JP2011519552A - 植物種子油 - Google Patents
植物種子油 Download PDFInfo
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- JP2011519552A JP2011519552A JP2011505517A JP2011505517A JP2011519552A JP 2011519552 A JP2011519552 A JP 2011519552A JP 2011505517 A JP2011505517 A JP 2011505517A JP 2011505517 A JP2011505517 A JP 2011505517A JP 2011519552 A JP2011519552 A JP 2011519552A
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- Prior art keywords
- acid
- oil
- plant seed
- seed oil
- plant
- Prior art date
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Images
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Abstract
【選択図】図1
Description
アラキドン酸(20:4 n−6) 約15
必須脂肪酸:
リノール酸(18:2 n−6) 約20〜25
α−リノレン酸(18:3 n−3) 約3〜7
乳児に有用な追加の脂肪酸:
γ−リノレン酸(GLA)(18:3 n−6) 約6〜11
ジホモ−γ−リノレン酸(DGLA)(20:3 n−6) 約4〜8
ステアリドン酸(SDA) 約1〜2
エイコサペンタエン酸(EPA) 約2〜4;
または
標的脂肪酸 %
アラキドン酸(20:4 n−6) 15
必須脂肪酸:
リノール酸(18:2 n−6) 20〜25
α−リノレン酸(18:3 n−3) 3〜7
乳児に有用な追加の脂肪酸:
γ−リノレン酸(GLA)(18:3 n−6) 6〜11
ジホモ−γ−リノレン酸(DGLA)(20:3 n−6) 4〜8
ステアリドン酸(SDA) 1〜2
エイコサペンタエン酸(EPA) 2〜4
乳児用人工乳は、食品法の用語法で、特に生まれて最初の6ヵ月の摂食のために意図され、かつ乳児が必要とするすべての栄養分を含む、すべての食品および製品として指定される。完成品の調製では、水をさらに加えることもある。
食品法における乳児用のフォローアップフードは、特に約4ヵ月目からの乳児用に意図され、かつ最初の人工乳(formula)と同様に液体粘度(liquid consistency)を有するが、より多量の炭水化物をデンプンの形式で含む、すべての食品および製品である。
補完食は、固形食への変化の準備をするために人工乳の代替物として乳児に使用されるすべての食品および調製物として指定される。
アレルギーへの高い遺伝的傾向を有するアレルギー体質のヒトの乳児および小児用。乳児の場合、腸粘膜は透過性のままであり、かつ異種タンパク質、例えば牛乳由来の異種タンパク質は食品アレルギーを引き起こすため、「低アレルゲン性(hypoallergenic)乳児食」が販売されている。特別食にもARAを補うべきである。
牛乳、ヤギ乳(例えば中国、オーストラリア)、カゼイン/カゼイン塩、脱塩ホエイ粉末、アミノ酸、タウリン、カルニチン;植物油(パーム油、ダイズ油、ヒマワリ油、高度に油を含むヒマワリ油、ベニバナ油、ヤシ油、アブラナ油)、乳脂肪、魚油(マグロ油、タラ肝油、オキアミ油)、卵脂質(オボチン(ovothin))、ARASCO(登録商標)、DHASCO(登録商標);炭水化物(ラクトース、バクガデキストリン、デンプン)、他のタイプの糖、オリゴ糖(プレバイオティクス)、細菌培養(プロバイオティクス);ビタミン、コリン、ミオ−イノシトール;ミネラル(Ca、Na、K、Mg、P、Cl)、微量元素(Fe、Zn、Cu、Mn、Cr、Se、F、I)、ヌクレオチド。
(a)配列番号15、16または17に示される核酸構築物を用いた形質転換によりトランスジェニック植物を作製するステップ、
(b)植物種子油の生合成を可能にする条件下でステップ(a)からのトランスジェニック植物を栽培するステップ、
(c)植物種子を回収し、植物種子油を抽出および精製するステップ
を含む、上記方法に関する。
(i) 好ましくは不活性雰囲気下で、0.2mm未満の粒径に砕くか圧縮することによって植物種子を粉砕するステップ;および
(ii) 圧力が少なくとも300barであり、40〜60℃の範囲の温度であり、抽出処理量がCO2 60kg/時であり、かつ30〜120分後に完了する、超臨界CO2抽出ステップ
を含む。
