プラスチック廃棄物が引き起こす環境汚染問題の解決策として，⽣分解性⾼分⼦が注⽬
されている。⼀⽅で，多くの⽣分解性⾼分⼦は，曝露環境によりその⽣分解速度が⼤きく異 なることが知られている。また，現在市販の⽣分解性⾼分⼦では，その⽣分解速度および分 解開始時期が制御されていない。これらのことは，⽣分解性⾼分⼦の普及を妨げている要因 でもある。理想的な⽣分解性⾼分⼦には，使⽤期間中，機械的強度を保ち，不要時あるいは 廃棄時に速やかに⽣分解されることが望まれる。本博⼠論⽂では，分解開始時期および分解 速度が⼗分に制御された⽣分解性⾼分⼦を，時限⽣分解性⾼分⼦と定義づけた。
本博⼠論⽂では，⼟壌，淡⽔中で⽣分解速度が⼤きく，海⽔中での⽣分解速度が極めて低 い化学合成⽣分解性⾼分⼦であるポリエチレンスクシネート(PESu)に着⽬した。第2章，第 3章では，PESuの環境分解性の環境依存性の原因を明らかにした。第4章では，第2章お よび第3章で得られた結果に基づき，PESuおよび芽胞を利⽤した，海洋中での時限⽣分解 性⾼分⼦の創製を検討した。
第2章では，PESuが微⽣物産⽣脂肪族ポリエステル(PHA)の分解酵素である，ポリ(3-ヒ ドロキシブタン酸)[P(3HB)]分解酵素によって酵素加⽔分解される性質に着⽬し，PESu と P(3HB)の環境分解性が異なる原因を，分解微⽣物の分布を調べ，分解微⽣物を単離解析す ることで，明らかにした。P(3HB)分解微⽣物は，環境を問わず存在しているものの，PESu 分解微⽣物は，海⽔中からは検出されなかった。また，P(3HB)分解細菌として単離された全 ての株が，PESuを分解しなかった。⼀⽅で，PESu分解細菌の⼤部分は，Bacillus属の細菌 であった。このBacillus属のPESu分解細菌は，P(3HB)を分解しなかった。これらのことか ら，PESuがP(3HB)と異なり海⽔中で⽣分解されない原因は，PESuがP(3HB)分解細菌にと
っての P(3HB)分解酵素誘導物質として機能せず，それに加えて PESu 分解細菌の属種およ
び環境中の分布の偏りがあるためとわかった。
第 3 章では，PESu が化学合成脂肪族ポリエステルである PCLと同⼀の酵素（クチナー ゼ）および微⽣物種により分解される性質に着⽬し，海洋環境から単離したPCLおよびPESu
分解細菌TKCM64株の特徴づけを⾏い，これに基づきPESuとPCLの海⽔中での環境分解
性の差が⽣じる原因を考察した。TKCM64株は，P. pachastrellaeに近縁な細菌種であった。
本株は，PCLを1.39 ± 0.09 mg·cm-2·day-1の速度で分解した。これに対して，本株は，PESu を，PCL分解速度の1/50の速度でしか分解できなかった。また，各種炭素源存在下での菌 体増殖レベルと，その培養上清のPCL およびPESu 分解活性との関係を調べたところ，本 株は，PCL，PCL 加⽔分解物(6-ヒドロキシヘキサン酸)および主要なクチン構成物(16-ヒド ロキシヘキサドデカン酸)の存在下で，菌体増殖とPCL加⽔分解酵素の⽣産誘導が⾒られた。
⼀⽅，PESuおよび PESu構成物であるエチレングリコールの存在下では，菌体増殖および 培養上清のPCL分解活性が⾒られなかった。また，各培養上清のPESu分解活性は，PCL分 解活性と⽐較して極めて⼩さかった。これらのことから，本株が⽣産するポリエステル加⽔
分解酵素は，PCL に対して基質特異性が⼤きく，他⽅ PESu に対して⼩さいことがわかっ
た。これらを総合的に判断すると，⾼いPCL分解活性を有するTKCM64株は，海洋中での PESuの分解には関与していないことが推定される(Fig. 30)。
Fig. 30. The inference of PCL and PESu-degrading mechanism in coastal environments.
