JP2582622B2 - Production of polyunsaturated fatty acids by filamentous fungi - Google Patents
Production of polyunsaturated fatty acids by filamentous fungiInfo
- Publication number
- JP2582622B2 JP2582622B2 JP63134354A JP13435488A JP2582622B2 JP 2582622 B2 JP2582622 B2 JP 2582622B2 JP 63134354 A JP63134354 A JP 63134354A JP 13435488 A JP13435488 A JP 13435488A JP 2582622 B2 JP2582622 B2 JP 2582622B2
- Authority
- JP
- Japan
- Prior art keywords
- fatty acids
- polyunsaturated fatty
- genus
- filamentous fungi
- 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.)
- Expired - Fee Related
Links
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 title claims description 13
- 241000233866 Fungi Species 0.000 title claims description 10
- 238000004519 manufacturing process Methods 0.000 title claims description 4
- 150000002632 lipids Chemical class 0.000 claims description 13
- 238000012258 culturing Methods 0.000 claims description 7
- 235000021122 unsaturated fatty acids Nutrition 0.000 claims description 6
- 150000004670 unsaturated fatty acids Chemical class 0.000 claims description 6
- 241000003482 Japonochytrium Species 0.000 claims description 4
- 241001578974 Achlya <moth> Species 0.000 claims description 2
- 241001278358 Thraustotheca Species 0.000 claims description 2
- 241000948838 Haliphthoros Species 0.000 claims 1
- 239000002609 medium Substances 0.000 description 12
- 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
- 235000014113 dietary fatty acids Nutrition 0.000 description 7
- 229930195729 fatty acid Natural products 0.000 description 7
- 239000000194 fatty acid Substances 0.000 description 7
- 150000004665 fatty acids Chemical class 0.000 description 7
- SBHCLVQMTBWHCD-METXMMQOSA-N (2e,4e,6e,8e,10e)-icosa-2,4,6,8,10-pentaenoic acid Chemical compound CCCCCCCCC\C=C\C=C\C=C\C=C\C=C\C(O)=O SBHCLVQMTBWHCD-METXMMQOSA-N 0.000 description 6
- 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
- HXWJFEZDFPRLBG-UHFFFAOYSA-N Timnodonic acid Natural products CCCC=CC=CCC=CCC=CCC=CCCCC(O)=O HXWJFEZDFPRLBG-UHFFFAOYSA-N 0.000 description 6
- 229940041514 candida albicans extract Drugs 0.000 description 6
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 description 6
- 239000008103 glucose Substances 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 6
- 239000012138 yeast extract Substances 0.000 description 6
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 235000021342 arachidonic acid Nutrition 0.000 description 4
- 229940114079 arachidonic acid Drugs 0.000 description 4
- MBMBGCFOFBJSGT-SFGLVEFQSA-N docosa-4,7,10,13,16,19-hexaenoic acid Chemical compound CC\C=C\C\C=C\C\C=C\C\C=C\C\C=C\C\C=C\CCC(O)=O MBMBGCFOFBJSGT-SFGLVEFQSA-N 0.000 description 4
- 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 4
- 229940045184 malt extract Drugs 0.000 description 4
- 244000005700 microbiome Species 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
- HOBAELRKJCKHQD-QNEBEIHSSA-N dihomo-γ-linolenic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/CCCCCCC(O)=O HOBAELRKJCKHQD-QNEBEIHSSA-N 0.000 description 3
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 239000013535 sea water Substances 0.000 description 3
- -1 F 1 Chemical compound 0.000 description 2
- YUFFSWGQGVEMMI-UHFFFAOYSA-N (7Z,10Z,13Z,16Z,19Z)-7,10,13,16,19-docosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCCCC(O)=O YUFFSWGQGVEMMI-UHFFFAOYSA-N 0.000 description 2
- HOBAELRKJCKHQD-UHFFFAOYSA-N (8Z,11Z,14Z)-8,11,14-eicosatrienoic acid Natural products CCCCCC=CCC=CCC=CCCCCCCC(O)=O HOBAELRKJCKHQD-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 235000021298 Dihomo-γ-linolenic acid Nutrition 0.000 description 2
- 241000235575 Mortierella Species 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
- 241000233667 Saprolegnia Species 0.000 description 2
- 241001278361 Thraustotheca clavata Species 0.000 description 2
- TWSWSIQAPQLDBP-UHFFFAOYSA-N adrenic acid Natural products CCCCCC=CCC=CCC=CCC=CCCCCCC(O)=O TWSWSIQAPQLDBP-UHFFFAOYSA-N 0.000 description 2
- TWSWSIQAPQLDBP-DOFZRALJSA-N all-cis-docosa-7,10,13,16-tetraenoic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCCCC(O)=O TWSWSIQAPQLDBP-DOFZRALJSA-N 0.000 description 2
