JP2022050546A - 微細藻類を含む組成物及びこの組成物を製造し使用する方法 - Google Patents
微細藻類を含む組成物及びこの組成物を製造し使用する方法 Download PDFInfo
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Abstract
Description
(a)偏性独立栄養微細藻類及び任意で偏性従属栄養微細藻類を、浮遊性要素を含む組成物へと調剤するステップであって、前記微細藻類の生育可能性を持続する条件下に行われるステップと、
(b)前記組成物中で前記微細藻類を培養し、これによって前記栄養組成物を製造するステップと
を含む方法である。
(a)少なくとも2つの区画を含む区画化済み組成物を製造するステップであって、前記少なくとも2つの区画のうち第1区画が偏性光独立栄養微細藻類を含み、前記少なくとも2つの区画のうち第2区画が偏性従属栄養又は混合栄養微細藻類を含み、前記区画が、前記偏性光独立栄養微細藻類と前記偏性従属栄養又は混合栄養微細藻類との間の共生を確実にする構造及び/又は組成から設計される、ステップと、
(b)粒子内で前記微細藻類を培養し、これによって前記栄養組成物を製造するステップと
を含む方法である。
(a)偏性独立栄養微細藻類及び任意で偏性従属栄養微細藻類を、浮遊性要素を含む組成物へと調剤するステップであって、微細藻類の生育可能性を持続する条件下に行われるステップと、
(b)粒子内で微細藻類を培養し、これによって栄養組成物を製造するステップと
を含む方法である。
(a)少なくとも2つの区画を含む区画化済み組成物を製造するステップであって、少なくとも2つの区画のうち第1区画が偏性光独立栄養微細藻類を含み、少なくとも2つの区画のうち第2区画が偏性従属栄養又は混合栄養微細藻類を含み、これらの区画が、偏性光独立栄養微細藻類と偏性従属栄養又は混合栄養微細藻類との間の共生を確実にする構造及び/又は組成から設計される、ステップと、
(b)粒子内で微細藻類を培養し、これによって栄養組成物を製造するステップと
を含む方法である。
「ゲルマイクロドロップ」(GMD)(本明細書ではゲルビーズ又はゲル粒子とも称される)は、ゲル(そして任意で液体)材料に含まれる非常に僅かな小滴、即ち非常に少量の存在物を参照するのであり、この存在物は、0個、1個、又は多数の生物学的存在物を含有することができる。例えば、アガロースGMD内に2種類以上の微細藻類の種をカプセル化することができる。特に、微細藻類と成長培地と液体アガロースとを含有する水滴をフッ素化オイル中で形成することができる。任意でGMDは、無機及び/又は有機化合物を含有することができる。これらの化合物は、任意で溶液中に存在させることができる。GMDは、別の液体、例えば非水液体に含まれる境界によって又は透水障壁、例えば膜によって規定することのできる容積を有する。膜は、対象である生物学的存在物(例えば微細藻類)をGMD内部に留めておくことができ、その他の生物学的存在物、例えば分子(微細藻類よりも小さいもの)を通過させることもできる。例えば、2つ以上の異なる微細藻類株の2つ以上の流れを生成し、その後、これらの流れを組み合わせて単一の小滴にし、その後、この小滴を重合してGMD(例えばアガロースGMD)にすることが可能であろう。この場合微細藻類株は区画化され、空間的に分離される。GMDは任意の形状とすることができるが、しばしばほぼ球形である。というのも、GMDの境界に関連する力が、変形可能なGMDをまとめようとする傾向をもつからである。その他の力として、例えば、GMD浮遊液の攪拌に関連する流体力学的剪断力、表面への付着力、又は重力に、球形状からの逸脱を引き起こす傾向がある。更に、その容積がGMD容積の比較的大部分である存在物をGMDが含有する又はGMDをこの存在物が占めることにより、GMDを非球形にすることができる。従って、例えば薄ゲル被膜により(任意で水溶液と共に)包囲される細胞又は細胞の集団がGMDであり、この薄ゲル被膜は非水液体により包囲される。同様に、薄ゲル被膜により(任意で水溶液と共に)包囲される非生物学的粒子もGMDであり、この薄ゲル被膜は非水液体により包囲される。
