JPH01199588A - Production of highly unsaturated aliphatic acid with filamentous fungus - Google Patents
Production of highly unsaturated aliphatic acid with filamentous fungusInfo
- Publication number
- JPH01199588A JPH01199588A JP63134354A JP13435488A JPH01199588A JP H01199588 A JPH01199588 A JP H01199588A JP 63134354 A JP63134354 A JP 63134354A JP 13435488 A JP13435488 A JP 13435488A JP H01199588 A JPH01199588 A JP H01199588A
- Authority
- JP
- Japan
- Prior art keywords
- genus
- highly unsaturated
- unsaturated aliphatic
- aliphatic acid
- filamentous fungus
- 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
- 241000233866 Fungi Species 0.000 title claims abstract description 14
- 238000004519 manufacturing process Methods 0.000 title claims description 4
- 239000002253 acid Substances 0.000 title abstract description 10
- 125000001931 aliphatic group Chemical group 0.000 title abstract 4
- 150000002632 lipids Chemical class 0.000 claims abstract description 15
- 241001278358 Thraustotheca Species 0.000 claims abstract description 5
- 238000012258 culturing Methods 0.000 claims abstract description 4
- 241000003482 Japonochytrium Species 0.000 claims abstract 2
- 235000021122 unsaturated fatty acids Nutrition 0.000 claims description 9
- 150000004670 unsaturated fatty acids Chemical class 0.000 claims description 9
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 claims description 8
- 241000354118 Achlya sp. (in: Oomycetes) Species 0.000 claims 1
- 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 abstract description 4
- 241001578974 Achlya <moth> Species 0.000 abstract description 2
- 241001278361 Thraustotheca clavata Species 0.000 abstract description 2
- 210000002966 serum Anatomy 0.000 abstract description 2
- 235000012000 cholesterol Nutrition 0.000 abstract 1
- 150000001875 compounds Chemical class 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 11
- 230000001580 bacterial effect Effects 0.000 description 9
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 8
- 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 6
- 229940041514 candida albicans extract Drugs 0.000 description 6
- 235000014113 dietary fatty acids Nutrition 0.000 description 6
- 229930195729 fatty acid Natural products 0.000 description 6
- 239000000194 fatty acid Substances 0.000 description 6
- 150000004665 fatty acids Chemical class 0.000 description 6
- 239000008103 glucose Substances 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 239000012138 yeast extract Substances 0.000 description 6
- 238000000034 method Methods 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 229940114079 arachidonic acid Drugs 0.000 description 4
- 235000021342 arachidonic acid Nutrition 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 238000004817 gas chromatography Methods 0.000 description 3
- 239000013535 sea water Substances 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 241000233614 Phytophthora Species 0.000 description 2
- 241000233639 Pythium Species 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 235000021323 fish oil Nutrition 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- XEYBRNLFEZDVAW-UHFFFAOYSA-N prostaglandin E2 Natural products CCCCCC(O)C=CC1C(O)CC(=O)C1CC=CCCCC(O)=O XEYBRNLFEZDVAW-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 239000010698 whale oil Substances 0.000 description 2
- GMVPRGQOIOIIMI-UHFFFAOYSA-N (8R,11R,12R,13E,15S)-11,15-Dihydroxy-9-oxo-13-prostenoic acid Natural products CCCCCC(O)C=CC1C(O)CC(=O)C1CCCCCCC(O)=O GMVPRGQOIOIIMI-UHFFFAOYSA-N 0.000 description 1
