JP2020141701A - バイオ燃料の生産に用いられる脂質の抽出プロセス - Google Patents
バイオ燃料の生産に用いられる脂質の抽出プロセス Download PDFInfo
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Abstract
Description
本出願は、2013年12月20日出願の米国仮特許出願第61/918,850号明細書の利益を主張する。
先行技術の判断のために、バイオ燃料の分野において、共同研究契約が、2008年12月18日付で、BP Biofuels UK LimitedとMartek Biosciences Corporationとの間で締結された。また、先行技術の判断のために、バイオ燃料の分野において、共同開発契約が、2009年8月7日付で、BP Biofuels UK LimitedとMartek Biosciences Corporationとの間で締結された。また、先行技術の判断のために、バイオ燃料の分野において、共同開発契約が、2012年9月1日付で、BP Biofuels UK LimitedとDSM Biobased Products and Services B.V.との間で締結された。
本発明は、バイオ燃料生産のための材料の抽出を対象とした方法および系に関する。本発明の態様は、油産生微生物(oleaginous microorganism)からの材料の抽出に関する。
フィードストックをバイオ燃料に変換するいくつかの技術が開発されてきた。しかしながら、これらの技術進歩をもってしても、再生可能炭素源の燃料への変換の経済的実行可能性を向上させる必要性および要求は残っている。
本発明は、材料を油産生微生物から抽出する方法およびシステム、ならびに抽出された材料からバイオ燃料を生産する方法およびシステムに関する。
本発明は、油産生微生物から材料を抽出する方法および系、ならびに抽出された材料からバイオ燃料を生産する方法および系を提供する。微生物からのバイオ燃料の生産は、植物(油糧種子を含む)からのバイオ燃料の生産に勝る、短い寿命サイクル、より少ない労働要件、季節および気候の独立性、ならびにより容易なスケールアップ等の多くの利点を有し得る。
1.産物にさらに変換され得る有機炭素の回収(利益5参照)
2.非脂質バイオマスおよび削減済み第1目的(reduced first intent)栄養分フィードからの有機養分および無機養分の少なくとも部分的な回収(例えばアンモニア)
3.バガスと混ぜ合わせることによる溶液からの未回収非脂質バイオマスの分離
4.バガスおよび未回収非脂質バイオマスを混ぜ合わせることによる付加的なボイラーフィードおよびエネルギーの発生
5.リサイクルによる有機炭素スリッページおよび回収をより大きくし得る発酵操作に対するストリンジェンシーの引下げ(利益1参照)
6.シード発酵槽および主発酵槽への特異的なフィード流として用いるための糖流106および120の最適化
7.吸収用のリサイクル水を用いることによる、フレッシュ水需要の引下げ
8.廃棄物処理のための資本および運営コストの削減
記載した微生物の性能を特徴付けるのに用いた一測定基準が、脂肪酸抽出率、すなわちFAEである。本開示による微生物のいずれのFAEも、以下の式にしたがって算出することができる:
式中、bは、細胞破断後の総バイオマスであり、典型的にはグラムで測定し;
Cbiomassは、細胞破断前のFAMEのパーセンテージであり、Cbiomassは、総グラムバイオマスあたりの総グラムFAMEとして算出し;本明細書中で用いる用語「FAME」は、脂肪酸メチルエステルを指し;
cbiomealは、細胞破断後のFAMEのパーセンテージであり、cbiomealは、総グラムバイオミールあたりの総グラムFAMEとして算出し;
lは、細胞破断後の油の総質量であるが、油回収工程の前であり、典型的にはグラムで測定する。微生物または発酵ブロスからこれらの値を得るのは、当業者の能力の範囲内である。