別の実施形態において、小滴は2つ以上のビーズを含有する。各ビーズは、同じ又は異なる種類、形状、及び寸法とすることができる。ビーズは接続することができる。ビーズはゲルビーズとすることができ、例えばアガロースビーズとすることができる。例えば、層状ビーズを生成することができ、その場合、各層は別々の種類の微細藻類を有することになる。幾つかの例において、ビーズは磁性にすることにより所望の領域に、例えば環境に分布させることができ、その後、ビーズがもはや不要である際には所望すれば、例えば電磁石又は永久磁石を使用することにより、容易に回収することができる。
小滴は、二相系(例えば有機相/水相、フルオラス相/水相)中、又は乳化剤/界面活性剤をもつ単相(例えば水性バルク溶液により包囲される水滴)中のいずれかに存在する液体(普通は水性液体)とすることができる。「プラグ」とは特定の種類の小滴のことである(Song et al., 2006, Angew. Chem. Int. Ed. 45: 7336-7356、Chen et al., 2006, Curr. Opin. Chem. Bio. 10: 226-231)。
第1ステップ―クロレラ藻類の従属栄養成長のための内側区画の製造
生きている単細胞の混合栄養クロレラ藻類を、アルギン酸の負に帯電した塩(約1/2、w/v)、例えば0.5~10%アルギン酸ナトリウム中で浮遊させる。この混合物の液滴を、淡水(約98%)及び食用の水溶性カルシウム塩(約2%)、例えば塩化カルシウム又は乳酸カルシウムを含有する硬化浴中に約2~30cmから滴下する。液滴(約0.1mm~3mm)を約1~30分の間浴中で放置すると、その後カプセルは硬くなり、破損することなく容易に取り扱われる。次に、カプセルを濾過により浴から除去して水で洗う。
生きている単細胞の偏性光独立栄養スピルリナ藻類を、アルギン酸の透過性塩(約1/2、w/v)、例えば0.5~10%アルギン酸ナトリウム中で浮遊させる。クロレラ藻類の入ったアルギン酸塩カプセル(第1ステップ)をスピルリナ藻類のアルギン酸塩浮遊液中に移す。アルギン酸塩中にクロレラのアルギン酸塩/キトサンカプセル及びスピルリナ藻類浮遊液を含有するこの混合物の液滴を、水(約98%)及び食用の水溶性カルシウム塩(約2%)、例えば塩化カルシウム又は乳酸カルシウムを含有する硬化浴に約2~30cmから滴下する。液滴(約0.5mm~20mm)を約1~30分の間浴中で放置すると、その後カプセルは硬くなり、破損することなく容易に取り扱われる。次に、カプセルを浴から除去して洗う。
従属栄養及び光独立栄養藻類を含有する区画化済みカプセルを、様々な塩及び固定炭素源(例えばグルコース)を含有する培地中、例えば水中で培養する。培地は、クロレラ藻類株に依存して、NH4.NO3(0.125g/L)、CaCl2.2H2O(0.025g/L)、MgSO4.7H2O(0.075g/L)、KQHPO4(0.075g/L)、KHZPO4(0.175g/L)、NaCl(0.025g/L)、及びグルコース(0.1g/L~2g/L)を含有する。
1.網等を使用する単純な方法により、浮遊カプセルを全部採取することができる。
2.微細藻類の異なる区画からの個別採取/単離は、様々な溶媒中での周辺区画及び中央区画(例えば、アルギン酸塩対キトサン)の溶解度が異なることに依拠する。
アルギン酸カルシウムは、中性pHであるポリリン酸ナトリウム及び炭酸ナトリウムの溶液中に可溶であるが、キトサンは酸性溶液(pH約5)のみに可溶である。
。