- 229910052799 carbon Inorganic materials 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
- 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
- VZCCETWTMQHEPK-UHFFFAOYSA-N gamma-Linolensaeure Natural products CCCCCC=CCC=CCC=CCCCCC(O)=O VZCCETWTMQHEPK-UHFFFAOYSA-N 0.000 description 2
- 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 description 2
- 235000020664 gamma-linolenic acid Nutrition 0.000 description 2
- 229960002733 gamolenic acid Drugs 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- YUFFSWGQGVEMMI-RCHUDCCISA-N (7e,10e,13e,16e,19e)-docosa-7,10,13,16,19-pentaenoic acid Chemical compound CC\C=C\C\C=C\C\C=C\C\C=C\C\C=C\CCCCCC(O)=O YUFFSWGQGVEMMI-RCHUDCCISA-N 0.000 description 1
- KSDMISMEMOGBFU-UHFFFAOYSA-N (all-Z)-7,10,13-Eicosatrienoic acid Natural products CCCCCCC=CCC=CCC=CCCCCCC(O)=O KSDMISMEMOGBFU-UHFFFAOYSA-N 0.000 description 1
- 241000233661 Achlya bisexualis Species 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
- 241001480517 Conidiobolus Species 0.000 description 1
- 241000195620 Euglena Species 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241000235395 Mucor Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- 241000206618 Porphyridium Species 0.000 description 1
- 241000233639 Pythium Species 0.000 description 1
- 241000235527 Rhizopus Species 0.000 description 1
- 241001125046 Sardina pilchardus Species 0.000 description 1
- NMZKLLJQNNTBRJ-OIXZZONUSA-N TXA3 Chemical compound OC(=O)CCC\C=C/C[C@@H]1[C@@H](/C=C/[C@@H](O)C\C=C/CC)O[C@@H]2O[C@H]1C2 NMZKLLJQNNTBRJ-OIXZZONUSA-N 0.000 description 1
- 238000005273 aeration Methods 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
- 238000004587 chromatography analysis 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
- 238000005516 engineering process Methods 0.000 description 1
- 235000004626 essential fatty acids Nutrition 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000007789 gas Substances 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
- 238000001727 in vivo Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- KQQKGWQCNNTQJW-UHFFFAOYSA-N linolenic acid Natural products CC=CCCC=CCC=CCCCCCCCC(O)=O KQQKGWQCNNTQJW-UHFFFAOYSA-N 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 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
- 239000002243 precursor Substances 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
- 150000003839 salts Chemical class 0.000 description 1
- 235000019512 sardine Nutrition 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000003756 stirring Methods 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
- 239000012137 tryptone Substances 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
- 239000010698 whale oil Substances 0.000 description 1
- DTOSIQBPPRVQHS-UHFFFAOYSA-N α-Linolenic acid Chemical compound CCC=CCC=CCC=CCCCCCCCC(O)=O DTOSIQBPPRVQHS-UHFFFAOYSA-N 0.000 description 1
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倍値に培養し、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。The present invention relates to a method for producing polyunsaturated fatty acids by culturing filamentous fungi at a nutrient doubling value and obtaining a lipid having a higher 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
18〜22、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γ−リノレン酸（9,12,15−オクタデカ
トリエン酸）、ジホモ−γ−リノレン酸（8,11,14−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）、EPA（5,8,11,14,17−エイコサペ
ンタエン酸）、DTA（7,10,13,16−ドコサテトラエン
酸）、DPA（4,7,10,13,16−または7,10,13,16,19−ドコ
サペンタエン酸）およびDHA（4,7,10,13,16,19−ドコサ
ヘキサエン酸）等を総称するものである。The polyunsaturated fatty acid produced in the present invention has a carbon number
18-22, fatty acids having 3 or more unsaturated double bonds, specifically, for example, γ-linolenic acid (9,12,15-octadecatrienoic acid), dihomo-γ-linolenic acid (8,11 , 14-eicosatrienoic acid), arachidonic acid (5,8,11,14-eicosatetraenoic acid), EPA (5,8,11,14,17-eicosapentaenoic acid), DTA (7,10, 13,16-docosatetraenoic acid), DPA (4,7,10,13,16- or 7,10,13,16,19-docosapentaenoic acid) and DHA (4,7,10,13,16 , 19-docosahexaenoic acid) and the like.