R1は、ヒドロキシル、補酵素A(チオエステル)、リゾホスファチジルコリン、リゾホスファチジルエタノールアミン、リゾホスファチジルグリセロール、リゾジホスファチジルグリセロール、リゾホスファチジルセリン、リゾホスファチジルイノシトール、スフィンゴ塩基(sphingo base)または式II:
R2は、水素、リゾホスファチジルコリン、リゾホスファチジルエタノールアミン、リゾホスファチジルグリセロール、リゾジホスファチジルグリセロール、リゾホスファチジルセリン、リゾホスファチジルイノシトールまたは飽和もしくは不飽和C2〜C24−アルキルカルボニルであり、
R3は、水素、飽和もしくは不飽和C2〜C24−アルキルカルボニルであるか、またはR2およびR3は、互いに独立して、式Ia:
nは2、3、4、5、6、7または9であり、mは2、3、4、5または6であり、およびpは0または3である、
に示される構造を有する物質を含む。
1. ラッカーおよびコーティング(酸化的乾燥油としての使用)
2. 床材(floor coverings)用ポリマーまたはプラスチック(酸化的乾燥油としての使用)
3. 他の化学的適用
4. 化粧品適用
5. 電子および半導体テクノロジー分野での適用。
クローニング手順、例えば制限切断、アガロースゲル電気泳動、DNA断片の精製、ニトロセルロースおよびナイロンメンブレンへの核酸のトランスファー、DNA断片の連結、大腸菌(Escherichia coli)細胞の形質転換、細菌の培養および組み換えDNAのシーケンス解析はSambrook et al. (1989) (Cold Spring Harbor Laboratory Press)に記載のように行った。
植物中の、または所望の化合物(例えば脂肪酸)の産生に対する遺伝子改変の影響は、適した条件(例えば上で記載される条件)下で改変植物を培養し、所望の生成物(すなわち脂質または脂肪酸)の生産の増加に関して培地および/または細胞成分を調査することによって決定することができる。これらの分析技術は当業者に公知であり、分光学、薄層クロマトグラフィー、種々のタイプの染色手順、酵素および微生物学的処理および分析用クロマトグラフィー、例えば高速液体クロマトグラフィーを含む(例えば、Ullman, Encyclopedia of Industrial Chemistry, Vol. A2, pp. 89-90 and pp. 443-613, VCH: Weinheim (1985); Fallon, A., et al., (1987) "Applications of HPLC in Biochemistry" in: Laboratory Techniques in Biochemistry and Molecular Biology, Vol. 17; Rehm et al. (1993) Biotechnology, Vol. 3, Chapter III: "Product recovery and purification", pp. 469-714, VCH: Weinheim; Belter, P.A., et al. (1988) Bioseparations: downstream processing for Biotechnology, John Wiley and Sons; Kennedy, J.F., and Cabral, J.M.S. (1992) Recovery processes for biological Materials, John Wiley and Sons; Shaeiwitz, J.A., and Henry, J.D. (1988) Biochemical Separations, in: Ullmann's Encyclopedia of Industrial Chemistry, Vol. B3; Chapter 11, pp. 1-27, VCH: Weinheim; およびDechow, F.J. (1989) Sep
aration and purification techniques in biotechnology, Noyes Publicationsを参照のこと)。
アブラナの種子でのLC−PUFAの合成のために、代謝経路で必要な遺伝子(表1)を、発現エレメント(プロモーター、ターミネーター、表2)と組み合わせて、形質転換ベクターに導入した。
