JP6947810B2 - 油糧微生物を含む発酵ブロスから多価不飽和脂肪酸を含む微生物油を抽出するための方法 - Google Patents
油糧微生物を含む発酵ブロスから多価不飽和脂肪酸を含む微生物油を抽出するための方法 Download PDFInfo
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Description
[0001]本出願は、その開示内容全体を本明細書に援用する、2016年7月13日に出願された米国仮特許出願第62/361,770号明細書の出願日の利益を主張する。
[0002]多くの有益な栄養素の食事摂取量を増やすことは望ましい。特に有益な栄養素として、オメガ3およびオメガ6長鎖多価不飽和脂肪酸(LC−PUFA)などの脂肪酸およびそれらのエステルが挙げられる。現在主に魚油または微生物油から得られる長鎖オメガ3およびオメガ6脂肪酸は、ヒトの食事の必須要素である。
[0007]本発明は、溶解した油糧微生物を含む発酵ブロスの解乳化を促進するための方法であって、a)発酵ブロスから水を除去する(溶解した油糧微生物を含む発酵ブロスの体積は、その当初の体積の60%未満である)こと;およびb)60℃〜110℃の温度に加熱することにより発酵ブロスを解乳化することを含む方法を対象とする。
[0040]適例として本明細書に示した実施形態は、例示を意図しており、限定を意図するものではない。
[実施例1]
[0089]図1および2に図示したように、微生物細胞懸濁液は、微生物細胞の溶解前、溶解中または溶解後のいずれで脱水してもよい。細胞溶解後の脱水の1つの特定の例を以下に説明する。
[0093]微生物細胞(シゾキトリウム・エスピー(Schizochytrium sp.))を含む未洗浄細胞ブロス(157.4kg)を60℃で1時間低温殺菌した。次いでブロスのpHを7.5に調整し、アルカラーゼ(登録商標)酵素(ノボザイムズ(フランクリントン,ノースカロライナ州)から入手可能)を細胞ブロスの重量に対して0.15%の量で加えた。ブロスを140RPMの速度で撹拌し、温度を60℃で2時間維持した。2時間後、溶解細胞組成物を90℃に加熱し、16.9%の当初の全固形分量から30.5%の最終全固形分量に蒸発させた。この結果、微生物油および細胞片を含む87.2kgの濃縮ブロスが得られた。体積減少率は44.5%であった。この溶解し濃縮した細胞組成物のpHを、2.6kgの50%NaOHを加えることにより10.5に調整した。ブロスを140RPMで撹拌し、24時間保持した。保持期間中、pHが9未満に下がったときpHを10に戻すため、NaOHを用いてさらに1回pH調整を行った。合一期間の終了時、2.8kgの3N H2SO4を用いてpHを9.7から8.0に調整し、温度を80℃に低下させた。形成された粗油相を、遠心(Alfa Laval Disc Stack Centrifuge、LAPX 404/Clara 20)により溶解細胞組成物から分離した。抽出収率は91.8%であった。
Claims (43)
- 溶解した油糧微生物を含む未洗浄の発酵ブロスの解乳化を促進するための方法であって、
a)前記発酵ブロスから水を除去して、前記溶解した油糧微生物を含む前記発酵ブロスの体積を、その当初の体積の60%未満に減少させること;および
b)前記発酵ブロスを60℃〜110℃の温度に加熱することにより解乳化すること
を含み、前記解乳化において塩を加えない方法。 - 前記解乳化は、解乳化の時間をステップa)が行われないときの解乳化に必要とされる時間の少なくとも1/3に短縮することにより促進される、請求項1に記載の方法。
- c)前記発酵ブロスから油を回収することをさらに含む、請求項1または請求項2に記載の方法。
- 油の前記回収は、溶媒の使用なしに行われる、請求項3に記載の方法。
- 前記回収される油の量は、ステップa)が行われないときの同じ方法と比較して少なくとも7%増加する、請求項4に記載の方法。
- ステップa)の溶解した油糧微生物を含む前記発酵ブロスの体積は、その当初の体積の70%未満、好ましくは80%未満に減少する、請求項1〜5のいずれか一項に記載の方法。
