Expanding the Substrate Scope of N - and O -Methyltransferases from Plants for Chemoselective Alkylation
Expanding the Substrate Scope of N - and O -Methyltransferases from Plants for Chemoselective Alkylation
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扩大植物中 N - 和 O - 甲基转移酶的底物范围以进行化学选择性烷基化
DOI：
10.1101/2023.07.21.549995
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Jockmann E
中科院分区：
文献类型：
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作者：
Jockmann E
在生成药物活性分子和其他应用的构建模块时，甲基化反应具有重要意义。合成甲基化试剂由于其高反应性通常有毒且无选择性。S-腺苷-L-甲硫氨酸 (SAM) 依赖性甲基转移酶 (MT) 提供了一种化学选择性且环境友好的替代品。芸香中的邻氨基苯甲酸 N-MT (RgANMT) 参与吖啶酮生物碱的生物合成，甲基化邻氨基苯甲酸。尽管已知底物仅在 N 位甲基化，但氨基酸序列相似性超过 60% 的最接近的亲戚是催化咖啡酸和仅含羟基的衍生物 (CaOMT) 甲基化反应的 O-MT。在这项研究中，我们使用同时具有氨基和羟基（氨基酚）的化合物作为可能的甲基受体，研究了 RgANMT 和桃红色 CaOMT（PpCaOMT）的底物范围。对于这两种酶，反应具有高度化学选择性。此外，原位生成辅因子衍生物可以将其他烷基链转移到氨基酚上，从而扩大产品库。选定的 MT 反应在体外和体内实验中以制备型生物催化规模进行，产率高达 62%。
Methylation reactions are of significant interest when generating pharmaceutically active molecules and building blocks for other applications. Synthetic methylating reagents are often toxic and unselective due to their high reactivity.S‐Adenosyl‐l‐methionine (SAM)‐dependent methyltransferases (MTs) present a chemoselective and environmentally friendly alternative. The anthranilateN‐MT fromRuta graveolens(RgANMT) is involved in acridone alkaloid biosynthesis, methylating anthranilate. Although it is known to methylate substrates only at theN‐position, the closest relatives with respect to amino acid sequence similarities of over 60 % areO‐MTs catalysing the methylation reaction of caffeate and derivatives containing only hydroxyl groups (CaOMTs). In this study, we investigated the substrate range ofRgANMT and a CaOMT fromPrunus persica(PpCaOMT) using compounds with both, an amino‐ and hydroxyl group (aminophenols) as possible methyl group acceptors. For both enzymes, the reaction was highly chemoselective. Furthermore, generating cofactor derivatives in situ enabled the transfer of other alkyl chains onto the aminophenols, leading to an enlarged pool of products. Selected MT reactions were performed at a preparative biocatalytic scale in in vitro and in vivo experiments resulting in yields of up to 62 %.