トランスジェニックアブラナ植物(Brassica napus)の作製では、適切に組み合わされた遺伝子を有する、上記でさらに記載されたpSUNプラスミドなどのバイナリーベクターをAgrobacterium tumefaciens C58C1:pGV2260中に形質導入した(Deblaere et al, 1984, Nucl. Acids. Res. 13, 4777-4788)。アブラナ植物の形質転換では、3%スクロースを含むMurashige−Skoog培地(Murashige and Skoog 1962 Physiol. Plant. 15, 473)(3MS培地)中の陽性に形質転換されたアグロバクテリウムコロニーの一晩培養物の1:50希釈物を使用した。新たに発芽させた滅菌アブラナ植物の葉柄または子葉下部(hypocotyledons)(それぞれ〜約1cm2)をペトリ皿で1:50アグロバクテリウム希釈液と5〜10分間インキュベートした。0.8%Bacto agarを含む3MS培地中で25℃で暗所で3日間の共インキュベーションを続けて行った。16時間の明期/8時間の暗期での3日間の後に培養を継続し、500mg/LのClaforan(セフォタキシムナトリウム)、50mg/Lのカナマイシン、20mMのベンジルアミノプリン(BAP)および1.6g/Lのグルコースを含むMS培地上で1週間の周期性で継続した。2%スクロース、250mg/LのClaforanおよび0.8%Bacto agarを含むMS培地に成長中の発芽を移した。3週間後に根が形成されなければ、発根の成長ホルモンとして2−インドール酪酸を培地に加えた。
トランスジェニックアマ植物の作製は、例えばBell et al., 1999, In Vitro Cell. Dev. Biol.-Plant. 35(6):456-465の方法にしたがって微粒子銃によってもたらすことができる。例えばMlynarova et al. (1994), Plant Cell Report 13: 282-285にしたがってアグロバクテリア媒介性形質転換を生じさせることができる。
実施例3で作製されたプラスミドを実施例4に記載のようにアブラナ(Brassica napus)に形質導入した。PCRによるトランスジェニック植物の選択後、これらを種子成熟(昼/夜サイクル:16時間,200mE,21℃,8時間暗,19℃)まで育て、種子を収穫した。
実施例4で作製されたトランスジェニックアブラナ植物および野生型アブラナ植物の種子の収穫、清浄化および空気乾燥(残留水分約7%)後、本発明の植物種子油および野生型油の取得のために、得られた種子を加工した。加工は、種子の粉砕および圧縮で始まり、抽出が続いた。抽出はヘキサンを用いて1回行い、他方、超臨界CO2抽出を用いて行った。次いで、ヘキサンで抽出された油の精製および安定化を行った。
本発明の植物種子油は、以下の重量パーセント(パーセント単位の、総脂肪酸含量のうちの脂肪酸の質量)の、乳児食に重要な脂肪酸を含む。
アラキドン酸(20:4 n−6) 15
必須脂肪酸:
リノール酸(18:2 n−6) 20〜25
α−リノレン酸(18:3 n−3) 3〜7
乳児に有用な追加の脂肪酸:
γ−リノレン酸(GLA)(18:3 n−6) 6〜11
ジホモ−γ−リノレン酸(DGLA)(20:3 n−6) 4〜8
ステアリドン酸(SDA) 1〜2
エイコサペンタエン酸(EPA) 2〜4
本明細書中に記載の典型的な乳児食中のARA含量を、授乳の最初の0〜12ヵ月間に母乳中で見出されたARAの総量に適合させた。追加の利点は、乳児乳にARAを補うために本発明の植物種子油を用いると、GLA、DGLA、SDAおよびEPAの値も母乳中の濃度範囲内になることにある。これは、本発明の植物種子油が3種の高級不飽和脂肪酸をほぼ母乳中でも見出された比で含むためである。ARA濃度を適合させるために本発明の植物種子油が乳児食の成分として同様に使用されると、GLA、DGLA、SDAおよびEPAは、特別な乳児食、ベビーフードおよびチャイルドフードに、対応する栄養分を利用可能にするために適当な濃度で供給される。この場合、例えば、さらにGLA、DGLA、SDAおよびEPAを含む油の例えば混合物のような油の変化は必要ではない。
本発明の植物種子油はまた、繁殖結果を向上させるための飼料適用のためのフードサプリメント製品として適している。それは、畜産(例えばサケ、ウシ、ブタ、ニワトリの畜産)結果を向上させるため、かつペット(例えばネコおよびイヌ)の健康のための飼料添加物として使用することができる。このために、本発明の植物種子油は、若い動物または母親動物への栄養摂取にARAを添加した場合に、生殖率を向上させるために適した濃度のアラキドン酸(ARA)を含む。
本発明の植物種子油の添加により、工業的適用のための重合成分として二重結合を含む特有の高濃度の不飽和脂肪酸を有する工業油が得られる。
1. ラッカーおよびコーティング(酸化的乾燥油としての使用)
2. 床材用ポリマーまたはプラスチック(酸化的乾燥油としての使用)
3. 他の化学的適用
4. 化粧品適用
5. 電子および半導体テクノロジー分野での適用。
Claims (15)
- 総脂肪酸含量のうち約7〜約26重量%を構成するアラキドン酸を含み、アラキドン酸とγ−リノレン酸との重量パーセント比が約1:1〜約5:1であり、アラキドン酸とジホモ−γ−リノレン酸との重量パーセント比が約1:1〜約5:1である植物種子油。
- リノール酸とα−リノレン酸との重量パーセント比が約3:1〜約12:1である、請求項1に記載の植物種子油。
- アラキドン酸とエイコサペンタエン酸との重量パーセント比が約3:1〜約7:1である、請求項1または2に記載の植物種子油。