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−リノレン酸はプロスタグランジンE1、F1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、I3、ト
ロンボキサン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、DHA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 in the body in mammals. Dihomo
etc. γ- linolenic acid prostaglandin E 1, F 1, arachidonic acid is prostaglandin E 2, F 2, I 2 , such as thromboxane A 2, EPA is prostaglandin E 3, F 3, I 3 , known as the respective precursor such as thromboxane a 3, at the same time is important, polyunsaturated fatty acids themselves, are known to exhibit a number of physiological activity in vivo.
For example, EPA, DHA, and the like have been noted for their effects on antithrombotic effects, heart disease, serum total cholesterol lowering effects, and the like.
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Conidiobolus属、Tricnothecium属、E
ntomophthora属、Phytophthora属、Saprolegnia属、Pyt
hium属、Mortierella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−γ−リノレ
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、Saprolegnia属、Pythium
属、Mortierella属が報告されている〔H.B.WHITE and
S.S.PWELL Biochim.Biophys.Acta116:391（1966）、D.T
yrell Can.J.Microbiol.13:755（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、Fusar
ium属、Penicillium属、Porphyridium属、Mucor属およ
びRhizopus属が知られている〔特開昭56−19231号公
報〕。Among these polyunsaturated fatty acids, microorganisms containing arachidonic acid include, in addition to protozoa Euglena, fungi such as Conidiobolus, Tricnothecium, and E.
Genus ntomophthora, Phytophthora, Saprolegnia, Pyt
The genus hium, the genus Mortierella and the genus Blastocladiella are known. These microorganisms simultaneously contain dihomo-γ-linolenic acid. Microorganisms containing EPA include filamentous fungi such as Phytophthora, Saprolegnia, and Pythium.
The genus Mortierella has been reported (HBWHITE and
SSPWELL Biochim. Biophys. Acta116: 391 (1966), DT
yrell Can. J. Microbiol. 13: 755 (1967) and the like]. Others
Arachidonic acid can be obtained by adding hydrocarbons during culturing, as filamentous fungi Aspergillus, Fusar
The genera genus Penicillium, Porphyridium, Mucor and Rhizopus are known (Japanese Patent Application Laid-Open No. 56-19231).
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、鰯油）中の該脂肪酸の含有量
が低いことや、高度不飽和脂肪酸は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, polyunsaturated fatty acids such as DHA are extracted from fish oil. However, the content of the fatty acids in fish oil (whale oil, sardine oil) is low, and polyunsaturated fatty acids themselves are easily oxidatively decomposed. It is difficult to obtain the desired highly unsaturated fatty acid 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American Type Culture Co
llection（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 ATCC 34112、Ac
hlya bisexualis ATCC 14524、Haliphthorous philippi
nensis ATCC 58303、Rizophydium littoreum ATCC 3610
0およびJaponochytrium sp.ATCC 28207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 such circumstances, the present inventors considered that it is most advantageous to produce polyunsaturated fatty acids using microorganisms, and as a result of intensive studies on filamentous fungi having the ability to produce lipids having a high content of polyunsaturated fatty acids. American Type Culture Co
thraustotheca clavata ATCC 34112, Ac, stored in the LLection (ATCC) and listed in the catalog
hlya bisexualis ATCC 14524, Haliphthorous philippi
nensis ATCC 58303, Rizophydium littoreum ATCC 3610
0 and Japonochytrium sp. ATCC 28207 produce polyunsaturated fatty acids, and the present invention has been made based on this finding.
