Synthetic bacterial vesicles to enhance recombinant protein production, delivery and isolation for Industrial Biotechnology applications.

The ability to reprogram a cell to direct the packaging of specific molecules into discrete membrane envelopes is a major objective for synthetic biology. This controlled packaging into membrane vesicles will allow biologists to create a plethora of new technologies, which could be applied in both biotechnology and medical industries. These include the generation of novel metabolic factories within a cell for energy production; for rapidly packaging toxic proteins into contained environments before they have a chance to harm any normal metabolic activities, so they can be purified for use in subsequent pharmaceutical applications; the creation of protective packages filled with difficult to isolate biomolecules, which can be kept in stable environment to allow their storage and purification; and also generate simple vehicles for delivery of drugs and vaccines to the patient. Here, we provide a simple and cost effective solution to the problem. We have discovered a method to program a simple cell to create membrane packages which can be filled with different molecules of interest. We have not only discovered a way to fine-tune the shape of the membrane package (e.g. into long tubular matrices or spherical vesicles), but we have also devised controllable mechanisms that either keep the package within, or secrete the package out of the cell. Thus we have therefore made a landmark breakthrough in synthetic biology research. A major aim of this BBSRC key strategic area is to design from new and improve on natural systems and exploit these for the production of commercially important chemicals and biotherapeutics, which is what we have achieved here. Our overall aim in this project is to make use of these exciting discoveries to modify cells, making them capable of creating membrane bound packages filled with any protein of interest, which can then either be secreted from the cell and isolated from the culture media using a simple one step filtration technique, or stored within the cell where it can be made to act as a metabolic micro-factories, producing useful and/or valuable molecules without intoxicating the cells. In this way we hope to develop new ways to produce fine and platform chemicals as well as biotherapeutics. Through the research described in this application we are certain that we will be able to contribute to the development of new sustainable approaches for generating biotherapeutics, which will be assimilated into production techniques by diverse bio-industries.