- -1 4,7,10,13,16,11- docosahexaenoic acid Chemical compound 0.000 description 1
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000235432 Blastocladiella Species 0.000 description 1
- YEBDWAHEIMUJQT-XVSDJDOKSA-N CCCCCC=CCC=CCC=CCC=CCCCC(O)=O.CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O Chemical compound CCCCCC=CCC=CCC=CCC=CCCCC(O)=O.CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YEBDWAHEIMUJQT-XVSDJDOKSA-N 0.000 description 1
- 241000195620 Euglena Species 0.000 description 1
- 240000002989 Euphorbia neriifolia Species 0.000 description 1
- 241000223218 Fusarium Species 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001558145 Mucor sp. Species 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 229960000711 alprostadil Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000002785 anti-thrombosis Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- XEYBRNLFEZDVAW-ARSRFYASSA-N dinoprostone Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1C\C=C/CCCC(O)=O XEYBRNLFEZDVAW-ARSRFYASSA-N 0.000 description 1
- 229960002986 dinoprostone Drugs 0.000 description 1
- MBMBGCFOFBJSGT-KUBAVDMBSA-N docosahexaenoic acid Natural products CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 description 1
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000004626 essential fatty acids Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- IQLUYYHUNSSHIY-UHFFFAOYSA-N icosa-2,4,6,8-tetraenoic acid Chemical compound CCCCCCCCCCCC=CC=CC=CC=CC(O)=O IQLUYYHUNSSHIY-UHFFFAOYSA-N 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 150000002823 nitrates Chemical class 0.000 description 1
- 125000001477 organic nitrogen group Chemical group 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- GMVPRGQOIOIIMI-DWKJAMRDSA-N prostaglandin E1 Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1CCCCCCC(O)=O GMVPRGQOIOIIMI-DWKJAMRDSA-N 0.000 description 1
- CBOMORHDRONZRN-QLOYDKTKSA-N prostaglandin E3 Chemical compound CC\C=C/C[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1C\C=C/CCCC(O)=O CBOMORHDRONZRN-QLOYDKTKSA-N 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 235000003441 saturated fatty acids Nutrition 0.000 description 1
- 150000004671 saturated fatty acids Chemical class 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- DSNBHJFQCNUKMA-SCKDECHMSA-N thromboxane A2 Chemical compound OC(=O)CCC\C=C/C[C@@H]1[C@@H](/C=C/[C@@H](O)CCCCC)O[C@@H]2O[C@H]1C2 DSNBHJFQCNUKMA-SCKDECHMSA-N 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- RROQIUMZODEXOR-UHFFFAOYSA-N triforine Chemical compound O=CNC(C(Cl)(Cl)Cl)N1CCN(C(NC=O)C(Cl)(Cl)Cl)CC1 RROQIUMZODEXOR-UHFFFAOYSA-N 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
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〕
本発明は糸状菌を栄養培地に培養し、培養物より高度不
飽和脂肪酸含量の高い脂質を得ることによる高度不飽和
脂肪酸の製造法に関する。DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a method for producing polyunsaturated fatty acids by culturing filamentous fungi in a nutrient medium and obtaining lipids with a high content of polyunsaturated fatty acids from the culture.
本発明において製造される高度不飽和脂肪酸は炭素数１
８〜２２、不飽和二重結合数３以上の脂肪酸であり、具
体的には、例えばＴ−リルン酸（９゜１２、１５−オク
タデカトリエン酸）、ジホモ−γ−リルン酸（８，ＩＣ
ｌ３−エイコサトリエン酸）、アラキドン酸（５，８，
１１，１４−エイコサテトラエン酸）、Ｅ Ｐ Ａ （
５，８，ＩＣｌ３，１７−ニイコサペンクエン酸）、Ｄ
Ｔ Ａ （７，１０，１３，１６−１’コサテトラエ
ン酸）、ＤＰＡ （４，７，１（１，１３，１６−また
は７．１０．１３．１６．１９−ドコサペンクエン酸）
およびＤ ＨＡ （４，７，１０，１３，１６，１１−
ドコサヘキサエン酸）等を総称するものである。The highly unsaturated fatty acid produced in the present invention has 1 carbon number.