式中、Bは、細胞破断前の総バイオマスであり、典型的にはグラムで測定し;
Cbiomassは、細胞破断前のFAMEのパーセンテージであり、Cbiomassは、総グラムバイオマスあたりの総グラムFAMEとして算出し;
Coilは、細胞破断および油回収後のFAMEのパーセンテージであり、Coilは、総グラム油あたりの総グラムFAMEによって算出し;
Lは、細胞破断および油回収後の油の総質量であり、典型的にはグラムで測定する。微生物または発酵ブロスからこれらの測定値を得るのは、当業者の能力の範囲内である。
[実施形態1]
全発酵ブロスからバイオ燃料の生産に適した脂質を抽出する方法であって:
(a)油産生微生物を含有する前記ブロスを、約90℃〜約150℃、または約100℃〜約150℃、または約110℃〜約150℃、または約120℃〜約150℃、または約130℃〜約150℃の温度に加熱することによって、前記全発酵ブロスをプレ処理することと;
場合により、
(i)前記全発酵ブロスを、約30分間〜約18時間、または3時間超〜約18時間、または3時間超〜約8時間にわたり加熱すること;
(ii)前記油産生微生物を含有する前記全発酵ブロスによって45℃〜80℃で費やされる時間を、前記油産生微生物を含有する前記全発酵ブロスを45℃から80℃に60分未満で加熱することによって、最小限にすること;および
(iii)前記全発酵ブロスを、毎分摂氏約0.1〜約80度の平均速度で加熱すること
のうちの少なくとも1つと、
(b)その後産物を前記油産生微生物から抽出することと
を含む、方法。
[実施形態2]
前記全発酵ブロスのpHを、酸または塩基のいずれかを加えることによって調整することをさらに含む、実施形態1に記載の方法。
[実施形態3]
さらなる等温プロセシングを可能にするために、前記全発酵ブロスを、約60℃超、または約70℃超、または約80℃超、または約85℃超、または約90℃超に冷却すること、および、場合により、前記全発酵ブロスを、毎分摂氏約0.2〜約80度の平均速度で冷却すること、をさらに含む、実施形態1または2に記載の方法。
[実施形態4]
前記さらなる等温プロセシングは、機械的破壊を適用することを含む、実施形態3に記載の方法。
[実施形態5]
(a)前記加熱の後、前記全発酵ブロスを乾燥させること;
(b)前記全発酵ブロスを、毎秒約10cm〜毎秒約240cmのインペラ先端速度で撹拌すること;および
(c)前記プレ処理中、前記全発酵ブロスを含有する系において、圧力を、約10psi〜約150psi、または約20psi〜約150psi、または約30psi〜約150psi、または約50psi〜約150psiに維持すること
のうちの少なくとも1つをさらに含む、実施形態1〜4のいずれか一項に記載の方法。
[実施形態6]
前記プレ処理中、前記全発酵ブロスを含有する系に塩が存在して、前記系中で約0.01M〜約2Mのイオン強度がもたらされる、実施形態1〜5のいずれか一項に記載の方法。
[実施形態7]
前記油産生微生物を溶解に曝して、小滴および破片の粒子サイズ分布をもたらすことをさらに含み、放出された産物油小滴および破片の少なくとも80容量%、好ましくは95容量%は、直径において0.1μmよりも大きいサイズを有する、実施形態1〜6のいずれか一項に記載の方法。
[実施形態8]
120cm/秒を超えるインペラ先端速度での混合によって、前記油および細胞破片小滴を連続相として回収することをさらに含む、実施形態7に記載の方法。
[実施形態9]
前記プレ処理を経た前記発酵ブロスにおいて、前記発酵ブロスを付加的に30分間〜8時間にわたり90℃超で加熱することによって、8時間未満の間に脂質が凝集する、実施形態1〜8のいずれか一項に記載の方法。
[実施形態10]
前記抽出プロセスは、前記全発酵ブロス中の金属を、油と比較して少なくとも2の比率で濃縮する、実施形態1〜9のいずれか一項に記載の方法。
[実施形態11]
