| 摘要: | 隨著人們對健康的關注和健康意識的提高，保健食品市場不斷增長，已成為人們
日常生活中的重要組成部分，廣泛應用於增強免疫力、促進新陳代謝、改善睡眠和延
緩衰老等方面。在這個背景下，煙酰胺單核苷酸（NMN）作為一種潛在的保健成分引
起了廣泛關注。NMN 作為煙酰胺（維生素 B3）的前體，在細胞代謝和能量生產中發
揮著重要作用，是合成煙酰胺腺嘌呤二核苷酸（NAD+）的重要生物合成途徑之一。研
究表明，NMN 的補充可以增加 NAD+水平，從而促進細胞活力、改善代謝健康、延緩
衰老和提高運動耐力等，因此被認為是一種具有潛力的保健食品成分。本研究旨在優
化核糖磷酸化反應步驟，以實現高效、經濟的NMN生產。首先使用核糖激酶將核糖與
三磷酸腺苷 (ATP) 反應生成核糖-5-磷酸 (ribose-5-phosphate) 與雙磷酸腺苷 (ADP) ，然
後再使用聚磷酸激酶 II（polyphosphatekinase, PPK2），將核糖磷酸化反應的副產物
ADP 與長鏈聚磷酸鹽（polyphosphate, polyP）反應，生成還原為 ATP，將上述兩個反
應偶聯，建立一種簡單高效的多酶組合催化體系，本研究藉由優化此反應的溫度、 pH
值、buffer 等條件來建立起一鍋法合成 NMN的基本反應條件，然後在合成 NMN中，
透過改變受質濃度、酵素濃度、離子強度等來增加NMN的產量，通過系統性的實驗設
計和數據分析，成功地優化了核糖磷酸化反應條件，實現了NMN的高效生產。;As people become more concerned about health and their health awareness increases, the
health food market continues to grow, becoming an important part of daily life. It is widely
used to boost immunity, promote metabolism, improve sleep, and delay aging. Against this
backdrop, nicotinamide mononucleotide (NMN) has attracted widespread attention as a
potential health supplement. NMN, a precursor of nicotinamide (vitamin B3), plays a crucial
role in cellular metabolism and energy production and is a key biosynthetic pathway for the
synthesis of nicotinamide adenine dinucleotide (NAD+). Research has shown that NMN
supplementation can increase NAD+ levels, thereby enhancing cellular vitality, improving
metabolic health, delaying aging, and increasing exercise endurance. Consequently, it is
considered a promising health food ingredient. This study aims to optimize the ribose
phosphorylation reaction steps to achieve efficient and economical NMN production. Initially,
ribokinase is used to react ribose with adenosine triphosphate (ATP) to generate ribose-5
phosphate and adenosine diphosphate (ADP). Subsequently, polyphosphate kinase II (PPK2)
is employed to react the by-product ADP from the ribose phosphorylation reaction with long
chain polyphosphate (polyP) to regenerate ATP. By coupling these two reactions, a simple and
efficient multi-enzyme catalytic system is established. This research optimizes the reaction
conditions such as temperature, pH, and buffer to establish a one-pot synthesis of NMN.
Additionally, in the synthesis of NMN, varying the substrate concentration, enzyme
concentration, and ionic strength increases the NMN yield. Through systematic experimental
design and data analysis, the conditions for the ribose phosphorylation reaction were
successfully optimized, achieving efficient NMN production. |