- ステップa)の水の除去は、前記発酵ブロスを110℃以下、好ましくは70℃〜100℃、一層好ましくは80℃〜90℃の温度で加熱することにより行われる、請求項1〜6のいずれか一項に記載の方法。
- ステップb)は、アルカリ化剤、好ましくは苛性ソーダを加えることを含む、請求項1〜7のいずれか一項に記載の方法。
- 前記発酵ブロスのpHは、5.5〜12、好ましくは7.0〜12.0、好ましくは9.5〜10.5、一層好ましくは9.7〜10.2のpH値に調整される、請求項8に記載の方法。
- ステップb)の前記温度は、85℃〜95℃、好ましくは約90℃である、請求項1〜9のいずれか一項に記載の方法。
- ステップb)の前記温度は、少なくとも1時間、少なくとも2時間、少なくとも3時間および少なくとも4時間維持される、請求項1〜10のいずれか一項に記載の方法。
- ステップb)の前記温度は、24〜72時間、好ましくは24〜36時間維持される、請求項8に記載の方法。
- 油糧微生物を含む未洗浄の発酵ブロスから1種または複数種の多価不飽和脂肪酸を含む微生物油を抽出するための方法であって、
(a)溶解細胞組成物を形成するため前記発酵ブロス中の前記油糧微生物を溶解すること;
(b)前記溶解細胞組成物から水を除去して、前記溶解細胞組成物の体積を、その当初の体積の60%未満に減少させること;
(c)ステップ(b)で得られた前記溶解細胞組成物を60℃〜110℃の温度に昇温すること;および
(d)前記微生物油を前記溶解細胞組成物から回収すること
を含み、前記抽出するための方法全体において塩が使用されない方法。 - ステップ(b)の前記溶解細胞組成物の体積は、その当初の体積の70%未満、好ましくは80%未満に減少する、請求項13に記載の方法。
- ステップ(b)の水の除去は、110℃以下、好ましくは70℃〜100℃、一層好ましくは80℃〜90℃の温度で前記溶解細胞組成物を加熱することにより行われる、請求項13または請求項14に記載の方法。
- ステップ(c)は、アルカリ化剤、好ましくは苛性ソーダを加えることを含む、請求項13〜15のいずれか一項に記載の方法。
- 前記溶解細胞組成物のpHは、5.5〜12、好ましくは7.0〜12.0、好ましくは9.5〜10.5、一層好ましくは9.7〜10.2のpH値に調整される、請求項16に記載の方法。
- ステップ(c)の前記温度は、85℃〜95℃、好ましくは約90℃である、請求項13〜17のいずれか一項に記載の方法。
- ステップ(c)の前記温度は、少なくとも1時間、少なくとも2時間、少なくとも3時間および少なくとも4時間維持される、請求項13〜18のいずれか一項に記載の方法。
- ステップ(c)の前記温度は、24〜72時間、好ましくは24〜36時間維持される、請求項19に記載の方法。
- 油糧微生物を含む未洗浄の発酵ブロスから1種または複数種の多価不飽和脂肪酸を含む微生物油を抽出するための方法であって、
(a)前記発酵ブロスから水を除去して、前記発酵ブロスの体積を、その当初の体積の60%未満に減少させること;
(b)溶解細胞組成物を形成するため前記発酵ブロス中の前記油糧微生物を溶解すること;
(c)ステップ(b)で得られた前記溶解細胞組成物を60℃〜110℃の温度に昇温すること;および
(d)前記微生物油を前記溶解細胞組成物から回収すること
を含み、前記抽出するための方法全体において塩が使用されない方法。 - ステップ(a)の前記発酵ブロスの体積は、その当初の体積の70%未満、好ましくは80%未満に減少する、請求項21に記載の方法。
- ステップ(a)の水の除去は、前記発酵ブロスを110℃以下、好ましくは70℃〜100℃、一層好ましくは80℃〜90℃の温度で加熱することにより行われる、請求項21または請求項22に記載の方法。
- ステップ(c)は、アルカリ化剤、好ましくは苛性ソーダを加えることを含む、請求項21〜23のいずれか一項に記載の方法。
- 前記溶解細胞組成物のpHは、5.5〜12、好ましくは7.0〜12.0、好ましくは9.5〜10.5、一層好ましくは9.7〜10.2のpH値に調整される、請求項24に記載の方法。
