Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.
Salaya Central Instrument Facility RSPG, Research Management and Development Division, Office of the President, Mahidol University, Nakhon Pathom 73170, Thailand.
Molecules. 2021 Oct 16;26(20):6264. doi: 10.3390/molecules26206264.
4-Hydroxycoumarin (4HC) has been used as a lead compound for the chemical synthesis of various bioactive substances and drugs. Its prenylated derivatives exhibit potent antibacterial, antitubercular, anticoagulant, and anti-cancer activities. In doing this, BL21(DE3)pLysS strain was engineered as the in vivo prenylation system to produce the farnesyl derivatives of 4HC by coexpressing the genes encoding aromatic prenyltransferase (AtaPT) and truncated 1-deoxy-D-xylose 5-phosphate synthase of (CstDXS), where 4HC was the fed precursor. Based on the high-resolution LC-ESI(±)-QTOF-MS/MS with the use of in silico tools (e.g., MetFrag, SIRIUS (version 4.8.2), CSI:FingerID, and CANOPUS), the first major prenylated product (named compound-1) was detected and ultimately elucidated as ferulenol, in which information concerning the correct molecular formula, chemical structure, substructures, and classifications were obtained. The prenylated product (named compound-2) was also detected as the minor product, where this structure proposed to be the isomeric structure of ferulenol formed via the tautomerization. Note that both products were secreted into the culture medium of the recombinant and could be produced without the external supply of prenyl precursors. The results suggested the potential use of this engineered pathway for synthesizing the farnesylated-4HC derivatives, especially ferulenol.
4-羟基香豆素（4HC）已被用作各种生物活性物质和药物的化学合成的先导化合物。其香叶基衍生物表现出强大的抗菌、抗结核、抗凝和抗癌活性。在这方面，BL21(DE3)pLysS 菌株被设计为体内香叶基化系统，通过共表达编码芳香基转移酶（AtaPT）和截断 1-脱氧-D-木酮糖 5-磷酸合酶的基因（CstDXS），以产生 4HC 的法呢基衍生物，其中 4HC 是进料前体。基于使用计算工具（例如 MetFrag、SIRIUS（版本 4.8.2）、CSI:FingerID 和 CANOPUS）的高分辨率 LC-ESI(±)-QTOF-MS/MS，首次检测到主要的香叶基化产物（命名为化合物-1），并最终阐明为阿魏醇，其中获得了有关正确分子公式、化学结构、亚结构和分类的信息。还检测到较小的香叶基化产物（命名为化合物-2），该结构被提议为通过互变异构形成的阿魏醇的异构结构。请注意，这两种产物都分泌到重组菌的培养液中，并且可以在没有外部提供香叶基前体的情况下生产。结果表明，该工程途径具有合成法呢基-4HC 衍生物的潜力，特别是阿魏醇。