粗糖に前記方法を実行することと、粗油を回収するために全乾燥バイオマスおよび/または溶媒を利用する抽出技術と比較して、金属および無機元素がより少ない粗油を回収することとを含む、実施形態1〜10のいずれか一項に記載の方法。
[実施形態12]
前記全発酵ブロスは、>0.05g/Lの濃度で塩およびイオンを伴う粗糖源を含み、
好ましくは、前記塩およびイオンは、Na、K、Ca、Mg、Zn、クロリド、サルフェート、ホスフェート、ニトレート、およびそれらの組合せからなる群から選択され、
より好ましくは、前記塩およびイオンは、カリウム、カルシウム、またはそれらの組合せを含む、実施形態1〜11のいずれか一項に記載の方法。
[実施形態13]
前記塩およびイオンは蓄積して、0.5〜40g/Lの濃度となる、実施形態12に記載の方法。
[実施形態14]
前記塩およびイオンは、
(a)ナトリウムよりも高い濃度のカリウム;
(b)1g/Lを超える濃度のカルシウム;および
(c)2.5g/Lを超える濃度のカリウム
のうちの少なくとも1つを含む、実施形態12または13に記載の方法。
[実施形態15]
前記塩およびイオンは、前記産物が前記油産生微生物から放出される場合に、凝集による油相の回収に役立つ、実施形態12〜14のいずれか一項に記載の方法。
[実施形態16]
前記発酵ブロスは、前記凝集の間、0.5〜40g/Lの濃度の塩およびイオンを含む、実施形態15に記載の方法。
[実施形態17]
前記凝集は、凝集脂質の粒子サイズ分布をもたらし、凝集脂質の少なくとも80容量%、好ましくは95容量%は、直径において40μmよりも大きいサイズを有する、実施形態15に記載の方法。
[実施形態18]
前記加熱の後に、前記発酵ブロスを減圧することと、さらなるプロセシング前に前記ブロス中で固体を濃縮するために前記全発酵ブロスを冷却することとをさらに含む、実施形態1〜17のいずれか一項に記載の方法。
[実施形態19]
混合物を形成するために、前記加熱の後に、溶媒を乾燥細胞または溶解発酵ブロスに加えることをさらに含み、
好ましくは、前記溶媒は、ヘキサン、ドデカン、デカン、ディーゼル、アルコール、およびそれらの組合せからなる群から選択される、実施形態1〜18のいずれか一項に記載の方法。
[実施形態20]
(a)接触させて油を前記油産生微生物から抽出するために、前記溶解発酵ブロスおよび前記溶媒の前記混合物を撹拌すること;
(b)前記溶媒および前記油を前記溶解発酵ブロスから分離すること;および
(c)前記溶媒および前記油を前記溶解発酵ブロスから分離するために遠心分離機を用いること
のうちの少なくとも1つをさらに含む、実施形態19に記載の方法。
[実施形態21]
前記油の少なくとも一部を燃料成分に変換するために、前記溶媒および前記油を反応させることをさらに含む、実施形態20に記載の方法。
[実施形態22]
前記溶媒および残りの前記油を、バイオ燃料を含む燃料に変換することをさらに含む、実施形態21に記載の方法。
[実施形態23]
前記消費されたブロスを、作物の肥料、動物の飼料、酵母抽出物、酵母加水分解物、または炭素/栄養源として用いることをさらに含む、実施形態20〜22のいずれか一項に記載の方法。
[実施形態24]
前記油産生微生物を含有する前記全発酵ブロスは、糖フィードストックを含み、
好ましくは、前記油産生微生物および前記糖フィードストックを含有する前記全発酵ブロスは、発酵槽ブロスのリットルあたり約50〜約250グラムの脂質、発酵槽ブロスのリットルあたり約0〜約50グラムの糖、発酵槽ブロスのリットルあたり約0〜約40グラムの塩、および発酵槽ブロスのリットルあたり約10〜約100グラムの脂質フリー乾燥バイオマスを含む、実施形態1〜23のいずれか一項に記載の方法。
[実施形態25]
前記油産生微生物は、少なくとも40重量%の脂肪を含む、実施形態1〜24のいずれか一項に記載の方法。
[実施形態26]
前記油産生微生物を含有する全発酵ブロスを低温殺菌することをさらに含み、
好ましくは、前記全発酵ブロスを約40℃〜約80℃に約1分間〜約3時間にわたり加熱することによって、前記全発酵ブロスを低温殺菌することをさらに含む、実施形態1〜25のいずれか一項に記載の方法。
