| 摘要: | 亞磷酸鹽（H₃PO₃）是一種重要的化學物質，在農業和工業領域皆具有廣泛應用。在農業上，亞磷酸鹽常作為殺菌劑與植物生長促進劑使用，能直接被植物吸收並迅速發揮功效，有效促進植物生長並增強植物對環境胁迫的抵抗能力。然而，在實際應用過程中，亞磷酸鹽若過度使用或處理不當，則可能導致環境污染問題，對生態系統造成負面影響。
本研究旨在透過酵素催化反應，以亞磷酸鹽作為受質，將β-煙醯胺單核苷酸（NMN）轉換成具有重要生理功能的還原型菸鹼醯胺腺嘌呤二核苷酸（NADH）及還原型菸鹼醯胺腺嘌呤二核苷酸磷酸（NADPH）。其中，NADH與NADPH在人體內是極為重要的輔酶，分別在細胞能量代謝與抗氧化防禦系統中扮演不可或缺的角色。然而，商業化的NADPH價格昂貴且化學性質不穩定，容易受到氧化而失去活性，因此透過酵素催化的方式合成NADPH，具有經濟及實用上的顯著優勢。
此外，本研究發現NMN轉換為NADH的效率幾乎完全，利用NADH在340 nm處具特殊吸光性的特點，即可快速且準確地定量NMN的濃度。與傳統利用高效液相層析儀（HPLC）進行檢測相比，此方法更加快速便捷，並可顯著降低有機溶液的使用量，有效減少分析過程中所產生的廢棄物，達到環保且永續的效益。
目前，亞磷酸鹽在土壤或水體中的去除方法主要分為物理、化學及生物三種方式，但使用物理或化學方法去除時，往往會產生二次污染問題。本研究提出以酵素工程的方式，將廢棄的亞磷酸鹽轉化為人體所需且昂貴的輔酶（NADH及NADPH），不僅可降低亞磷酸鹽對環境的負擔，亦能避免HPLC分析所需有機流動相造成的環境污染，符合聯合國永續發展目標（SDGs）第12項指標「Responsible Consumption and Production」的理念，兼具經濟與環保的雙重效益。
;Phosphite (H₃PO₃) is an important chemical compound with extensive applications in both agricultural and industrial fields. In agriculture, phosphite is frequently utilized as a fungicide and plant growth promoter due to its ability to be directly absorbed by plants and rapidly exert its beneficial effects, enhancing growth and improving plant resistance against environmental stress. However, improper handling or excessive use of phosphite can lead to environmental contamination, negatively impacting ecosystems.
This study aims to enzymatically catalyze the conversion of β-nicotinamide mononucleotide (NMN) into biologically essential reduced coenzymes, nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH), using phosphite as a substrate. NADH and NADPH are critical coenzymes within the human body, significantly involved in cellular metabolism and antioxidative defense systems, respectively. Commercially available NADPH, however, is costly and chemically unstable due to its susceptibility to oxidation, resulting in loss of activity. Therefore, enzymatic synthesis of NADPH presents significant economic and practical advantages.
Furthermore, this study demonstrates that the conversion efficiency from NMN to NADH is nearly complete. By exploiting the distinctive absorbance characteristic of NADH at 340 nm, this method allows for rapid and accurate quantification of NMN concentration. Compared to traditional detection methods using high-performance liquid chromatography (HPLC), this enzymatic approach is quicker, more convenient, and significantly reduces the use of organic solvents, thereby minimizing analytical waste and achieving environmental sustainability.Currently, the removal of phosphite from soil or water typically employs physical, chemical, or biological methods. However, physical and chemical methods often generate secondary pollutants. This research proposes an enzymatic engineering approach to transform waste phosphite into valuable and costly coenzymes (NADH and NADPH), effectively reducing environmental impact. Additionally, this method avoids the environmental issues associated with organic solvents used in HPLC analysis, aligning with the United Nations Sustainable Development Goals (SDGs), particularly Goal 12: "Responsible Consumption and Production," achieving both economic and environmental benefits. |
