Our AMP delivery system is designed in such a way that it gets activated only at temperatures above 42°C. Since B.subtilis can withstand extreme environmental conditions unlike E.coli, we decided to use it as the host organism our plasmid.
Provision of subtilis in its vegetative form have a few disadvantages such as
- Continual supply of nutrients for its survival.
- No control on Bacillus count.
- Death of cells due to exhaustion of nutrients,thus all the three affecting the production of our desired product AMPs.
These disadvantages can be overcome by B.subtilis spores. Bacillus subtilis under stress, forms spores which are highly resistant to extreme temperature and pH and can grow back to vegetative form on provision of favourable conditions.
Hence we decided to exploit these properties of these spores for the efficient production of AMPS by our temperature activated system.Thereby maintaining the cell count and conversion to vegetative form at the required time.
Our final product design is a double membrane system, with the inner membrane containing the B.subtilis spores that will be converted to vegetative cells on contact with the medium so that AMPs can be produced at temperatures above 42°C.