第2章の結果から，PESuの主たる分解者がBacillus属の細菌であることがわかった。ま た，第2章および第3章で，海⽔中でのPESu分解速度が極めて低い原因を明らかにした。
そこで，第4章では，Bacillus属細菌の芽胞と海洋環境中での潜在的⽣分解性⾼分⼦である PESuに着⽬し，これらを⽤いて海洋環境中での時限⽣分解性⾼分⼦の創製を検討した。海 砂中から，PESu分解活性を有する芽胞形成細菌，YKCMOA1株を単離した。本株は，⼈⼯
海⽔中で，PESuを，141 µg·cm-2·day-1の速度で分解した。遺伝系統学的および⽣化学的解析
の結果，YKCMOA1株はBacillus pumilusに近縁な種であることがわかった。本株の芽胞は，
110 ˚C（PESuの融点は104 ˚C）でのD値が27.8分であり，YEおよびアスパラギンとカラ
メルの混合物を含む⼈⼯海⽔中で発芽することがわかった。本株の芽胞を含有したPESuフ ィルムは，⼈⼯海洋環境で分解することがわかった。また，芽胞含有PESuフィルムは，実 際の海⽔中でも，重量減少および表⾯形態の変化が⾒られた。以上の結果から，芽胞を利⽤
することで，海洋中での PESu の分解を促進させることが可能であることが⽰された(Fig.
Fig. 31. Timing biodegradable polymers by spores.
本博⼠論⽂で得られた結論を踏まえ，今後の展開について述べる。本博⼠論⽂では，⽣分 解性が環境依存的である⾼分⼦を「潜在的⽣分解性⾼分⼦」と定義づけた。潜在的⽣分解性
⾼分⼦は，ある環境下に存在する時のみ，⽣分解が開始する。すなわち，潜在的⽣分解性⾼
分⼦では，⼈為的に⽣分解を開始させる環境を再現することで，その分解を制御できる可能 性が⾼い。本博⼠論⽂では，海洋における潜在的⽣分解性⾼分⼦である PESuに注⽬した。
PESu 以外にも，潜在的⽣分解性⾼分⼦は存在している。例えば，PBSuやPBAT の⽣分解 する環境は限定的である[35,64]。これら潜在的⽣分解性⾼分⼦の環境分解性の原因を明らか にすることは，これを利⽤した時限⽣分解性⾼分⼦の創製に繋がる。そこで，各ポリエステ ルの環境分解性を，その環境中の微⽣物菌叢やこれら微⽣物によるポリエステル分解活性 と共に考察する必要がある。
本博⼠論⽂では，潜在的⽣分解性⾼分⼦に対する分解開始時期および分解速度の制御⼿
法として，芽胞の利⽤を提案した。芽胞を⽤いた⽅法で，⽣分解性⾼分⼦の分解開始時期 を制御するためには，芽胞の発芽時期を制御する必要がある。ここでは，物理的破損を発 芽開始のトリガーとする⼿法を提案する。芽胞および発芽誘起物質を，芽胞を埋設する⾼
分⼦あるいは環境中で容易に⽣分解される⾼分⼦でカプセル化し，⽔と接触しないように する（芽胞カプセル化⾼分⼦）。これを潜在的⽣分解性⾼分⼦にブレンドする（芽胞含有
⾼分⼦）。物理的破損時にのみ，芽胞あるいは芽胞包括⾼分⼦が環境と接触するように設 計する。この⼿法によって，芽胞を利⽤した時限分解性⾼分⼦の創製が可能となる。
⼀⽅で，芽胞による⾼分⼦の⽣分解速度は，その芽胞形成細菌や⽣分解性⾼分⼦が流出
案する。すなわち流出環境中に存在する微⽣物の活性を利⽤し，その⽣分解を開始させ る。⽣分解性⾼分⼦の中には，その加⽔分解物および加⽔分解物の代謝産物が加⽔分解酵 素の誘導物質となるものがある。PCLのモノマーおよびオリゴマーは，クチン分解物にそ の構造が類似しており，クチン分解酵素(クチナーゼ)の誘導物質として機能する[88]。そこ で，これらの⽣分解性⾼分⼦加⽔分解酵素誘導物質を，潜在的⽣分解性⾼分⼦へ導⼊させ ることで，時限⽣分解性⾼分⼦創製を実現する。これら⽣分解性⾼分⼦加⽔分解酵素誘導 物質を詳細に特定するためには，ポリエステル分解酵素の遺伝⼦発現に関する制御機構を 明らかにする必要があると考えられる(Fig. 32)。
Fig. 32. Timing biodegradable polymers.
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