JP5804666B2 - バイオマスの処理および利用における別の供給流れの集中 - Google Patents
バイオマスの処理および利用における別の供給流れの集中 Download PDFInfo
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Description
本発明は、エネルギー省によって授与された契約番号04−03−CA−70224の下で米国政府後援でなされたものであった。政府は本発明における特定の権利を有する。
a)バイオマスを提供する工程、
b)a)のバイオマスに少なくとも1つの別の供給流れを添加して集中供給原料を生産する工程、
c)b)の集中供給原料をアンモニアを含んでなる水溶液と接触させ、集中供給原料−水性アンモニア混合物を形成することで、前処理された集中供給原料生産物を生産する工程であって、ここでアンモニアは集中供給原料−水性アンモニア混合物のアルカリ性pHを維持するための少なくとも十分な濃度で存在するが、アンモニアは集中供給原料の乾燥重量に対して約12重量パーセント未満で存在し、かつさらに乾燥重量の集中供給原料は集中供給原料−水性アンモニア混合物の重量に対して少なくとも約15重量パーセントの高固体濃度で存在する、工程、
d)c)の生産物を適切な条件下で糖化酵素共同体と接触させ、発酵可能な糖生産物を生産する工程、
を含んでなる方法で処理される。
a)バイオマスを提供する工程と、
b)a)のバイオマスに前処理方法を施すことで前処理されたバイオマス生産物を生産する工程と、
c)b)の前処理されたバイオマス生産物に少なくとも1つの別の供給流れを添加して第1または第2の集中供給原料を生産する工程と、
d)c)の第1または第2の集中供給原料を適切な条件下で糖化酵素共同体と接触させ、発酵可能な糖生産物を生産する工程と、
を含んでなる方法で処理および糖化される。本方法の態様では、b)の前処理方法は、バイオマスをアンモニアを含んでなる水溶液に接触させてバイオマス−水性アンモニア混合物を形成する工程を含み、ここでアンモニアはバイオマス−水性アンモニア混合物のアルカリ性pHを維持するための少なくとも十分な濃度で存在するが、アンモニアはバイオマスの乾燥重量に対して約12重量パーセント未満で存在し、かつさらにバイオマスの乾燥重量はバイオマス−水性アンモニア混合物の重量に対して少なくとも約15重量パーセントの高固体濃度にある。本方法のさらなる態様では、b)の前処理されたバイオマス生産物に添加される別の供給流れは、種子加工の間に生産されるスチラーゲ(stillage)を含んでなる。さらに別の態様では、a)のバイオマスは、本明細書で記載の集中供給原料である場合もあればそうでない場合もある。
に
本開示では多数の用語が用いられる。以下の定義が提供される。
「発酵可能な糖」という用語は、発酵方法中の微生物による炭素源として使用可能なオリゴ糖および単糖を示す。
本方法の態様は、任意のバイオマス供給原料と、廃棄物流れなどの低価値の副産物または工業方法流れよりなる少なくとも1つの別の供給流れとを複合させる工程を含む。
本方法における集中供給原料の前処理にて使用されるアンモニアの濃度は、最低では集中供給原料−水性アンモニア混合物のアルカリ性のpHを維持するための十分な濃度であり、かつ最大では集中供給原料の乾燥重量に対して約12重量パーセント未満である。アンモニアのこの低濃度は前処理にとって十分であり、かつ低濃度は集中供給原料の乾燥重量に対して約10重量パーセント未満でもありうる。集中供給原料の乾燥重量に対して6パーセントもしくはそれ以下の極めて低濃度のアンモニアもまた前処理において使用されうる。アルカリ性は7.0より大きいpHを意味する。特に適切なのは、pHが8より大きい集中供給原料−水性アンモニア混合物である。一実施形態では、アンモニアは集中供給原料の乾燥重量に対して約8重量パーセント未満で存在する。一実施形態では、アンモニアは集中供給原料の乾燥重量に対して約10重量パーセント未満で存在する。特に適切なのは、集中供給原料の乾燥重量に対して約6重量パーセント未満のアンモニアである。
