One-Pot Biocatalytic In Vivo Methylation-Hydroamination of Bioderived Lignin Monomers to Generate a Key Precursor to L-DOPA.
One-Pot Biocatalytic In Vivo Methylation-Hydroamination of Bioderived Lignin Monomers to Generate a Key Precursor to L-DOPA.
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DOI：
10.1002/anie.202112855
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发表时间：
2022-02-14
影响因子：
16.6
通讯作者：
Turner, Nicholas J.
中科院分区：
文献类型：
--
作者：
Galman, James L.;Parmeggiani, Fabio;Seibt, Lisa;Birmingham, William R.;Turner, Nicholas J.
富电子的酚类底物可以衍生自木质素原料的解聚。所获得的羟基肉桂酸单体的直接生物转化可用于生产高价值的化学品，例如α-氨基酸，然而该反应通常受到碱性或苛刻反应介质中的化学自氧化的阻碍。区域选择性O-甲基转移酶（OMT）是天然次级代谢途径中普遍存在的酶，利用昂贵的共底物S-腺苷-L-甲硫氨酸（SAM）作为甲基化试剂，改变羟基肉桂酸的理化性质。在这项研究中，我们设计了一种OMT，以接受各种富电子的酚底物，修饰了一种商业大肠杆菌菌株BL 21（DE 3），以在体内再生SAM，并将其与工程化的氨裂解酶结合，以参与一锅法，两个全细胞酶级联反应，从木质素衍生的阿魏酸中产生l-多巴前体l-藜芦基甘氨酸。 蛋白质和菌株工程相结合。已经开发了两种工程化酶（甲基转移酶和氨裂解酶）和工程化大肠杆菌菌株（用于SAM辅因子的再生）的组合，以实现完全生物催化的一锅甲基化-加氢胺化级联。例如，已经证明了从可再生的木质素衍生的阿魏酸合成l-藜芦基甘氨酸的高产率和优异的ee。
Electron‐rich phenolic substrates can be derived from the depolymerisation of lignin feedstocks. Direct biotransformations of the hydroxycinnamic acid monomers obtained can be exploited to produce high‐value chemicals, such as α‐amino acids, however the reaction is often hampered by the chemical autooxidation in alkaline or harsh reaction media. Regioselective O‐methyltransferases (OMTs) are ubiquitous enzymes in natural secondary metabolic pathways utilising an expensive co‐substrate S‐adenosyl‐l‐methionine (SAM) as the methylating reagent altering the physicochemical properties of the hydroxycinnamic acids. In this study, we engineered an OMT to accept a variety of electron‐rich phenolic substrates, modified a commercial E. coli strain BL21 (DE3) to regenerate SAM in vivo, and combined it with an engineered ammonia lyase to partake in a one‐pot, two whole cell enzyme cascade to produce the l‐DOPA precursor l‐veratrylglycine from lignin‐derived ferulic acid. Protein and strain engineering combined. The combination of two engineered enzymes (a methyltransferase and an ammonia lyase) and an engineered E. coli strain (for regeneration of the SAM cofactor) has been developed to enable a fully biocatalytic one‐pot methylation–hydroamination cascade. As an example, the synthesis of l ‐veratrylglycine from renewable lignin‐derived ferulic acid has been demonstrated, in high yield and excellent ee.
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影响因子：
16.6
作者：
Mallinson SJB;Machovina MM;Silveira RL;Garcia-Borràs M;Gallup N;Johnson CW;Allen MD;Skaf MS;Crowley MF;Neidle EL;Houk KN;Beckham GT;DuBois JL;McGeehan JE
通讯作者：
McGeehan JE
影响因子：
4.5
作者：
Aleku GA;Prause C;Bradshaw-Allen RT;Plasch K;Glueck SM;Bailey SS;Payne KAP;Parker DA;Faber K;Leys D
通讯作者：
Leys D
影响因子：
5.3
作者：
Itoh N;Iwata C;Toda H
通讯作者：
Toda H
影响因子：
37.8
作者：
Liao, Cangsong;Seebeck, Florian P.
通讯作者：
Seebeck, Florian P.
影响因子：
16.6
作者：
Mordhorst, Silja;Siegrist, Jutta;Andexer, Jennifer N.
通讯作者：
Andexer, Jennifer N.