JP2017513455A - 高温で成熟胞子接種及び鉄イオン媒介ハーバー・ワイス反応によるヘマトコッカスプルビアリス内アスタキサンチン生産量増進方法 - Google Patents
高温で成熟胞子接種及び鉄イオン媒介ハーバー・ワイス反応によるヘマトコッカスプルビアリス内アスタキサンチン生産量増進方法 Download PDFInfo
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Abstract
Description
本発明に使用された菌株は、Haematococcus pluvialis NIES−144であり、National Institute for Environmental Studies,Tsukuba,Japanから購入して使用した。
下記の表1で、
−NIES−C培地:autotrophic medium(独立栄養培地、目的:生長)
−NIES−N培地:autotrophic medium(独立栄養培地、目的:生育抑制、光誘起、アスタキサンチン生産)
−CO2 supply:3%(v/v)
−光条件:20μE/m2/s(for vegetative growth);150μE/m2/s(for inductive growth)
−FeSO4:450μM
−MV(methyl viologen):10−11M、10−9M、10−7M(artificial O2−generator)
−H2DCFDA(carboxy−2’,7’−dichlorofluorescein diacetate)5μM
−DCF:H2DCFDAが活性酸素(O2 −、H2O2)により酸化されて生成される物質
−SOD(superoxide dismuase):SODは、細胞内でO2 −をH2O2に転換する酵素
−MDA(malondialdehyde):MDAは、細胞内PUFA(polyunsaturated fatty acids)が酸素(O2)及び活性酸素(O2 −、H2O2及びOH・)により酸化されて転換される2次代謝物質であり、細胞内MDA量を通して細胞の酸化程度を間接的に確認できる一種の酸化指標である。
独立栄養条件で多様な高温の培養条件及び鉄イオン媒介Haber−Weiss反応がヘマトコッカスプルビアリスのアスタキサンチン蓄積に及ぼす影響を調べてるために、下記の実験を行った。
図2(A)に示したように、独立栄養条件で温度の増加によるヘマトコッカスプルビアリスのバイオマス生産能力は、23℃に比べて30℃で30%、36℃で57%減少することが確認された反面、培養温度が30℃で、Fe2+を添加してHaber−Weiss反応を誘導した場合、ヘマトコッカスプルビアリスのバイオマス生産能力はFe2+を添加せず、培養温度が23℃である場合に比べて9%増加したことが確認され、Fe2+を添加せず、培養温度が30℃である場合に比べて41%増加したことが確認された。
図2(B)に示したように、独立栄養の条件で温度の増加によるヘマトコッカスプルビアリスのアスタキサンチン生産能力は、培養温度が23℃である場合に比べて30℃で23%、36℃で42%減少することが確認された反面、培養温度が30℃で、Fe2+を添加してHaber−Weiss反応を誘導した場合、ヘマトコッカスプルビアリスのアスタキサンチン生産能力は、Fe2+を添加せず、培養温度が23℃である場合に比べて17%増加したことが確認され、Fe2+を添加せず、培養温度が30℃である場合に比べて66%増加したことが確認された。
高温の独立栄養条件でアスタキサンチン合成時、ヘマトコッカスプルビアリスのアスタキサンチン生合成阻害現象を解明するために、下記の実験を行った。
室温(23℃)の独立栄養条件でアスタキサンチン合成時ヘマトコッカスプルビアリス細胞内LROS(O2 −、H2O2)の増加によるアスタキサンチン生合成阻害現象を解明するために、有機炭素源が欠乏したNIES−C培地で低光度の光(20μE/m2/s)とただ二酸化炭素のみを唯一炭素源として生育して対数期に達した細胞培養液(OD680=約0.8)をNIES−N培地に移して増殖を抑制して、強い光(150μE/m2/s)と共に様々な濃度(10−11〜10−7M)のMethyl viologenを添加してO2 −を人為的に強度毎で発生させて、これと共に通常の室温の培養温度(23℃)で鉄イオン媒介Haber−Weiss反応を誘導する物質FeSO4(450μM)を添加した場合としなかった場合を分けてアスタキサンチン合成条件を作った後、培養4日目細胞内アスタキサンチン蓄積程度を分析した。
4−1:グリーンステージで成熟胞子(cyst)の接種
本実施例では、二段(two−stage)光培養工程で、ヘマトコッカスプルビアリス内アスタキサンチンの生産量が増進されることを確認した。
実施例4−1の結果により、夏場高温の独立栄養条件のグリーンステージで成熟胞子(red cyst)接種及びレッドステージで鉄イオン添加を通して、ヘマトコクス細胞のアスタキサンチン生産量を調べた。グリーンステージ培養は、15日間行って、鉄イオン添加後レッドステージ培養は63日間行った。
Claims (8)
- 下記のステップを含む、ヘマトコッカスプルビアリス(Haematococcus pluvialis)の培養によるアスタキサンチンの生産量を増進させる方法:
(a)ヘマトコッカスプルビアリスの成熟胞子(cyst)を接種して増殖(vegetative growth)させるステップ;及び
(b)窒素が欠乏して鉄イオンが添加された独立栄養条件で、100〜300μE/m2/sの光度を照射してヘマトコッカスプルビアリス内アスタキサンチンの生成を誘導するステップ。 - 前記培養温度は25〜40℃であることを特徴とする請求項1に記載のアスタキサンチンの生産量を増進させる方法。