本方法の別の態様では、種子方法流れなどの別の供給流れを前処理されたバイオマスと結合させることで、糖化される第2の集中供給原料がもたらされる。前処理されたバイオマスは、本明細書に記載のように前処理されている材料でバイオマスの代わりに集中供給原料を用いたものであり、あるいは、上記を例とする別の供給流れである種子方法流れを前処理された集中供給原料と複合させることで、糖化される第2の集中供給原料が提供される。
本方法では、以下の供給原料が糖化されうる。それは、1)前処理された集中供給原料;2)前処理後に少なくとも1つの別の供給流れと複合されることで糖化される第2の集中供給原料を形成する前処理された集中供給原料;または3)前処理後に少なくとも1つの別の供給流れと複合されることで第2の集中供給原料を形成する、前処理された集中されていないバイオマスである。本方法では、1〜3の番号が付与された上記の1つもしくはそれ以上の集中供給原料を糖化酵素共同体の存在下で加水分解することで、加水分解物中でオリゴ糖および/または単糖が放出される。本方法における糖化は、前処理に先立ち、前処理後、またはそれら双方において融合が行われるか否かにかかわらず、集中供給原料に関するものである。バイオマス処理のための糖化酵素および方法は、リンド L.R.(Lynd L.R.)ら(Microbiol.Mol.Biol.Rev.(2002年)66:506−577頁)でレビューされている。
る
他の発酵成分から分離されうる。生産物は、イオン交換、溶媒抽出、または電気透析により抽出されうる。凝集剤を使用することで、生産物の分離が促進される。具体例として、バイオ生産された1−ブタノールは、ABE発酵についての当該技術分野で既知の方法を用いて発酵培地から単離されうる(例えば、デュレ(Durre)、Appl.Microbiol.Biotechnol.49:639−648頁(1998年)、グルート(Groot)ら、Process.Biochem.27:61−75頁(1992年)、およびそれらの中の参考文献を参照)。例えば、遠心分離、濾過、デカンテーションまたは同類のものにより、固体が発酵培地から除去されうる。次いで、1−ブタノールは、蒸留、共沸蒸留、液体−液体抽出、吸着、ガスストリッピング、膜蒸発、または透析蒸発などの方法を用いて発酵培地から単離されうる。発酵媒体からの1,3プロパンジオールの精製が、例えば、反応混合物に有機溶媒、蒸留、およびカラムクロマトグラフィーによる抽出を施すことにより実施されうる(米国特許第5,356,812号明細書)。この方法にとって特に良好な有機溶媒はシクロヘキサンである(米国特許第5,008,473号明細書)。アミノ酸は、イオン交換樹脂吸着および/または結晶化などの方法により発酵培地から回収されうる。
以上、本発明を要約すれば以下のとおりである。
1.a)バイオマスを提供する工程と、
b)a)のバイオマスに少なくとも1つの別の供給流れを添加して集中供給原料を生産する工程、
c)b)の集中供給原料をアンモニアを含んでなる水溶液と接触させ、集中供給原料−水性アンモニア混合物を形成することで、前処理された集中供給原料生産物を生産する工程であって、ここで、アンモニアは集中供給原料−水性アンモニア混合物のアルカリ性pHを維持するための少なくとも十分な濃度で存在するが、前記アンモニアは集中供給原料の乾燥重量に対して約12重量パーセント未満で存在し、かつさらに乾燥重量の集中供給原料は集中供給原料−水性アンモニア混合物の重量に対して少なくとも約15重量パーセントの高固体濃度で存在する工程、
d)c)の生産物を適切な条件下で糖化酵素共同体と接触させ、発酵可能な糖生産物を生産する工程、
を含んでなる、集中供給原料より構成されたバイオマスの処理方法。
