- 學位論文
腸菌素衍生物的簡易和通用化學酶合成法及其在細菌檢測中的應用
Facile and Versatile Chemoenzymatic Synthesis of Enterobactin Analogues and their Application in Bacterial Detection
摘要
鐵載體，如腸菌素，是可以與被螯合的鐵一起通過特殊受體膜蛋白被選擇性地導入到細菌內的小分子。因此，將功能性分子連接到鐵載體上，從而運送功能性試劑進入細菌是很被看好的藥物運送戰略。為了實現這一目標，合成具有特殊官能化以作為連接其他分子的連接點的鐵載體是成功的關鍵。在這碩士論文中，我們開發了一種易於執行的功能化單葡萄醣基化腸菌素的一鍋煮化學酶反應合成。從可以容易的合成的起始物腸菌素和2-硝基-4-氯苯酚葡萄糖苷類似物開始，我們可以執行雙葡糖轉移酶反應將腸菌素修飾上具有不同特殊化官能基的葡萄糖類似物，得到多種功能化單葡萄醣基化腸菌素。當我們將羅丹明B熒光團綴合到功能化單葡萄醣基化腸菌素上，從而合成羅丹明B-葡萄糖-腸菌素，它可以有選擇地標記會利用腸菌素的革蘭氏陰性菌。這包括一些大腸桿菌菌株和綠膿桿菌。會利用單水解的線性腸菌素的細菌霍亂弧菌，也可以被微弱的偵測到。此外，此螢光偵測法是只有在限鐵環境下會有效，而不會在含鐵豐富的條件下有效。我們的研究結果表明，羅丹明B-葡萄糖-腸菌素探針的菌種偵測不僅是對物種差異有分辨姓，而且對環境中的鐵的含量也有分辨性。我們在此份論文中開發的功能化單葡萄醣基化腸菌素的方便的酶法合成將促進鐵載體結合物的應用與發展。
並列摘要
Siderophores, such as enterobactin (Ent), are small molecules that can be selectively imported into bacteria along with iron via cognate transporters. Siderophore conjugates are thus a promising strategy for delivering functional reagents into bacteria. To achieve this, synthesizing functionalized siderophores is the key. In this work, we present an easy-to-perform, one-pot chemoenzymatic synthesis of functionalized monoglucosylated enterobactin (MGE). Starting from Enterobactin and 2-nitro-4-chlorophenol (CNP) glucoside analogues, both can be easily synthesized, we can perform a double glucosyl transfer enzymatic reaction to achieve the modification of enterobactin with different glucose analogues. When functionalized MGE is conjugated to a rhodamine fluorophore, thereby affording RhB-Glc-Ent, it can selectively label Gram-negative bacteria that utilize Ent, including some E. coli strains and P. aeruginosa. V. cholera, a bacterium that utilizes linearized Ent, can also be weakly targeted. Moreover, the targeting is only effective under iron-limiting and not iron-rich conditions. Our results suggest that the RhB-Glc-Ent probe is sensitive not only to strain differences but also to the iron levels in the environment. In conclusion, we have provided a convenient chemoenzymatic synthesis of functionalized MGE that would facilitate the application of siderophore conjugates.
參考文獻
延伸閱讀
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