8 to 22, unsaturated double bonds of 3 or more, specifically, for example, T-lylunic acid (9°12,15-octadecatrienoic acid), dihomo-γ-lylunic acid (8, IC
l3-eicosatrienoic acid), arachidonic acid (5,8,
11,14-eicosatetraenoic acid), EP A (
5,8,ICl3,17-nicosapencitric acid), D
T A (7,10,13,16-1'cosatetraenoic acid), DPA (4,7,1 (1,13,16- or 7.10.13.16.19-docosapencitric acid)
and D HA (4,7,10,13,16,11-
docosahexaenoic acid), etc.
これら高度不飽和脂肪酸は、哺乳動物では体内において
合成することのできない必須脂肪酸である。ジホモ−γ
−リルン酸はプロスタグランジンＥ１、Ｐ、など、アラ
キドン酸はプロスタグランジンＥ２、Ｆ２、Ｉ２、トロ
ンボキサンＡ２など、ＥＰＡはプロスタグランジンＥ３
、Ｆ３、＋３、Ｉ−ロンホキサンＡ３などのそれぞれ前
駆体として知られており、重要であると同時に、高度不
飽和脂肪酸自身、生体内において多くの生理活性を示す
ことが知られている。例えば、ＥＰＡ、ＤＭＡ等は抗血
栓作用、心臓病、血清全コレステロールの低下作用等に
対する効果が注目されている。These highly unsaturated fatty acids are essential fatty acids that cannot be synthesized within the body of mammals. Dihomo-γ
- Rilunic acid is prostaglandin E1, P, etc., arachidonic acid is prostaglandin E2, F2, I2, thromboxane A2, etc., EPA is prostaglandin E3, etc.
, F3, +3, I-lomphoxane A3, etc., and are important, and at the same time, highly unsaturated fatty acids themselves are known to exhibit many physiological activities in vivo. For example, EPA, DMA, and the like are attracting attention for their antithrombotic effects, their effects on heart disease, and their effects on lowering serum total cholesterol.
これら高度不飽和脂肪酸中、アラキドン酸を含む微生物
としては、これまでに、原生動物のユウグレナのほかに
、糸状菌であるＣｏｎ１ｄｉｏｂｏｌｕｓ属、Ｔｒｉｃ
ｈｏｔｈｅｃｉｕｍ属、Ｅｎ ｔｏｍｏｐｈ ｔｈｏｒ
ａ属、Ｐｈｙｔｏｐｈｔｈ−ｏｒａ属、Ｓａｐｒｏｌｅ
ｇｎｉａ属、Ｐｙｔｈｉｕｍ属、Ｍｏｒｔｉｅｒｅｌ
ｌａ属およびＢｌａｓｔｏｃｌａｄｉｅｌｌａ属が知ら
れている。これらの微生物は同時にジホモ−Ｔ−リルン
酸を含んでいる。また、ＥＰＡを含む微生物としても糸
状菌であるＰｈｙｔｏｐｈｔｈｏｒａ属、Ｓａｐｒｏｌ
ｅｇｎｉａ属、Ｐｙｔｈｉｕｍ属、Ｍｏｒｔｉｅｒｅｌ
ｌａ属が報告されている〔１１゜Ｂ、ＷＩＩＩＴＥ
ａｎｄ Ｓ、Ｓ、ＰＯＷＥＬＬ 旧ｏｃｈｉｍ、Ｂ
ｉｏｐｈｙｓ、 八ｃｔａ１１６：３９１ （１９６
６）、Ｄ、Ｔｙｒｅｌｌ Ｃａｎ、Ｊ、Ｍｉｃｒｏｂｉ
ｏｌ、 １３ニア５５（１９６７）等）。その他、培養
の際に炭化水素を添加することにより、アラキドン酸が
得られる菌として、糸状菌のＡｓｐｅｒｇｉｌｌｕｓ属
、Ｆｕｓａｒｉｕｍ属、Ｐｅｎ１ｃ目１ｉｕｍ属、Ｐｏ
ｒｐｈｙｒｉｄｉｕｍ属、Ｍｕｃｏｒ属およびＲｈ１ｚ
ｏｐｕｓ属が知られている〔特公昭５６−１９２３１号
公報〕。Microorganisms that contain arachidonic acid among these highly unsaturated fatty acids include the protozoan Euglena, the filamentous fungi Con1diobolus genus, and Tri.
