Dimer-assisted mechanism of (un)saturated fatty acid decarboxylation for alkene production.
Dimer-assisted mechanism of (un)saturated fatty acid decarboxylation for alkene production.
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DOI：
10.1073/pnas.2221483120
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发表时间：
2023-05-30
影响因子：
11.1
通讯作者：
Zanphorlin, Leticia M.
中科院分区：
文献类型：
--
作者：
Rade, Leticia L.;Generoso, Wesley C.;Das, Suman;Souza, Amanda S.;Silveira, Rodrigo L.;Avila, Mayara C.;Vieira, Plinio S.;Miyamoto, Renan Y.;Lima, Ana B. B.;Aricetti, Juliana A.;de Melo, Ricardo R.;Milan, Natalia;Persinoti, Gabriela F.;Bonomi, Antonio M. F. L. J.;Murakami, Mario T.;Makris, Thomas M.;Zanphorlin, Leticia M.
关键词：
由于化学和物理上的相似性，插入式生物碳氢化合物有可能直接取代基于化石的产品，并与现有的石油加工、分配和组件基础设施完全兼容。在这里，我们报道了细胞色素P450的结构和功能，它是一种末端(多不饱和)产生烯烃的脱羧酶，能够利用不饱和脂肪酸作为原料来生产烯烃。与迄今鉴定的CYP-脱羧酶相比，该酶在底物结合和化学选择性方面采用了不同的分子机制。代谢的底物范围广，反应条件限制较少，代表着朝着开发生产可持续碳氢化合物的生物路线的方向迈进，这是减缓气候变化和减少对石化产品的依赖的迫切需要。脂肪酸(FAs)的酶促脱羧化代表了生物路线生产插入式碳氢化合物的进展。目前P450催化脱酸的机制主要是从细菌细胞色素P450 OleTJE建立起来的。在这里，我们描述了OleTPRN，一种产生多不饱和烯烃的脱羧酶，它超越了模型酶的功能性质，并利用了一种独特的底物结合和化学选择性的分子机制。除了从各种饱和脂肪酸转化为烯烃的高转化率而不依赖于高盐浓度外，OleTPRN还可以有效地从不饱和(油酸和亚油酸)酸中生成烯烃，不饱和脂肪酸是自然界中发现的最丰富的脂肪酸。OleTPRN通过催化路线进行碳-碳裂解，其中涉及血红素-铁基中间化合物I的氢原子转移，并在底物结合口袋的远端区域具有疏水摇篮，这在OleTJE中没有，这被认为在长链FAs的生产性结合中发挥作用，并有利于短链FAs代谢产物的快速释放。此外，OleTPRN的二聚体构型参与了A-A‘螺旋基序的稳定，A-A’螺旋基序是底物的第二配位球体，有助于脂肪族尾巴在远端和内侧活性部位口袋中的适当调节。这些发现为P450过氧化酶产生烯烃提供了另一种分子机制，为生物生产可再生碳氢化合物创造了新的机会。
Drop-in biohydrocarbons have the potential to directly replace fossil-based products due to their chemical and physical similarity, and are fully compatible with the existing petroleum processing, distribution, and components infrastructure. Herein, we report the structure and function of a cytochrome P450, a terminal (poly-unsaturated) olefin-producing decarboxylase, which enables the use of unsaturated fatty acids as feedstocks for alkene production. The enzyme exploits a distinct molecular mechanism for substrate binding and chemoselectivity comparing to the CYP-decarboxylases characterized thus far. The broad spectrum of substrates metabolized, and less-restrictive reaction conditions represent an advance toward the development of biological routes for the production of sustainable hydrocarbons, which is an urgent need for the mitigation of climate change and reduction of the dependence on petrochemicals. The enzymatic decarboxylation of fatty acids (FAs) represents an advance toward the development of biological routes to produce drop-in hydrocarbons. The current mechanism for the P450-catalyzed decarboxylation has been largely established from the bacterial cytochrome P450 OleTJE. Herein, we describe OleTPRN, a poly-unsaturated alkene-producing decarboxylase that outrivals the functional properties of the model enzyme and exploits a distinct molecular mechanism for substrate binding and chemoselectivity. In addition to the high conversion rates into alkenes from a broad range of saturated FAs without dependence on high salt concentrations, OleTPRN can also efficiently produce alkenes from unsaturated (oleic and linoleic) acids, the most abundant FAs found in nature. OleTPRN performs carbon–carbon cleavage by a catalytic itinerary that involves hydrogen-atom transfer by the heme-ferryl intermediate Compound I and features a hydrophobic cradle at the distal region of the substrate-binding pocket, not found in OleTJE, which is proposed to play a role in the productive binding of long-chain FAs and favors the rapid release of products from the metabolism of short-chain FAs. Moreover, it is shown that the dimeric configuration of OleTPRN is involved in the stabilization of the A-A’ helical motif, a second-coordination sphere of the substrate, which contributes to the proper accommodation of the aliphatic tail in the distal and medial active-site pocket. These findings provide an alternative molecular mechanism for alkene production by P450 peroxygenases, creating new opportunities for biological production of renewable hydrocarbons.
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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：
4.6
作者：
Bharadwaj VS;Kim S;Guarnieri MT;Crowley MF
通讯作者：
Crowley MF
影响因子：
6.3
作者：
Liu Y;Wang C;Yan J;Zhang W;Guan W;Lu X;Li S
通讯作者：
Li S
影响因子：
15
作者：
Grant, Job L.;Hsieh, Chun H.;Makris, Thomas M.
通讯作者：
Makris, Thomas M.
P450 OleT(JE)的分子基础：底物结合机制和主要途径的研究
影响因子：
3.5
作者：
Du, Juan;Liu, Lin;Yang, Jian Ming
通讯作者：
Yang, Jian Ming