A bicyclic S-adenosylmethionine regeneration system applicable with different nucleosides or nucleotides as cofactor building blocks.
A bicyclic S-adenosylmethionine regeneration system applicable with different nucleosides or nucleotides as cofactor building blocks.
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DOI：
10.1039/d1cb00033k
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发表时间：
2021-03-22
影响因子：
4.1
通讯作者：
Andexer JN
中科院分区：
文献类型：
--
作者：
Popadić D;Mhaindarkar D;Dang Thai MHN;Hailes HC;Mordhorst S;Andexer JN
无处不在的辅助因子s -腺苷-l-蛋氨酸（SAM）是代谢、表观遗传学和癌症发展中许多生化反应的一部分。由于甲基化通常会改善与药物用途相关的化合物的理化性质，因此在生物技术应用中持续利用SAM作为甲基供体是一个重要的目标。因此，sam依赖的甲基转移酶是一种新兴的环境友好型和选择性烷基化的生物催化工具。然而，SAM表现出在温和条件下降解等不良特性，其化学计量使用在经济上不合理。在这里，我们报道了一个优化的SAM和SAM类似物再生的仿生系统，包括有效的三磷酸核苷形成和一个额外的l-蛋氨酸再生循环，没有副产物积累。双环系统使用7种酶，s -甲基蛋氨酸作为甲基供体和剩余的无机聚磷酸盐，以及催化量的l-蛋氨酸和辅助因子构建块，转化率高达99%（高达200次周转）。我们还表明，该循环可以与含有不同嘌呤和嘧啶核碱基的辅因子构建块一起运行，这些辅因子可以在核苷或核苷酸阶段输入。这些替代辅因子依次转化为相应的SAM类似物，这被认为是生物正交系统发展的关键。除了纯化酶外，双环系统还可以与粗裂解物一起使用，突出了其广泛的生物催化适用性。多磷酸盐驱动的双环s -腺苷蛋氨酸（SAM）再生系统使用s -甲基蛋氨酸作为“2合1”的甲基供体而不产生副产物，并且可以与SAM核碱基类似物如s -胞基和s -氨基蛋氨酸一起运行。
The ubiquitous cofactor S-adenosyl-l-methionine (SAM) is part of numerous biochemical reactions in metabolism, epigenetics, and cancer development. As methylation usually improves physiochemical properties of compounds relevant for pharmaceutical use, the sustainable use of SAM as a methyl donor in biotechnological applications is an important goal. SAM-dependent methyltransferases are consequently an emerging biocatalytic tool for environmentally friendly and selective alkylations. However, SAM shows undesirable characteristics such as degradation under mild conditions and its stoichiometric use is economically not reasonable. Here, we report an optimised biomimetic system for the regeneration of SAM and SAM analogues consisting of effective nucleoside triphosphate formation and an additional l-methionine regeneration cycle without by-product accumulation. The bicyclic system uses seven enzymes, S-methylmethionine as methyl donor and a surplus of inorganic polyphosphate, along with catalytic amounts of l-methionine and cofactor building block reaching conversions of up to 99% (up to 200 turnovers). We also show that the cycle can be run with cofactor building blocks containing different purine and pyrimidine nucleobases, which can be fed in at the nucleoside or nucleotide stage. These alternative cofactors are in turn converted to the corresponding SAM analogues, which are considered to be a key for the development of bioorthogonal systems. In addition to purified enzymes, the bicyclic system can also be used with crude lysates highlighting its broad biocatalytic applicability. The polyphosphate-driven bicyclic S-adenosylmethionine (SAM) regeneration system uses S-methylmethionine as a ‘2-in-1’ methyl donor without producing by-products and can be run with SAM nucleobase analogues such as S-cytidyl- and S-inosylmethionine.
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DOI：
10.1002/anie.201608625
发表时间：
2017-05-02
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Deen J;Vranken C;Leen V;Neely RK;Janssen KPF;Hofkens J
通讯作者：
Hofkens J
影响因子：
3.9
作者：
Castro, Carmen;Millian, Norman S.;Garrow, Timothy A.
通讯作者：
Garrow, Timothy A.
影响因子：
8.4
作者：
Law BJC;Struck AW;Bennett MR;Wilkinson B;Micklefield J
通讯作者：
Micklefield J
影响因子：
37.8
作者：
Liao, Cangsong;Seebeck, Florian P.
通讯作者：
Seebeck, Florian P.
影响因子：
4.8
作者：
Lee, JE;Cornell, KA;Howell, PL
通讯作者：
Howell, PL