Genus hothecium, En tomoph thor
a genus, Phytophth-ora genus, Saprole
genus gnia, genus Pythium, Mortierel
The genera La and Blastocladiella are known. These microorganisms also contain dihomo-T-lylunic acid. In addition, EPA-containing microorganisms include the filamentous fungi Phytophthora genus and Saprol.
Genus egnaia, Genus Pythium, Mortierel
The genus la has been reported [11°B, WIIITE
and S, S, POWELL old ochim, B
iophys, octa116:391 (196
6), D. Tyrell Can, J. Microbi.
ol, 13 Near 55 (1967), etc.). Other fungi that can produce arachidonic acid by adding hydrocarbons during culture include filamentous fungi of the genus Aspergillus, the genus Fusarium, the genus Pen1c, the genus Po
rphyridium sp., Mucor sp. and Rh1z
The genus Opus is known [Japanese Patent Publication No. 19231/1983].
また、ＤＨＡなどの高度不飽和脂肪酸は魚油から抽出さ
れているが、魚油（鯨油、鯨油）中の該脂肪酸の含有量
が低いことや、高度不飽和脂肪酸ばそれ自身酸化分解さ
れ易い等により、高純度に目的とする高度不飽和脂肪酸
を得ることは困難である。In addition, highly unsaturated fatty acids such as DHA are extracted from fish oil, but the content of these fatty acids in fish oil (whale oil, whale oil) is low, and highly unsaturated fatty acids themselves are easily oxidized and degraded. It is difficult to obtain the desired polyunsaturated fatty acids with high purity.
このような状況下、本発明者らは、高度不飽和脂肪酸を
微生物を用い製造することが最も有利と考え、高度不飽
和脂肪酸含量の高い脂質を生産する能力を有する糸状菌
について鋭意検討した結果へｍｅｒｉｃａｎ Ｔｙｐｅ
Ｃｕ１ｔｕｒｅ Ｃｏ１１ｅｃｔｉｏｎ （ＡＴＣＣ
） に保存され、カタログに記載されている糸状菌であ
るＴｈｒａｕｓｔｏｔｈｅｃａ ｃｌａｖａｔａ
ＡＴＣＣ３４１１２、八ｃｈｌｙａ ｂｉ−ｓｅｘｕ
ａｌｉｓ ＡＴＣＣ１４５２４、１ｌａｌｉｐｈｔｈ
ｏｒｏｕｓ ｐｈｉｌｉｐｌｉｎｅｎｓｉｓ ＡＴ
ＣＣ５８３０３、Ｒｉｚｏｐｈｙｄｉｕｍ Ｉｉｔｔ
ｏｒｅｕｍ ＡＴＣＣ３６１００およびＪａｐｏｎｏ
ｃｈｙｔｒｉｕｍ ｓｐ、＾ＴＣＣ２８２０７が、高度
不飽和脂肪酸を生産することを見出し、本発明はこの知
見によりなされたものである。Under these circumstances, the present inventors believed that it would be most advantageous to produce highly unsaturated fatty acids using microorganisms, and as a result of intensive study on filamentous fungi that have the ability to produce lipids with a high content of polyunsaturated fatty acids. Hemerican Type
Culture Co11ection (ATCC
Thraustotheca clavata, a filamentous fungus stored in
ATCC34112, 8chlya bi-sexu
alis ATCC14524, 1laiphth
orous philiplinensis AT
CC58303, Rizophydium Iitt
oreum ATCC36100 and Japono
It was discovered that chytrium sp. ^TCC28207 produces highly unsaturated fatty acids, and the present invention was made based on this finding.
