臺灣博碩士論文加值系統

Shewanella屬為具有異化金屬還原能力的兼氣厭氧菌群，於環境復育上極具應用潛力。本研究自新竹客雅溪及高雄後勁溪底泥分別篩選出代號KR-12與HC-12的Shewanella屬本土菌株。分離株KR-12及HC-12可於含有6 %的鹽分下生長且能氧化醋酸、檸檬酸、甲酸及乳糖做為厭氧生長時的能量來源。此外，兩分離株可還原硝酸根、亞硝酸根與Fe3+，而做為無氧呼吸的最終電子接受者。比對16S rDNA序列，分離株KR-12與菌株Shewanella baltica OS185最為相似，序列相似度為94 %；而HC-12與最相似菌株Shewanella putrefaciens CN-32有著98 % 序列相似度。綜合保守基因序列、脂肪酸組成與含有酵素等試驗結果，推測分離株KR-12為新種。
而具有金屬還原能力是該菌屬的特點，因此試驗添加不同金屬離子與分離株在厭氧環境共同培養，以測試分離株移除溶液中金屬及蓄積在菌體上的能力。結果顯示分離株KR-12對Ag2+, Zn2+, Cu2+, Pb2+有顯著移除結果；而分離株HC-12則對Ag2+及Pb2+有較佳的移除效果。以穿透式電子顯微鏡觀測金屬蓄積試驗後的菌株，顯示分離株皆可透過還原Ag2+形成銀顆粒的方式而蓄積於菌體上；相對於以Zn2+, Cu2+的金屬蓄積試驗，分離株KR-12及HC-12皆雖可移除溶液中的金屬離子，卻無觀察到任何顆粒產生。值得注意的是，以Pb2+進行金屬蓄積時，分離株KR-12移除Pb2+的過程中，會產生排列呈直線或是曲線的奈米顆粒；對照KR-12，分離株HC-12、模式菌株Shewnella oneidensis MR-1、E.coli及滅菌後的KR-12在相同的培養條件下都不會有類似的顆粒產生。除此之外，KR-12所形成的顆粒直徑會隨著時間增加，並在2小時內由2~3 nm增長至6~8 nm，顯示顆粒的形成與菌體代謝有關；而能量色散光譜結果顯示顆粒的主要組成為Pb與P。此外，同時菌體會延伸出類似菌毛的構造，而這個構造上也出現奈米顆粒。與文獻比較類菌毛構造可能與能傳遞電子功能的奈米線相似。
有機物降解部分，兩株分離株皆無法在96小時內分解或以除草劑草脫淨做為能量來源。然而，兩分離株卻可有效使偶氮染料於厭氧下脫色，而最適當進行脫色反應的條件為pH 7與37 ℃。分離株KR-12在最佳環境下，6小時內可使Methyl Red、Reactive Red 120與Direct Blue 71顏色減少至少75 %；而分離株HC-12擁有更高的反應活性，脫色速率達89.85 % 以上。
綜觀以上試驗結果，顯示分離株KR-12對於金屬蓄積的效果極為顯著，並擁有產生生物性奈米顆粒的潛力；而分離株HC-12則展現相對較佳的偶氮染料脫色能力。若能持續研究，應可找尋更具有生物復育潛力與商業價值的本土菌株。

前言 1
一、 重金屬與偶氮染料 1
二、 異化性金屬還原與奈米線 8
三、 Shewanella 菌屬文獻回顧 10
四、 Shewanella 菌屬應用於環境復育潛力 11
五、 生物奈米材料 18
六、 研究目的 23
材料與方法 24
一、 試驗流程 24
二、 試驗材料 25
三、 微生物篩選 25
四、 微生物生理分析 31
五、 微生物生化分析 32
六、 重金屬蓄積試驗 39
七、 有機物降解試驗 42
八、 統計分析 45
結果與討論 46
一、 本土Shewanella菌種分離與形態觀察 46
二、 菌種序列分析與演化樹建構 50
三、 菌種生理與生化特性 56
四、 重金屬蓄積試驗 63
五、 有機物降解試驗 85
結論與建議 90
參考文獻 92
附錄 108

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