2.集中供給原料−水性アンモニア混合物のpHが8より大きい前記1に記載の方法。
3.前記乾燥重量の集中供給原料が少なくとも約15%〜約80%の初期濃度で存在する前記1に記載の方法。
4.前記乾燥重量の集中供給原料が少なくとも約15%〜約60%の初期濃度で存在する前記3に記載の方法。
5.前記アンモニアが集中供給原料の乾燥重量に対して約10重量パーセント未満で存在する前記1に記載の方法。
6.前記アンモニアが集中供給原料の乾燥重量に対して約6%もしくはそれ以下の重量パーセントで存在する前記1に記載の方法。
7.工程(c)が約4℃〜約200℃の温度で実施される前記1に記載の方法。
8.工程(c)が約75℃〜約150℃の温度で実施される前記7に記載の方法。
9.工程(c)が約90℃より高く約150℃以下の温度で実施される前記8に記載の方法。
10.工程(c)が最大約8時間の期間実施される前記1に記載の方法。
11.a)バイオマスを提供する工程と、
b)a)のバイオマスに前処理方法を施すことで前処理されたバイオマス生産物を生産する工程と、
c)b)の前処理されたバイオマス生産物に少なくとも1つの別の供給流れを添加して第1または第2の集中供給原料を生産する工程と、
d)c)の第1または第2の集中供給原料を適切な条件下で糖化酵素共同体と接触させ、発酵可能な糖生産物を生産する工程と、
を含んでなる、集中供給原料より構成されたバイオマスの処理方法。
12.(b)の前処理方法が、バイオマスをアンモニアを含んでなる水溶液に接触させてバイオマス−水性アンモニア混合物を形成する工程を含み、ここでアンモニアはバイオマス−水性アンモニア混合物のアルカリ性pHを維持するための少なくとも十分な濃度で存在するが、前記アンモニアはバイオマスの乾燥重量に対して約12重量パーセント未満で存在し、かつさらにバイオマスの乾燥重量はバイオマス−水性アンモニア混合物の重量に対して少なくとも約15重量パーセントの高固体濃度にある、前記11に記載の方法。
13.バイオマス−水性アンモニア混合物のpHが8より大きい前記12に記載の方法。14.前記バイオマスの乾燥重量が少なくとも約15%〜約80%の初期濃度にある前記12に記載の方法。
15.前記バイオマスの乾燥重量が少なくとも約15%〜約60%の初期濃度にある前記12に記載の方法。
16.前記アンモニアがバイオマスの乾燥重量に対して約10重量パーセント未満で存在する前記12に記載の方法。
17.前記アンモニアがバイオマスの乾燥重量に対して約6%もしくはそれ以下の重量パーセントで存在する前記16に記載の方法。
18.工程(b)が約4℃〜約200℃の温度で実施される前記12に記載の方法。
19.工程(b)が約75℃〜約150℃の温度で実施される前記18に記載の方法。
20.工程(b)が90℃より高く約150℃以下の温度で実施される前記19に記載の方法。
21.工程(b)が最大約8時間の期間実施される前記12に記載の方法。
22.工程(a)のバイオマスが集中供給原料である前記11に記載の方法。
23.別の供給流れがスチラーゲよりなる前記11に記載の方法。
24.バイオマスが、バイオエネルギー作物、農業残渣、都市固体廃棄物、産業固体廃棄物、工場廃棄物、木材および林業廃棄物よりなる群から選択される前記1または11に記載の方法。
25.バイオマスが、スイッチグラス、紙くず、製紙汚泥、トウモロコシ粒、トウモロコシ穂軸、トウモロコシの皮、コーンストーバー、草、小麦、小麦のわら、まぐさ、稲わら、サトウキビバガス、モロコシ、大豆、木、枝、根、葉、ウッドチップ、おがくず、低木およびブッシュ、野菜、果物、花ならびに動物糞尿よりなる群から選択される前記1または11に記載の方法。
26.バイオマスが、トウモロコシ穂軸、コーンストーバー、トウモロコシの皮、サトウキビバガス、スイッチグラス、小麦のわら、まぐさ、大麦、大麦のわら、稲わら、および草よりなる群から選択される前記1または11に記載の方法。