すなわち、本発明はＴｈｒａｕｓｔｏｔｈｅｃａ属、Ａ
ｃｈｌｙａ属、１ｌａｌｉｐｈｔｈｏｒｏｕｓ属、Ｒｉ
ｚｏｐｈｙｄｉｕｍ属またはＪａｐ−ｏｎｏｃｈｙｔｒ
ｉｕｍ属に属し、高度不飽和脂肪酸含量の高い脂質生産
能を有する糸状菌を培養し、培養物＝ ３−
より高度不飽和脂肪酸含量の高い脂質を採取することを
特徴とする糸状菌による高度不飽和脂肪酸の製造法であ
る。That is, the present invention relates to the genus Thraustotheca, A.
Chlya spp., 1laliphthorous spp., Ri
Genus zophydium or Jap-onochytr
Cultivate filamentous fungi belonging to the genus P. umum and having the ability to produce lipids with a high content of polyunsaturated fatty acids, and culture = 3- A highly unsaturated fungus characterized by collecting lipids with a higher content of polyunsaturated fatty acids. This is a method for producing saturated fatty acids.
上記糸状菌を培養する培地組成については特に規定する
ものではないが、炭素源としては、グルコース、エタノ
ール、グリ七ロールなどが好ましく、その使用量は通常
０．５〜２０重量％の範囲である。また、窒素源として
はイースト・エキス、マルタ・エキス、ペプトンなどの
有機窒素源の使用が好ましいが、ほかに硝酸塩などの無
機窒素も使用でき、その使用量は通常０．１〜１０重量
％の範囲である。また、これらの成分のほかにアミノ酸
、ビタミン類などの微量要素や無機塩類の添加は菌の生
育を促進させるのに効果的である。The composition of the medium for culturing the filamentous fungi is not particularly defined, but the carbon source is preferably glucose, ethanol, glycerol, etc., and the amount used is usually in the range of 0.5 to 20% by weight. . In addition, as a nitrogen source, it is preferable to use organic nitrogen sources such as yeast extract, malta extract, and peptone, but inorganic nitrogen sources such as nitrates can also be used, and the amount used is usually 0.1 to 10% by weight. range. In addition to these ingredients, addition of trace elements such as amino acids and vitamins and inorganic salts is effective in promoting the growth of bacteria.
培地のｐｌ＋は４〜１０、好ましくは５〜８、培養温度
は１０〜３５°Ｃの範囲が良く、培養時間は２〜１５日
間程度必要である。The pl+ of the medium is 4 to 10, preferably 5 to 8, the culture temperature is preferably in the range of 10 to 35°C, and the culture time is required to be about 2 to 15 days.
培養は、通常液体培養培地で、静置培養、振とう培養、
通気撹拌培養などの方法により行う。Culture is usually done in a liquid culture medium, such as static culture, shaking culture,
This is done by methods such as aerated agitation culture.
このようにして得られた培養物より、濾過、遠心分離な
どの方法で菌体を集め、その菌体より脂質抽出を行う。From the culture thus obtained, bacterial cells are collected by methods such as filtration and centrifugation, and lipids are extracted from the bacterial cells.
脂質の抽出は常法に従い、例えば溶媒抽出などによって
行われる。Lipids are extracted according to conventional methods, such as by solvent extraction.
実施例１゜
下記の有機栄養培地１１にＴｈｒａｕｓｔｏｔｈｅｃａ
ｃｌａ−ｖａｔａ ＡＴＣＣ３４１１２を接種し、２
５°Ｃで７日間振とう培養後、濾過にて菌体を回収し凍
結乾燥した。Example 1 Thraustotheca was added to the organic nutrient medium 11 described below.
cla-vata ATCC34112 and 2
After shaking culture at 5°C for 7 days, the bacterial cells were collected by filtration and freeze-dried.