すなわち、本発明はThraustotheca属、Achlya属、Hal
iphthorous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である。That is, the present invention relates to the genera Thraustotheca, Achlya, Hal
A filamentous fungus belonging to the genus iphthorous, Rizophydium or Japonochytrium, culturing a filamentous fungus having a high unsaturated fatty acid content and having a high lipid-producing ability, and collecting a lipid having a high unsaturated fatty acid content from the culture. Is a method for producing polyunsaturated 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、炭素源ととしては、グルコース、エ
タノール、グリセ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、マルツ・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 fungus is not particularly limited. However, as the carbon source, glucose, ethanol, glycerol and the like are preferable, and the amount used is usually in the range of 0.5 to 20% by weight. As the nitrogen source, it is preferable to use an organic nitrogen source such as yeast extract, malt extract, peptone, etc. In addition, inorganic nitrogen such as nitrate can also be used, and the amount used is usually in the range of 0.1 to 10% by weight. is there. Further, in addition to these components, the addition of trace elements such as amino acids and vitamins and inorganic salts are effective in promoting the growth of bacteria.
培地のpHは４〜10、好ましくは５〜８、培養温度は10
〜35℃の範囲が良く、培養時間は２〜15日間程度必要で
ある。The pH of the medium is 4-10, preferably 5-8, and the culture temperature is 10
The temperature is preferably in the range of ~ 35 ° C, and the culturing time is required 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、通気攪拌培養などの方法により行う。The cultivation is usually performed in a liquid culture medium by a method such as stationary culture, shaking culture, or aeration and stirring 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。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。Cells are collected from the thus obtained culture by filtration, centrifugation, or the like, and lipids are extracted from the cells. Extraction of lipids is performed by a conventional method, for example, by solvent extraction.
実施例1. 下記の有機栄養培地１にThraustotheca clavata AT
CC 34112を接種し、25℃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。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。Example 1. Thraustotheca clavata AT in the following organic nutrient medium 1
After inoculating CC 34112 and culturing with shaking at 25 ° C. for 7 days, the cells were collected by filtration and freeze-dried. As a result, 5.8 g of dried cells were obtained. After the dried cells were pulverized and extracted and purified by the Folch method, the solvent was distilled off under reduced pressure, and the total lipid was measured by a gravimetric method. As a result, 0.5 g of crude lipid was obtained.
培地 グルコース 10g/ イースト・エキス 3g/ マルツ・エキス 3g/ トリプトン 5g/ 蒸留水 １ pH 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。Medium Glucose 10g / Yeast extract 3g / Malt's extract 3g / Tryptone 5g / Distilled water 1 pH 7.0 Lipids extracted and purified from cells were partially methylesterified, and 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。ガスクロマトフラフィー分析条件 カラム充填剤 Diasolid ZF カラム温度200℃ キャリアーガス N2 検出器温度250℃ 検出器 FID 注入温度 250℃ カラム 1.5m×３φ 乾燥菌体当りのアラキドン酸含量は21.6mg/gである。In addition, each peak obtained by gas chromatography was identified by mass spectrometry. Gas chromatographic analysis conditions Column packing Diasolid ZF Column temperature 200 ° C Carrier gas N 2 Detector temperature 250 ° C Detector FID injection temperature 250 ° C Column 1.5m × 3φ The arachidonic acid content per dry cell is 21.6 mg / g.
乾燥菌体当りのEPA含量は4.6mg/gである。 The EPA content per dry cell is 4.6 mg / g.
実施例2. 下記の有機栄養培地１にAchlya bisexualis ATCC 1
4524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 bisexualis ATCC 1 in the following organic nutrient medium 1
After inoculating 4524, the same operation as in Example 1 was performed, and the fatty acid composition was analyzed.
結果を表−２に示した。 The results are shown in Table-2.
