Print Email Facebook Twitter Cell-Free Phospholipid Biosynthesis by Gene-Encoded Enzymes Reconstituted in Liposomes Title Cell-Free Phospholipid Biosynthesis by Gene-Encoded Enzymes Reconstituted in Liposomes Author Scott, A.D.D. (TU Delft BN/Christophe Danelon Lab; Kavli institute of nanoscience Delft) Noga, M.J. (TU Delft BN/Greg Bokinsky Lab; Kavli institute of nanoscience Delft) de Graaf, P. (Kavli institute of nanoscience Delft; Student TU Delft) Westerlaken, I. (TU Delft BN/Technici en Analisten; Kavli institute of nanoscience Delft) Yildirim, E. (TU Delft BN/Technici en Analisten; Kavli institute of nanoscience Delft) Danelon, C.J.A. (TU Delft BN/Christophe Danelon Lab; Kavli institute of nanoscience Delft) Date 2016 Abstract The goal of bottom-up synthetic biology culminates in the assembly of an entire cell from separate biological building blocks. One major challenge resides in the in vitro production and implementation of complex genetic and metabolic pathways that can support essential cellular functions. Here, we show that phospholipid biosynthesis, a multiple-step process involved in cell membrane homeostasis, can be reconstituted starting from the genes encoding for all necessary proteins. A total of eight E. coli enzymes for acyl transfer and headgroup modifications were produced in a cell-free gene expression system and were co-translationally reconstituted in liposomes. Acyl-coenzyme A and glycerol-3-phosphate were used as canonical precursors to generate a variety of important bacterial lipids. Moreover, this study demonstrates that two-step acyl transfer can occur from enzymes synthesized inside vesicles. Besides clear implications for growth and potentially division of a synthetic cell, we postulate that gene-based lipid biosynthesis can become instrumental for ex vivo and protein purification-free production of natural and non-natural lipids. Subject LipidsLiposomesVesiclesEnzymesGene expressionLiquid chromatography-mass spectrometryPhospholipidsBiosynthesisOA-Fund TU Delft To reference this document use: http://resolver.tudelft.nl/uuid:b9e24dde-0d88-4346-9fd6-8d5da128d0a8 DOI https://doi.org/10.1371/journal.pone.0163058 ISSN 1932-6203 Source PLoS ONE, 11 (10) Part of collection Institutional Repository Document type journal article Rights © 2016 A.D.D. Scott, M.J. Noga, P. de Graaf, I. Westerlaken, E. Yildirim, C.J.A. Danelon Files PDF journal.pone.0163058.pdf 1.58 MB Close viewer /islandora/object/uuid:b9e24dde-0d88-4346-9fd6-8d5da128d0a8/datastream/OBJ/view