- 前記独立栄養条件は、光合成のための無機炭素源として3〜4%の二酸化炭素を供給することを含む請求項1に記載のアスタキサンチンの生産量を増進させる方法。
- 前記ステップ(a)は、35μE/m2/s以下の光度を照射することを含む請求項1に記載のアスタキサンチンの生産量を増進させる方法。
- 前記鉄イオンは、Fe2SO4、FeCl2、FeCl3及びFe2(SO4)3からなる群から選択される1種以上であることを特徴とする請求項1に記載のアスタキサンチンの生産量を増進させる方法。
- 前記鉄イオンの濃度は、40〜80μMであることを特徴とする請求項1に記載のアスタキサンチンの生産量を増進させる方法。
- 前記鉄イオンは、活性酸素O2 −及びH2O2含量に対して100〜600モル比で添加されることを特徴とする請求項1に記載のアスタキサンチンの生産量を増進させる方法。
- 前記アスタキサンチンの含量は、ヘマトコッカスプルビアリス細胞内活性酸素O2 −及びH2O2が活性酸素O2及びOH・に転換することによって増進されることを特徴とする請求項1に記載のアスタキサンチンの生産量を増進させる方法。
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2001061466A (ja) * | 2000-07-31 | 2001-03-13 | Higashimaru Shoyu Co Ltd | アスタキサンチンの製造方法 |
JP2004033070A (ja) * | 2002-07-01 | 2004-02-05 | Yamaha Motor Co Ltd | 緑藻ヘマトコッカスへの外来遺伝子導入方法 |
WO2007029627A1 (ja) * | 2005-09-06 | 2007-03-15 | Yamaha Hatsudoki Kabushiki Kaisha | アスタキサンチン含量の高い緑藻抽出物およびその製造方法 |
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CN1181184C (zh) * | 2002-07-26 | 2004-12-22 | 中国科学院武汉植物研究所 | 一种培养雨生红球藻生产虾青素的方法 |
CN101144058A (zh) * | 2007-08-22 | 2008-03-19 | 厦门大学 | 一种虾青素的微藻培养基 |
KR101046310B1 (ko) * | 2008-03-04 | 2011-07-07 | 서희동 | 해양 심층수를 이용하여 해마토코쿠스 조류를 배양하는 방법과 이를 이용한 아스타크산틴을 생산하는 방법 |
CN101586140B (zh) * | 2009-06-09 | 2011-09-07 | 宁波大学 | 一种培养雨生红球藻生产虾青素的简易方法 |
CN101974599B (zh) * | 2010-10-14 | 2011-11-09 | 山东理工大学 | 利用油菜素内酯刺激雨生红球藻快速生产虾青素的方法 |
CN101974598A (zh) * | 2010-10-14 | 2011-02-16 | 山东理工大学 | 利用茉莉酸促进雨生红球藻生产虾青素的方法 |
KR101356477B1 (ko) * | 2011-06-27 | 2014-01-29 | 고려대학교 산학협력단 | 광민감성 증가로 인한 아스타잔틴 생산력이 향상된 고광유발 헤마토코쿠스 돌연변이체 및 이의 선별방법 |
CN102337215A (zh) * | 2011-10-20 | 2012-02-01 | 烟台华融生物科技有限公司 | 培养雨生红球藻及生产虾青素的方法 |
CN103571906B (zh) * | 2012-07-27 | 2018-12-11 | 上海泽元海洋生物技术有限公司 | 一种利用微藻高效生产虾青素的新方法 |
CN102994603B (zh) * | 2012-12-21 | 2014-09-10 | 丽江程海保尔生物开发有限公司 | 一种培养雨生红球藻生产虾青素的转化方法 |
CN103044304B (zh) * | 2012-12-21 | 2014-12-03 | 宁波红龙生物科技有限公司 | 从雨生红球藻粉中制备虾青素提取物的方法 |
CN103114121A (zh) * | 2013-01-31 | 2013-05-22 | 宁波大学 | 一种雨生红球藻生产虾青素的方法 |
CN103232375B (zh) * | 2013-04-03 | 2015-04-29 | 大连医诺生物有限公司 | 雨生红球藻中虾青素的高效提取工艺 |
CN103695314B (zh) * | 2013-12-26 | 2015-11-04 | 宁波大学 | 一种雨生红球藻营养细胞的保存方法 |
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2001061466A (ja) * | 2000-07-31 | 2001-03-13 | Higashimaru Shoyu Co Ltd | アスタキサンチンの製造方法 |
JP2004033070A (ja) * | 2002-07-01 | 2004-02-05 | Yamaha Motor Co Ltd | 緑藻ヘマトコッカスへの外来遺伝子導入方法 |
WO2007029627A1 (ja) * | 2005-09-06 | 2007-03-15 | Yamaha Hatsudoki Kabushiki Kaisha | アスタキサンチン含量の高い緑藻抽出物およびその製造方法 |
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