- ステアリドン酸をさらに含む、請求項1〜3のいずれか1項に記載の植物種子油。
- ステアリドン酸が総脂肪酸含量の約0.1〜約1重量%を構成して存在する、請求項4に記載の植物種子油。
- 配列番号15、16または17に示される核酸構築物を用いて形質転換されたトランスジェニック植物から取得可能である、請求項1〜5のいずれか1項に記載の植物種子油。
- パルミチン酸、ステアリン酸、オレイン酸、リノール酸、γ−リノレン酸、α−リノレン酸、ステアリドン酸、ジホモ−γ−リノレン酸、アラキドン酸およびエイコサペンタエン酸を含む脂肪酸スペクトルを含む、植物種子油。
- 総脂肪酸含量に基づき、約3.2〜5.3%のパルミチン酸、約2.2〜5.3%のステアリン酸、約10〜25%のオレイン酸、約22〜36%のリノール酸、約4〜12%のγ−リノレン酸、約3〜8%のα−リノレン酸、約0.2〜1%のステアリドン酸、約3〜9%のジホモ−γ−リノレン酸、約12〜25%のアラキドン酸および約1〜4%のエイコサペンタエン酸を含む、請求項7に記載の植物種子油。
- 請求項1〜8のいずれか1項に記載の植物種子油と、植物油、微生物油および魚油からなる群より選択される少なくとも1種のさらなる油とを含む配合物または混合油であって、該植物油、微生物油または魚油はドコサヘキサエン酸を含む、上記配合物または混合油。
- 請求項1〜8のいずれか1項に記載の植物種子油または請求項9に記載の配合物もしくは混合油を含む食品。
- 請求項1〜8のいずれか1項に記載の植物種子油または請求項9に記載の配合物もしくは混合油を含むベビーフード。
- 以下のステップ:
(a)配列番号15、16または17に示される核酸構築物を用いた形質転換によりトランスジェニック植物を作製するステップ、
(b)植物種子油の生合成を可能にする条件下でステップ(a)からのトランスジェニック植物を栽培するステップ、および
(c)植物種子を回収し、植物種子油を抽出および精製するステップ
を含む、請求項1〜8のいずれか1項に記載の植物種子油の製造方法。 - 前記植物種子油を、油、脂質または脂肪酸組成物として配合するステップ(d)をさらに含む、請求項12に記載の方法。
- 前記油、脂質または脂肪酸組成物をさらに配合して食品、好ましくはベビーフードを提供する、請求項13に記載の方法。
- 食品、好ましくはベビーフード、化粧品、および飼料、好ましくは魚飼料、または医薬品の製造のための、請求項1〜8のいずれか1項に記載されるかもしくは請求項12〜14のいずれか1項に記載の方法により製造される植物種子油の、または請求項11に記載の配合物もしくは混合油の使用。
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JP2018075003A (ja) * | 2012-06-15 | 2018-05-17 | コモンウェルス サイエンティフィック アンド インダストリアル リサーチ オーガナイゼーション | 植物細胞における長鎖多価不飽和脂肪酸の産生 |
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JP7254744B2 (ja) | 2012-06-15 | 2023-04-10 | コモンウェルス サイエンティフィック アンド インダストリアル リサーチ オーガナイゼーション | 植物細胞における長鎖多価不飽和脂肪酸の産生 |
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JP2017503053A (ja) * | 2013-12-18 | 2017-01-26 | コモンウェルス サイエンティフィック アンド インダストリアル リサーチ オーガナイゼーション | 長鎖多価不飽和脂肪酸を含む脂質 |
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JP7337499B2 (ja) | 2017-12-19 | 2023-09-04 | 日本ニュートリション株式会社 | 豚用飼料添加物および哺乳子豚の発育改善方法 |
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US8455035B2 (en) | 2013-06-04 |
WO2009130291A3 (de) | 2010-05-06 |
AU2009239949A1 (en) | 2009-10-29 |
US20110039010A1 (en) | 2011-02-17 |
EP2271223A2 (de) | 2011-01-12 |
CA2721879A1 (en) | 2009-10-29 |
WO2009130291A2 (de) | 2009-10-29 |
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