27.バイオマスが、トウモロコシ穂軸、コーンストーバーおよびサトウキビバガスよりなる群から選択される前記26に記載の方法。
28.アンモニアが、アンモニアガス、水酸化アンモニウム、尿素、およびこれらの組み合わせよりなる群から選択される前記1または12に記載の方法。
29.アンモニアを含んでなる前記水溶液が少なくとも1つの追加塩基をさらに含んでなる前記1または12に記載の方法。
30.アンモニアが再生される前記1または12に記載の方法。
31.別の供給流れが、副産物、中間流れ、および廃棄物流れよりなる群から選択される前記1または11に記載の方法。
32.別の供給流れが、トウモロコシ、オート、小麦、大麦、米、カノーラ、ヒマワリ、綿花、エンドウ豆、または大豆、および他のマメ科植物からの種子方法流れである前記31に記載の方法。
33.前記糖化酵素共同体が少なくとも1つのグリコシダーゼを含んでなる前記1または11に記載の方法。
34.前記糖化酵素共同体が、セルロースを加水分解するグリコシダーゼ、ヘミセルロースを加水分解するグリコシダーゼ、澱粉を加水分解するグリコシダーゼ、ペプチダーゼ、リパーゼ、リグニナーゼおよびフェルロイルエステラーゼよりなる群から選択される少なくとも1つの酵素を含んでなる前記1または11に記載の方法。
35.前記糖化酵素共同体が、セルラーゼ、エンドグルカナーゼ、エキソグルカナーゼ、セロビオヒドロラーゼ、β−グルコシダーゼ、キシラナーゼ、エンドキシラナーゼ、エキソキシラナーゼ、β−キシロシダーゼ、アラビノキシラナーゼ、マンナーゼ、ガラクターゼ、ペクチナーゼ、グルクロニダーゼ、アミラーゼ、α−アミラーゼ、β−アミラーゼ、グルコアミラーゼ、α−グルコシダーゼ、イソアミラーゼよりなる群から選択される少なくとも1つの酵素を含んでなる前記1または11に記載の方法。
36.工程(d)が約15℃〜約100℃の温度および約2〜約11のpHで実施される前記1または11に記載の方法。
37.a)のバイオマスが少なくとも1つの別の供給流れよりなる前記11に記載の方法。
38.前記スチラーゲが利用されることで前処理された材料のpHが低下し、それにより前記物質が糖化の作用をより受けやすくなる前記23に記載の方法。
39.前記発酵可能な糖類が用いられることで、付加価値のある化学物質、燃料または他の高価値の生産物が生産される前記1または11に記載の方法。
以下の略語が用いられる。
「HPLC」は高性能液体クロマトグラフィー、「C」は摂氏、「kPa」はキロパスカル、「m」はメートル、「mm」はミリメートル、「kW」はキロワット、「μm」はマイクロメートル、「μL」はマイクロリットル、「mL」はミリリットル、「L」はリットル、「min」は分、「mM」はミリモル、「cm」はセンチメートル、「g」はグラム、「kg」はキログラム、「wt」は重量、「hr」は時間、「temp」または「T」は温度、「theoret」は理論、「pretreat」は前処理、「DWB」はバイオマスの乾燥重量である。
図2に示すような寸法および特徴を有しかつ本明細書中、上記のバイオマス処理装置はPEHReactorと称されるものであり、以下の実施例にてそれを使用した。つまり、9LのPEHReactor(NREL、ゴールデン(Golden)、コロラド州にて構築;同時係属出願CL3447に詳細に記載)は、処理反応体を導入するための注入ランスを具備する約15cm×51cmのステンレス鋼製の反応容器を有する。注入ランスは、容器の一端部上でカバー内のポートに回転ジョイントを用いて接続され、それは容器に接近するための追加ポートを有する。4つのバッフルが容器壁の全長にわたり、壁に垂直に取り付けられる。バッフルおよび容器内で浮遊している3.2cm×3.2cmの22個のセラミック製の摩擦媒体シリンダー(アドバンスド・セラミックス(Advanced Ceramics)、イーストパレスタイン(East Palestine)、オハイオ州)は、容器回転時でのバイオマスと反応物との機械的混合を適用したものであり、それにより反応物のバイオマスへの吸収が促進される。