区画化済みアルギン酸塩ビーズ中の光独立栄養及び混合栄養微細藻類の共カプセル化(実施例1に記載したようなもの)により、恐らくは固定化した微細藻類の種の抗酸化特性と抗菌特性との相乗効果に起因する、共培養した微細藻類の種の一方又は両方の保護も与えられる。図3A、図3Bに示す画像は、区画化済みアルギン酸塩ビーズ中の光独立栄養スピルリナ及び混合栄養クロレラ微細藻類の共カプセル化が、貯蔵寿命を延ばし、暗所にて室温での保存中、開発された製品の市場性のある特徴を数週間の間維持できることを明示する。図3Aは、クロレラ藻類と共カプセル化されたスピルリナ細胞の暗所にて室温での良好に維持された構造を示す。対照的に、同じ条件で保存されたスピルリナ微細藻類単独のものは脱色し崩壊した(図3B)。
Claims (17)
- 少なくとも2つの区画を含む区画化済み組成物であって、
前記少なくとも2つの区画のうち第1区画が、偏性光独立栄養微細藻類を含み、
前記少なくとも2つの区画のうち第2区画が、偏性従属栄養又は混合栄養微細藻類を含み、
前記区画が、前記偏性光独立栄養微細藻類と前記偏性従属栄養又は混合栄養微細藻類との間の共生を確実にする構造及び/又は組成から設計される
区画化済み組成物であって、
前記少なくとも2つの区画のうち前記第1区画が、光に対して透過性であり、
前記第2区画が、混合栄養微細藻類を含む場合に光に対して不透過性である
区画化済み組成物。 - 請求項1に記載の組成物であって、前記少なくとも2つの区画間での小分子、ミネラル、及びガスの自由拡散を可能にする組成物。
- 請求項1~2のいずれか1項に記載の組成物であって、カプセルとして調剤された組成物。
- 請求項1~3のいずれか1項に記載の組成物であって、生物体により摂取可能である組成物。
- 請求項4に記載の組成物であって、前記生物体が、人間である組成物。
- 請求項1~5のいずれか1項に記載の組成物であって、前記従属栄養微細藻類又は前記混合栄養微細藻類が、前記偏性光独立栄養微細藻類の成長率よりも速い成長率を特徴とする組成物。
- 請求項1~6のいずれか1項に記載の組成物であって、前記混合栄養微細藻類が、クロレラの種及びクラミドモナスの種から成る群から選択される組成物。
- 請求項1~6のいずれか1項に記載の組成物であって、
前記混合栄養微細藻類が、クロレラの種の群からのものであり、
前記偏性光独立栄養微細藻類が、スピルリナの種の群からのものである
組成物。 - 請求項1~7のいずれか1項に記載の組成物であって、前記微細藻類が、遺伝子改変される組成物。
- 請求項1~9のいずれか1項に記載の組成物であって、前記第2区画が、濁度に影響を及ぼす添加剤を含む組成物。
- 栄養組成物を製造する方法であって、
(a)少なくとも2つの区画を含む、カプセルとして調剤された区画化済み組成物を製造するステップであって、前記少なくとも2つの区画のうち第1区画が、偏性光独立栄養微細藻類を含み、前記少なくとも2つの区画のうち第2区画が、偏性従属栄養又は混合栄養微細藻類を含み、前記区画が、前記偏性光独立栄養微細藻類と前記偏性従属栄養又は混合栄養微細藻類との間の共生を確実にする構造及び/又は組成から設計されるステップと、
(b)粒子内で前記微細藻類を培養し、これによって前記栄養組成物を製造するステップと
を含む方法。 - 請求項11に記載の方法であって、前記カプセルが、繊維又は球体として成形される方法。
- 請求項11~12のいずれか1項に記載の方法であって、前記カプセル中の前記偏性光独立栄養微細藻類の濃度が、106~1010細胞/cm3である方法。
- 請求項11~13のいずれか1項に記載の方法であって、前記組成物が、食用である方法。
- 請求項11に記載の方法であって、前記培養のステップに続いて、前記微細藻類を単離するステップを更に含む方法。
- 請求項11~15のいずれか1項に記載の方法であって、前記培養のステップが、屋外を含む方法。
- 請求項11~16のいずれか1項に記載の方法であって、前記培養のステップが、室内を含む方法。
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