RT Journal Article
SR Electronic
T1 Chemoenzymatic Generation of Phospholipid Membranes Mediated by Type I Fatty Acid Synthase
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.04.21.440816
DO 10.1101/2021.04.21.440816
A1 Satyam Khanal
A1 Roberto J. Brea
A1 Michael D. Burkart
A1 Neal K. Devaraj
YR 2021
UL http://biorxiv.org/content/early/2021/04/22/2021.04.21.440816.2.abstract
AB 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 non-canonical amidophospholipid that self-assembles into micron-sized 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.Competing Interest StatementThe authors have declared no competing interest.