#16 - Bottom-Up Construction of a Minimal System for Cellular Respiration and Energy Regeneration
Olivier Biner (University of Cambridge)
Monday, 30 Nov 21:15 - 22:00 CET
Access to the BigBlueButton rooom for this Mini Talk via the lists for Monday and Tuesday.
Please make yourself familiar with BigBlueButton before you join the Mini Talk - read the instructions.
Title: Bottom-Up Construction of a Minimal System for Cellular Respiration and Energy Regeneration
Author(s): Olivier Binera
Affiliations: aUniversity of Cambridge
Abstract: Adenosine triphosphate (ATP), the cellular energy currency, is essential for life. The ability to provide a constant supply of ATP is therefore crucial for the construction of artificial cells in synthetic biology. Here, we describe the bottom-up assembly and characterization of a minimal respiratory system that uses NADH as a fuel to produce ATP from ADP and inorganic phosphate, and is thus capable of sustaining both upstream metabolic processes that rely on NAD+, and downstream energy-demanding processes that are powered by ATP hydrolysis. A detergent-mediated approach was used to co-reconstitute respiratory mitochondrial complex I and an F-type ATP synthase into nanosized liposomes. Addition of the alternative oxidase to the resulting proteoliposomes produced a minimal artificial "organelle" that reproduces the energy-converting catalytic reactions of the mitochondrial respiratory chain: NADH oxidation, ubiquinone cycling, oxygen reduction, proton pumping, and ATP synthesis. As a proof-of-principle, we demonstrate that our nanovesicles are capable of using an NAD+-linked substrate to drive cell-free protein expression. Our nanovesicles are both efficient and durable and may be applied to sustain artificial cells in future work.