- ステップ(c)の前記温度は、85℃〜95℃、好ましくは約90℃である、請求項21〜24のいずれか一項に記載の方法。
- ステップ(c)の前記温度は、少なくとも1時間、少なくとも2時間、少なくとも3時間および少なくとも4時間維持される、請求項21〜25のいずれか一項に記載の方法。
- ステップ(c)の前記温度は、24〜72時間、好ましくは24〜36時間維持される、請求項27に記載の方法
- 前記油糧微生物は、1種または複数種の多価不飽和脂肪酸を含む微生物油を産生する、請求項1〜28のいずれか一項に記載の方法。
- 前記多価不飽和脂肪酸は、オメガ3脂肪酸、オメガ6脂肪酸およびそれらの混合物を含む、請求項29に記載の方法。
- 前記多価不飽和脂肪酸は、ドコサヘキサエン酸(DHA)、エイコサペンタエン酸(EPA)、ドコサペンタエン酸(DPA)、アラキドン酸(ARA)、ガンマリノレン酸(GLA)、ジホモガンマリノレン酸(DGLA)、ステアリドン酸(SDA)およびそれらの混合物を含む、請求項29に記載の方法。
- 前記多価不飽和脂肪酸は、ドコサヘキサエン酸(DHA)である、請求項31に記載の方法。
- 前記多価不飽和脂肪酸は、エイコサペンタエン酸(EPA)である、請求項31に記載の方法。
- 前記多価不飽和脂肪酸は、アラキドン酸(ARA)である、請求項31に記載の方法。
- 前記微生物細胞は、藻類細胞、酵母細胞、真菌細胞、プロテスト細胞または細菌細胞である、請求項1〜34のいずれか一項に記載の方法。
- 前記微生物細胞は、クリプテコディニウム(Crypthecodinium)属、モルティエレラ(Mortierella)属またはヤブレツボカビ(Thraustochytriales)目由来である、請求項1〜35のいずれか一項に記載の方法。
- 前記微生物細胞は、ヤブレツボカビ(Thraustochytriales)目由来である、請求項36に記載の方法。
- 前記微生物細胞は、トラウストキトリウム(Thraustochytrium)属、シゾキトリウム(Schizochytrium)属またはそれらの混合物由来である、請求項37に記載の方法。
- 前記微生物細胞は、モルティエレラ・アルピナ(Mortierella alpina)由来である、請求項36に記載の方法。
- 前記溶解細胞組成物は、液体、細胞片および微生物油を含む、請求項13〜39のいずれか一項に記載の方法。
- 前記油は、少なくとも15重量%エイコサペンタエン酸を含む、請求項40に記載の方法。
- 前記油は、少なくとも30重量%ドコサヘキサエン酸を含む、請求項40または請求項41に記載の方法。
- 前記油は、少なくとも30重量%アラキドン酸を含む、請求項42に記載の方法。
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CA3048289C (en) | 2016-12-27 | 2023-09-26 | Evonik Degussa Gmbh | Method of isolating lipids from a lipids containing biomass |
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WO2019121752A1 (en) * | 2017-12-20 | 2019-06-27 | Evonik Degussa Gmbh | Method of isolating lipids from a lipids containing biomass |
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KR102552228B1 (ko) * | 2013-12-20 | 2023-07-05 | 디에스엠 아이피 어셋츠 비.브이. | 미생물 세포로부터의 미생물 오일의 수득 방법 |
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