Methods Enzymol. 2026;728:1-21. doi: 10.1016/bs.mie.2026.01.013. Epub 2026 Jan 20.

ABSTRACT

Phospholipid biogenesis couples fatty acid synthesis to glycerol acylation and headgroup installation, producing amphiphilic lipids that self-assemble into membranes. Reconstituting this process in the absence of pre-existing bilayers remains a central challenge for bottom-up synthetic biology and origins-of-life research. Here we describe a chemoenzymatic strategy that constructs phospholipids and membranes de novo in aqueous solution. A soluble type I fatty acid synthase generates palmitoyl-CoA from acetyl-CoA and malonyl-CoA, which is then coupled to cysteine-modified lysophospholipids through native chemical ligation to yield amidophospholipids that spontaneously assemble into bilayers. The approach is modular, permitting independent validation of enzymatic and chemical phases, yet also functions in one pot under uniform reaction conditions. This methodology provides a tractable platform for de novo membrane construction and offers a path toward fully synthetic lipid systems that link composition, growth, and function.

PMID:41895902 | DOI:10.1016/bs.mie.2026.01.013