これにより、５．８ｇの乾燥菌体を得た。この乾燥菌体
を粉砕してＦｏｌｃｈ法により抽出精製した後、溶媒を
減圧留去し、重量法により全脂質を測定した。その結果
、０．５ｇの粗脂質が得られた。As a result, 5.8 g of dried bacterial cells were obtained. After pulverizing the dried bacterial cells and extracting and purifying them by the Folch method, the solvent was distilled off under reduced pressure, and the total lipids were measured by gravimetric method. As a result, 0.5 g of crude lipid was obtained.
培地 グルコース １０ｇ／ ｆ！。Medium Glucose 10g/f! .
イースト・エキス ３ｇ／ １
マルタ・エキス ３ｇ／ １
ト リ ブ ト ン ５ｇ／ １
蒸留水 １乏
ｐｌ＋７．０
菌体から抽出精製した脂質は一部をメチルエステル化し
た後、ガスクロマトグラフィーにより脂肪酸組成の分析
を行った。その結果を表−１に示した。Yeast Extract 3g/1 Malta Extract 3g/1 Tributone 5g/1
Distilled water 1 pl+7.0 The lipid extracted and purified from the bacterial cells was partially methyl esterified, and then the fatty acid composition was analyzed by gas chromatography. The results are shown in Table-1.
尚、ガスクロマトグラフィーにより得られた各ピークは
、マススペクトロメトリーにより同定を行った。In addition, each peak obtained by gas chromatography was identified by mass spectrometry.
ガスクロマトフラフィーハ 条件
カラム充填剤 １１ｉａｓｏｌｉｄ ＺＦ カラム温
度２００’ＣキヤリアーガスＮ２ 検出器温
度２５０°Ｃ検出器 ＦＩＤ 注入温
度 ２５０’Ｃカラム １．５ｍ Ｘ３ φ
表−１
乾燥菌体当りのアラキドン酸含量は２１．６■／ｇであ
る。Gas chromatography condition Column packing material 11iasolid ZF Column temperature 200'C Carrier gas N2 Detector temperature 250°C Detector FID Injection temperature 250'C Column 1.5 m X3 φ Table 1 Arachidonic acid content per dry bacterial cell is It is 21.6■/g.
乾燥菌体当りのＥＰＡ含量は４．６■／ｇである。The EPA content per dry bacterial cell was 4.6 /g.
実施例２゜
下記の有機栄養培地１１にＡｃｈｌｙａ ｂｉｓｅｘｕ
ａｌｉｓＡＴＣＣ１４５２４を接種し、以後、実施例１
と同様の操作を行い、脂肪酸組成を分析した。Example 2 Achlya bisexu was added to the following organic nutrient medium 11.
alisATCC14524 was inoculated, and hereafter, Example 1
The fatty acid composition was analyzed using the same procedure as above.
結果を表−２に示した。The results are shown in Table-2.
培地 グルコース １０ｇ／ １イースト・エ
キス ３ｇ／ ｊ２
マルク・エキス ３ｇ／！
ポリペプトン ５ｇ／ Ｅ
蒸留水 １！
ｐＨ６，５
表−２
実施例３゜
下記の有機栄養培地１１にＨａｌｉｐｈｔｈｏｒｏｕｓ
ｐｈ−１１ｉｐｐｉｎｅｎｓｉｓ ＡＴＣＣ５８３０
３を接種し、以後、実施例１と同様の操作を行い、脂肪
酸組成を分析し、結果を表−３に示した。Medium Glucose 10g/1 Yeast Extract 3g/j2 Marc Extract 3g/! Polypeptone 5g/E Distilled water 1! pH 6,5 Table 2 Example 3
ph-11ippinensis ATCC5830
After that, the same operations as in Example 1 were performed to analyze the fatty acid composition, and the results are shown in Table 3.
