を構成する
脂肪酸の分析を行った。その結果を表1に示す。表1に
おいて炭素数18.二重結合数3の脂肪酸はγ−リノレ
ン酸を示す。This was methyl esterified using a conventional method, and the fatty acids constituting the oil and fat were analyzed. The results are shown in Table 1. In Table 1, the number of carbon atoms is 18. A fatty acid with three double bonds indicates γ-linolenic acid.
表1
(^)カルスの誘導および培II:実施例1 (^)項
と同様である。Table 1 (^) Callus induction and culture II: Example 1 Same as section (^).
(B) γ−リノレン酸エステル含有油脂を含むカル
スの培養および油脂の抽出ニオ−キシンとしてIAAを
ippm、そしてサイトカイニンとしてベンジルアデニ
ンをtpp−の割合で含有する一hiteの液体培地を
調製し、実施例1 (1)項と同様に殺菌を行った。こ
の培地2501Illを500m1の三角フラスコに入
れ、これに本実施例(A)項で得られたカルスを加えて
暗所にて25℃で往復式振盪機(90ストローク/si
n、)により振盪培養を行った。(B) Cultivation of callus containing γ-linolenic acid ester-containing oil and fat extraction A one-hite liquid medium containing IAA as nioxin at ippm and benzyladenine as cytokinin at tpp was prepared and carried out. Example 1 Sterilization was performed in the same manner as in section (1). 2501 Ill of this medium was placed in a 500 ml Erlenmeyer flask, the callus obtained in Example (A) was added thereto, and the mixture was heated in the dark at 25°C using a reciprocating shaker (90 strokes/si).
Shaking culture was performed using the following method.
4週間培養を続けた後、カルスを濾別し、凍結乾燥を行
い0.9gの乾燥カルスを得た。この乾燥カルスにクロ
ロホルム−メタノール混液(2:1(v/v))を加え
てホモジナイザーにかけ、抽出を行った。得られた油脂
は0.04gであった。この油脂を実施例1と同様の方
法でメチルエステル化し。After continuing the culture for 4 weeks, the callus was filtered and freeze-dried to obtain 0.9 g of dry callus. A chloroform-methanol mixture (2:1 (v/v)) was added to the dried callus and extracted using a homogenizer. The amount of oil and fat obtained was 0.04 g. This fat and oil was methyl esterified in the same manner as in Example 1.
脂肪酸の分析を行った。その結果を表2に示す。Fatty acid analysis was performed. The results are shown in Table 2.
表2
(発明の効果)
本発明方法によれば、このように、γ−リノレン酸エス
テルを高い割合で含有するγ−リノレン酸エステル含有
油脂が効果的に得られる。植物の組織培養を利用するた
め、季節に左右されずかつ栽培のための広大な土地を必
要とせずに短期間で大量のγ−リルン酸エステル含有油
脂を得ることが可能となる。このようなγ−リルン酸エ
ステル含有油脂は9例えば、不飽和化酵素(Δ−6−d
esaturase)の活性低下によって生じる各種健
康障害の解消などを目的とした健康食品に用いられる。Table 2 (Effects of the Invention) According to the method of the present invention, a γ-linolenic acid ester-containing fat or oil containing a high proportion of γ-linolenic acid ester can be effectively obtained. Since plant tissue culture is used, it is possible to obtain a large amount of γ-lyrunic acid ester-containing oil and fat in a short period of time without being affected by the season and without requiring a large area of land for cultivation. Such γ-lylunic acid ester-containing fats and oils contain, for example, desaturase (Δ-6-d
It is used in health foods for the purpose of resolving various health disorders caused by a decrease in the activity of esaturase.
さらに医薬品などの工業用原料として、広い分野で利用
されうる。Furthermore, it can be used in a wide range of fields as an industrial raw material for pharmaceuticals and other products.
