[DIYbio] Re: Exocytosis in Bacteria

The problem with this Jacob is that you are fighting against thermodynamics and metabolism. The amount of energy cost to export proteins is huge, in the folding, transport, generation of transport proteins. Also, one needs to think about ligands, which many proteins contain or need to fold. I think the reason that this avenue has not been explored thoroughly is that you will be trying to optimize many different processes with many bottlenecks. Also, disrupting cells using some detergent (SDS or something) and maybe some freeze-thawing is darn easy. It would probably be extremely difficult to match normal expression methods for the wide variety of proteins that people want to purify. This could be useful for some niche cases but usually that just involves slapping a signal peptide on the proteint.


Josiah

On Thursday, February 5, 2015 at 2:40:18 PM UTC-8, Jacob Palumbo wrote:

Hey all,

I'm working on a project where it would be extremely useful to have bacteria (E. Coli specifically) exocytose proteins continuously, rather than grow a culture and lyse it the old fashion way. I've found a few papers on this so far (1, 2, 3), but they are all for Gram-positive bacteria, which E. Coli is not. If need be, I could switch over, but it would be nice to not have to. I do have some information the Sec pathway in Gram-negative bacteria, but it is somewhat general.

Anyone have an experience/insight into this?

Thanks,
Jake