For our amphiphiles, we use a succinic acid-based fatty acid. In previous work, we found that this molecule is readily converted to anhydride using chemical energy. Specifically, we used 2-decen-1yl-succinic acid (C10) at 65 mM in 200 mM MES buffer, the condi-tions throughout the work. We found that the behavior of C10 varies drastically with pH, which was investigated by confocal microscopy and turbidity measurements (Fig. ) . At pH below 4.0, we observed that C10 predominantly forms oil droplets, whereas at pH above 4.5, unilamellar and multilamellar vesicles emerge clearly in solution. We measured the pKa for C10 to be 5.41 (Fig. ) and can therefore conclude that C10 transitions from vesicles to oil droplets when its carboxylates become protonated. This behavior is in line with other fatty acids that tend to form oil droplets at pH values below their pKa and transition to form vesicles near their pKa. . Following our confocal microscopy findings, we focused on working at a pH of 4.9 to ensure a constant presence of vesicles. Additionally, we found that the critical vesicle concentration is above 1 mM (Fig ). Additional confocal experiments were conducted to determine the optimal concentration of C10 at which vesicles were formed that were predominantly greater than 1 µm in size form (Fig S4 ). Our reaction cycle uses a condensing agent (1-ethyl-3-(3dimethylaminopropyl)carbodiimide hydrochloride (EDC or fuel) as a chemical fuel to convert C10 to its corresponding anhydride (activation). The anhydride is unstable in the aqueous medium and hydrolyzes to the precursor (deactivation). Therefore, when a finite amount of fuel is added, the anhydride emerges and disappears again as the fuel is depleted. Using confocal microscopy, we observed remarkable transformations upon adding EDC to a vesicle solution of 65 mM C10-in the first 60 seconds after adding 23 mM EDC, we observed a marked increase in the fluorescence intensity of the vesicles. This increase is attributed to the solvatochromic dye Nile Red, whose quantum yield is greater in oily phases than in lipid bilayers. Moreover, minutes after the addition of EDC, the lumen of the vesicles was no longer observed (Fig. ). We conclude that the vesicles were almost all completely converted to oil droplets as a result of the partial conversion of the C10 into its corresponding anhydride. In the following minutes, the fluorescence intensity decreased again, and new membranous structures formed on the surface of the oil droplets. These new "daughter" vesicles were initially elongated and attached to the mother vesicle, then began to grow and, after 15 min, adopted unilamellar and multilamellar forms. However, these new vesicles remained attached to the mother vesicle until, finally, the original multilamellar vesicles reduced in size and disappeared completely, leaving a set of separate, individual daughter vesicles (Fig. , Fig. , Movie S1-S2). Likely, the new vesicles formed as the reaction cycle was deactivating the anhydride state and thus, locally, producing C10 amphiphiles. In addition, we monitored the pH of the reaction cycle throughout the experimental timeline to ensure that pH fluctuations did not influence our observations. Consistently, the pH remained stable at around 4.95 after adding EDC (Supplementary Table ) We quantified the production of daughter vesicles using confocal microscopy, by imaging the C10 vesicles before and after adding varying concentrations of EDC, with at least 25 images captured per condition. We observed that the number of vesicles increases significantly with higher EDC concentrations, evidencing a dramatic increase when the EDC concentration is raised from 16 mM to 65 mM (Fig. ). For example, with 16 mM EDC, the number of vesicles doubles compared to the initial amount observed before fuel addition. This upward trend suggests that the efficiency of EDC-induced vesicle division is markedly enhanced as its concentration increases. On the other hand, we also observed a significant reduction in the size of vesicles formed in response to EDC compared to the parent vesicles (Fig. ). Specifically, while the average diameter of the stock vesicles was approximately 15.8 + 5.6 µm, the vesicles formed after EDC addition showed an average diameter of only 3.6 + 2.0 µm (Fig. ). This demonstrates the efficacy of EDC in increasing the number of vesicles and its ability to promote structural cleavage of vesicles, resulting in smaller and more numerous units. This effect was more pronounced at higher EDC concentrations, where the decrease in vesicle size was even more evident. These changes reflect a dynamic process of vesicle reorganization and division driven by the addition of EDC. To investigate whether this mechanism of EDC action depends on the initial vesicle size, we carried out experiments with C10 vesicles prepared by extrusion through a 400 nm filter. Before the addition of fuel, cryo-TEM micrographs reveal vesicles that were polydisperse in size ranging from a diameter of 50 nm to hundreds of nm, despite the extrusion at 400 nm (Fig. ). We hypothesize that the vesicles rapidly fuse due to their fatty acid nature and their high concentration. Furthermore, DLS confirmed the polydisperse nature with a broad peak centered around 400 nm (Fig. ). Nevertheless, 1.5 hours after adding 23 mM EDC, cryo-TEM micrographs show a marked decrease in vesicle size (Fig. ). DLS analysis further corroborates these observations-following the EDC exposure, the DLS data reveals a significant shift in the size distribution towards smaller vesicle sizes, around 100 nm. Additionally, a secondary peak around 1000 nm suggests the presence of larger vesicular structures pointing towards the rapid fusion of these vesicles. These findings generally align with recent studies suggesting membrane phase transitions driven by environmental fluctuations can generate daughter protocells with reorganized contents. In contrast to these studies, where phase transitions are induced by temperature and pH fluctuations, our research demonstrates that fuel-driven chemical conversion can effectively induce vesicle division and daughter vesicle formation.