以上that's all
Claims (1)
組織培養し細胞塊を得る工程、および 該細胞塊を植物ホルモンを含有する有機培地で培養し、
γ−リノレン酸エステル含有油脂を生産させる工程。 を包含するマツヨイグサの組織培養によるγ−リノレン
酸エステル含有油脂の製造方法。 2、前記植物ホルモンがオーキシン類および/またはサ
イトカイニン類を含有する特許請求の範囲第1項に記載
の製造方法。 3、前記オーキシン類がナフタレン酢酸、2・4−ジク
ロロフェノキシ酢酸およびインドール酢酸の少なくとも
1種であり、前記サイトカイニン類がカイネチンおよび
/またはベンジルアデニンである特許請求の範囲第1項
に記載の製造方法。 4、前記γ−リノレン酸エステル含有油脂を前記培養細
胞塊から抽出する特許請求の範囲第1項に記載の製造方
法。 5、γ−リノレン酸エステルを構成するγ−リノレン酸
が、総油脂の脂肪酸組成の20重量%以上の割合で含有
される特許請求の範囲第1項に記載の製造方法。[Scope of Claims] 1. A step of tissue culturing a plant of the genus Oenothera to obtain a cell mass, and culturing the cell mass in an organic medium containing a plant hormone,
A process of producing γ-linolenic acid ester-containing oil and fat. A method for producing a γ-linolenic acid ester-containing oil and fat by tissue culture of evening primrose. 2. The production method according to claim 1, wherein the plant hormone contains auxins and/or cytokinins. 3. The production method according to claim 1, wherein the auxin is at least one of naphthaleneacetic acid, 2,4-dichlorophenoxyacetic acid, and indoleacetic acid, and the cytokinin is kinetin and/or benzyladenine. . 4. The manufacturing method according to claim 1, wherein the γ-linolenic acid ester-containing fat or oil is extracted from the cultured cell mass. 5. The manufacturing method according to claim 1, wherein the γ-linolenic acid constituting the γ-linolenic acid ester is contained in a proportion of 20% by weight or more of the fatty acid composition of the total oil and fat.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP61036390A JPS62195291A (en) | 1986-02-20 | 1986-02-20 | Production of fats and oils containing gamma-linolenic ester by tissue culture of evening primrose |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP61036390A JPS62195291A (en) | 1986-02-20 | 1986-02-20 | Production of fats and oils containing gamma-linolenic ester by tissue culture of evening primrose |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS62195291A true JPS62195291A (en) | 1987-08-28 |
JPH0322158B2 JPH0322158B2 (en) | 1991-03-26 |
Family
ID=12468521
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP61036390A Granted JPS62195291A (en) | 1986-02-20 | 1986-02-20 | Production of fats and oils containing gamma-linolenic ester by tissue culture of evening primrose |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS62195291A (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS62210995A (en) * | 1986-03-12 | 1987-09-17 | Nisshin Oil Mills Ltd:The | Production of gamma-linolenic acid |
WO2010062236A1 (en) * | 2008-11-28 | 2010-06-03 | Maria Brodelius | Fatty acids and methods for producing them from cultures derived from plants of genus portulaca |
-
1986
- 1986-02-20 JP JP61036390A patent/JPS62195291A/en active Granted
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS62210995A (en) * | 1986-03-12 | 1987-09-17 | Nisshin Oil Mills Ltd:The | Production of gamma-linolenic acid |
WO2010062236A1 (en) * | 2008-11-28 | 2010-06-03 | Maria Brodelius | Fatty acids and methods for producing them from cultures derived from plants of genus portulaca |
Also Published As
Publication number | Publication date |
---|---|
JPH0322158B2 (en) | 1991-03-26 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Kikowska et al. | Accumulation of rosmarinic, chlorogenic and caffeic acids in in vitro cultures of Eryngium planum L. | |
Sugla et al. | Micropropagation of Pongamia pinnata through enhanced axillary branching | |
Park et al. | Medicinal orchids: production of bioactive compounds and biomass | |
Kochan et al. | Rosmarinic acid and other phenolic acids in hairy roots of Hyssopus officinalis | |
JPS62195291A (en) | Production of fats and oils containing gamma-linolenic ester by tissue culture of evening primrose | |
Liang et al. | Selection of cell lines for the production of rosmarinic acid from cell suspension cultures of Orthosihon stamineus benth | |
EKHTERAEI et al. | Enhanced production of valerenic acids and valepotriates by in vitro cultures of Valeriana officinalis L | |
JPH03262488A (en) | Production of podophyllotoxin compound | |
Asad et al. | 32. In vitro callus induction and plantlet regeneration of sesame (Sesamum Indicum L.) | |
CN101120654B (en) | Tissue culturing method for chia | |
US20050255569A1 (en) | Method for producing triterpene composition | |
Ghasemnezhad et al. | A study on evening-primrose (Oenothera biennis L.) callus regeneration and somatic embryogenesis | |
Kumar et al. | Development of resin canals during somatic embryogenesis in callus cultures of Commiphora wightii | |
Fiasal et al. | Effect of some plant hormones on growth of Mentha piperita L. in vitro. | |
Priya et al. | In vitro effect of Colchiploidy on andrographolide enhancement in Andrographis paniculata (Burm. f.) Wall. ex. Nees. | |
Dimitrova et al. | Influence of 6-benzylaminopurine and indole-3-butyric acid on in vitro propagation and secondary metabolites accumulation in Lamium album from Lozen mountain | |
Boo et al. | In vitro plant regeneration of Aster scaber via somatic embryogenesis | |
Heidaeifar et al. | Establishment of callus and suspension culture in Sesamum indicum L | |
JPS6387989A (en) | Production of vegetable oil and fat | |
EP2370588A1 (en) | Fatty acids and methods for producing them from cultures derived from plants of genus portulaca | |
Ahmad et al. | In-vitro propagation of Chamomilla recutita from capitulum inflorescence: A medicinal plant with multiple therapeutic applications | |
JP2873023B2 (en) | Method for producing podophyllotoxin compounds | |
JP2545359B2 (en) | Plant culture cells | |
Mousa | In vitro, callus induction and estimation of some active constituents in three different medicinal plants | |
KR20060123377A (en) | Process for producing triterpene composition |