培地 グルコース ５ｇ／ １イースト・エ
キス １ｇ／！
ポリペプトン １ｇ／！
人工海水 １！
ｐｌ＋６．５
表−３
実施例４゜
下記の有機栄養培地１１．にＲｒｚｏｐｈｙｄｌｕｍ
］ｉｔＬ−ｏｒｅｕｍ ＡＴＣＣ３６１００を接種し、
以後、実施例１と同様の操作を行い、脂肪酸組成を分析
し、結果を表−４に示した。Medium Glucose 5g/1 Yeast Extract 1g/! Polypeptone 1g/! Artificial seawater 1! pl+6.5 Table-3 Example 4゜The following organic nutrient medium 11. Rrzophydlum
]itL-oreum ATCC36100 was inoculated,
Thereafter, the same operations as in Example 1 were performed to analyze the fatty acid composition, and the results are shown in Table 4.
培地 グルコース ５ｇ／！イースト・エキ
ス Ｉｇ／ｊ２
ポリペプトン １ｇ／２
人工海水 １！
ｐｉ ６．５
表−４
実施例５゜
下記の有機栄養培地１１にＪａｐｏｎｏｃｈｙｔｒｉｕ
ｍ ｓｐ。Medium glucose 5g/! Yeast extract Ig/j2 Polypeptone 1g/2 Artificial seawater 1! pi 6.5 Table 4 Example 5
m sp.
ＡＴＣＣ２８２０７を接種し、以後、実施例１と同様の
操作を行い、１．７ｇの乾燥菌体、次いで０．１４ｇの
粗脂質が得られた。さらに、実施例１と同様にして、こ
の脂質の脂肪酸組成を分析した。ATCC28207 was inoculated, and the same operations as in Example 1 were performed to obtain 1.7 g of dried bacterial cells and then 0.14 g of crude lipid. Furthermore, in the same manner as in Example 1, the fatty acid composition of this lipid was analyzed.
結果を表−５に示した。The results are shown in Table-5.
培地 グルコース １０ｇ／ｆイースト・エキ
ス ３ｇ／ ｊ２
マルタ・エキス ３ｇ／ １
ペプトン １ｇ／Ｉ！。Medium Glucose 10g/f Yeast Extract 3g/j2 Malta Extract 3g/1 Peptone 1g/I! .
人工海水 １ρ ｐｎ ７．０ 表−５Artificial seawater 1ρ pn 7.0 Table-5
Claims (1)
ｌｉｐｈｔｈｏｒｏｕｓ属Ｒｉｚｏｐｈｙｄｉｕｍ属ま
たはＪａｐｏｎｏｃｈｙｔｒｉｕｍ属に属し、高度不飽
和脂肪酸含量の高い脂質生産能を有する糸状菌を培養し
、培養物より高度不飽和脂肪酸含量の高い脂質を採取す
ることを特徴とする糸状菌による高度不飽和脂肪酸の製
造法。Thraustotheca sp., Achlya sp., Ha
By culturing a filamentous fungus belonging to the genus Liphthorous, the genus Rizophydium, or the genus Japonochytrium and having the ability to produce lipids with a high content of polyunsaturated fatty acids, and collecting lipids with a high content of polyunsaturated fatty acids from the culture. Method for producing highly unsaturated fatty acids.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63134354A JP2582622B2 (en) | 1987-10-27 | 1988-06-02 | Production of polyunsaturated fatty acids by filamentous fungi |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP27062487 | 1987-10-27 | ||
| JP62-270624 | 1987-10-27 | ||
| JP63134354A JP2582622B2 (en) | 1987-10-27 | 1988-06-02 | Production of polyunsaturated fatty acids by filamentous fungi |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH01199588A true JPH01199588A (en) | 1989-08-10 |
| JP2582622B2 JP2582622B2 (en) | 1997-02-19 |
Family
ID=26468485
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63134354A Expired - Fee Related JP2582622B2 (en) | 1987-10-27 | 1988-06-02 | Production of polyunsaturated fatty acids by filamentous fungi |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2582622B2 (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998029558A1 (en) * | 1996-12-27 | 1998-07-09 | Suntory Limited | Media for culturing microorganisms and process for producing unsaturated fatty acids or lipids containing the same |
| LT4550B (en) | 1996-07-23 | 1999-10-25 | Nagase Biochemicals, Ltd | Process for preparing docosahexaenoic acid and docosapentaenoic acid |
