Chemoenzymatic Generation of Phospholipid Membranes Mediated by Type I Fatty Acid Synthase
Chemoenzymatic Generation of Phospholipid Membranes Mediated by Type I Fatty Acid Synthase
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DOI：
10.1021/jacs.1c02121
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发表时间：
2021-05-12
影响因子：
15
通讯作者：
Devaraj, Neal K.
中科院分区：
文献类型：
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作者：
Khanal, Satyam;Brea, Roberto J.;Devaraj, Neal K.
从最小反应性前体从头形成脂质膜是合成细胞研究的主要目标。在自然界中，膜磷脂的合成是由许多酶协调的，包括脂肪酸脱氢酶和膜结合酰基转移酶。然而，这些酶途径难以在体外完全再现。因此，从简单的代谢结构单元重建磷脂膜合成仍然是一个挑战。在这里，我们描述了一种化学酶的策略，利用可溶性细菌脂肪酸合成酶（CGFAS I）的脂质膜的产生，在原位合成棕榈酰辅酶A的乙酰辅酶A和丙二酰辅酶A。脂肪酸衍生物通过天然化学连接（NCL）自发地与半胱氨酸修饰的溶血磷脂反应，提供自组装成微米级膜结合囊泡的非规范酰胺磷脂。据我们所知，这是直接从乙酰辅酶A和丙二酰辅酶A前体重建磷脂膜形成的第一个例子。我们的研究结果表明，结合的特异性和效率的I型脂肪酸合酶与高度选择性的生物共轭反应提供了一个仿生途径从头形成膜结合囊泡。
The de novo formation of lipid membranes from minimal reactive precursors is a major goal in synthetic cell research. In nature, the synthesis of membrane phospholipids is orchestrated by numerous enzymes, including fatty acid synthases and membrane-bound acyltransferases. However, these enzymatic pathways are difficult to fully reproduce in vitro. As such, the reconstitution of phospholipid membrane synthesis from simple metabolic building blocks remains a challenge. Here, we describe a chemoenzymatic strategy for lipid membrane generation that utilizes a soluble bacterial fatty acid synthase (cgFAS I) to synthesize palmitoyl-CoA in situ from acetyl-CoA and malonyl-CoA. The fatty acid derivative spontaneously reacts with a cysteine-modified lysophospholipid by native chemical ligation (NCL), affording a noncanonical amidophospholipid that self-assembles into micronsized membrane-bound vesicles. To our knowledge, this is the first example of reconstituting phospholipid membrane formation directly from acetyl-CoA and malonyl-CoA precursors. Our results demonstrate that combining the specificity and efficiency of a type I fatty acid synthase with a highly selective bioconjugation reaction provides a biomimetic route for the de novo formation of membrane-bound vesicles.