培地 グルコース 10g/ イースト・エキス 3g/ マルツ・エキス 3g/ ポリペプトン 5g/ 蒸留水 １ pH 6.5 実施例3. 下記の有機栄養培地１にHaliphthorous philippine
nsis ATCC 58303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 / Yeast extract 3g / Malt's extract 3g / Polypeptone 5g / Distilled water 1 pH 6.5 Example 3. Haliphthorous philippine was added to the following organic nutrient medium 1.
nsis ATCC 58303 was inoculated, and thereafter the same operation as in Example 1 was performed to analyze the fatty acid composition. The results are shown in Table-3.
培地 グルコース 5g/ イースト・エキス 1g/ ポリペプトン 1g/ 人工海水 １ pH 6.5 実施例4. 下記の有機栄養培地１にRizophydium littoreum AT
CC 36100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 / Yeast extract 1g / Polypeptone 1g / Artificial seawater 1 pH 6.5 Example 4. Rizophydium littoreum AT in the following organic nutrient medium 1
After inoculating CC 36100, the same operation as in Example 1 was performed, and the fatty acid composition was analyzed. The results are shown in Table-4.
培地 グルコース 5g/ イースト・エキス 1g/ ポリペプトン 1g/ 人工海水 １ pH 6.5 実施例5. 下記の有機栄養培地１にJaponochytrium sp.ATCC 2
8207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。Medium Glucose 5g / Yeast extract 1g / Polypeptone 1g / Artificial seawater 1 pH 6.5 Example 5. Japonochytrium sp. ATCC 2 was added to the following organic nutrient medium 1.
8207, and the same operation as in Example 1 was carried out.
7 g of dried cells were obtained, followed by 0.14 g of crude lipid. Further, the fatty acid composition of this lipid was analyzed in the same manner as in Example 1.
結果を表−５に示した。 The results are shown in Table-5.
培地 グルコース 10g/ イースト・エキス 3g/ マルツ・エキス 3g/ ペプトン 1g/ 人工海水 １ pH 7.0 Medium Glucose 10g / Yeast extract 3g / Malt's extract 3g / Peptone 1g / Artificial seawater 1 pH 7.0
Claims (1)
us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。1. The genus Thraustotheca, the genus Achlya, the Haliphthoro
By culturing a filamentous fungus belonging to the genus Rizophydium or Japonochytrium and having a lipid-producing ability with a high content of highly unsaturated fatty acids, and collecting a lipid with a high content of a highly unsaturated fatty acid from the culture, a filamentous fungus characterized by A method for producing polyunsaturated 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 |
|---|---|---|---|
| JP62-270624 | 1987-10-27 | ||
| JP27062487 | 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 JPH01199588A (en) | 1989-08-10 |
| JP2582622B2 true JP2582622B2 (en) | 1997-02-19 |
Family
ID=26468485
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| 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) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1092352A3 (en) * | 1991-01-24 | 2003-08-27 | Martek Corporation | Microbial oil mixtures and uses thereof |
| WO1998003671A1 (en) | 1996-07-23 | 1998-01-29 | Nagase Biochemicals, Ltd. | Process for preparing docosahexaenoic acid and docosapentaenoic acid |
| EP0960943B1 (en) * | 1996-12-27 | 2014-02-12 | Suntory Holdings Limited | Media for culturing microorganisms and process for producing unsaturated fatty acids or lipids containing the same |
| JP4088097B2 (en) | 2002-04-26 | 2008-05-21 | サントリー株式会社 | Method for producing highly unsaturated fatty acid-containing lipid |
| JP3840459B2 (en) * | 2003-03-20 | 2006-11-01 | 裕司 島田 | Glyceride and method for producing the same |
| US7588931B2 (en) | 2004-11-04 | 2009-09-15 | E. I. Du Pont De Nemours And Company | High arachidonic 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 |
| EP2603092A1 (en) | 2010-08-11 | 2013-06-19 | E.I. Du Pont De Nemours And Company | Improved aquaculture meat products |
| US20120040076A1 (en) | 2010-08-11 | 2012-02-16 | E. I. Du Pont De Nemours And Company | Aquaculture feed compositions |
-
1988
- 1988-06-02 JP JP63134354A patent/JP2582622B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPH01199588A (en) | 1989-08-10 |
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