| KR100321543B1 (en) * | 1991-01-24 | 2002-09-12 | 마르텍 코포레이션 | Microbial oil mixtures and uses thereof |
| JP2004285182A (en) * | 2003-03-20 | 2004-10-14 | Yuji Shimada | Glyceride and its manufacturing process |
| US7550286B2 (en) | 2004-11-04 | 2009-06-23 | E. I. Du Pont De Nemours And Company | Docosahexaenoic acid producing strains of Yarrowia lipolytica |
| US7863024B2 (en) | 2002-04-26 | 2011-01-04 | Suntory Holdings Limited | Process for producing highly unsaturated fatty acid-containing lipid |
| WO2011133610A1 (en) | 2010-04-22 | 2011-10-27 | E. I. Du Pont De Nemours And Company | Method for obtaining polyunsaturated fatty acid-containing compositions from microbial biomass |
| WO2012021700A1 (en) | 2010-08-11 | 2012-02-16 | E. I. Du Pont De Nemours And Company | Improved aquaculture feed compositions |
| WO2012021686A1 (en) | 2010-08-11 | 2012-02-16 | E. I. Du Pont De Nemours And Company | Improved aquaculture meat products |
-
1988
- 1988-06-02 JP JP63134354A patent/JP2582622B2/en not_active Expired - Fee Related
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100321543B1 (en) * | 1991-01-24 | 2002-09-12 | 마르텍 코포레이션 | Microbial oil mixtures and uses thereof |
| EP2213750A1 (en) | 1996-07-23 | 2010-08-04 | Nagase ChemteX Corporation | Process for preparing docosahexaenoic acid and docosapentaenoic acid |
| LT4550B (en) | 1996-07-23 | 1999-10-25 | Nagase Biochemicals, Ltd | Process for preparing docosahexaenoic acid and docosapentaenoic acid |
| US6746857B2 (en) | 1996-12-27 | 2004-06-08 | Suntory Limited | Media for culturing microorganisms and process for producing unsaturated fatty acids or lipids containing the same |
| WO1998029558A1 (en) * | 1996-12-27 | 1998-07-09 | Suntory Limited | Media for culturing microorganisms and process for producing unsaturated fatty acids or lipids containing the same |
| US7863024B2 (en) | 2002-04-26 | 2011-01-04 | Suntory Holdings Limited | Process for producing highly unsaturated fatty acid-containing lipid |
| JP2004285182A (en) * | 2003-03-20 | 2004-10-14 | Yuji Shimada | Glyceride and its manufacturing process |
| US7550286B2 (en) | 2004-11-04 | 2009-06-23 | E. I. Du Pont De Nemours And Company | Docosahexaenoic acid producing strains of Yarrowia lipolytica |
| US8685682B2 (en) | 2004-11-04 | 2014-04-01 | E I Du Pont De Nemours And Company | Docosahexaenoic acid producing strains of yarrowia lipolytica |
| WO2011133610A1 (en) | 2010-04-22 | 2011-10-27 | E. I. Du Pont De Nemours And Company | Method for obtaining polyunsaturated fatty acid-containing compositions from microbial biomass |
| WO2012021700A1 (en) | 2010-08-11 | 2012-02-16 | E. I. Du Pont De Nemours And Company | Improved aquaculture feed compositions |
| WO2012021686A1 (en) | 2010-08-11 | 2012-02-16 | E. I. Du Pont De Nemours And Company | Improved aquaculture meat products |
| WO2012021711A1 (en) | 2010-08-11 | 2012-02-16 | E.I. Du Pont De Nemours And Company | Improved aquaculture feed compositions |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2582622B2 (en) | 1997-02-19 |
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