PEHReactorは回転のための機構を提供するベルコ・セル−プロダクション・ローラー・アパラタス(Bellco Cell−Production Roller Apparatus)(ベルコ・テクノロジー(Bellco Technology)、ヴァインランド(Vineland)、ニュージャージー州)上に設けられ、ローラー装置を具備する反応器は熱を供給する温度制御チャンバー内に収容される。温度制御チャンバーは、ベルコ・セル・プロダクション・アパラタス(Bellco Cell Production Apparatus)を囲むコルク製の絶縁パッドを支持するためのアルミニウムフレームよりなり、それに注入ランスの中央を通して挿入される熱電対により制御されるヒーターが取り付けられる。真空および圧力を、カバー内でランスに接続されたポートに外部ソースを取り付けることにより、反応容器に適用可能である。
糖、アセトアミド、乳酸および酢酸含有量の測定
糖化液中の可溶性糖(グルコース、セロビオース、キシロース、ガラクトース、アラビノースおよびマンノース)を、適切なガードカラムを有するバイオ・ラッド(Bio−Rad)HPX−87Pおよびバイオ・ラッド(Bio−Rad)HPX−87Hカラム(バイオ・ラッドラボラトリーズ(Bio−Rad Laboratories)、ハーキュリーズ(Hercules)、カリフォルニア州)を用い、HPLC(アジレント(Agilent)モデル1100、アジレント・テクノロジーズ(Agilent Technologies)、パロアルト(Palo Alto)、カリフォルニア州)によって測定した。試料のpHを測定し、必要に応じ硫酸を用いて5〜6に調節した。次いで、試料を直接に0.2μmのシリンジフィルターを通してHPLCバイアルに通過させた。HPLCの稼動条件は以下の通りであった。
HPX−87P(炭水化物用):
注入容量:10〜50μL、濃度および検出器の限界に依存
移動相:HPLCグレードの水、0.2μmに濾過および脱気
流速:0.6mL/分
カラム温度:80〜85℃、ガードカラム温度<60℃
検出器温度:可能な限り主カラム温度に近い
検出器:屈折率
実行時間:35分間のデータ収集に加え、実行後の15分間(後の溶出化合物のために可能な調節を行う)
バイオ・ラッド(Biorad)アミネックス(Aminex)HPX−87H(炭水化物を対象)
注入容量:5〜10μL、濃度および検出器の限界に依存
移動相:0.01N硫酸、0.2μmに濾過および脱気
流速:0.6mL/分
カラム温度:55℃
検出器温度:可能な限りカラム温度に近い
検出器:屈折率
実行時間:25〜75分間のデータ収集
PEHReactor内でのトウモロコシ穂軸および異なる使用済み穀物試料を含有する複合バイオマスの前処理および糖化
使用済み穀物試料を次のように調製した。
1.#2黄色全粒デントコーン粒(アグウェイ(Agway)から購入)
2.イリノイ大学(University of Illinois)で開発されたクイック・ジャーム(Quick Germ)方法(シン(Singh)およびエックホフ(Eckoff)(1996年) Cereal Chem.74:462−466頁)により、トウモロコシ粒からジャームを除去した(degermed)。出発材料をイリノイ大学(University of Illinois)のヴィジャイ・シン(Vijay Singh)から入手した。
3.クイック・ファイバー(Quick Fiber)方法によるトウモロコシ粒方法により、ジャームおよび外皮繊維を除去した(米国特許第6,254,914号明細書)。出発材料をイリノイ大学(University of Illinois)のヴィジャイ・シン(Vijay Singh)から入手した。