[実施形態27]
前記全発酵ブロスを、約90℃〜約150℃、または約100℃〜約150℃、または約110℃〜約150℃、または約120℃〜約150℃、または約130℃〜約150℃の温度にて、約30分間〜約18時間、または3時間超〜約18時間、または3時間超〜約8時間にわたり保持することをさらに含む、実施形態26に記載の方法。
[実施形態28]
(a)前記全発酵ブロスを、加熱インターバル中に撹拌すること;
(b)酸を前記全発酵ブロスに加えること;および
(c)塩基を前記全発酵ブロスに加えること
のうちの少なくとも1つをさらに含む、実施形態27に記載の方法。
[実施形態29]
ビーズミル、ホモジナイザー、オリフィスプレート、高剪断ミキサー、プレス、押出機、圧力破壊、湿式ミリング、乾式ミリング、または他の剪断もしくは機械破壊装置に少なくとも1回、好ましくは少なくとも2回、前記全発酵ブロスを通過させることをさらに含む、実施形態27または28に記載の方法。
[実施形態30]
前記溶解発酵ブロスを、ベッセル内で約70℃〜約100℃にて約1〜約60時間にわたり撹拌することをさらに含む、実施形態29に記載の方法。
[実施形態31]
塩を前記ベッセル内の前記溶解発酵ブロスに加えることをさらに含む、実施形態30に記載の方法。
[実施形態32]
最大約2重量%の前記塩を加えることを含み、
好ましくは、前記塩は、NaCl、KCl、K2SO4、Na2SO4であるか、またはH2SO4をプラスした少なくとも1つのNaOHおよびKOHの組合せに由来する、実施形態31に記載の方法。
[実施形態33]
前記ベッセル内の前記溶解発酵ブロスのpHを約3〜約11に調整するために、塩基を加えることをさらに含む、実施形態30〜32のいずれか一項に記載の方法。
[実施形態34]
20%未満の遊離脂肪酸である油を、遠心分離により、前記全発酵ブロスから分離することをさらに含む、実施形態30〜33のいずれか一項に記載の方法。
[実施形態35]
前記油産生微生物は、油産生酵母細胞である、実施形態1〜34のいずれか一項に記載の方法。
[実施形態36]
前記油産生微生物の油産生細胞壁を破壊するために、アミラーゼ、1−4マンノシダーゼ、および1−3マンノシダーゼを含む酵素の組合せを用いることをさらに含み、
好ましくは、酵素の前記組合せはさらに、スルファターゼ、プロテアーゼ、およびキチナーゼからなる群から選択される少なくとも1つの補助酵素を含む、実施形態1〜35のいずれか一項に記載の方法。
[実施形態37]
前記アミラーゼは、α1−4結合グルコースに特異的である、実施形態36に記載の方法。
[実施形態38]
酵素の前記組合せは、約5重量%〜約30重量%のアミラーゼを含む、実施形態36または37のいずれか一項に記載の方法。
[実施形態39]
酵素の前記組合せは、約5重量%〜約45重量%の1−4マンノシダーゼを含む、実施形態36〜38のいずれか一項に記載の方法。
[実施形態40]
酵素の前記組合せは、約5重量%〜約45重量%の1−3マンノシダーゼを含む、実施形態36〜39のいずれか一項に記載の方法。
[実施形態41]
前記油産生細胞壁を破壊した後に前記油産生細胞壁から細胞内代謝物質を収穫することをさらに含む、実施形態36〜40のいずれか一項に記載の方法。
[実施形態42]
前記細胞内代謝物質は、脂質を含む、実施形態41に記載の方法。
[実施形態43]
前記細胞内代謝物質をバイオ燃料に変換することをさらに含む、実施形態41または42に記載の方法。
[実施形態44]
前記細胞内代謝物質を収穫した後に残る水抽出廃水をリサイクルすることをさらに含む、実施形態41〜43のいずれか一項に記載の方法。
[実施形態45]
前記リサイクルされた抽出水を、糖を抽出するためにプロセスフィードストックを洗浄する吸収水として用いることをさらに含む、実施形態44に記載の方法。
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