4.ブリューワーズ・グリット(Brewers’grits)をカーギル(Cargill)(ミネアポリス(Minneapolis)、ミネソタ州)から入手した。
PEHReactor内でのトウモロコシ穂軸、使用済み穀物、および追加成分を含有する複合バイオマスの前処理および糖化
実施例1に記載のように調整した粉砕トウモロコシ穂軸およびウイスキーの使用済み穀物を、実施例1に記載のようにPEHReactor内で複合させた。さらに、他の穀物成分を添加した。一方の試料では、澱粉(シグマ(Sigma)S4126、ロット番号093K0033)を5g/バイオマスの全乾燥重量100gで添加した。もう一方の試料では、トウモロコシ油(シスコクラシック(Sysco Classic)製トウモロコシ油、ロット番号4119095)を約2g/乾燥バイオマス全体100gの濃度で添加した。実施例1に記載のように試料を前処理して糖化した。結果を表3に示す。これらの結果はまた、表2におけるトウモロコシ穂軸のみの対照データと有利に比較したものである。
PEHReactor内でのトウモロコシ穂軸およびトウモロコシ繊維を含有する複合バイオマスの前処理および糖化
粉砕トウモロコシ穂軸およびカーギル(Cargill)製ブラン80(Bran 80)(カーギル(Cargill)、ミネトンカ(Minnetonka)、ミネソタ州)トウモロコシ繊維を、繊維が乾燥バイオマス全体の約10%であるように複合させた。実施例1に記載のように、複合バイオマスを前処理して糖化した。生産された糖の収率を表4に示す。トウモロコシ穂軸とトウモロコシ繊維とを複合させたバイオマスの収率は、トウモロコシ穂軸単独試料の収率に類似していた。
Claims (2)
- a)バイオマスを提供する工程と、
b)a)のバイオマスに少なくとも1つの別の供給流れを添加して集中供給原料を生産する工程、
c)b)の集中供給原料をアンモニアを含んでなる水溶液と接触させ、集中供給原料−水性アンモニア混合物を形成することで、前処理された集中供給原料生産物を生産する工程であって、ここで、アンモニアは集中供給原料−水性アンモニア混合物のアルカリ性pHを維持するための少なくとも十分な濃度で存在するが、前記アンモニアは集中供給原料の乾燥重量に対して12重量パーセント未満で存在し、かつさらに乾燥重量の集中供給原料は集中供給原料−水性アンモニア混合物の重量に対して少なくとも15重量パーセントの高固体濃度で存在する工程、
d)c)の生産物を適切な条件下で糖化酵素共同体と接触させ、発酵可能な糖生産物を生産する工程、
を含んでなり、ここで、工程c)が90℃より高い温度で最大8時間の期間実施される、集中供給原料より構成されたバイオマスの処理方法。 - a)バイオマスを提供する工程と、
b)a)のバイオマスに前処理方法を施すことで前処理されたバイオマス生産物を生産する工程であって、ここで、前処理方法は、a)のバイオマスをアンモニアを含んでなる水溶液と接触させ、バイオマス−水性アンモニア混合物を形成する工程であって、アンモニアはバイオマス−水性アンモニア混合物のアルカリ性pHを維持するための少なくとも十分な濃度で存在するが、前記アンモニアはバイオマスの乾燥重量に対して12重量パーセント未満で存在し、かつさらに乾燥重量のバイオマスはバイオマス−水性アンモニア混合物の重量に対して少なくとも15重量パーセントの高固体濃度で存在する工程を包含する、工程と、
c)b)の前処理されたバイオマス生産物に少なくとも1つの別の供給流れを添加して第1または第2の集中供給原料を生産する工程と、
d)c)の第1または第2の集中供給原料を適切な条件下で糖化酵素共同体と接触させ、発酵可能な糖生産物を生産する工程と、
を含んでなり、ここで、工程b)が90℃より高い温度で最大8時間の期間実施される、集中供給原料より構成されたバイオマスの処理方法。
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